Elephant
Bibliographic
Database

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Elephant Bibliographic Database
www.elephantcare.org

References updated October 2009 by date of publication, most recent first.

Behr, B., Rath, D., Hildebrandt, T.B., Goeritz, F., Blottner, S., Portas, T.J., Bryant, B.R., Sieg, B., Knieriem, A., de Graaf, S.P., Maxwell, W.M., Hermes, R., 2009. Germany/Australia index of sperm sex sortability in elephants and rhinoceros. Reprod. Domest. Anim 44, 273-277.
Abstract: Flow cytometric sexing of spermatozoa followed by application in artificial insemination or in vitro fertilization provides a unique opportunity to predetermine the sex of offspring and might enhance the conservation management of endangered species in captivity such as the elephant and rhinoceros. To obtain an indication of the sortability of spermatozoa from these species, the relative DNA differences between X and Y chromosome bearing spermatozoa (fresh, frozen thawed, epididymal) from three rhinoceros species [white (Ceratotherium simum), black (Diceros bicornis), Indian (Rhinoceros unicornis)] and both elephant species, the Asian and the African elephant (Elephas maximus, Loxodonta Africana), were determined through separation of spermatozoa into X and Y chromosome bearing populations, using a modified high speed flow cytometer. The head profile areas of spermatozoa from all five species were measured using light microscopy. By multiplying the relative DNA differences and the head profile areas, the sperm sorting indices were calculated to be 47, 48 and 51 for white, black and Indian rhinoceros respectively. The calculated sorting index for the Asian elephant was 66. In the African elephant, we determined the highest sorting index of 76. These results indicate the practicability of flow cytometric sex sorting of spermatozoa from the tested rhinoceros species and both elephant species. The lower sorting indices in rhinos indicate that sex sorting of spermatozoa from the rhinoceros will be more challenging than in elephants

Brodie, J.F., Helmy, O.E., Brockelman, W.Y., Maron, J.L., 2009. Bushmeat poaching reduces the seed dispersal and population growth rate of a mammal-dispersed tree. Ecological Applications 19, 854-863.
Abstract: Myriad tropical vertebrates are threatened by overharvest. Whether this harvest has indirect effects on nonhunted organisms that interact with the game species is a critical question. Many tropical birds and mammals disperse seeds. Their overhunting in forests can cause zoochorous trees to suffer from reduced seed dispersal. Yet how these reductions in seed dispersal influence tree abundance and population dynamics remains unclear. Reproductive parameters in long-lived organisms often have very low elasticities; indeed the demographic importance of seed dispersal is an open question. We asked how variation in hunting pressure across four national parks with seasonal forest in northern Thailand influenced the relative abundance of gibbons, muntjac deer, and sambar deer, the sole dispersers of seeds of the canopy tree Choerospondias axillaris. We quantified how variation in disperser numbers affected C. axillaris seed dispersal and seedling abundance across the four parks. We then used these data in a structured population model based on vital rates measured in Khao Yai National Park (where poaching pressure is minimal) to explore how variation in illegal hunting pressure might influence C. axillaris population growth and persistence. Densities of the mammals varied strongly across the parks, from relatively high in Khao Yai to essentially zero in Doi Suthep-Pui. Levels of C. axillaris seed dispersal and seedling abundance positively tracked mammal density. If hunting in Khao Yai were to increase to the levels seen in the other parks, C. axillaris population growth rate would decline, but only slightly. Extinction of C. axillaris is a real possibility, but may take many decades. Recent and ongoing extirpations of vertebrates in many tropical forests could be creating an extinction debt for zoochorous trees whose vulnerability is belied by their current abundance.

Brown, J.L., Kersey, D.C., Freeman, E.W., Wagener, T., 2009. Assessment of diurnal urinary cortisol excretion in Asian and African elephants using different endocrine methods. Zoo. Biol.
Abstract: Longitudinal urine samples were collected from Asian and African elephants to assess sample processing and immunoassay techniques for monitoring adrenal activity. Temporal profiles of urinary cortisol measured by RIA and EIA, with and without dichloromethane extraction, were similar; all correlation coefficients were >0.90. However, based on regression analyses, cortisol immunoactivity in extracted samples was only 72-81% of that of unextracted values. Within assay technique, RIA values were only 74-81% of EIA values. Collection of 24-hr urine samples demonstrated a clear diurnal pattern of glucocorticoid excretion, with the lowest concentrations observed just before midnight and peak concentrations occurring around 0600-0800 hr. These results indicate that elephants fit the pattern of a diurnal species, and that glucocorticoid production is affected by a sleep-wake cycle similar to that described for other terrestrial mammals. Cortisol can be measured in both extracted and unextracted urine using RIA and EIA methodologies. However, unexplained differences in quantitative results suggest there may be sample matrix effects and that data generated using different techniques may not be directly comparable or interchangeable. Zoo Biol 28:1-10, 2009. (c) 2009 Wiley-Liss, Inc

Freeman, E.W., Schulte, B.A., Brown, J.L., 2009. Investigating the impact of rank and ovarian activity on the social behavior of captive female African elephants . Zoo Biology 0, 1-14.
Abstract: Over a third of captive female African elephants in North America fail to exhibit normal estrous cycles based on long-term serum progestagen analyses. Why acyclicity occurs is unknown; however, the majority of noncycling females are ranked by keepers as the dominant individual within the group. To investigate the relationship between ovarian cyclicity status and keeper-determined social rank, observations were conducted on 33 female African elephants (18 cycling, 15 noncycling). Based on keeper evaluations, five cycling elephants were ranked dominant, seven in the middle and six as subordinate. In contrast, 10 noncycling elephants were ranked as dominate and five as subordinate with none ranked as middle. When comparing the behavior of the elephants by their keeper-determined rank, the dominant females dominant were significantly more likely to approach, displace and push. Similarly, keeper-determined subordinate females more frequently presented their hind end and held their ears erect. Behaviors initiated by one elephant toward another did not vary between cycling and noncycling females, except when the interaction with social rank was tested. Dominant, noncycling females initiated a higher percentage of approach and displace behaviors than both cycling and noncycling, subordinate elephants. Subordinate, noncycling elephants displayed the highest percentage of ears erect. Social rank drives the interactions of ex situ female African elephants more than ovarian cyclicity status. Thus, behavioral interactions cannot be used to predict which cycling elephants are most likely to become acyclic.

Freeman, E.W., Schulte, B.A., Brown, J.L., 2009. Investigating the impact of rank and ovarian activity on the social behavior of captive female African elephants . Zoo Biology 0, 1-14.
Abstract:
Over a third of captive female African elephants in North America fail to exhibit normal estrous cycles based on long-term serum progestagen analyses. Why acyclicity occurs is unknown; however, the majority of noncycling females are ranked by keepers as the dominant individual within the group. To investigate the relationship between ovarian cyclicity status and keeper-determined social rank, observations were conducted on 33 female African elephants (18 cycling, 15 noncycling). Based on keeper evaluations, five cycling elephants were ranked dominant, seven in the middle and six as subordinate. In contrast, 10 noncycling elephants were ranked as dominate and five as subordinate with none ranked as middle. When comparing the behavior of the elephants by their keeper-determined rank, the dominant females dominant were significantly more likely to approach, displace and push. Similarly, keeper-determined subordinate females more frequently presented their hind end and held their ears erect. Behaviors initiated by one elephant toward another did not vary between cycling and noncycling females, except when the interaction with social rank was tested. Dominant, noncycling females initiated a higher percentage of approach and displace behaviors than both cycling and noncycling, subordinate elephants. Subordinate, noncycling elephants displayed the highest percentage of ears erect. Social rank drives the interactions of ex situ female African elephants more than ovarian cyclicity status. Thus, behavioral interactions cannot be used to predict which cycling elephants are most likely to become acyclic.

Freeman, E.W., Whyte, I., Brown, J.L., 2009. Reproductive evaluation of elephants culled in Kruger National Park, South Africa between 1975 and 1995. African Journal of Ecology 47, 192-201.
Abstract: To reduce elephant densities and preserve biological diversity, 14,629 elephants were culled from Kruger National Park, South Africa (1967-1999). Data were catalogued between 1975 and 1996 on 2737 male and female elephants, including pregnancy and lactational status for 1620 females (>= 5 years of age) and, uterine and/or ovarian characteristics for 1279. This study used these data to investigate the effects of age and precipitation on reproduction. The youngest age of conception was 8 years (n = 6) and by 12 years of age all females were sexually mature. From the age of 14 years, the percentage of reproductively active females (pregnant and/or lactating) was > 90%; however, this percentage declined when females reached 50 years of age. Overall, one-tenth of females were nonreproductive (not pregnant or lactating) at any given time, mostly in the youngest (< 15 years) and oldest (> 50 years) age classes. Eighteen (3.3%) of the nonpregnant females had reproductive tract pathologies, including endometrial, uterine or ovarian cysts. There was a seasonal distribution of mating activity that correlated with the rainy season. As has been demonstrated in other populations of free-ranging African elephants, most of the females in Kruger National Park were reproductively active; however, age and climate affected reproductive activity.

Freeman, E.W., Guagnano, G., Olson, D., Keele, M., Brown, J.L., 2009. Social factors influence ovarian acyclicity in captive African elephants (Loxodonta africana). Zoo. Biol. 28, 1-15.
Abstract: Nearly one-third of reproductive age African elephants in North America that are hormonally monitored fail to exhibit estrous cycle activity, which exacerbates the nonsustainability of the captive population. Three surveys were distributed to facilities housing female African elephants to determine how social and environmental variables contribute to cyclicity problems. Forty-six facilities returned all three surveys providing information on 90% of the SSP population and 106 elephants (64 cycling, 27 noncycling and 15 undetermined). Logistic analyses found that some physiological and social history variables were related to ovarian acyclicity. Females more likely to be acyclic had a larger body mass index and had resided longer at a facility with the same herdmates. Results suggest that controlling the weight of an elephant might be a first step to helping mitigate estrous cycle problems. Data further show that transferring females among facilities has no major impact on ovarian activity. Last, social status appears to impact cyclicity status; at 19 of 21 facilities that housed both cycling and noncycling elephants, the dominant female was acyclic. Further studies on how social and environmental dynamics affect hormone levels in free-living, cycling elephants are needed to determine whether acyclicity is strictly a captivity-related phenomenon

Freeman, E.W., Schulte, B.A., Brown, J.L., 2009. Using behavioral observations and keeper questionnaires to assess social relationships among captive female African elephants
60. Zoo. Biol. 28, 1-14.
Abstract: Free-ranging African elephants are highly social animals that live in a society where age, size, kinship, and disposition all contribute to social rank. Although captive elephant herds are small and largely comprises of unrelated females, dominance hierarchies are common. The goal of this study was to delineate how the behavior of captive female African elephants varies with respect to age and social rank based on a combination of keeper questionnaires and behavioral observations. "Body movements" and "trunk to" behaviors of 33 nonpregnant female African elephants housed at 14 North American zoos were recorded over 8 hr. Keepers at each facility also rated each elephant based on a series of questions about interactions with herdmates. The assessment of social rank based on observations correlated strongly with ranks assigned by keepers via the questionnaires. Observations and questionnaire responses indicated that body weight of the female, and to a lesser extent age, were significantly related to rates and types of "body movements" and that these demographic factors dictate the captive elephant hierarchy, similar to that observed in the wild. Many of the observed "body movements," such as back away, displace, push, and present, were correlated with keeper questionnaire responses about elephant interactions. However, none of the "trunk to" behaviors were related to age, size, or questionnaire responses even though they occurred frequently. In conclusion, we demonstrated that short-term behavioral observations and keeper questionnaires provided similar behavioral profiles for female African elephants housed in North American zoos. Zoo Biol 28:1-14, 2009. (c) 2009 Wiley-Liss, Inc

Fulka, J., Jr., Loi, P., Ptak, G., Fulka, H., John, J.S., 2009. Hope for the mammoth? Cloning Stem Cells 11, 1-4.

Hakeem, A.Y., Sherwood, C.C., Bonar, C.J., Butti, C., Hof, P.R., Allman, J.M., 2009. Von Economo neurons in the elephant brain. Anat. Rec. (Hoboken. ) 292, 242-248.
Abstract: Von Economo neurons (VENs), previously found in humans, all of the great ape species, and four cetacean species, are also present in African and Indian elephants. The VENs in the elephant are primarily found in similar locations to those in the other species. They are most abundant in the frontoinsular cortex (area FI) and are also present at lower density in the anterior cingulate cortex. Additionally, they are found in a dorsolateral prefrontal area and less abundantly in the region of the frontal pole. The VEN morphology appears to have arisen independently in hominids, cetaceans, and elephants, and may reflect a specialization for the rapid transmission of crucial social information in very large brains

Hermes, R., Behr, B., Hildebrandt, T.B., Blottner, S., Sieg, B., Frenzel, A., Knieriem, A., Saragusty, J., Rath, D., 2009. Sperm sex-sorting in the Asian elephant (Elephas maximus). Anim Reprod. Sci. 112, 390-396.
Abstract: In captive Asian elephants, there is a strong need for production of female offspring to enhance reproduction, counter premature aging processes in female animals and reduce challenging management situations derived from husbandry of several bulls in one institution. Artificial insemination of flow cytometrically sex-sorted spermatozoa offers the possibility to predetermine the sex of offspring with high accuracy. The aims of this study were to determine a suitable semen extender and basic parameters for flow cytometrical sex-sorting of Asian elephant spermatozoa. In total 18 semen samples were collected by manual rectal stimulation from one bull. Sperm quality parameters and sex sortability of spermatozoa were evaluated after dilution in three semen extenders (MES-HEPES-skim milk, MES-HEPES, TRIS-citric acid) and DNA staining. MES-HEPES-skim milk was the only semen extender found suitable to sex Asian elephant spermatozoa. From 18 ejaculates collected, 12 were successfully sorted with a purity of 94.5+/-0.7% at an average sort rate of 1945.5+/-187.5 spermatozoa per second. Sperm integrity, progressive and total motility were 42.6+/-3.9%, 48.1+/-3.3%, 59.4+/-3.8% after DNA labelling, and 64.8+/-3.2%, 58.0+/-5.0%, 70.8+/-4.4% after sorting, respectively. After liquid storage of sorted spermatozoa for 12h at 4 degrees C, sperm integrity, progressive and total motility were 46.4+/-5.2%, 32.2+/-4.2% and 58.2+/-3.9%, respectively. The obtained results provide a promising base to inseminate Asian elephants with sexed semen

Leighty, K.A., Soltis, J., Wesolek, C.M., Savage, A., Mellen, J., Lehnhardt, J., 2009. GPS determination of walking rates in captive African elephants (Loxodonta africana)
79. Zoo. Biol. 28, 16-28.
Abstract: The movements of elephants in captivity have been an issue of concern for animal welfare activists and zoological professionals alike in recent years. In order to fully understand how movement rates reflect animal welfare, we must first determine the exact distances these animals move in the captive environment. We outfitted seven adult female African elephants (Loxodonta africana) at Disney's Animal Kingdom with collar-mounted global positioning recording systems to document their movement rates while housed in outdoor guest viewing habitats. Further, we conducted preliminary analyses to address potential factors impacting movement rates including body size, temperature, enclosure size, and social grouping complexity. We found that our elephants moved at an average rate of 0.409+/-0.007 km/hr during the 9-hr data collection periods. This rate translates to an average of 3.68 km traveled during the observation periods, at a rate comparable to that observed in the wild. Although movement rate did not have a significant relationship with an individual's body size in this herd, the movements of four females demonstrated a significant positive correlation with temperature. Further, females in our largest social group demonstrated a significant increase in movement rates when residing in larger enclosures. We also present preliminary evidence suggesting that increased social group complexity, including the presence of infants in the herd, may be associated with increased walking rates, whereas factors such as reproductive and social status may constrain movements

Mason, G.J., Veasey, J.S., 2009. How should the psychological well-being of zoo elephants be objectively investigated?
47. Zoo. Biol.
Abstract: Animal welfare (sometimes termed "well-being") is about feelings - states such as "suffering" or "contentment" that we can infer but cannot measure directly. Welfare indices have been developed from two main sources: studies of suffering humans, and of research animals deliberately subjected to challenges known to affect emotional state. We briefly review the resulting indices here, and discuss how well they are understood for elephants, since objective welfare assessment should play a central role in evidence-based elephant management. We cover behavioral and cognitive responses (approach/avoidance; intention, redirected and displacement activities; vigilance/startle; warning signals; cognitive biases, apathy and depression-like changes; stereotypic behavior); physiological responses (sympathetic responses; corticosteroid output - often assayed non-invasively via urine, feces or even hair; other aspects of HPA function, e.g. adrenal hypertrophy); and the potential negative effects of prolonged stress on reproduction (e.g. reduced gametogenesis; low libido; elevated still-birth rates; poor maternal care) and health (e.g. poor wound-healing; enhanced disease rates; shortened lifespans). The best validated, most used welfare indices for elephants are corticosteroid outputs and stereotypic behavior. Indices suggested as valid, partially validated, and/or validated but not yet applied within zoos include: measures of preference/avoidance; displacement movements; vocal/postural signals of affective (emotional) state; startle/vigilance; apathy; salivary and urinary epinephrine; female acyclity; infant mortality rates; skin/foot infections; cardio-vascular disease; and premature adult death. Potentially useful indices that have not yet attracted any validation work in elephants include: operant responding and place preference tests; intention and vacuum movements; fear/stress pheromone release; cognitive biases; heart rate, pupil dilation and blood pressure; corticosteroid assay from hair, especially tail-hairs (to access endocrine events up to a year ago); adrenal hypertrophy; male infertility; prolactinemia; and immunological changes. Zoo Biol 28:1-19, 2009. (c) 2009 Wiley-Liss, Inc

Owens, M.J., Owens, D., 2009. Early age reproduction in female savanna elephants (Loxodonta africana) after severe poaching. African Journal of Ecology 47, 214-222.
Abstract: A 10-year study revealed that after severe poaching (> 93% killed) of elephants (Loxodonta africana) in Zambia's North Luangwa National Park (NLNP) during the 1970s and 1980s, the age of reproduction in females was greatly reduced. Fifty-eight per cent of births were delivered by females aged 8.5-14 years, an age at which elephants were reported to be sexually immature in nearby South Luangwa National Park (SLNP) before poaching. The mean age of females at first birth (AFB) (1993, 1994) was 11.3 years. Prior to poaching, the mean age AFB in SLNP was 16 years. The NLNP age structure and sex ratio were skewed, mean family unit size was reduced, and 37% of family units contained no females older than 15 years. Twenty-eight per cent of family units were comprised entirely of a single mother and her calf, and 8% of units consisted only of orphans who would have been considered sexually immature prior to poaching. Only 6% of survivors were older than 20 years, the age at which females in little-poached populations generally become most reproductively active. After a community-based conservation programme and the UN-CITES ban on the ivory trade were introduced, no elephants were recorded killed. In spite of a high reproductive rate, 6 years after poaching decreased, the density of the NLNP population had not increased, supporting predictions that the removal of older matriarchs from family units will have serious consequences on the recovery of this species.

Pinter-Wollman, N., Isabell, L.A., Hart, L.A., 2009. Assessing translocation outcome: Comparing behavioral and physiological aspects of translocated and resident African elephants (Loxodonta africana). Biological Conservation 142, 1116-1124.
Abstract: Evaluating translocation outcomes is important for improving wildlife management and conservation actions. Often, when quick decisions need to be made and long-lived animals with slow reproductionrates are translocated, traditional assessment methods such as long-term survival and reproductive successcannot be used for assessing translocation outcomes. Thus, alternative, seldom used, measures suchas comparing the behavior and physiology of translocated animals to those of local residents should beemployed to assess the translocated animals' acclimation to their new home. Here we monitored the survival,physiology, and behavior of translocated African elephants (Loxodonta africana) and comparedthese measures to the local resident population at the release site. Adult male and female translocatedelephants' death rates were higher than those of the local population. Furthermore, the mortality rateof translocated adult males and calves was greater than expected based on their proportion in the translocatedelephant population. No difference was found in stress hormone levels between the two populations,but the body condition of the translocated elephants was significantly poorer than that of the localpopulation throughout the study period. The behavioral time budgets of the translocated elephants convergedwith those of the local population over time. Finally, translocated elephants utilized habitat thatwas similar to their source site (hills and permanent rivers) more than did the local population. Based on these findings we recommend careful consideration of timing, release location, and individuals targetedin future elephant translocations. More broadly, we introduce and explore seldom used translocation assessment techniques.

Saragusty, J., Hermes, R., Goritz, F., Schmitt, D.L., Hildebrandt, T.B., 2009. Skewed birth sex ratio and premature mortality in elephants. Anim Reprod. Sci. 115, 247-254.
Abstract: Sex allocation theories predict equal offspring number of both sexes unless differential investment is required or some competition exists. Left undisturbed, elephants reproduce well and in approximately even numbers in the wild. We report an excess of males are born and substantial juvenile mortality occurs, perinatally, in captivity. Studbook data on captive births (CB, n=487) and premature deaths (PD, <5 years of age; n=164) in Asian and African elephants in Europe and North America were compared with data on Myanmar timber (Asian) elephants (CB, n=3070; PD, n=738). Growth in CB was found in three of the captive populations. A significant excess of male births occurred in European Asian elephants (ratio: 0.61, P=0.044) and in births following artificial insemination (0.83, P=0.003), and a numerical inclination in North American African elephants (0.6). While juvenile mortality in European African and Myanmar populations was 21-23%, it was almost double (40-45%) in all other captive populations. In zoo populations, 68-91% of PD were within 1 month of birth with stillbirth and infanticide being major causes. In Myanmar, 62% of juvenile deaths were at >6 months with maternal insufficient milk production, natural hazards and accidents being the main causes. European Asian and Myanmar elephants PD was biased towards males (0.71, P=0.024 and 0.56, P<0.001, respectively). The skewed birth sex ratio and high juvenile mortality hinder efforts to help captive populations become self-sustaining. Efforts should be invested to identify the mechanism behind these trends and seek solutions for them.

Saragusty, J., Hildebrandt, T.B., Behr, B., Knieriem, A., Kruse, J., Hermes, R., 2009. Successful cryopreservation of Asian elephant (Elephas maximus) spermatozoa. Anim Reprod. Sci. 115, 255-266.
Abstract: Reproduction in captive elephants is low and infant mortality is high, collectively leading to possible population extinction. Artificial insemination was developed a decade ago; however, it relies on fresh-chilled semen from just a handful of bulls with inconsistent sperm quality. Artificial insemination with frozen-thawed sperm has never been described, probably, in part, due to low semen quality after cryopreservation. The present study was designed with the aim of finding a reliable semen freezing protocol. Screening tests included freezing semen with varying concentrations of ethylene glycol, propylene glycol, trehalose, dimethyl sulfoxide and glycerol as cryoprotectants and assessing cushioned centrifugation, rapid chilling to suprazero temperatures, freezing extender osmolarity, egg yolk concentration, post-thaw dilution with cryoprotectant-free BC solution and the addition of 10% (v/v) of autologous seminal plasma. The resulting optimal freezing protocol uses cushioned centrifugation, two-step dilution with isothermal 285 m Osm/kg Berliner Cryomedium (BC) with final glycerol concentration of 7% and 16% egg yolk, and freezing in large volume by the directional freezing technique. After thawing, samples are diluted 1:1 with BC solution. Using this protocol, post-thaw evaluations results were: motility upon thawing: 57.2+/-5.4%, motility following 30 min incubation at 37 degrees C: 58.5+/-6.0% and following 3h incubation: 21.7+/-7.6%, intact acrosome: 57.1+/-5.2%, normal morphology: 52.0+/-5.8% and viability: 67.3+/-6.1%. With this protocol, good quality semen can be accumulated for future use in artificial inseminations when and where needed

Sherwood, C.C., Stimpson, C.D., Butti, C., Bonar, C.J., Newton, A.L., Allman, J.M., Hof, P.R., 2009. Neocortical neuron types in Xenarthra and Afrotheria: implications for brain evolution in mammals. Brain Struct. Funct. 213, 301-328.
Abstract: Interpreting the evolution of neuronal types in the cerebral cortex of mammals requires information from a diversity of species. However, there is currently a paucity of data from the Xenarthra and Afrotheria, two major phylogenetic groups that diverged close to the base of the eutherian mammal adaptive radiation. In this study, we used immunohistochemistry to examine the distribution and morphology of neocortical neurons stained for nonphosphorylated neurofilament protein, calbindin, calretinin, parvalbumin, and neuropeptide Y in three xenarthran species-the giant anteater (Myrmecophaga tridactyla), the lesser anteater (Tamandua tetradactyla), and the two-toed sloth (Choloepus didactylus)-and two afrotherian species-the rock hyrax (Procavia capensis) and the black and rufous giant elephant shrew (Rhynchocyon petersi). We also studied the distribution and morphology of astrocytes using glial fibrillary acidic protein as a marker. In all of these species, nonphosphorylated neurofilament protein-immunoreactive neurons predominated in layer V. These neurons exhibited diverse morphologies with regional variation. Specifically, high proportions of atypical neurofilament-enriched neuron classes were observed, including extraverted neurons, inverted pyramidal neurons, fusiform neurons, and other multipolar types. In addition, many projection neurons in layers II-III were found to contain calbindin. Among interneurons, parvalbumin- and calbindin-expressing cells were generally denser compared to calretinin-immunoreactive cells. We traced the evolution of certain cortical architectural traits using phylogenetic analysis. Based on our reconstruction of character evolution, we found that the living xenarthrans and afrotherians show many similarities to the stem eutherian mammal, whereas other eutherian lineages display a greater number of derived traits

Soltis, J., 2009. Vocal communication in African Elephants (Loxodonta africana)
61. Zoo. Biol. 28, 1-18.
Abstract: Research on vocal communication in African elephants has increased in recent years, both in the wild and in captivity, providing an opportunity to present a comprehensive review of research related to their vocal behavior. Current data indicate that the vocal repertoire consists of perhaps nine acoustically distinct call types, "rumbles" being the most common and acoustically variable. Large vocal production anatomy is responsible for the low-frequency nature of rumbles, with fundamental frequencies in the infrasonic range. Additionally, resonant frequencies of rumbles implicate the trunk in addition to the oral cavity in shaping the acoustic structure of rumbles. Long-distance communication is thought possible because low-frequency sounds propagate more faithfully than high-frequency sounds, and elephants respond to rumbles at distances of up to 2.5 km. Elephant ear anatomy appears designed for detecting low frequencies, and experiments demonstrate that elephants can detect infrasonic tones and discriminate small frequency differences. Two vocal communication functions in the African elephant now have reasonable empirical support. First, closely bonded but spatially separated females engage in rumble exchanges, or "contact calls," that function to coordinate movement or reunite animals. Second, both males and females produce "mate attraction" rumbles that may advertise reproductive states to the opposite sex. Additionally, there is evidence that the structural variation in rumbles reflects the individual identity, reproductive state, and emotional state of callers. Growth in knowledge about the communication system of the African elephant has occurred from a rich combination of research on wild elephants in national parks and captive elephants in zoological parks. Zoo Biol 28:1-18, 2009. (c) 2009 Wiley-Liss, Inc

Steinetz, B., Lasano, S., de Haas van, D.F., Glickman, S., Bergfelt, D., Santymire, R., Songsassen, N., Swanson, W., 2009. Relaxin concentrations in serum and urine of endangered and crazy mixed-up species
66. Ann. N. Y. Acad. Sci. 1160, 179-185.
Abstract: The human population explosion has pushed many mammalian wildlife species to the brink of extinction. Conservationists are increasingly turning to captive breeding as a means of preserving the gene pool. We previously reported that serum immunoactive relaxin provided a reliable means of distinguishing between true and pseudopregnancy in domestic dogs, and this method has since been found to be a reliable indicator of true pregnancy in endangered Asian and African elephants and Sumatran rhinoceroses. Our canine relaxin radioimmunoassay (RIA) has now been adapted and validated to measure relaxin in the serum and urine of felids, including domestic and wild species. Moreover, a commercially available canine serum relaxin kit (Witness) Relaxin Kit; Synbiotics, San Diego, CA), has been adapted for reliable detection of relaxin in urine of some felid species. Our porcine relaxin RIA has also been utilized to investigate the role of relaxin in reproductive processes of the spotted hyena, a species in which the female fetuses are severely masculinized in utero. Indeed, this species might well now be extinct were it not for the timely secretion of relaxin to enable copulation and birth of young through the clitoris. Additional studies have suggested relaxin may be a useful marker of pregnancy in the northern fur seal and the maned wolf (the former species has been designated as "depleted" and the latter as "near threatened"). Given appropriate immunoassay reagents, relaxin determination in body fluids thus provides a powerful tool for conservationists and biologists investigating reproduction in a wide variety of endangered and exotic species

Thitaram, C., Chansitthiwet, S., Pongsopawijit, P., Brown, J.L., Wongkalasin, W., Daram, P., Roongsri, R., Kalmapijit, A., Mahasawangkul, S., Rojanasthien, S., Colenbrander, B., van der Weijden, G.C., van Eerdenburg, F.J., 2009. Use of genital inspection and female urine tests to detect oestrus in captive Asian elephants
116. Anim Reprod. Sci. 115, 267-278.
Abstract: Captive Asian elephant (Elephas maximus) populations are decreasing due to low birth rates compared to wild elephants. Improving oestrous detection in female elephants is required to ensure successful mating in captive and semi-captive herds. Responsive behaviours of eight semi-captive bull elephants to the uro-genital area (genital inspection test) or urinary pheromones (urine test) of 14 female elephants throughout the oestrous cycle were evaluated. Weekly blood samples were collected for 27 consecutive months (14 months for the genital inspection test and 13 months for the urine test) from female elephants to characterize the patterns of circulating progestagen. Responsive behaviours of bulls were compared between females in the follicular versus the luteal phase of the cycle. The sensitivity and specificity of the genital inspection test were 65% and 68%, while those of the urine test were 52% and 61%, respectively. The bulls showed significantly higher "genital inspection", "flehmen from genital area" and "trunk on back" behaviours during the genital inspection test, and "flehmen" behaviours during the urine test in oestrous than in non-oestrous females. In sum, this study showed that monitoring sexual behaviours of Asian elephant bulls towards females or their urine can be used to detect the oestrous period. Although the sensitivity and specificity of both tests were not as high as expected, still, these methods appear to be more efficient at detecting oestrous than traditional methods based on mahout estimations of female receptivity. The use of genital inspection and urine tests may lead to more successful matings and thus to creating self-sustaining populations of captive elephants in range countries

Thitaram, C., Pongsopawijit, P., Chansitthiwet, S., Brown, J.L., Nimtragul, K., Boonprasert, K., Homkong, P., Mahasawangkul, S., Rojanasthien, S., Colenbrander, B., van der Weijden, G.C., van Eerdenburg, F.J., 2009. Induction of the ovulatory LH surge in Asian elephants (Elephas maximus): a novel aid in captive breeding management of an endangered species
49. Reprod. Fertil. Dev. 21, 672-678.
Abstract: A unique feature of the reproductive physiology of Asian elephants (Elephas maximus) is the occurrence of two LH surges before ovulation, instead of one. An anovulatory LH (anLH) surge, the function of which is unknown, occurs consistently 3 weeks before the ovulatory LH (ovLH) surge that induces ovulation. Thus, the ability to induce an ovLH surge would be useful for scheduling natural mating or artificial insemination. The present study tested the efficacy of a gonadotrophin-releasing hormone agonist (GnRH-Ag) to induce LH surges during the follicular phase of the oestrous cycle, which resulted in varied LH responses, but generally none were as high as previously documented natural surges. Thus, for the ovulation-induction trials, nine females were administered 80 microg GnRH-Ag intravenously at three time periods during the oestrous cycle, namely the anovulatory follicular phase, the ovulatory follicular phase and the luteal phase. During the late anovulatory follicular phase, nine of 10 females (90%) responded with an immediate LH surge followed 15-22 days later by an ovLH surge or a post-ovulatory increase in progestagens. In contrast, despite responding to the GnRH-Ag with an immediate increase in LH, none of the females treated during other periods of the oestrous cycle exhibited subsequent ovLH surges. One cow got pregnant from natural mating following the induced ovLH surge. In conclusion, ovLH induction is possible using a GnRH-Ag, but only during a specific time of the anovulatory follicular phase

Thongtip, N., Mahasawangkul, S., Thitaram, C., Pongsopavijitr, P., Kornkaewrat, K., Pinyopummin, A., Angkawanish, T., Jansittiwate, S., Rungsri, R., Boonprasert, K., Wongkalasin, W., Homkong, P., Dejchaisri, S., Wajjwalku, W., Saikhun, K., 2009. Successful artificial insemination in the Asian elephant (Elephas maximus) using chilled and frozen-thawed semen. Reprod. Biol. Endocrinol. 7, 75.
Abstract: BACKGROUND: Artificial insemination (AI) using frozen-thawed semen is well established and routinely used for breeding in various mammalian species. However, there is no report of the birth of elephant calves following AI with frozen-thawed semen. The objective of the present study was to investigate the fertilizing ability of chilled and frozen-thawed semen in the Asian elephant following artificial insemination (AI). METHODS: Semen samples were collected by from 8 bulls (age range, 12-to 42-years) by manual stimulation. Semen with high quality were either cooled to 4 degrees C or frozen in liquid nitrogen (-196 degrees C) before being used for AI. Blood samples collected from ten elephant females (age range, 12-to 52-years) were assessed for estrus cycle and elephants with normal cycling were used for AI. Artificial insemination series were conducted during 2003 to 2008; 55 and 2 AI trials were conducted using frozen-thawed and chilled semen, respectively. Pregnancy was detected using transrectal ultrasonography and serum progestagen measurement. RESULTS: One female (Khod) inseminated with chilled semen became pregnant and gave birth in 2007. The gestation length was 663 days and the sex of the elephant calf was male. One female (Sao) inseminated with frozen-thawed semen showed signs of pregnancy by increasing progestagen levels and a fetus was observed for 5 months by transrectal ultrasonography. CONCLUSION: This is the first report showing pregnancy following AI with frozen-thawed semen in the Asian elephant. Successful AI in the Asian elephant using either chilled or frozen-thawed semen is a stepping stone towards applying this technology for genetic improvement of the elephant population.

Trimble, M.J., Ferreira, S.M., van Aarde, R.J., 2009. Drivers of megaherbivore demographic fluctuations: inference from elephants. Journal of Zoology 1-9.
Abstract: Environmentally induced variation in survival and fecundity generates demographic fluctuations that affect population growth rate. However, a general pattern of the comparative influence of variation in fecundity and juvenile survival on elephant population dynamics has not been investigated at a broad scale. We evaluated the relative importance of conception, gestation, first year survival and subsequent survivorship for controlling demographic variation by exploring the relationship between past environmental conditions determined by integrated normalized difference vegetation index (INDVI) and the shape of age distributions at 17 sites across Africa. We showed that, generally, INDVI during gestation best explained anomalies in age structure. However, in areas with low mean annual rainfall, INDVI during the first year of life was critical. The results challenge Eberhardt's paradigm for population analysis that suggests that populations respond to limited resource availability through a sequential decrease in juvenile survival, reproductive rate and adult survival. Contrastingly, elephants appear to respond first through a reduction in reproductive rate. We conclude that this discrepancy is likely due to the evolutionary significance of extremely large body size - an adaptation that increases survival rate but decreases reproductive potential. Other megaherbivores may respond similarly to resource limitation due to similarities in population dynamics. Knowing how vital rates vary with changing environmental conditions will permit better forecasts of the trajectories of megaherbivore populations.

Tripp, K.M., Dubois, M., Delahaut, P., Verstegen, J.P., 2009. Detection and identification of plasma progesterone metabolites in the female Florida manatee (Trichechus manatus latirostris) using GC/MS/MS
54. Theriogenology 72, 365-371.
Abstract: Florida manatees (Trichechus manatus latirostris) have relatively low peripheral concentrations of progesterone (P4). The objective of this study was to determine if these relatively low P4 concentrations are associated with a high ratio of progestin metabolites and to document metabolite concentrations from individual blood samples obtained from manatees during diestrus or pregnancy. Metabolites known to exist in elephants-terrestrial manatee relatives-were targeted. These included 5alpha-reduced progestins (5alpha-pregnane-3,20-dione [5alpha-DHP] and 3alpha-hydroxy-5alpha-pregnan-20-one [5alpha-P3-OH]) and 17alpha-hydroxyprogesterone (17alpha-OHP), which occurs in Asian elephants. An additional, inactive metabolite, 20alpha-hydroxyprogesterone (20alpha-OHP), indicative of P4 overproduction, was also targeted. Progesterone itself was the predominant progestin detected in pregnant and nonpregnant manatee plasma (n = 10) using gas chromatography-mass spectrometry with tandem quadrupole detectors (GC/MS/MS). Progesterone concentrations in pregnant females varied from early (moderate to high) through mid and late (low) pregnancy. Progesterone concentrations ranged from low to high in nonpregnant, nonlactating females. The most commonly detected metabolite was 5alpha-P3-OH (n = 7), which occurred in pregnant (lower limit of detection [LLOD] to high) and nonpregnant (trace to high) females. The 5alpha-DHP metabolite was also detected in pregnant (LLOD to moderate) and nonpregnant (low) females. The 17alpha-OHP metabolite was not detected in any tested female. The 20alpha-OHP metabolite was detected in one nonpregnant, nonlactating, captive female (LLOD). Metabolites were most prevalent during early pregnancy, concurrent with maximum P4 concentrations. Based on their concentrations in peripheral circulation, we inferred that these metabolites may have, opposite to elephants, a limited physiologic role during luteal, pregnant, and nonpregnant phases in the manatee

Wallis, M., 2009. Prolactin in the Afrotheria: characterization of genes encoding prolactin in elephant (Loxodonta africana), hyrax (Procavia capensis) and tenrec (Echinops telfairi). J. Endocrinol. 200, 233-240.
Abstract: Pituitary prolactin shows an episodic pattern of molecular evolution, with occasional short bursts of rapid change imposed on a generally rather slow evolutionary rate. In mammals, episodes of rapid change occurred in the evolution of primates, cetartiodactyls, rodents and the elephant. The bursts of rapid evolution in cetartiodactyls and rodents were followed by duplications of the prolactin gene that gave rise to large families of prolactin-related proteins including placental lactogens, while in primates the burst was followed by corresponding duplications of the related GH gene. The position in elephant is less clear. Extensive data relating to the genomic sequences of elephant and two additional members of the group Afrotheria are now available, and have been used here to characterize the prolactin genes in these species and explore whether additional prolactin-related genes are present. The results confirm the rapid evolution of elephant (Loxodonta africana) prolactin - the sequence of elephant prolactin is substantially different from that predicted for the ancestral placental mammal. Hyrax (Procavia capensis) prolactin is even more divergent but tenrec (Echinops telfairi) prolactin is strongly conserved. No evidence was obtained from searches of public databases for additional genes encoding prolactin-like proteins in any of these species. Detailed analysis of evolutionary rates, and other factors, indicates that the episode of rapid change in hyrax, and probably elephant, was adaptive, though the nature of the associated biological change(s) is not clear

Weissenbock, N.M., Schwammer, H.M., Ruf, T., 2009. Estrous synchrony in a group of African elephants (Loxodonta africana) under human care. Anim Reprod. Sci. 113, 322-327.
Abstract: Synchrony of estrous, and consequently of conception and birth of young, may be of adaptive significance for certain mammals. Among the species in which estrous synchrony has been suspected several times are elephants, but clear evidence is still missing. We determined estrous cycles of African elephants (Loxodonta africana) (n=4) at the Vienna Zoo, Austria, between June 2003 and January 2006 by measuring serum progesterone levels from weekly blood samples. Except for the dominant female when she was intensively lactating, all animals showed clear cycles or progesterone release with a mean period of 105.3+/-15.37 days. For most of the study period, estrous cycles were asynchronous between females. However, after re-occurrence of the progesterone cycle in the dominant female following the first period of lactation, all four females showed high synchrony of progesterone release over the two subsequent cycles. Large changes in individual period lengths indicated that synchronization was due to the adjustment of cycle length in subdominants to that of the dominant female. We used a bootstrap procedure, based on resampling measured times of progesterone peaks, to determine if this apparent synchrony could have been caused by chance alone. This statistical analysis indicated that between-individual variances of the timing of progesterone peaks were much smaller that to be expected by chance (P=0.009). This finding represents the first evidence for estrous synchrony between elephants. We discuss various hypotheses to explain the biological function of cycle synchrony in elephants

Archie, E.A., Maldonado, J.E., Hollister-Smith, J.A., Poole, J.H., Moss, C.J., Fleischer, R.C., Alberts, S.C., 2008. Fine-scale population genetic structure in a fission-fusion society. Mol. Ecol. 17, 2666-2679.
Abstract: Nonrandom patterns of mating and dispersal create fine-scale genetic structure in natural populations - especially of social mammals - with important evolutionary and conservation genetic consequences. Such structure is well-characterized for typical mammalian societies; that is, societies where social group composition is stable, dispersal is male-biased, and males form permanent breeding associations in just one or a few social groups over the course of their lives. However, genetic structure is not well understood for social mammals that differ from this pattern, including elephants. In elephant societies, social groups fission and fuse, and males never form permanent breeding associations with female groups. Here, we combine 33 years of behavioural observations with genetic information for 545 African elephants (Loxodonta africana), to investigate how mating and dispersal behaviours structure genetic variation between social groups and across age classes. We found that, like most social mammals, female matrilocality in elephants creates co-ancestry within core social groups and significant genetic differentiation between groups (Phi(ST) = 0.058). However, unlike typical social mammals, male elephants do not bias reproduction towards a limited subset of social groups, and instead breed randomly across the population. As a result, reproductively dominant males mediate gene flow between core groups, which creates cohorts of similar-aged paternal relatives across the population. Because poaching tends to eliminate the oldest elephants from populations, illegal hunting and poaching are likely to erode fine-scale genetic structure. We discuss our results and their evolutionary and conservation genetic implications in the context of other social mammals

Aupperle, H., Reischauer, A., Bach, F., Hildebrandt, T., Goritz, F., Jager, K., Scheller, R., Klaue, H.J., Schoon, H.A., 2008. Chronic endometritis in an Asian elephant (Elephas maximus). J. Zoo. Wildl. Med. 39, 107-110.
Abstract: A 48-yr-old female Asian elephant with a history of pododermatitis developed recurrent hematuria beginning in 2002. Transrectal ultrasonography and endoscopic examination in 2004 identified the uterus as the source of hematuria and excluded hemorrhagic cystitis. Treatment with Desloreline implants, antibiotics, and homeopathic drugs led to an improved general condition of the elephant. In July 2005, the elephant was suddenly found dead. During necropsy, the severely enlarged uterus contained about 250 L of purulent fluid, and histopathology revealed ulcerative suppurative endometritis with high numbers of Streptococcus equi ssp. zooepidemicus and Escherichia coli identified on aerobic culture. Additional findings at necropsy included: multifocal severe pododermatitis, uterine leiomyoma, and numerous large calcified areas of abdominal fat necrosis. Microbiologic culture of the pododermatitis lesion revealed the presence of Streptococcus agalactiae, Streptococcus equi ssp. zooepidemicus, Staphylococcus sp., Corynebacterium sp., and Entercoccus sp

Drews, B., Hermes, R., Goritz, F., Gray, C., Kurz, J., Lueders, I., Hildebrandt, T.B., 2008. Early embryo development in the elephant assessed by serial ultrasound examinations. Theriogenology 69, 1120-1128.
Abstract: The elephant has an extraordinary long pregnancy, lasting 21 months. However, knowledge on embryo development is limited. To date, only single morphological observations of elephant embryo development associated with placentation are available, all lacking correlation to gestational age. The present study describes morphological characteristics of early embryo development in the elephant with exact biometric staging. Six pregnancies in five Asian and one African elephants with known conception dates were followed by 2D and 3D ultrasound, covering the embryonic period from ovulation to day 116 post-ovulation. The embryonic vesicle was earliest observed was on day 50 p.o. The proper embryo was not detected until day 62 p.o. Embryonic heartbeat was first observed on day 71 p.o. The allantois, which became visible as a single sacculation on day 71 p.o. was subdivided in four compartments on day 76 p.o. By day 95 p.o., head, rump, front and hind legs were clearly distinguished. Between days 95 and 103 p.o. the choriovitelline placenta was replaced by the chorioallantoic placenta. A physiological midgut herniation was transiently present between days 95 and 116 p.o. On the basis of the late appearance of the embryonic vesicle, delayed implantation in the elephant is discussed. The study provides a coherent description of elephant embryonic development, formation of the extraembryonic organs and their role in placenta formation, all of which are of interest for both comparative evolutionary studies and the improvement of assisted reproduction techniques

Evans, K., Harris, S., 2008. Adolescence in male African elephants, Loxodonta africana, and the importance of sociality. Animal Behavior 76, 779-787.
Abstract: The degree of sociality during an animal's life changes as it modulates its behaviour to reflect different lifestages. Only a few species of mammal undergo a period of adolescence, but for these species it is probablyone of their most important life stages. It is when individuals acquire skills and develop relationships thatare of both immediate and long-term benefit to their survival and reproductive success, particularly in polygynous males in which sexual selection favours size and dominance. We collected focal and observationaldata on male African elephants in the Okavango Delta, Botswana, to assess behaviour and socialinteractions during adolescence. Adolescent males (10-15 and 16-20 years of age) were the most sociableage group, showing preferences for larger social groupings and being in closer proximity to other elephants;later adolescent males (ages 16-20) showed a tendency for higher social levels. Males of all agespreferred to have males 36 years of age as their nearest neighbour. We argue that this proximity to oldermales provides opportunities for males to learn from more experienced individuals. It has long been recognized that matriarchs are the repositories of social and ecological knowledge within elephant breedingherds: we suggest that mature males are reservoirs for such knowledge within bull society.

Gobush, K.S., Mutayoba, B.M., Wasser, S.K., 2008. Long-term impacts of poaching on relatedness, stress physiology, and reproductive output of adult female african elephants. Conserv. Biol. 22, 1590-1599.
Abstract: Widespread poaching prior to the 1989 ivory ban greatly altered the demographic structure of matrilineal African elephant (Loxodonta africana) family groups in many populations by decreasing the number of old, adult females. We assessed the long-term impacts of poaching by investigating genetic, physiological, and reproductive correlates of a disturbed social structure resulting from heavy poaching of an African elephant population in Mikumi National Park, Tanzania, prior to 1989. We examined fecal glucocorticoid levels and reproductive output among 218 adult female elephants from 109 groups differing in size, age structure, and average genetic relatedness over 25 months from 2003 to 2005. The distribution in group size has changed little since 1989, but the number of families with tusked old matriarchs has increased by 14.2%. Females from groups that lacked an old matriarch, first-order adult relatives, and strong social bonds had significantly higher fecal glucocorticoid values than those from groups with these features (all females R(2)= 0.31; females in multiadult groups R(2)= 0.46). Females that frequented isolated areas with historically high poaching risk had higher fecal glucocorticoid values than those in low poaching risk areas. Females with weak bonds and low group relatedness had significantly lower reproductive output (R(2)[U]=0.21). Females from disrupted groups, defined as having observed average group relatedness 1 SD below the expected mean for a simulated unpoached family, had significantly lower reproductive output than females from intact groups, despite many being in their reproductive prime. These results suggest that long-term negative impacts from poaching of old, related matriarchs have persisted among adult female elephants 1.5 decades after the 1989 ivory ban was implemented

Hermes, R., Saragusty, J., Schaftenaar, W., Goritz, F., Schmitt, D.L., Hildebrandt, T.B., 2008. Obstetrics in elephants. Theriogenology 70, 131-144.
Abstract: Obstetrics, one of the oldest fields in veterinary medicine, is well described and practiced in domestic and exotic animals. However, when providing care during elephant birth or dystocia, veterinary intervention options differ greatly from any domestic species, and are far more limited due to the dimensions and specific anatomy of the elephant reproductive tract. In addition, aging of captive elephant populations and advanced age of primiparous females make active birth management increasingly important. Intrauterine infection, uterine inertia and urogenital tract pathologies are emerging as major causes for dystocia, often leading to foetal and dam death. This paper reviews the current knowledge on elephant birth and the factors associated with dystocia. It then summarises recommendations for birth and dystocia management. As Caesarean section, the most common ultima ratio in domestic animal obstetrics, is lethal and therefore not an option in the elephant, non-invasive medical treatment, induction of the Fergusson reflex or the conscious decision to leave a retained foetus until it is expelled voluntarily, are key elements in elephant obstetrics. Surgical strategies such as episiotomy and foetotomy are sometimes inevitable in order to try to save the life of the dam, however, these interventions result in chronic post-surgical complications or even fatal outcome. Limited reliable data on serum calcium concentrations, and pharmacokinetics and effect of exogenous oestrogen, oxytocin, and prostaglandins during birth provide the scope of future research, necessary to advance scientific knowledge on obstetrics in elephants

Hollister-Smith, J.A., Alberts, S.C., Rasmussen, L.E.L., 2008. Do male African elephants, Loxodonta africana, signal musth via urine dribbling? Animal Behavior 76, 1829-1841.
Abstract: The phenomenon of musth in male elephants involves increased sexual activity, heightened aggression and nearly continuous dribbling of pungent smelling urine. Urine chemistry during musth is altered, suggesting that urine may signal the musth status of the individual. Signalling musth remotely may benefit individuals if it reduces the likelihood of physical confrontation between males, which can lead to injury and even death. Few studies, however, have asked whether and how male elephants respond to urine of other males. We tested two predictions of the hypothesis that urine signals musth status to male conspecifics: (1) that male African elephants differentiate musth and nonmusth urine, and (2) that males differentiate between urine dribbled during early and late musth. The second prediction stems from the observation that males lose weight and presumably body condition during musth. We conducted two related bioassays with 26 captive nonmusth males ranging from 13 to 52 years of age. In each assay, subjects were simultaneously presented with three urine samples (nonmusth, early musth, late musth), each from a different donor male, and a control. We found that subjects differentiated between musth and nonmusth samples using their vomeronasal organ system, but did not discriminate between the samples using their main olfactory system. Males did not differentiate early from late musth. In addition, we found that subject contextual factors, specifically age, dominance status and social grouping, significantly predicted response. We discuss these results within the framework of male elephant longevity and social relationships and their importance to reproductive success.

Lotfy, W.M., Brant, S.V., DeJong, R.J., Le, T.H., Demiaszkiewicz, A., Rajapakse, R.P., Perera, V.B., Laursen, J.R., Loker, E.S., 2008. Evolutionary origins, diversification, and biogeography of liver flukes (Digenea, Fasciolidae). American Journal of Tropical Medicine and Hygiene 79, 248-255.
Abstract: Fasciolid flukes are among the largest and best known digenetic trematodes and have considerable historical and veterinary significance. Fasciola hepatica is commonly implicated in causing disease in humans. The origins, patterns of diversification, and biogeography of fasciolids are all poorly known. We have undertaken a molecular phylogenetic study using 28S, internal transcribed spacer 1 and 2 (ITS-1 and ITS-2) of nuclear ribosomal DNA, and mitochondrial nicotinamide dehydrogenase subunit 1 (nad1) that included seven of the nine recognized species in the family. The fasciolids examined comprise a monophyletic group with the most basal species recovered from African elephants. We hypothesize fasciolids migrated from Africa to Eurasia, with secondary colonization of Africa. Fasciolids have been conservative in maintaining relatively large adult body size, but anatomical features of their digestive and reproductive systems are available. These flukes have been opportunistic, with respect to switching to new snail (planorbid to lymnaeid) and mammalian hosts and from intestinal to hepatic habitats within mammals

Lynch, V.J., Tanzer, A., Wang, Y., Leung, F.C., Gellersen, B., Emera, D., Wagner, G.P., 2008. Adaptive changes in the transcription factor HoxA-11 are essential for the evolution of pregnancy in mammals. Proc. Natl. Acad. Sci. U. S. A 105, 14928-14933.
Abstract: Evolutionary change in gene regulation can result from changes in cis-regulatory elements, leading to differences in the temporal and spatial expression of genes or in the coding region of transcription factors leading to novel functions or both. Although there is a growing body of evidence supporting the importance of cis-regulatory evolution, examples of protein-mediated evolution of novel developmental pathways have not been demonstrated. Here, we investigate the evolution of prolactin (PRL) expression in endometrial cells, which is essential for placentation/pregnancy in eutherian mammals and is a direct regulatory target of the transcription factor HoxA-11. Here, we show that (i) endometrial PRL expression is a derived feature of placental mammals, (ii) the PRL regulatory gene HoxA-11 experienced a period of strong positive selection in the stem-lineage of eutherian mammals, and (iii) only HoxA-11 proteins from placental mammals, including the reconstructed ancestral eutherian gene, are able to up-regulate PRL from the promoter used in endometrial cells. In contrast, HoxA-11 from the reconstructed therian ancestor, opossum, platypus, and chicken are unable to up-regulate PRL expression. These results demonstrate that the evolution of novel gene expression domains is not only mediated by the evolution of cis-regulatory elements but can also require evolutionary changes of transcription factor proteins themselves

Meyer, J., Goodwin, T., Schulte, B., 2008. Intrasexual chemical communication and social responses ofcaptive female African elephants. Animal Behavior 76, 163-174.
Abstract: In matrilineal societies, competition between females can occur within and between social units. Dominance hierarchies reduce costly conflicts when reliable cues of status are available, and reproductive condition may alter individual or group status. Female African elephants live in matriarchal groups with linear dominance hierarchies occurring within and between groups; elephants use chemical signals to mediate social interactions. If reproductive condition has important implications for inter- or intragroup behaviour, then females should discriminate between chemical signal sources that reveal reproductive condition. We examined whether trunk-tip contacts between females within a social group were related to phase of oestrus. Observations were conducted on 21 reproductively viable females at nine zoological facilities in North America. Females in the follicular phase received contacts to the urogenital region at a higher rate than did luteal phase females, and contacts increased with approaching ovulation. This supports the existence of an oestrous signal. We also examined whether an oestrous signal was evident by female investigation of urine collected from the luteal and follicular phases of unfamiliar conspecifics. Elephants responded to unfamiliar urine more than to the control, but response rates to the urine types did not differ. Females within a social unit detected differences in oestrus, but they did not show such discrimination to urinary signals from unfamiliar females. Further evaluation of the existence of a female-to-female oestrous pheromone requires assessing responses to urine from familiar individuals. Understanding the relationship between oestrous condition and dominance status can shed light on the adaptive value of sociality.

Meyers, D.A., Isaza, R., MacNeill, A. Evaluation of acute phase proteins for diagnosis of inflammation in Asian elephants ( Elephas maximus). Proc American Associaton of Zoo Veterinarians and Assoc of Reptile and Amphibian Veterinarians.  128. 2008. 11-10-2008.
Ref Type: Conference Proceeding
Abstract: In many domestic species, routine hematology assays are useful diagnostic tools to diagnose inflammatory conditions. Unlike other species, these hematologic tests apparently are insensitive indicators of inflammation in elephants.1 We studied a novel group of blood proteins, called acute phase proteins, which increase during inflammatory conditions, for their usefulness in diagnosing elephants with inflammatory diseases. Although these proteins currently are useful in humans and domestic animals, each species has a different set of important proteins that must be individually investigated.2 We tested several acute phase proteins (C-reactive protein, alpha-1 glycoprotein, alpha-1 antitrypsin, serum amyloid A, haptoglobin, fibrinogen, ceruloplasmin, and albumin) as well as complete blood counts, chemistry panels, serum protein electrophoresis, and 3-D gel electrophoresis to determine their usefulness for diagnosing different types of inflammatory conditions in Asian elephants (Elephas maximus). Animals with inflammatory conditions were classified as those individuals with known illnesses such as mycobacteriosis, arthritis, nail bed abscesses, and malignant tumors. Control animals were thoseanimals that were suspected to not have any inflammation and be healthy at the time of testing as determined by physical examination and obtaining a thorough medical history.
LITERATURE CITED
1. Lyashchenko, K., R. Greenwald, J. Esfandiari, J. Olsen, R. Ball, G. Dumonceaux, F. Dunker, C. Buckley, M.
Richard, S. Murray, J.B. Payeur, P. Anderson, J.M. Pollock, S. Mikota, M. Miller, D. Sofranko, and W.R.
Waters. 2006. Tuberculosis in Elephants: Antibody responses to defined antigens of Mycobacterium
tuberculosis, potential for early diagnosis, and monitoring of treatment. Clin. Vacc. Immunol. 13: 722-732.
2. Murata H., N. Shimada, M. Yoshioka. 2004. Current research on acute phase proteins in veterinary diagnosis:
an overview. Vet J. 168: 28-40.

Miller, J., McClean, M. Pharmacokinetics of enrofloxacin in African elephants (Loxodonta africana) after a single rectal dose. Proc American Associaton of Zoo Veterinarians and Assoc of Reptile and Amphibian Veterinarians.  224-225. 2008. 11-10-2008.
Ref Type: Conference Proceeding
Abstract: Captive African elephants (Loxodonta Africana) are susceptible to many types of gram negative bacterial infections such as Escherichia coli, Mycoplasma  spp., Salmonella spp., Klebsiella spp., Pseudomonas spp., and Proteus spp. Enrofloxacin (Baytril®, Bayer Health Care, Animal Health Division, P.O. Box 390, Shawnee Mission, KS 66201) is a potentially effective antibiotic for
treatment of these bacterial infections in elephants. Very limited data exists on the pharmacokinetics of enrofloxacin in elephants2 and most of the dosage regimes for gastrointestinal absorption are based on horse dosages since they share a similar  gastrointestinal tract. Three African elephants from Wildlife Safari in Winston, Oregon, two females both 37-yr-old and one male 26-yr-old, were used to determine whether therapeutic levels of enrofloxacin could be achieved thru rectal administration of liquid injectable enrofloxacin (Baytril 100®, 100 mg/ml, Bayer Health Care, Animal Health Division, P.O. Box 390, Shawnee Mission, KS 66201) at a dosage of 2.5 mg/kg. A pretreatment baseline blood sample was collected. Following administration, blood samples were collected at 45 min, 1.5hr, 2.5hr, 5hr, 9hr, 23hr, 36hr to determine plasma enrofloxacin levels. Plasma enrofloxacin levels were measured at North Carolina State University, College of Veterinary Medicine using high performance liquid chromatography (HPLC) analysis. Plasma ciprofloxacin levels were measured concurrently. Results indicate plasma concentrations of enrofloxacin did not reach adequate bacteriocidal levels for any of the the following common bacterial isolates in captive elephants: Mycoplasma
spp., Escherichia coli, Salmonella spp., Klebsiella spp., Pseudomonas spp., and Proteus spp. The study determined that a rectally administered dosage of 2.5 mg/kg of liquid injectable enrofloxacin was insufficient to obtain therapeutic levels in African elephants. The low plasma levels of enrofloxacin in all three elephants may be a result of poor absorption in the distal large intestine. A future study will determine if oral administration will provide a more efficient mode of drug delivery and absorption in African elephants. It is also possible that the current dosage of 2.5 mg/kg is too low to achieve adequate therapeutic levels.
ACKNOWLEDGMENTS
I would like to thank the elephant and veterinary staff at Wildlife Safari for their participation in conducting this study. Thanks to Doctors: Modesto McClean, Jason Bennett, Andi Chariffe, Tessa Lohe, Benji Alacantar. Also thanks to Dinah Wilson, Carol Matthews, Anthony Karels, Mary Iida, Shawn Finnell, Patches Stroud, Katie Alayan.
LITERATURE CITED
1. Haines, G.R., et. al. 2000. Serum concentrations and pharmacokinetics of enrofloxacin after intravenous and intragastric administration to mares. Can. J.Vet. Res. 64(3):171-177.
2. Sanchez, C.R., et. al. 2005. Pharmacokinetics of a single dose of enrofloxacin administered orally to captive Asian elephants (Elephas maximus). Am. J. Vet. Res. 66:1948-1953.

Nicholls, H., 2008. Darwin 200: Let's make a mammoth. Nature 456, 310-314.

Rasmssen, H.B., Ganswindt, A., Douglas-Hamilton, I., Vollrath, F., 2008. Endocrine and behavioral changes in male African elephants: Linking hormone changes to sexual state and reproductive tactics. Hormones and Behavior.
Abstract: Endocrine and behavioral changes in male African elephants: Linking hormone changes to sexual state and reproductive tactics.Henrik B Rasmussen, Andre Ganswindt, Iain Douglas-Hamilton, and Fritz VollrathHormones and Behavior, May 22, 2008                     
Hormones play a crucial role in mediating genetic and environmental effects into morphological and behavioral phenotypes. In systems with alternative reproductive tactics (ART) shifts between tactics are hypothesized to be under proximate hormonal control. Most studies of the underlying endocrine changes behind ART have focused on fish and amphibians rather than mammals and few have investigated the potential interaction between different endocrine axes in regulating shifts between conditional dependent tactics. Using a combination of endocrine and behavioral data from male African elephants we expand on our previously published analysis and show that the initial increase in androgens predates the behavioral shifts associated with reproductively active periods, supporting the role of androgens in activating sexually active periods in males. A strong interactive effect between androgens and glucocorticoids was found to determine the presence or absence of temporal gland secretion and urine dribbling, signals associated with the competitive reproductive tactic of musth, with elevated glucocorticoids levels suppressing the occurrence of musth signals. In addition external environmental conditions affected hormone levels. The presence of receptive females resulted in elevated androgens in dominant musth males but increased glucocorticoids in subordinate non-musth males. The presented data on hormones, behavior and reproductive tactics strongly support an underlying endocrine mechanism for mediating the translation of intrinsic as well as extrinsic local conditions into the conditional dependent reproductive tactics in male elephants via interactions between the hypothalamic-pituitary-gonadal and -adrenal axes.

Rasmussen, H.B., Ganswindt, A., Douglas-Hamilton, I., Vollrath, F., 2008. Endocrine and behavioral changes in male African elephants: linking hormone changes to sexual state and reproductive tactics. Horm. Behav. 54, 539-548.
Abstract: Hormones play a crucial role in mediating genetic and environmental effects into morphological and behavioral phenotypes. In systems with alternative reproductive tactics (ART) shifts between tactics are hypothesized to be under proximate hormonal control. Most studies of the underlying endocrine changes behind ART have focused on fish and amphibians rather than mammals and few have investigated the potential interaction between different endocrine axes in regulating shifts between conditional dependent tactics. Using a combination of endocrine and behavioral data from male African elephants we expand on our previously published analysis and show that the initial increase in androgens predates the behavioral shifts associated with reproductively active periods, supporting the role of androgens in activating sexually active periods in males. A strong interactive effect between androgens and glucocorticoids was found to determine the presence or absence of temporal gland secretion and urine dribbling, signals associated with the competitive reproductive tactic of musth, with elevated glucocorticoids levels suppressing the occurrence of musth signals. In addition external environmental conditions affected hormone levels. The presence of receptive females resulted in elevated androgens in dominant musth males but increased glucocorticoids in subordinate non-musth males. The presented data on hormones, behavior and reproductive tactics strongly support an underlying endocrine mechanism for mediating the translation of intrinsic as well as extrinsic local conditions into the conditional dependent reproductive tactics in male elephants via interactions between the hypothalamic-pituitary-gonadal and -adrenal axes

Schmitt, D., Charmason, S., Wiedner, E. Use of luteinizing hormone ELISAs  in breeding elephants. Proc American Associaton of Zoo Veterinarians and Assoc of Reptile and Amphibian Veterinarians.  120-121. 2008. 11-10-2008.
Ref Type: Conference Proceeding
Abstract: Successful artificial insemination (AI) of elephants depends heavily on determining the unique luteinizing hormone (LH) surges that occur during the follicular phase of the elephant's estrous cycle. Natural breeding of elephants also can benefit from a rapid and accurate determination of the two LH surges found in elephants. There are three ELISAs available for determining the LH
surge; two are commercially-available assays and one is a laboratory in-house assay. Each vary in their cost, time to complete the assay, and ease of performing the procedures. Detection of the initial non-ovulatory peak in luteinizing hormone (LH1) is best accomplished by use of an in house LH assay, or use of the LH assay available from Dr. Nancy Dahl (UC-Davis, Davis, CA
95616 USA), both of which are quantitative assays for detection of LH. For cow-side use during estrus, the qualitative ELISA Witness® LH Ovulation Timing Test Kit (Symbiotics Corporation, Kansas City, MO 64163 USA) detects LH in elephants within 20 min. This assay requires a minimum of laboratory precision to detect the ovulatory LH peak (LH2).
Introduction
Elephants are the only species known to exhibit a double LH peak during a single estrous cycle.2,4 Increased success of artificial insemination in elephants occurred partly in response to the ability to detect the LH1 surge about 21 days prior to the ovulatory LH2 surge that occurs at the end of a two to three day estrus.1 The first reports regarding detection of the double LH
surges were performed in laboratories using custom ELISA technology that require exacting procedures and two days to complete the quantitative assays.2,4 A semi-quantitative elephant LH ELISA that can be performed in the field in about 2.5 hr was developed at UC-Davis.3 A qualitative LH assay was developed for use in dogs and cats that uses a latex strip ELISA. The time for development of the test is 20 min and detects a LH surge greater than 1 ng/ml using serum. Elephants have LH1 and LH2 surges in the 4-16 ng/ml range,2,4 well within the detectable range for all of the assays described. The detection of the LH1 peak usually is from daily samples submitted weekly; this allows some efficiency of assay resources and provides at least a two-wk notice of LH2. However, accurate and timely detection of LH2 is needed at least daily and at times twice daily during estrus. The use of an LH assay which can be performed 'cow-side' and accurately detect LH2 is essential for successful AI and can be helpful in determining estrus status for natural breeding. The Witness® LH Ovulation Timing Test Kit from Symbiotics was developed for use in dogs and cats, but is effective in other species, including elephants, and meets these requirements.
Discussion
Detection of LH1 provides information for predicting the LH2 surge and performance of assays that require more laboratory time and precision are useful since detection of LH1 is not as timesensitive as LH2 detection. Both of the quantitative assays have unique advantages. An inhouse assay can be set up, but requires greater preparation time, precision of laboratory procedures is more demanding, often takes two days to perform, and is more susceptible to environmental variables. The assay developed by UC-Davis costs about $5.00 per well, takes about 2.5 hr to perform and is more stable. However, for quantitative results the overhead costs of the standard curve requires about 16 wells ($90), plus two wells for each unknown sample. The UC-Davis assay can be set up as a qualitative test with high and low controls and no standard curve. This requires from three to six wells for a single sample. The Witness® LH Ovulation Timing Test Kit has a control built into each test strip and costs about $25.00 per sample. Because 'cow-side' testing possible using the Witness® LH Ovulation Timing Test Kit, I recommend its use for detection of LH2, although the UC-Davis Elephant ELISA is competitively priced and can be performed in a nearby temporary laboratory. Because timing is
critical in detecting LH2 and performing subsequent AI, I recommend using the Witness® LH Ovulation Timing Test Kit at the time of estrus, preceded by either one of the other assays for detecting LH1, depending on availability of laboratory labor and equipment.
LITERATURE CITED
1. Brown, J. L., F. Goritz, N. Pratt-Hawkes, R. Hermes, M. Galloway, L. H. Graham, C. Gray, S. L. Walker, A. Gomez, R. Moreland, S. Murray, D. L. Schmitt, J. G. Howard, J. Lehnhardt, B. Beck, A. Bellem, R. Montali, and T. B. Hildebrandt. 2004. Successful artificial insemination of an Asian elephant at the National Zoological Park. Zoo Biol. 23: 45-63.
2. Brown, J. L., D. L. Schmitt, A. Bellem, L. H. Graham, and J. Lehnhardt. 1999. Hormone secretion in the Asian elephant (Elephas maximus): Characterization of ovulatory and anovulatory luteinizing hormone surges. Biol. Reprod. 61: 1294-1299.
3. Dahl, N. J., D. Olson, D. L. Schmitt, D. R. Blasko, R. S. Kristipati, and J. F. Roser. 2004. Development of an enzyme-linked immunosorbent assay (ELISA) in the elephant (Loxodonta africana and Elephas maximus). Zoo Biol. 23: 65-78.
4. Kapustin, N., J. K. Critser, D. Olson, and P. V. Malven. 1996. Nonluteal estrous cycles of 3-week duration are initiated by anovulatory luteinizing hormone peaks in African elephants. Biol. Reprod. 55:1147-1154.

Slade-Cain, B.E., Rasmussen, L.E., Schulte, B.A., 2008. Estrous state influences on investigative, aggressive, and tail flicking behavior in captive female Asian elephants
78. Zoo. Biol. 27, 167-180.
Abstract: Females of species that live in matrilineal hierarchies may compete for temporally limited resources, yet maintain social harmony to facilitate cohesion. The relative degree of aggressive and nonaggressive interactions may depend on the reproductive condition of sender and receiver. Individuals can benefit by clearly signaling and detecting reproductive condition. Asian elephants (Elephas maximus) live in social matrilineal herds. Females have long estrous cycles (14-16 weeks) composed of luteal (8-12 weeks) and follicular (4-8 weeks) phases. In this study, we observed the behavior of four captive Asian elephant females during multiple estrous cycles over 2 years. We evaluated whether investigative, aggressive, and tail flicking behaviors were related to reproductive condition. Investigative trunk tip contacts showed no distinct pattern by senders, but were more prevalent toward female elephants that were in their follicular compared with their luteal phase. The genital area was the most frequently contacted region and may release reproductively related chemosignals. Aggression did not differ significantly with estrus; however, rates of aggression were elevated when senders were approaching ovulation and receivers were in the luteal phase. Females in the follicular phase may honestly advertise their condition. Contacts by conspecifics may serve to assess condition and reduce aggression. A behavior termed "tail flicking" was performed mainly during the mid-follicular phase when estrogen and luteinizing hormone levels are known to spike. Tail flicking may disperse chemical signals in urine or mucus as well as act as a tonic signal that could provide a means of anticipating forthcoming ovulation by elephants and also for human observers and caretakers. Zoo Biol 27:167-180, 2008. (c) 2008 Wiley-Liss, Inc

Steinmetz, H.W., Eulenberger, U., Hatt, J.M. Daily clinical examinations in a herd of captive asian elephants. Proc American Associaton of Zoo Veterinarians and Assoc of Reptile and Amphibian Veterinarians.  124. 2008. 11-10-2008.
Ref Type: Conference Proceeding
Abstract: The captive population of Asian elephants (Elephas maximus) is not self-sustaining.2 Poor reproduction and high juvenile mortality are key factors in the decreasing population. Infection with endotheliotropic elephant herpes virus (EEHV) is one of the major causes of death in the captive population, and has resulted in the loss of at least 40 captive animals.1 EEHV has been
responsible for the peracute death of two juvenile males at Zurich Zoo, Switzerland. Mortality due to peracute infection with EEHV mainly is seen in juveniles. Early detection of characteristic clinical signs of EEHV and immediate initiation of therapy are of crucial
importance due to its rapid progression. Based on past fatal EEHV experiences, Zurich Zoo modified its daily clinical health monitoring program to increase staff awareness of EEHV infection. Examinations have been incorporated into the daily routine and include daily evaluation of behaviour, appetite, colour of mucosal membranes and the measurement of body temperature; these examinations are performed by keepers. In our experiences, characteristic signs of acute EEHV infection are lethargy, anorexia, mild
colic, and cyanosis of the mucosal membranes. Results of temperature measurements have shown that best estimations of body temperature are done by measurement of the temperature in the centre of a fecal ball 5-9 min after defecation. Mean values of 36.5°C (± 0.2°C SD) are within published reference values, although adult elephants have shown significantly lower body temperature than juveniles. Establishment of individual reference values for each elephant is essential to detect unusual temperature peaks that may indicate possible EEHV viremia. The present study has shown that daily health examinations increase the awareness of keepers for
early signs of EEHV infection (e.g., peaks in body temperature and cyanotic mucosal membranes).
ACKNOWLEDGMENTS
The authors thank B. Aeschbach and all elephant keepers for taking special care of our elephants. The work and organization of Ms. G. Hürlimann is gratefully appreciated.
LITERATURE CITED
1. Mikota, S. 2007. Endotheliotropic Herpesvirus (EEHV). http://www.elephantcare.org/herpes.htm. cited: 10.04.2008:
2. Wiese, R. J. 2000. Asian elephants are not self-sustaining in North America. Zoo Biol. 19: 299-309.

Thongtip, N., Saikhun, J., Mahasawangkul, S., Kornkaewrat, K., Pongsopavijitr, P., Songsasen, N., Pinyopummin, A., 2008. Potential factors affecting semen quality in the Asian elephant (Elephas maximus). Reprod. Biol. Endocrinol. 6, 9.
Abstract: BACKGROUND: One of the major obstacles in using artificial insemination to manage genetics of elephant population in captivity is the large variations in semen quality among ejaculates within the same and among individuals. The objectives of this study were to determine the influences of (1) age (2) seasonality (3) and circulating testosterone (SrTest), triiodothyronine (SrT3) and tetraiodothyronine (SrT4), as well as seminal (4) testosterone (SpTest), zinc (SpZn) and protein (SpTP) on semen quality in the Asian elephant METHODS: Analyses, including motility, viability and morphology were performed in semen samples collected twice monthly from 13 elephant bulls (age range, 10-to 72-years) by manual stimulation between July 2004 and June 2005. Serum samples obtained monthly were assessed for SrTest, SrT3, SrT4, and seminal plasma samples were evaluated for, SpTest, SpZn and SpTP. RESULTS: The highest semen quality was observed at age 23 to 43 years. Percentages of progressive motility and viable sperm were lowest at age 51 to 70 years (P < 0.05); on the other hand, sperm concentration was lowest at age 10 to 19 years (P < 0.05). Percentage of sperm with normal morphology was highest at age 23 to 43 years. The levels of SrT3, SrTest, SpTest and SpZn were lowest at age 51 to 70 years, whereas SrT4 was lowest at age 23 to 43 years. Seasonality significantly affected semen characteristics in which percentage of viable sperm and cell concentration were highest during rainy season and lowest during summer months (P < 0.05). However, percentage of sperm with normal morphology was highest in summer and lowest in rainy season (P < 0.05). Seasonality significantly influenced SrTest with elevated concentrations observed in rainy season and winter (P < 0.05). CONCLUSION: This study indicates that age and seasonality had influence on semen characteristics in the Asian elephant. The knowledge obtained in this study will improve our understanding of the reproductive biology of this species

Turner, J.W., Rutberg, A.T., Naugle, R.E., Kaur, M.A., Flamagan, D.R., Bertschinger, H.J., Liu, I.K.M., 2008. Controlled-release components of PZP contraceptive vaccine extend duration of infertility. Wildlife Research 35, 555-562.
Abstract: Successful immunocontraception of wildlife relying on repeated access to individuals for boosters has highlighted the need to incorporate primer and booster immunisations into one injection. We have investigated use of controlled-release polymers (lactide-glycolide) in small pellets to provide delayed in vivo delivery of booster porcine zona pellucida (PZP) antigen and adjuvant. This report reviews pellet-making methodology, in vitro testing of controlled-release pellets and in vivo effects of controlled-release PZP vaccine. We assessed 3 different manufacturing approaches for producing reliable, cost-effective pellets: (1) polymer melting and extrusion; (2) solvent evaporation from polymer solution; and (3) punch and die polymer moulding. In vitro testing of release patterns of controlled-release formulations, towards development of a 3-year duration vaccine, provided estimates for in vivo use of pellet preparations. These in vitro studies demonstrated protein release delay up to 22 months using 100% l-lactide or polycaprolactone polymers. For in vivo tests, pellets (1-, 3-, and 12-month release delay) serving as boosters were administered intramuscularly with PZP/adjuvant liquid primer to wild horses (Equus caballus), white-tailed deer (Odocoileus virginanus) and African elephants (Loxodonta africana). Horse. field studies assessed fertility via offspring counts and/or faecal-hormone pregnancy testing. Treatment decreased fertility 5.3-9.3-fold in Year 1 and 3.6-fold in Year 2. In preliminary testing in deer, offspring counts revealed treatment-associated fertility reduction of 7.1-fold Year 1 and 3.3-fold Year 2. In elephants, treatment elevated anti-PZP titres 4.5-6.9- fold from pretreatment (no fertility data).

van der Kolk, J.H., van Leeuwen, J.P., van den Belt, A.J., van Schaik, R.H., Schaftenaar, W., 2008. Subclinical hypocalcaemia in captive Asian elephants (Elephas maximus). Veterinary Record 162, 475-479.
Abstract: The hypothesis that hypocalcaemia may play a role in dystocia in captive Asian elephants (Elephas maximus) was investigated. The objectives of the study were to measure the total calcium concentration in elephant plasma; assess the changes in parameters of calcium metabolism during a feeding trial; investigate a possible relationship between calcium metabolism and dystocia; and assess bone mineralisation in captive Asian elephants in vivo. The following parameters were measured: total and ionised calcium, inorganic phosphorous and magnesium, the fractional excretions of these minerals, intact parathyroid hormone, 25-OH-D(3) and 1,25-OH-D(3). Radiographs were taken from tail vertebrae for assessment of bone mineralisation. The mean (sd) heparinised plasma total calcium concentration was 2.7 (0.33) mmol/l (n=43) ranging from 0.84 to 3.08 mmol/l in 11 Asian elephants. There was no significant correlation between plasma total calcium concentration and age. Following feeding of a calcium rich ration to four captive Asian elephant cows, plasma total and ionised calcium peaked at 3.6 (0.24) mmol/l (range 3.4 to 3.9 mmol/l) and 1.25 (0.07) mmol/l (range 1.17 to 1.32 mmol/l), respectively. Plasma ionised calcium concentrations around parturition in four Asian elephant cows ranged from 0.37 to 1.1 mmol/l only. The present study indicates that captive Asian elephants might be hypocalcaemic, and that, in captive Asian elephants, the normal plasma concentration of total calcium should actually be around 3.6 mmol/l and normal plasma concentration of ionised calcium around 1.25 mmol/l. Given the fact that elephants absorb dietary calcium mainly from the intestine, it could be concluded that elephants should be fed calcium-rich diets at all times, and particularly around parturition. In addition, normal values for ionised calcium in captive Asian elephants should be reassessed

Viijoen, J.J., Ganswindt, A., du Toit, J.T., Langbauer, W.R., 2008. Translocation stress and faecal glucocorticoid metabolite levels in free-ranging African savanna elephants. South African Journal of Wildlife Research 38, 146-152.
Abstract: There are local populations of African elephants (Loxodonta africana) which have increased to levels where they are implicated in altering vegetation types. The local reduction of elephant numbers for wildlife management objectives can involve contraception, killing excess animals, or translocation to alternative habitats. The effects these management decisions can have on the physiological stress response of free-ranging African savanna elephants are still not fully understood. We examined the effect of translocation on faecal glucocorticoid metabolite levels of an African elephant family group, which was translocated within the Kruger National Park, South Africa. We found that translocation resulted in a significant increase in faecal glucocorticoid metabolite levels (up to 646 ng/g wet weight) compared to (1) pre-translocation levels in this group, (2) post-translocation levels in this group, and (3) levels measured in undisturbed 'control' groups in the area. However, the faecal glucocorticoid metabolite levels had returned to <100 ng/g by the time the translocated animals had navigated their way back to their previous home range, covering 300 km in 23 days.

von, A., I, Nimzyk, R., Klemke, M., Bullerdiek, J., 2008. A microRNA encoded in a highly conserved part of the mammalian HMGA2 gene. Cancer Genet. Cytogenet. 187, 43-44.
Abstract: The high mobility group protein HMGA2 plays an important role as a chromatin component of stem cells and as a protein causally related to the development of a variety of benign tumors (e.g., uterine leiomyomas, lipomas, and pleomorphic adenomas of the salivary glands). Herein, the existence of a highly conserved region within intron 3 of HMGA2 encoding a microRNA is described. The co-expression with HMGA2 suggests that as an intronic microRNA, this microRNA may cooperate with HMGA2 in its physiological and/or aberrant functions

Wittemyer, G., Polansky, L., Douglas-Hamilton, I., Getz, W.M., 2008. Disentangling the effects of forage, social rank, and risk on movement autocorrelation of elephants using Fourier and wavelet analyses. Proc. Natl. Acad. Sci. U. S. A 105, 19108-19113.
Abstract: The internal state of an individual-as it relates to thirst, hunger, fear, or reproductive drive-can be inferred by referencing points on its movement path to external environmental and sociological variables. Using time-series approaches to characterize autocorrelative properties of step-length movements collated every 3 h for seven free-ranging African elephants, we examined the influence of social rank, predation risk, and seasonal variation in resource abundance on periodic properties of movement. The frequency domain methods of Fourier and wavelet analyses provide compact summaries of temporal autocorrelation and show both strong diurnal and seasonal based periodicities in the step-length time series. This autocorrelation is weaker during the wet season, indicating random movements are more common when ecological conditions are good. Periodograms of socially dominant individuals are consistent across seasons, whereas subordinate individuals show distinct differences diverging from that of dominants during the dry season. We link temporally localized statistical properties of movement to landscape features and find that diurnal movement correlation is more common within protected wildlife areas, and multiday movement correlations found among lower ranked individuals are typically outside of protected areas where predation risks are greatest. A frequency-related spatial analysis of movement-step lengths reveal that rest cycles related to the spatial distribution of critical resources (i.e., forage and water) are responsible for creating the observed patterns. Our approach generates unique information regarding the spatial-temporal interplay between environmental and individual characteristics, providing an original approach for understanding the movement ecology of individual animals and the spatial organization of animal populations

Yon, L., Chen, J., Moran, P., Lasley, B., 2008. An analysis of the androgens of musth in the Asian bull elephant (Elephas maximus). Gen. Comp Endocrinol. 155, 109-115.
Abstract: During musth in bull elephants, the androgens testosterone (T), dihydrotestosterone (DHT), and androstenedione all increase significantly. Given the unusual endocrine physiology that has been discovered in female elephants, it is also possible that bull elephants produce some unusual androgens. A cell-based androgen receptor assay was used to explore this possibility using two different methods. The first method compared the level of T measured by radioimmunoassay (RIA) with the level of androgen receptor (AR) activity measured in the serum of eight bull elephants during musth and non-musth periods. A ratio was calculated for T/AR activity for non-musth and musth, to determine if there was a change in the ratio between these two states. The second method used HPLC to separate two pooled serum samples (one non-musth and one musth) into fractions using a protocol which separates known androgens into specific, previously identified fractions. Each fraction was then tested with the AR assay to determine the androgenicity of any compounds present. This was done to determine if there were any fractions which had androgenic activity but did not contain any previously identified androgens. Results from the first analysis indicated no change in the T/AR ratio between non-musth and musth states. Clearly whatever active androgens are present during musth, they increase proportionately with T. Findings from the second analysis suggested that the only bioactive androgen present in the serum of non-musth Asian bulls is a low level of T. During musth, the only bioactive androgens detected were T and DHT; of these, T was by far the predominant active androgen present. Taken together, these two analyses suggest that T is by far the predominant active androgen present during musth in Asian bull elephants, and that no previously unidentified bioactive androgen is present

Archie, E.A., Hollister-Smith, J.A., Poole, J.H., Lee, P.C., Moss, C.J., Maldonado, J.E., Fleischer, R.C., Alberts, S.C., 2007. Behavioural inbreeding avoidance in wild African elephants. Molecular Ecology 16, 4138-4148.
Abstract: The costs of inbreeding depression, as well as the opportunity costs of inbreeding avoidance, determine whether and which mechanisms of inbreeding avoidance evolve. In African elephants, sex-biased dispersal does not lead to the complete separation of male and female relatives, and so individuals may experience selection to recognize kin and avoid inbreeding. However, because estrous females are rare and male-male competition for mates is intense, the opportunity costs of inbreeding avoidance may be high, particularly for males. Here we combine 28 years of behavioural and demographic data on wild elephants with genotypes from 545 adult females, adult males, and calves in Amboseli National Park, Kenya, to test the hypothesis that elephants engage in sexual behaviour and reproduction with relatives less often than expected by chance. We found support for this hypothesis: males engaged in proportionally fewer sexual behaviours and sired proportionally fewer offspring with females that were natal family members or close genetic relatives (both maternal and paternal) than they did with nonkin. We discuss the relevance of these results for understanding the evolution of inbreeding avoidance and for elephant conservation.

Brown, J.L., Somerville, M., Riddle, H.S., Keele, M., Duer, C.K., Freeman, E.W., 2007. Comparative endocrinology of testicular, adrenal and thyroid function in captive Asian and African elephant bulls. Gen. Comp Endocrinol. 151, 153-162.
Abstract: Concentrations of serum testosterone, cortisol, thyroxine (free and total T4), triiodothyronine (free and total T3) and thyroid stimulating hormone (TSH) were measured to assess adrenal and thyroid function as they relate to testicular activity and musth in captive elephants. Blood samples were collected approximately weekly from Asian (n=8) and African (n=12) bulls at seven facilities for periods of 4 months to 9.5 years. Age ranges at study onset were 8-50 years for Asian and 10-21 years for African elephants. Based on keeper logs, seven Asian and three African bulls exhibited behavioral and/or physical (temporal gland secretion, TGS, or urine dribbling, UD) signs of musth, which lasted 2.8+/-2.5 months in duration. Serum testosterone was elevated during musth, with concentrations often exceeding 100 ng/ml. Patterns of testosterone secretion and musth varied among bulls with no evidence of seasonality (P>0.05). Only three bulls at one facility exhibited classic, well-defined yearly musth cycles. Others exhibited more irregular cycles, with musth symptoms often occurring more than once a year. A number of bulls (1 Asian, 9 African) had consistently low testosterone (<10 ng/ml) and never exhibited significant TGS or UD. At facilities with multiple bulls (n=3), testosterone concentrations were highest in the oldest, most dominant male. There were positive correlations between testosterone and cortisol for six of seven Asian and all three African males that exhibited musth (range, r=0.23-0.52; P<0.05), but no significant correlations for bulls that did not (P>0.05). For the three bulls that exhibited yearly musth cycles, TSH was positively correlated (range, r=0.22-0.28; P<0.05) and thyroid hormones (T3, T4) were negatively correlated (range, r=-0.25 to -0.47; P<0.05) to testosterone secretion. In the remaining bulls, there were no clear relationships between thyroid activity and musth status. Overall mean testosterone and cortisol concentrations increased with age for all bulls combined, whereas thyroid activity declined. In summary, a number of bulls did not exhibit musth despite being of adequate physical maturity. Cortisol and testosterone were correlated in most bulls exhibiting musth, indicating a possible role for the adrenal gland in modulating or facilitating downstream responses. Data were generally inconclusive as to a role for thyroid hormones in male reproduction, but the finding of discrete patterns in bulls showing clear testosterone cycles suggests they may facilitate expression or control of musth in some individuals

Brown, J.L., Somerville, M., Riddle, H.S., Keele, M., Duer, C.K., Freeman, E.W., 2007. Comparative endocrinology of testicular, adrenal and thyroid function in captive Asian and African elephant bulls. General and Comparative Endocrinology 151, 153-162.
Abstract: Concentrations of serum testosterone, cortisol, thyroxine (free and total T4), triiodothyronine (free and total T3) and thyroid stimulating hormone (TSH) were measured to assess adrenal and thyroid function as they relate to testicular activity and musth in captive elephants. Blood samples were collected approximately weekly from Asian (n = 8) and African (n = 12) bulls at seven facilities for periods of 4 months to 9.5 years. Age ranges at study onset were 8-50 years for Asian and 10-21 years for African elephants. Based on keeper logs, seven Asian and three African bulls exhibited behavioral and/or physical (temporal gland secretion, TGS, or urine dribbling, UD) signs of musth, which lasted 2.8 +/- 2.5 months in duration. Serum testosterone was elevated during musth, with concentrations often exceeding 100 ng/ml. Patterns of testosterone secretion and musth varied among bulls with no evidence of seasonality (P > 0.05). Only three bulls at one facility exhibited classic, well-defined yearly musth cycles. Others exhibited more irregular cycles, with musth symptoms often occurring more than once a year. A number of bulls (I Asian, 9 African) had consistently low testosterone (< 10 ng/ml) and never exhibited significant TGS or UD. At facilities with multiple bulls (n = 3), testosterone concentrations were highest in the oldest, most dominant male. There were positive correlations between testosterone and cortisol for six of seven Asian and all three African males that exhibited musth (range, r = 0.23-0.52; P < 0.05), but no significant correlations for bulls that did not (P > 0.05). For the three bulls that exhibited yearly musth cycles, TSH was positively correlated (range, r = 0.22-0.28; P < 0.05) and thyroid hormones (T3, T4) were negatively correlated (range, r = -0.25 to -0.47; P < 0.05) to testosterone secretion. In the remaining bulls, there were no clear relationships between thyroid activity and musth status. Overall mean testosterone and cortisol concentrations increased with age for all bulls combined, whereas thyroid activity declined. In summary, a number of bulls did not exhibit musth despite being of adequate physical maturity. Cortisol and testosterone were correlated in most bulls exhibiting musth, indicating a possible role for the adrenal gland in modulating or facilitating downstream responses. Data were generally inconclusive as to a role for thyroid hormones in male reproduction, but the finding of discrete patterns in bulls showing clear testosterone cycles suggests they may facilitate expression or control of musth in some individuals.

Dehnhard, M., 2007. Characterisation of the sympathetic nervous system of Asian (Elephas maximus) and African (Loxodonta africana) elephants based on urinary catecholamine analyses. Gen. Comp Endocrinol. 151, 274-284.
Abstract: Assessing the welfare status of captive animals using non-invasive measurements of hormones is of growing interest because this can serve as an effective tool to facilitate the optimization of environmental and husbandry conditions. Both the African elephant (Loxodonta africana) and the Asian elephant (Elephas maximus) exhibit extremely low breeding success in captivity, and because elevated levels of stress may negatively influence reproductive functions, this study sought to establish a method for assessing sympathoadrenal activity in captive female elephants. We found a circadian variation in urinary noradrenaline (norepinephrine, NE), adrenaline (epinephrine, Epi) and dopamine (DA) under short day length. Peak activity of noradrenaline and dopamine was noted at 3 a.m. Adrenaline showed a biphasic pattern with a minor peak recorded at 3 a.m. and a major peak 9 a.m. Under long-day photoperiodic conditions, simultaneous peaks of noradrenaline and adrenaline were again noted at 3 a.m. whereas dopamine does not appear to have a distinct circadian pattern under long-day length. A transfer of two elephant cows resulted in a marked increase in urinary adrenaline and noradrenaline levels, confirming that the transfer represented a stressful event. During the peripartal period, noradrenaline concentrations increased and maximum concentrations were obtained at delivery. Daily measurements of urinary dopamine throughout the follicular phase revealed an increase in dopamine secretion close to ovulation. This increase might indicate a role of dopamine in the ovulatory mechanisms. These results suggest that changes in urinary catecholamine excretion reflect fluctuations in sympathoadrenal activity and may be a useful indicator of stress

Duer, C., Carden, M., Tomasi, T., 2007. Detection of fetal gender differences in maternal serum progesterone concentrations of Asian elephants (Elephas maximus)
486. Anim Reprod. Sci. 97, 278-283.
Abstract: Previous studies have analyzed total testosterone concentrations in maternal serum for a reliable method of fetal gender determination in Asian elephants (Elephas maximus). The present study investigated the possibility that progesterone concentrations in maternal serum may reflect these testosterone patterns. Weekly serum samples were collected from 17 pregnancies in captive Asian elephants and analyzed via radioimmunoassay (RIA) for progesterone concentrations. Nine and eight cows carried male and female calves, respectively. Mean progesterone concentrations in maternal serum of elephants carrying male calves were greater than in those carrying female calves (P<0.01). Mean progesterone concentrations (based on 5-week means) in maternal serum were greater at weeks 20-55 (P<0.01) and 60-65 (P<0.05) for elephants carrying male calves

Hildebrandt, T., Drews, B., Gaeth, A.P., Goeritz, F., Hermes, R., Schmitt, D., Gray, C., Rich, P., Streich, W.J., Short, R.V., Renfree, M.B., 2007. Foetal age determination and development in elephants. Proc. Biol. Sci. 274, 323-331.
Abstract: Elephants have the longest pregnancy of all mammals, with an average gestation of around 660 days, so their embryonic and foetal development have always been of special interest. Hitherto, it has only been possible to estimate foetal ages from theoretical calculations based on foetal mass. The recent development of sophisticated ultrasound procedures for elephants has now made it possible to monitor the growth and development of foetuses of known gestational age conceived in captivity from natural matings or artificial insemination. We have studied the early stages of pregnancy in 10 captive Asian and 9 African elephants by transrectal ultrasound. Measurements of foetal crown-rump lengths have provided the first accurate growth curves, which differ significantly from the previous theoretical estimates based on the cube root of foetal mass. We have used these to age 22 African elephant foetuses collected during culling operations. Pregnancy can be first recognized ultrasonographically by day 50, the presumptive yolk sac by about day 75 and the zonary placenta by about day 85. The trunk is first recognizable by days 85-90 and is distinct by day 104, while the first heartbeats are evident from around day 80. By combining ultrasonography and morphology, we have been able to produce the first reliable criteria for estimating gestational age and ontological development of Asian and African elephant foetuses during the first third of gestation.

Hollister-Smith, J.A., Poole, J.H., Archie, E.A., Vance, E.A., Georgiadis, N.J., Moss, C.J., Alberts, S.C., 2007. Age, musth and paternity success in wild maleAfrican elephants, Loxodonta africana. Animal Behaviour 74, 287-296.
Abstract: Male African elephants experience intense intrasexual selection in gaining access to oestrous females, who represent a very scarce and highly mobile resource. An unusual combination of behavioural and physiologica ltraits in males probably reflects this intense selection pressure. Males show prolonged growth, growing throughout much or perhaps all of their long life span (ca. 60-65 years), and they show musth,a physiological and behavioural condition exclusive to elephants, which is manifested by bouts of elevated testosterone and aggression and heightened sexual activity. Most observed matings are by males over 35years of age and in musth, suggesting that age and musth are both important factors contributing to male reproductive success. Here we report the results of a genetic paternity analysis of a well-studied population of wild African elephants. Patterns of paternity for 119 calves born over a 22-year period showed significant effects of both age and musth on paternity success. Among males in musth, paternity success increased significantly with age until the very oldest age classes, when it modestly declined. When not inmusth, males experienced relatively constant, low levels of paternity success at all ages. Thus, despite the importance of both musth and age in determining male paternity success, adult males both in and out ofmusth, and of all ages, produced calves. In general, however, older males had markedly elevated paternitysuccess compared with younger males, suggesting the possibility of sexual selection for longevity in this species.

Kirkpatrick, J.F., 2007. Measuring the effects of wildlife contraception: the argument for comparing apples with oranges. Reprod. Fertil. Dev. 19, 548-552.
Abstract: There are few wildlife populations existing today that can be supported without some form of management. Wildlife fertility control, as one option, has moved from the research stage to actual application with a number of species, including wild horses, urban deer, captive exotic species and even African elephants, but this approach remains controversial in many quarters. Strident debate has arisen over the possible effects of contraception on behaviour, genetics, stress and even management economics, among other parameters. Part of the debate arises from the fact that critics often fail to recognise that some form of alternative management will be applied, and a second problem arises when critics fail to identify and demand the same concern for the consequences of the alternative management approaches. Thus, any rational debate on the merits or possible effects of contraceptive management of wildlife must also recognise all alternative management approaches and apply the same concern and questions to these alternative approaches--including 'no management'--as are currently being applied to fertility control. Only then will the stewards of wildlife be in a position to make wise and informed decisions about management options

Kusuda, S., Wakimoto, T., Nishimura, K., Kawakami, S., Okuda, K., Saito, E., Shimado, T., Sakamoto, H., Yanagimoto, H., Wada, S., Nishio, K., Fuji, H., Suzuki, T., Hashikawa, H., Kusunoki, H., Doi, O., 2007. Relationship between body temperature and ovarian cycle in Asian and African elephants. J Reprod Dev 53, 1099-1105.
Abstract: The aim of the present study was to investigate whether changes in body temperature are related to the ovarian cycle in elephants. Rectal, tongue or fecal temperature was measured for 2 Asian and 5 African elephants using an electric thermometer. Evaluation of ovarian cycles was based on the changes in serum or fecal progestin. The mean  SD values of the rectal, tongue, and fecal temperatures were 36.3  0.3 (2 Asian), 36.2  0.5 (1 African) and 36.5 0.3 C (4 African), respectively; the fecal temperature was the highest of the 3 temperatures (p<0.01). The longitudinal changes in body temperatures correlated with the ovarian cycle, with higher temperatures occurring during the luteal phase. The fecal temperatures of one acyclic African elephant did not change cyclically. These results suggest that measurement of body temperature can be used to easily evaluate the ovarian cyclicity of an individual animal, although
it might not be able to determine the ovarian cycle length.

Portas, T., Bryant, B., Goritz, F., Hermes, R., Keeley, T., Evans, G., Maxwell, M., Hildebrand, T., 2007. Semen collection in an Asian elephant (Elephas maximus) under combined physical and chemical restraint. Aust Vet J 85, 425-427.

Smit, I.P.J., Grant, C.C., Whyte, I.J., 2007. Landscape-scale sexual segregation in the dry season distribution and resource utilization of elephants in Kruger National Park, South Africa. Diversity And Distributions 13, 225-236.
Abstract:  This study compared the dry season distribution of elephant bull groups and mixed herds and the resources driving these distributions within Kruger National Park, South Africa. It is important to understand what resources drive the distribution of elephants as this may be of relevance to understanding and managing their impact. It is also important to distinguish between resource use by bull groups and mixed herds because their impact on the habitat may differ. Our results indicated that sexual segregation, both in space and in resource selection, did occur in Kruger Park. Bulls roamed more widely in the park, and although their distribution and resource use overlapped with mixed herds, they also occurred in areas that mixed herds apparently did not, or could not, utilize in the dry season. This gave rise to areas used exclusively by bulls but no areas used exclusively by mixed herds. Lower collective feeding requirements as a result of smaller group size, wider habitat tolerance, and increased mobility as a result of bigger body size, as well as conflict avoidance with musth bulls in areas with mixed herds, might have been some of the reasons for bull groups roaming more widely and for the establishment of separate bull areas. Rivers were an important resource driving both the distribution of the mixed herds and bull groups, but with the mixed herds occurring closer to these resource hot spots than the bull groups. Tree cover proved important for mixed herds, probably because of the importance of shade and the higher nutritional requirements of the smaller-sized cows and calves. Artificial waterholes might have opened up previously unutilized areas for bulls in the dry season, especially on the grassy basaltic plains in the north of the park. However, the distribution of the mixed herds suggested that they did not occur in higher densities in areas surrounding waterholes.

Wittemyer, G., Ganswindt, A., Hodges, K., 2007. The impact of ecological variability on the reproductive endocrinology of wild female African elephants. Hormones and Behavior 51, 346-354.
Abstract: Non-invasive endocrine methods enable investigation of the relationship between ecological variation and ovarian activity and how this impacts on demographic processes. The underlying physiological factors driving high variation in inter-calving intervals among multi-parous African elephants offer an interesting system for such an investigation. This study investigates the relationship between Normalized Differential Vegetation Index (NDVI), an ecosystem surrogate measure of primary productivity, and fecal progestin concentrations among wild female elephants. Matched fecal samples and behavioral data on reproductive activity were collected from 37 focal individuals during the two-year study. Linear mixed models were used to explore the relationship between fecal 5 alpha-pregnane-3-ol-20-one concentrations and the independent variables of NDVI, calf sex, female age, gestation day, and time since last parturition. Among both non-pregnant and pregnant females, fecal 5 alpha-pregnane-3-ol-20-one concentrations were significantly correlated with time-specific NDVI indicating a strong relationship between ecological conditions and endocrine activity regulating reproduction. In addition, the age of a female and time since her last parturition impacted hormone concentrations. These results indicate that the identification of an individual's reproductive status from a single hormone sample is possible, but difficult to achieve in practice since numerous independent factors, particularly season, impact fecal hormone concentrations. Regardless of season, however, fecal 5 alpha-pregnane-3-ol-20-one concentrations below 1 mu g/g were exclusively collected from non-pregnant females, which could be used as a threshold value to identify non-pregnant individuals. Collectively the information generated contributes to a better understanding of environmental regulation of reproductive endocrinology in wild elephant populations, information salient to the management and manipulation of population dynamics in this species.

Wittemyer, G., Rasmussen, H.B., Douglas-Hamilton, I., 2007. Breeding phenology in relation to NDVI variability in free-ranging African elephant. Ecography 30, 42-50.
Abstract: The phenology of reproduction is often correlated with resource availability and is hypothesized to be shaped by selective forces in order to maximize lifetime reproductive success. African elephants have the distinctive life history traits of a 22 month gestation and extended offspring investment, necessitating a long-term strategy of energy acquisition and reproductive expenditure to ensure successful offspring recruitment. We investigated the relationship between the reproductive phenology of a wild elephant population and resource availability using remotely sensed Normalized Differential Vegetation Index (NDVI) data as a measure of time-specific primary productivity and hence forage quality. The initiation of female elephants' 3+yr reproductive bout was dependent on conditions during the season of conception but timed so parturition occurred during the most likely periods of high primary productivity 22 months later. Thus, the probability of conception is linked to the stochastic variation in seasonal quality and the phenology of parturition is related to the predictable seasonality of primary productivity, indicating elephants integrate information on known current and expected future conditions when reproducing. Juvenile mortality was not correlated with ecological variability, hence female fecundity rather than calf mortality appears to drive demographic processes in the study population. Extreme climatic events, such as those associated with the El Niño-Southern-Oscillation (ENSO), acted to synchronize female fecundity in the population. This study suggests that the relationship between fecundity and ecological variability instigates the characteristic demographic fluctuations in elephant populations, rather than the mortality-driven fluctuations observed in many ungulate populations.

Wittemyer, G., Ganswindt, A., Hodges, K., 2007. The impact of ecological variability on the reproductive endocrinology of wild female African elephants. Horm. Behav. 51, 346-354.
Abstract: Non-invasive endocrine methods enable investigation of the relationship between ecological variation and ovarian activity and how this impacts on demographic processes. The underlying physiological factors driving high variation in inter-calving intervals among multi-parous African elephants offer an interesting system for such an investigation. This study investigates the relationship between Normalized Differential Vegetation Index (NDVI), an ecosystem surrogate measure of primary productivity, and fecal progestin concentrations among wild female elephants. Matched fecal samples and behavioral data on reproductive activity were collected from 37 focal individuals during the two-year study. Linear mixed models were used to explore the relationship between fecal 5alpha-pregnane-3-ol-20-one concentrations and the independent variables of NDVI, calf sex, female age, gestation day, and time since last parturition. Among both non-pregnant and pregnant females, fecal 5alpha-pregnane-3-ol-20-one concentrations were significantly correlated with time-specific NDVI indicating a strong relationship between ecological conditions and endocrine activity regulating reproduction. In addition, the age of a female and time since her last parturition impacted hormone concentrations. These results indicate that the identification of an individual's reproductive status from a single hormone sample is possible, but difficult to achieve in practice since numerous independent factors, particularly season, impact fecal hormone concentrations. Regardless of season, however, fecal 5alpha-pregnane-3-ol-20-one concentrations below 1 microg/g were exclusively collected from non-pregnant females, which could be used as a threshold value to identify non-pregnant individuals. Collectively the information generated contributes to a better understanding of environmental regulation of reproductive endocrinology in wild elephant populations, information salient to the management and manipulation of population dynamics in this species

Witter, K., Egger, G.F., Boeck, P., 2007. Renaut bodies in nerves of the trunk of the African elephant, Loxodonta africana. J. Morphol. 268, 414-422.
Abstract: Renaut bodies are loosely textured, cell-sparse structures in the subperineurial space of peripheral nerves, frequently found at sites of nerve entrapment. The trunk of the elephant is a mobile, richly innervated organ, which serves for food gathering, object grasping and as a tactile organ. These functions of the trunk lead to distortion and mechanical compression of its nerves, which can therefore be expected to contain numerous Renaut bodies. Samples of the trunk wall of an adult African elephant (Loxodonta africana) were examined histologically using conventional staining methods, immunohistochemistry, and lectin histochemistry. Architecture of nerve plexuses and occurrence of Renaut bodies in the elephant trunk were compared with those in tissues surrounding the nasal vestibule of the pig. Prominent nerve plexuses were found in all layers of the elephant trunk. Almost all (81%) nerve profiles contained Renaut bodies, a basophilic, discrete subperineurial layer resembling cushions around the nerve core. In contrast, Renaut bodies were seen in only 15% of nerve profiles in the porcine nasal vestibule. Within Renaut bodies, fusiform fibroblasts and round, ruff-like cells were placed into a matrix of acidic glycosaminoglycans with delicate collagen and very few reticular fibers. The turgor of this matrix is thought to protect nerves against compression and shearing strain. Renaut bodies are readily stained with alcian blue (pH 2.5) favorably in combination with immunohistochemical markers of nerve fibers. They should be regarded as a physiological response to repeated mechanical insults and are distinct from pathological alterations. alterations

Yon, L., Kanchanapangka, S., Chaiyabutr, N., Stanczyk, F., Meepan, S., Lasley, B., 2007. ACTH stimulation in four Asian bull elephants (Elephas maximus): an investigation of androgen sources in bull elephants. Gen. Comp Endocrinol. 151, 246-251.
Abstract: The phenomenon of musth is a very stressful event, both behaviorally and physiologically. An ACTH stimulation test was conducted in four adult Asian bull elephants to investigate the possibility that the classical hypothalamic-pituitary-adrenal (HPA) axis is active during musth, resulting in an increase in adrenally produced steroids. Serum cortisol, testosterone (T), androstenedione (A4), androstenediol (A5), and dehydroepiandrosterone (DHEA) were measured. Cortisol increased 3-10 times above baseline in response to ACTH stimulation, and DHEA doubled. A4 and A5 were erratic, while testosterone decreased significantly in all bulls. The pattern of results suggests that the adrenal steroid increase which occurs during musth results from some mechanism other than the classical HPA axis

Yon, L., Chen, J., Moran, P., Lasley, B., 2007. An analysis of the androgens of musth in the Asian bull elephant (Elephas maximus). Gen Comp Endocrinol. Mar 24; [Epub ahead of print].
Abstract: During musth in bull elephants, the androgens testosterone (T), dihydrotestosterone (DHT), and androstenedione all increase significantly. Given the unusual endocrine physiology that has been discovered in female elephants, it is also possible that bull elephants produce some unusual androgens. A cell-based androgen receptor assay was used to explore this possibility using two different methods. The first method compared the level of T measured by radioimmunoassay (RIA) with the level of androgen receptor (AR) activity measured in the serum of eight bull elephants during musth and non-musth periods. A ratio was calculated for T/AR activity for non-musth and musth, to determine if there was a change in the ratio between these two states. The second method used HPLC to separate two pooled serum samples (one non-musth and one musth) into fractions using a protocol which separates known androgens into specific, previously identified fractions. Each fraction was then tested with the AR assay to determine the androgenicity of any compounds present. This was done to determine if there were any fractions which had androgenic activity but did not contain any previously identified androgens. Results from the first analysis indicated no change in the T/AR ratio between non-musth and musth states. Clearly whatever active androgens are present during musth, they increase proportionately with T. Findings from the second analysis suggested that the only bioactive androgen present in the serum of non-musth Asian bulls is a low level of T. During musth, the only bioactive androgens detected were T and DHT; of these, T was by far the predominant active androgen present. Taken together, these two analyses suggest that T is by far the predominant active androgen present during musth in Asian bull elephants, and that no previously unidentified bioactive androgen is present.

Yon, L., Kanchanapangka, S., Chaiyabutr, N., Meepan, S., Stanczyk, F.Z., Dahl, N., Lasley, B., 2007. A longitudinal study of LH, gonadal and adrenal steroids in four intact Asian bull elephants (Elephas maximus) and one castrate African bull (Loxodonta africana) during musth and non-musth periods. Gen. Comp Endocrinol. 151, 241-245.
Abstract: During their annual musth cycle, adult African and Asian bull elephants have increased gonadal androgens (testosterone [T], dihydrotestosterone [DHT], androstenedione [A4]). Because musth is a physiologically and psychologically stressful time, this study was conducted to investigate whether the adrenal glands (stimulated by stress) increase production of both glucocorticoids and androgens during musth. Weekly serum samples were taken for 11-15 months from four intact adult Asian bull elephants, and from a castrate African bull elephant who exhibits musth. Testosterone, androstenediol (A5), A4, luteinizing hormone (LH), cortisol, and dehydroepiandrosterone (DHEA) were measured in each sample. In three of the four intact bulls, all hormones measured increased during musth. Adrenal androgens were strongly correlated with LH and testicular androgens, though not to cortisol. None of the hormones measured in the castrate bull increased during his musth cycles. While the significance of adrenal activity in the elephant during musth has yet to be determined, this study provides evidence that the adrenal gland actively produces both glucocorticoids and androgens during musth in the Asian elephant

Allen, W.R., 2006. Ovulation, pregnancy, placentation and husbandry in the African elephant (Loxodonta africana)
470. Philos. Trans. R. Soc. Lond B Biol. Sci. 361, 821-834.
Abstract: The African elephant reproduces so efficiently in the wild that overpopulation is now a serious problem in some game parks in Zimbabwe, Botswana and South Africa. The female reaches puberty between 10 and 12 years of age in the wild and, when in captivity, shows oestrous cycles of 14-15 weeks duration. She readily conceives a singleton in the wild yet her uterus has the capacity for twins. She shows a gestation length of 22 months and, in the wild, shows a population density and feed dependent intercalving interval of 4-8 years. The trophoblast erodes the lumenal epithelium of the endometrium and stimulates upgrowths of blood vessel-containing stromal villi, which develop eventually into the broad, tightly folded lamellae of the zonary, endotheliochorial placenta. Significant quantities of leaked maternal erythrocytes and ferric iron are phagocytosed by specialized trophoblast cells in the haemophagous zones at the lateral edges of the placental band. Although the placenta itself is endocrinologically inert, the foetal gonads, which enlarge greatly during the second half of pregnancy can synthesize 5alpha-dihydryoprogesterone and other 5alpha pregnane derivatives from cholesterol and pregnenolone. These products may synergize with progestagens secreted by the 2-8 large corpora lutea which are always present in the maternal ovaries throughout gestation to maintain the pregnancy state

Bagley, K.R., Goodwin, T.E., Rasmussen, L.E.L., Schulte, B.A., 2006. Male African elephants, Loxodonta africana, can distinguish oestrous status via urinary signals. Animal Behaviour 71, 1445.
Abstract: African elephants are a polygynous species that raise offspring in a matriarchal society. Unlike females, males disperse, spend time in mate groups and search for mates when mature. Urinary chemical signals aid males in detecting reproductively active females. A preovulatory pheromone has been identified in Asian elephants, Elephas maximus, but has not yet been experimentally identified in African elephants. In this study, the goal was to determine whether adult captive male African elephants can distinguish between urine from conspecific females in luteal and periovulatory oestrous stages as an indication that a preovulatory pheromone is released in the urine. Urine was collected from seven different female African elephants during their luteal and periovulatory periods of oestrus. Bioassays were conducted with nine adult male elephants housed at six different facilities. Males were presented with the two urine types and a control sample once a day over 3 days to reduce sample novelty, which can result in misleadingly high responses. All mates showed greater chemosensory responses to the periovulatory urine by trial 3 with the ability to distinguish the urines increasing over the 3 days. This is the first experimental behavioural evidence that African elephants release an oestrous pheromone in the urine. The ability of the captive male elephants to discern between the two urine types bolsters the hypothesis that there is a preovulatory pheromone in African elephants and encourages efforts to identify it.

Ball, R., Fad, O. Serum cortisols in captive Asian elephants (Elephas maximus) in different management systems at Busch Gardens Tampa Bay.  2006 Proceedings American Association of Zoo Veterinarians.  177-180. 2006.
Ref Type: Conference Proceeding
Abstract: Introduction:Cortisol is a widely accepted measure of stress in wild and captive animals.  In the past, captive elephant management systems have been criticized as potential stress inducers. The analysis of fecal cortisols is non-invasive and has been used to give long term evalutions of social and ecologic pressures in elephants and other species.  Salivary cortisols have also been used as a minimally invasive technique to measure social stress in captive elephants. The herd of Asian elephants at Busch Gardens Tampa Bay (BGT) changed from a traditional contact managemnt (free ccontact, FC) to a protected contact (PC) system utilizing positive-reinforcement based operant conditioning in 2004.  Serum cortisols were measured after the change and evaluated along wth banked samples from before. Long term sampling will be utilized to measure this transition but evaluating a single process will hopefully reflect the overall changes that can be expected with this change in management. While the individual variations are notable and other issues potentially confound the issue, it appears that this transition has lowered the serum cortisols in this herd.  In addition to serum cortisol measurements, the actual process of collecting the samples appears to be less stressful behaviorally. Pathologic processes should not be discounted when considering cortisol levels in evalauting stress in captive elephants.

Methods and Materials: Six female Asian elephants (Studbook numbers 30, 32, 304, 34, 35, 3) had been managed in a free contact system for many years.  Studbook number 304 was captive born and the others were wild born. Serum was collected intermittently during this management system to bank and for reproductive hormone analysis.  The elephants were placed in lateral recumbency by the handlers and blood collected from the ear vein on the caudal aspect of the down ear.  Reproductively sound animals were bled more frequently than the others.  Serum was frozen at -80°C until analyzed.  In August 2004, the first group of three animals was moved to the new barn and started the new positive-reinforcement, PC management system.  Within 5 wk, all animals had been moved over. All animals had been trunkwashed and were culture negative for Mycobacterium tuberculosis and negative on the newly developed MultiAntigen Print ImmunoAssay (MAPIA) and lateral-flow technology (Rapid Test) developed to detected antigen to M. tuberculosis.  As the caudal aspect of the ear was used for sampling, each elephant was asked to station in a static chute designed to allow training of voluntary ear-presentation for manipulation and blood collection. Handler safety and creating an effective learning environment for the elephants required training each to proceed to the chute solo and station there calmly. General desensitization techniques were applied as session durations were increased. Within the chute,individual elephants had significant room to maneuver. Since no physical restraint or sedation was utilized,animals were trained to cooperate fully and voluntarily allowing for blood sampling and other husbandry procedures. By May 2005, training for voluntary bloods draws was firmly established on all six animals.  The first approximately 20 samples collected under this new system were matched against the samples collected in the previous system.  Samples were selected against if the animal had an active problem or was on therapy for any reason.  Several animals had undergone a drug trial and these samples were selected against as well.  Serum was again stored in -80°C freezer until analyzed at Conservation and Research Center (CRC) Endocrine Research Laboratory, Smithsonian Institution, National Zoological Park, Front Royal, VA.  T-tests were utilized to discern any statistically significant results in the mean serum cortisols collected from animals before and after the implementation of the new husbandry systems.  Results were considered significant at alpha levels <0.05.

Results: The results and simple means of serum cortisols are listed in Table 1. Elephant No. 34 had essentially the same level of cortisol in both systems.  Elephant No. 32 had a reduction in the mean cortisol level of approximately 32% (20.84 versus 14.28 ng/ml) from the FC to the PC system.  Elephant No. 304 had a similar reduction of 37% in the mean cortisol (22.59 versus 14.29 ng/ml).  Statistical analyses results are reported here (means, standard deviations, t-test results).

Discussion: Serum was chosen over salivary and fecal sampling as a means to measure cortisol for several reasons. While fecal and salivary cortisol changes can reflect stresses within a reasonable period after the stressor (approximately 24 hr), serum cortisols is more likely to be reflective of the stressors closer to the moment of sampling.  The methodology is straightforward and less subject to the hazards for sample storage.  Timeliness of the sample result is also a benefit to serum sampling.  Blood sampling is a required husbandry practice in all elephant holding facilities belonging to the American Zoo and Aquarium Association (AZA).  While fecal cortisol samples may be useful to look at over a long term period to evaluate the transition from FC to PC, we choose to additionally look at how one specific task, blood collection, was affected by making this transition.  Fecal cortisols have been used to measure stress in transportation and environmental stress in some species, but are not thought to be reflective of the stress in a diagnostic procedure itself.  For this evaluation, the lag time period between the potential stressor (blood collection) and the means to measure the stressor are same.  Elephants No. 304 and 32 both had significant reductions in the mean serum cortisol levels.  Both are in good health and had no apparent inflammatory problems.  The logical deduction here is that the sampling process itself is less stressful in the PC management than the FC management.  Elephant 34 and 30 had essentially the same level of serum cortisol as measured by the mean in the different management systems.  Elephant 34 has developed significant uterine leiomyomas during the time period measured.  Elephant 30 has recently had clinical bouts of anterior enteritis and is suspected of having a dietary hypersensitivity to wheat.  Even with these two pathologic processes, the serum cortisol did not rise.  Elevations in cortisol are quite often explained as resulting from social, behavioral, or environmental causes and little attention is paid to inflammatory causes.  Associations between infections and elevated cortisols  have been noted in wild animals.  It is reasonable to assume that if these two processes did not exist, these levels would indeed be lower. Based on the other two elephants, a reduction of approximately 30% could be expected. Overall it appears that collecting blood from the elephants at BGT in the PC system is less stressful that the FC system.  As this is an example of how the routine husbandry and medical husbandry is now conducted, it can be expected that the overall net effect is going to be lowered stress in the elephants at BGT. ……………………………………………………………………………

Ball, R.L., Brown, J. Preliminary results of a cabergoline trial in captive elephant with hyperprolactinemia.  2006 Proceedings American Association of Zoo Veterinarians.  174-176. 2006.
Ref Type: Conference Proceeding
Abstract: Introduction: An Asian elephant (Elephas maximus) at Busch Gardens Tampa Bay (BGT) was diagnosed with hyperprolactinemia, with a persistently elevated serum prolactin concentration greater than 15 ng/ml, by the Conservation & Research Center (CRC) laboratory in January 1996.  She also had a number of other problems, including uterine disorders that resulted in consistently elevated progesterone. In March 2002, she was given cabergoline orally at a dose of 1 mg twice weekly p.o. for 6 mo.  Cabergoline is a long-acting dopamine receptor agonist with a high affinity for D2 receptors.  It exerts a direct inhibitory effect on the secretion of prolactin.  Cabergoline (Dostinex®, Pfizer Inc. Kalamazoo, Michigan 49007 USA) was purchased from a local pharmacy.  Serum prolactin concentrations declined almost immediately after treatment initiation, followed about 1 mo later by a drop in progesterone to baseline.  Progesterone secretion remained low until November 2002 when she resumed cycling based on the observation of a normal luteal phase based on serum progesterone profile.  From November 2002 through January 2004 she exhibited four normal estrous cycles.  Prolactin secretion also remained within the normal range for elephants, over 1 yr after treatment withdrawal. This female suffered no adverse effects due to the cabergoline treatment.  There were no behavioral changes noted or changes in appetite. Given the need to increase reproductive rates of African elephants (Loxodonta africana) to prevent captive extinction, it might be efficacious to treat genetically valuable females with cabergoline in the hope it will reinitiate reproductive cyclicity. Nearly 1/3 of African elephants with hormone data are not cycling normally, and in an earlier study 1/3 of these (11 of 30) were found to have increased serum prolactin levels.
 
Methods and Materials:
A clinical trial was undertaken with six captive African elephant females that were identified as good candidates for a cabergoline treatment study (i.e., they are acylic and had mean prolactin concentrations of >15 ng/ml).  The treatment consisted of 1 mg cabergoline given twice weekly p.o. for 6 mo.  Serum was banked and then analyzed at the CRC for progesterone and prolactin. All elephants were thought to be otherwise healthy.  Because prolactin is known to be an inflammatory marker, all candidates were required to have a negative lateral flow immunochromatograpy (Rapid Test) and multiple antigen immunoassay (MAPIA) for Mycobacteria tuberculosis.

Results:
A summary of the results is given in Table 1.  The treatment period is complete for three elephants, all of which showed a decrease in prolactin levels.  Elephant 1 showed a good response while on treatment, but did not cycle and serum prolactin has subsequently risen to pretreatment levels.  Increasing the dose in Elephant 2 and 3 reduced prolactin to baseline levels, but again did not result in a return to ovarian cyclicity.  Elephant 4 was taken off the study after only a few doses due to increased aggressive behaviors.  This is believed to be due to changes in the group social dynamics and not related to the cabergoline, as this behavior has continued after withdrawal of the drug. Based on these findings, the two newest candidates, Elephant 5 and 6, with very high prolactin concentrations have been placed on 2 mg/twice weekly for 1 yr pending continuation of this project.

Discussion:
Normalization of prolactin levels facilitated the return of normal cycles in an Asian elephant, but none of the African elephants have resumed cycling so far.  Thus, while the use of cabergoline shows promise in reducing elevated prolactin levels in both Asian and African elephants, other factors may need to be considered or a longer course at higher doses may be required for treatment to be successful in reinitiating ovarian activity.  The latter suggestion is supported by two of the animals (Elephants 2 and 3) in this limited trial, in which a decline in prolactin occurred after the dose was increased.  Understanding the etiology of hyperprolactinemia in elephants may also help in returning females to normal cycling.Relapse of hyperprolactinemia is more common in humans with micro- or macroprolactinomas.  Chronic estrogen stimulation is also known to increase prolactin levels.  A proposed pathophysiology is that elevated estrogen levels from persistent cycling will lead to elevated prolactin levels and acyclicity.  A difference between the two species in the causes of and potential treatment options for hyperprolactinemia should also be evaluated more closely.

1Busch Gardens Tampa Bay, 3605 Bougainvillea Drive, Tampa, FL 33674 USA;2Smithsonian Institution, National Zoological Park, Conservation & Research Center, Front Royal, VA 22630 USA
ACKNOWLEDGMENTS
We would like to thank the participating zoos for their cooperation and patience during this trial.
LITERATURE CITED
1Brown, J.L., S.L. Walker and T. Moeller. 2004.  Comparative endocrinology of cycling and noncycling Asian (Elephas maximus) and African (Loxodonta africana) elephants.  Gen. Comp. Endocrinol. 136:360-370.
2 Colao, A., A. Di Sarno, P. Cappabianca, C. Di Somma, R. Pivonello, and G. Lombardi. 2003. Withdrawal of long-term cabergoline therapy for tumoral and non-tumoral hyperprolactinemia.  New Engl. J. Med. 349:20232033.
3 Ismail, M.S., G.I. Serour, U. Torsten, H. Weitzel, and H.P. Berlien. 1998. Elevated serum prolactin level with high-dose estrogen contraceptive pills. .Eur. J. Contracept. Reprod. Health Care. 3(1):45-50.
4 Montero, A.M., O.A. Bottasso, M.R.Luraghi, A.G. Giovannoni, and L. Sen. 2001. Association between high serum prolactin and concomitant infections in HIV-infected patients. Human Immunol.62: 191-196.
5 Lyashchenko, K., M. Miller, and W.R. Waters. 2005. Application of multiple antigen print immunoassay and rapid lateral flow technology for tuberculosis testing of elephants. .  Proc. Am. Assoc. Zoo Vet. Annu. Meet. Pp. 64-65

Bertschinger, H., Delsink, A., Kirkpatrick, J.F., Human, A., Grobler, D., van Altena, J.J. Management of elephant populations in private South African game reserves with porcine zona pellucida vaccine.  2006 Proceedings American Association of Zoo Veterinarians.  283-285. 2006.
Ref Type: Conference Proceeding
Abstract: Control of African elephant populations has become an absolute necessity in a number of game reserves in southern Africa.  The two main methods used to control populations so far are culling and translocation. Culling, besides being regarded as inhumane and unacceptable in many quarters, is not suitable for smaller populations.  It requires that whole family units are culled simultaneously which could mean that in reserves with 10 to 50 elephants a considerable portion, if not the entire population, is killed.  As far as translocation is concerned, limited new space is available for elephants. The only alternative to the two above options is to control the rate of reproduction. The porcine zona pellucida (pZP) vaccine has been used to successfully contracept wild horses and other wildlife species.  Work on the contraception of African elephants was initiated in the Kruger National Park in 1995 when the potential for using the porcine zona pellucida (pZP) was investigated. Subsequently the first field trials on wild elephants were carried out in Kruger and the results clearly showed that elephants could be contracepted with the pZP vaccine, although the efficacy achieved was 80%. During these field trials safety and reversibility werecould be demonstrated. In 2000 an elephant contraceptive program was initiated at Makalali Private Game Reserve, RSA, which has become the flagship model for immunocontrol in African elephants. The preliminary findings have been reported in three publications.During the first year, all 18 cows that were individually identified and older than 12 yr of age were treated.  During the next 4 yr the number of cows contracepted increased to 23 as young animals were added to the program. The standard vaccination procedure during the first year consisted of a primary vaccination (600 μg or 400 μg pZP with 0.5 ml Freund's modified complete adjuvant) followed by boosters (200 μg pZP with 0.5 ml Freund's incomplete adjuvant) at 3 to 6-wk intervals. Annual boosters to maintain antibody titers and contraceptive effect followed.  To date, the success rate on cows that have passed reserve-specific intercalving period of 56 mo has been 100%. The population stabilized within 3 yr by which time when all cows that had been pregnant at the time of first vaccination in 2000 had calved. Once again safety during pregnancy (14 cows pregnant at 2-21 mo gestation when first treated gave birth to normal healthy calves) as well as side effects that were limited to occasional lumps at the site of vaccination could be shown. Following ground darting, behavioral patterns returned to pre-darting status within 2 days. During 2003 and 2004 most boosters were administered from a helicopter; whereas, previously they had been done from a vehicle or on foot.  In all cases, drop-out darts were used. Time taken for vaccination from helicopter take-off to landing was about 30 min (1.5 min per cow; 30 min for total time). This required prior knowledge of the locations of family units or that an individual in each unit is radio-collared. Herds settled down much more quickly (1-2 days) than if darted from the ground. Since then we have vaccinated another 107 elephant cows in eight game reserves.  The cow populations have ranged from 4 to 43. In one of the reserves, Mabula, RSA, two of the four cows vaccinated have passed the mean intercalving intervals of the reserve with neither of them producing a calf. Treatment at the remaining reserves was initiated in 2004 or 2005 and it is too early to evaluate results.  The most difficult reserve in terms of the vaccination process was Welgevonden, RSA, (35 000 ha) with 43 cows.  The reserve is mountainous and heavily wooded. None of the elephants were collared and individuals could not be easily identified on the day of primary vaccination.  The total flying time during which individuals were identified and vaccinated was 4.5 hr.  Administration of the first booster took about 2 hr to locate and vaccinate each cow. Between the first and second booster the first rains occurred, followed by the spring flush of the vegetation. By the time the second booster was attempted late in November, the trees all had foliage. Only half the cows were located and darted because the elephants were very difficult to spot under the tree canopies.  The valuable lessons we learned from this were: 1) that helicopter vaccinations should be performed when most trees are bare, and 2) when larger populations are vaccinated repeatedly during the first year, one cow in each family unit should be radio-collared. This makes rapid location of each unit possible and cuts down on the major cost factor that is flying time. Elephant behavior is being monitored in all eight reserves where contraception is being applied. Because most of them have been contracepted recently, only the data from Makalali is available. The elephants at Makalali have been monitored intensively almost on a daily basis. To date, no anomalies in terms of aggressive or indifferent behavior with regards to nursing time, nursing behavior and calf proximity have been noted. No change in the cows' social hierarchy has been noted. Since January 2003, a total of 15 heats were observed in 10 cows (nine in 2003 and six in 2004) with four mating episodes. For the same period, 38 musth occasions were seen in five bulls (26 in 2003 and 12 in 2004). These occasions include musth displayed in the same bull during consecutive days or within the same musth cycle. The greatest occurrence of musth was recorded in the largest, dominant bull. Bulls were not observed harassing or separating cows off from their herds or calves as a result of increased estrous frequency. Thus, the Makalali program demonstrates that pZP does not cause herd fragmentation, harassment by bulls, change in rank and other negative behaviors normally associated with hormonal contraceptives. In conclusion we feel that it is important to emphasize the following points: The pZP vaccine can be used successfully to contracept African elephants The vaccine is safe during pregnancy and has no negative effect on birth or calf raising It has no side effects other than occasional swelling at the site of vaccination It is reversible Other than an increased incidence of heat no behavioral side effects were seen.

Brown, J.L., Somerville, M., Riddle, H.S., Keele, M., Duer, C. Comparative endocrinology of testicular and thyroid function in captive Asian and African elephant bulls. Proceedings International Elephant Conservation & Research Symposium.  58-75. 2006. 2006.
Ref Type: Conference Proceeding

Brown, J.L., Freeman, E., Duce, C. Update on the reproductive status of female Asian and African elephants in the SSP population of North America. Proceedings International Elephant Conservation & Research Symposium.  48-57. 2006. 2006.
Ref Type: Conference Proceeding

Cooper, D.W., Larsen, E., 2006. Immunocontraception of mammalian wildlife: Ecological and immunogenetic issues. Reproduction 132, 821-828.
Abstract: Immunocontraception involves stimulating immune responses against gametes or reproductive hormones thus preventing conception. The method is being developed for the humane control of pest and overabundant populations of mammalian wildlife. This paper examines three fundamental issues associated with its use: (1) the difficulties of obtaining responses to self-antigens, (2) the likely evolution of genetically based non-response to immunocontraceptive agents, and (3) the possible changes in the array of pathogens possessed by the target species after generations of immunocontraception. Our review of the literature demonstrates that the barriers to an effective immunocontraceptive are at present very basic. Should they be overcome, the effects of immunocontraception on the immunogenetic constitution of wildlife populations through the selection for nonresponders must be examined. We suggest that the attempt to use the animal's own immune system to modulate reproduction may be incompatible with the basic biological function of protection against infectious disease. Research programs on mammalian immunocontraception should involve measurement of the heritability of non-response and an assessment of the likely change in the response of the contracepted population to possible pathogens.

Delsink, A.K., van Alten, J.J., Grobler, D., Bertschinger, H., Kirkpatrick, J., Slotow, R., 2006. Regulation of a small, discrete African elephant population through immunocontraception in the Makalali Conservancy, Limpopo, South Africa. South African Journal of Science 102, 403-405.
Abstract: Populations of the African elephant, Loxodonta africana, are growing rapidly in southern Africa, to the extent that population control has become essential. The management option of translocation is no longer realistically available, whilst culling has become ethically unacceptable, especially to the general public. Previous immunocontraception trials on elephants with Porcine Zona Pellucida (PZP) vaccine demonstrated that it is safe, effective, reversible, remotely deliverable, and has had no evident adverse side effects. We demonstrate effective contraceptive management of a discrete, small population of free-roaming elephants in the Makalali Conservancy, Limpopo province, South Africa. Complete reproductive control has been demonstrated in all 18 original targeted females, who have by now passed the population's average intercalving interval of 56 months without giving birth. A zero population growth rate has been maintained within this target group since August 2002. On the basis of this small sample over a short period, immunocontraception should be considered a viable means of population management as an alternative to long-term culling strategies in small populations

Drews, B., Göritz, F., Hermes, R., Streich, J.W., Rich, P., Schmitt, D., Lung, N., Renfree, M.B., Gaeth, A.P., Short, R.V., Hildebrandt, T.B. Morphological and ultrasonographic characterization of the embryonic development in elephants. Proceedings International Elephant Conservation & Research Symposium.  82-83. 2006. 2006.
Ref Type: Conference Proceeding

Druce, H., Pretorius, K., Druce, D., Slotow, R., 2006. The effect of mature elephant bull introductions on resident bull's group size and musth periods: Phinda Private Game Reserve, South Africa. South African Journal of Wildlife Research 36, 133-137.
Abstract: African elephants have been reintroduced into small, enclosed reserves in South Africa,many populations being established with orphans <10 years old.This has resulted in abnormal behaviour in some elephant populations, which was corrected in Pilanesberg National Park by introducing older bulls and culling certain problem elephants.In July 2003, three older bulls (29-41 years old) were introduced into Phinda Private Game Reserve, KwaZulu-Natal, South Africa, in order to normalize the bull age structure and in an attempt to reduce the abnormally long musth period of one particular resident bull. These introduced bulls were monitored intensively after release, as was the resident bull population, both before and after introduction of the older bulls.The introduced bulls all came into musth within eleven months postrelease.The older bulls do not appear to have had any influence on the musth periods of the oldest resident bull (36 years old at introduction). Detailed behavioural studies of the effects of management actions on elephant populations, within small, enclosed reserves provide information and resources for future management decisions.This study demonstrates that old bulls can be successfully introduced to very small areas provided that electrification of the entire perimeter is secure. Further, the introduction has no detectable medium-term (one year) effect on the behaviour of a relatively dense population of resident elephants, and the welfare of the elephants was not greatly affected.

Freeman, E.W., Brown, J.L., Whyte, I. Reproductive success of elephants in Kruger National Park, South Africa (1976-1995). Proceedings International Elephant Conservation & Research Symposium.  87-91. 2006. 2006.
Ref Type: Conference Proceeding

Hildebrandt, T., Goeritz, F., Hermes, R., Schaftenaar, W., Drews, B. Dystocia - an increasing problem in captive breeding programs: Causes and treatment. Proceedings International Elephant Conservation & Research Symposium.  92. 2006. 2006.
Ref Type: Conference Proceeding

Larke, A., Crews, D.E., 2006. Parental investment, late reproduction, and increased reserve capacity are associated with longevity in humans. J Physiol Anthropol 25, 119-131.
Abstract: Throughout the living world trade-offs between reproductive success and longevity have been observed. In general, two extremes of life history patterning are reported, r- and K-selected species. The latter tend toward larger body sizes, few offspring from any one pregnancy, few offspring over the female reproductive span, longer life spans, and greater parental investment (PI: all efforts and expenses associated with the production, gestation, post-natal care, feeding, and protection of young) (e.g., whales, elephants, hominids). r-selected species tend toward smaller body size, multiple births/litters per pregnancy, female production of many gametes and offspring over the life span, and low levels of PI (e.g., most plants, insects, mice). These differences have significant influences on physiological variation among human populations.Across human samples, reproductive success (RS: the number of offspring successfully birthed and reared to reproductive age) has been reported to vary positively, negatively, and not at all with longevity of women. This complexity may be in part due to the fact that both early-life and late-life fecundity are associated with longevity in women, while total parity seems a poor gauge of female longevity in humankind. Large variations in associations of RS with longevity in women suggest that multiple factors may confound this association. One confounding factor is that among women, RS is largely determined not by fecundity, but by the quality of PI available to offspring. Among modern humans, PI is more complex, longer lasting (both relatively and absolutely), and extensive than for any other ammal. This suggests that modern human life history is a reflection of the co-evolution of longevity and extensive PI as part of our species' biocultural evolution. The need for long-term PI has greatly shaped human physiological variation and patterns of longevity.

Oerke, A.-K., Heistermann, M.A., Hodges, K. Duration of pregnancy and its relation to sex of calf and age of cow in the European population of Asian and African elephants. Proceedings International Elephant Conservation & Research Symposium.  125-131. 2006. 2006.
Ref Type: Conference Proceeding

Riley, L.W., 2006. Of mice, men, and elephants: Mycobacterium tuberculosis cell envelope lipids and pathogenesis
454. J. Clin. Invest 116, 1475-1478.
Abstract: Mycolic acids and structures attached to them constitute a major part of the protective envelope of Mycobacterium tuberculosis, and for this reason, their role in tuberculosis pathogenesis has been extensively studied. In this issue of the JCI, Rao et al. examine the effect of trans-cyclopropanation of oxygenated mycolic acids attached to trehalose dimycolate (TDM) on the murine immune response to infection (see the related article beginning on page 1660). Surprisingly, they found that an M. tuberculosis mutant lacking trans-cyclopropane rings was hypervirulent in mice. The recent recognition of a hypervirulence phenotype in mice associated with laboratory and clinical M. tuberculosis strains with altered cell wall components has provided new insights into how M. tuberculosis may establish persistent infection. However, to date, characterization of these bioactive products in pathogenesis has been largely reductionistic; the relationship of their effects observed in mice to the persistent infection and tuberculosis caused by M. tuberculosis observed in humans remains obscure

Sa-Ardrit, M., Saikhun, J., Thongtip, N., Damyang, M., Mahasawangkul, S., Angkawanish, T., Jansittiwate, S., Faisaikarm, T., Kitiyanant, Y., Pavasuthipaisit, K., Pinyopummin, A., 2006. Ultrastructural alterations of frozen-thawed Asian elephant (Elephas maximus) spermatozoa
491. Int. J. Androl 29, 346-352.
Abstract: Intact plasma and acrosome membranes and functional mitochondria following cryopreservation are important attributes for the fertilizing ability of spermatozoa. In the present study, functional and ultrastructural changes of Asian elephant spermatozoa after cryopreservation either in TEST + glycerol or HEPT + dimethyl sulphoxide (DMSO) were evaluated by fluorescent techniques and electron microscopy. Sperm frozen in TEST + glycerol had higher proportion of sperm with intact plasma (49.1 +/- 9.2% vs. 30.9 +/- 3.9%) and acrosomal (53.7 +/- 4.9% vs. 35.8 +/- 6.1%) membranes, as well as active mitochondria (57.0 +/- 7.2% vs. 42.0 +/- 5.0%) than those cryopreserved in HEPT + DMSO. The results obtained from electron microscopy were similar to those obtained by fluorescence microscopy. The percentage of normal spermatozoa was higher when spermatozoa were frozen in TEST + glycerol than those frozen in HEPT + DMSO (31.8 +/- 5.6 vs. 28.5 +/- 6.4). The ultrastructural alterations revealed by transmission electron microscopy could be classified as (i) distension of plasma membrane, while the acrosome was swollen; (ii) disruption or loss of plasma membrane, while acrosome was swollen with distended outer acrosomal membrane; (iii) disruption or loss of plasma and outer acrosomal membrane with leakage of acrosome content; (iv) extensive vesiculation of plasma and outer acrosomal membrane and leakage of acrosome content; (v) a complete loss of both plasma membrane and outer acrosomal membrane; and (vi) swelling of mitochondria. These findings suggest that the freezing and thawing procedure caused structural damage to elephant spermatozoa, especially in the plasma membrane, acrosome and mitochondria. Fluorescence and electron microscopic evaluations are potentially a powerful tool in the analysis of elephant spermatozoa after freezing and thawing

Saikhun, J., Thongtip, N., Kornkaewrat, K., Mahasawangkul, S., Angkawanish, T., Boonprasert, K., Pinyopummin, A. Osmotic stress on motility and membrane integrity of Asian elephant spermatozoa analyzed by computer-assisted semen analysis. Proceedings International Elephant Conservation & Research Symposium.  163. 2006. 2006.
Ref Type: Conference Proceeding

Shannon, G., Page, B.R., Duffy, K.J., Slotow, R., 2006. The role of foraging behaviour in the sexual segregation of the African elephant
419. Oecologia. 150, 344-354.
Abstract: Elephants (Loxodonta africana) exhibit pronounced sexual dimorphism, and in this study we test the prediction that the differences in body size and sociality are significant enough to drive divergent foraging strategies and ultimately sexual segregation. Body size influences the foraging behaviour of herbivores through the differential scaling coefficients of metabolism and gut size, with larger bodied individuals being able to tolerate greater quantities of low-quality, fibrous vegetation, whilst having lower mass-specific energy requirements. We test two distinct theories: the scramble competition hypothesis (SCH) and the forage selection hypothesis (FSH). Comprehensive behavioural data were collected from the Pongola Game Reserve and the Phinda Private Game Reserve in South Africa over a 2.5-year period. The data were analysed using sex as the independent variable. Adult females targeted a wider range of species, adopted a more selective foraging approach and exhibited greater bite rates as predicted by the body size hypothesis and the increased demands of reproductive investment (lactation and pregnancy). Males had longer feeding bouts, displayed significantly more destructive behaviour (31% of observations, 11% for females) and ingested greater quantities of forage during each feeding bout. The independent ranging behaviour of adult males enables them to have longer foraging bouts as they experience fewer social constraints than females. The SCH was rejected as a cause of sexual segregation due to the relative abundance of low quality forage, and the fact that feeding heights were similar for both males and females. However, we conclude that the differences in the foraging strategies of the sexes are sufficient to cause spatial segregation as postulated by the FSH. Sexual dimorphism and the associated behavioural differences have important implications for the management and conservation of elephant and other dimorphic species, with the sexes effectively acting as distinct "ecological species"

Shannon, G., Page, B.R., Duffy, K.J., Slotow, R., 2006. The consequences of body size dimorphism: Are African elephants sexually segregated at the habitat scale? Behaviour 143, 1145-1168.
Abstract: Sexual segregation is a commonly observed phenomenon in dimorphic ungulates, which has been categorised into two distinct components: social segregation and habitat segregation. In this study we investigated whether elephants were sexually segregated at the habitat scale. The locations of 12 family groups and 16 males, in three distinct populations were recorded over a period of 2.5 years. Selection ratios were calculated for each habitat type and a Kendall's coefficient of concordance was used for the analyses. The habitat and foraging preferences were firstly tested for concordance within sex, and then between the sexes. Female habitat preferences showed significant concordance across all reserves and they also exhibited strong concordance in their summer foraging preferences. Their weakest association with habitat and foraging preference was during winter, which may be related to resource scarcity. Males exhibited significant concordance in their habitat preferences in two out of the three reserves. They had their weakest associations in the summer months and this may be linked to avoidance of other males in musth and the abundance of forage. There were no significant differences in habitat preference between males and females and it is likely that individual preferences vary as much within sex as between sexes. Differential habitat utilisation does not appear to be driving sexual segregation in elephants and it is postulated that sociality, divergent reproductive strategies and foraging behaviour at the plant scale play a more significant role. The results of this study highlight the importance of scale in elucidating the mechanisms involved in sexual segregation.

Shrader, A.M., McElveen, M.E., Lee, P.C., Moss, C.J., van Aarde, R.J., 2006. Growth and age determination of African savanna elephants. Journal of Zoology, London 270, 40-48.
Abstract: Understanding the population dynamics of savanna elephants depends on estimating population parameters such as the age at first reproduction, calving interval and age-specific survival rates. The generation of these parameters, however, relies on the ability to accurately determine the age of individuals, but a reliable age estimation technique for free-ranging elephants is presently not available. Shoulder heights of elephants were measured in 10 populations in five countries across southern and eastern Africa. Data included shoulder height measurements from two populations where the age of each individual was known (i.e. Addo Elephant National Park, South Africa and Amboseli National Park, Kenya). From the known-age data, Von Bertalanffy growth functions were constructed for both male and female elephants. Savanna elephants were found to attain similar asymptotic shoulder heights in the 10 populations, while individuals in the two known-age populations grew at the same rate. The Von Bertalanffy growth curves allowed for the accurate age estimation of females up to 15 years of age and males up to 36 years of age. The results indicate that shoulder height can serve as an indicator of chronological age for elephants below 15 years of age for females and 36 years of age for males. Ages derived from these growth curves can then be used to generate age-specific population variables, which will help assess the demographic status of savanna elephant populations across Africa.

Thitaram, C., Pongsopawijit, P., Thongtip, N., Angkavanich, T., Chansittivej, S., Wongkalasin, W., Somgird, C., Suwankong, N., Prachsilpchai, W., Suchit, K., Clausen, B., Boonthong, P., Nimtrakul, K., Niponkit, C., Siritepsongklod, S., Roongsri, R., Mahasavankul, S., 2006. Dystocia following prolonged retention of a dead fetus in an Asian elephant (Elephas maximus)
458. Theriogenology 66, 1284-1291.
Abstract: A 32-year-old nulliparous female Asian elephant (Elephas maximus) showed signs of parturition 8 months later than predicted from the breeding records. However, while serosanguineous fluid, necrotic tissue and pieces of amnion were expelled, second-stage labor did not progress. Since the fetus was not found during an endoscopic examination of the vestibule, it was assumed that the elephant had calved unseen and she was returned to the forest to recuperate. Twelve months later, the elephant showed clear signs of second-stage labor accompanied by a bulge in the perineum and passage of keratinized nail through the vulva. A 35 cm episiotomy incision was made in the perineum just below the anus, via which chains were attached to the forelimbs of the fetus. Traction on the forelimbs alone proved insufficient to achieve delivery because the fetal head kept rotating and impacting in the pelvis. However, traction applied via a hook inserted behind the mandibular symphysis allowed the head to be elevated and extended, and the fetus to be delivered. The episiotomy wound was sutured in two layers and although the skin did not heal during primary closure it subsequently healed uneventfully by second intention. Retrospective evaluation of the elephant's serum progestagens profile demonstrated a fall to baseline at the suspected onset of parturition, supporting the supposition that the fetus was retained in the uterus for 12 months after parturition began. It is suggested that serum progestagens concentrations should be monitored regularly in mated elephant cows to verify the establishment of pregnancy and to better estimate the expected timing, and the onset of calving

Thitaram, C., Thongtip, N., Somgird, C., Colenbrander, B., Van Boxtel, D.C.J., Lenstra, J.A. Molecular tool for genetic management and parentage test to control poaching in Asian elephants. Proceedings International Elephant Conservation & Research Symposium.  205-209. 2006.
Ref Type: Conference Proceeding

Thongtip, N., Mahasawangkul, S., Kornkaewrat, K., Saikhun, J., Angkwanish, T., Jansittiwate, S., Boonprasert, K., Wajjwalku, W., Songsasen, N., Pinyopummintr, T., Pinyopummin, A. Potential factors affecting Asian elephant semen quality in Thailand. Proceedings International Elephant Conservation & Research Symposium.  210. 2006. 2006.
Ref Type: Conference Proceeding

Thongtip, N., Mahasawangkul, S., Thitaram, C., Pongsopavijitr, P., Kornkaewrat, K., Saikhun, J., Angkawanish, T., Jansittiwate, S., Rungsri, R., Boonprasert, K., Somkird, C., Wongkalasinh, W., Wajjwalku, W., Songsasen, N., Pinyopummintr, T., Pinyopummin, A. First report of pregnancy by artificial insemination with chilled semen of an Asian elephant in Thailand. Proceedings International Elephant Conservation & Research Symposium.  273. 2006. 2006.
Ref Type: Conference Proceeding

Vinogradov, I.V., Kochneva, G.V., Shchelkunov, S.N., Riabchikova, E.I., 2006. [Reproduction of cowpox virus strain EP-2 isolated from an elephant in primary fibroblast cultures and chorion-allantoic chick embryos]
451. Vopr. Virusol. 51, 44-48.
Abstract: Electron microscopy was used to study the reproduction of cowpox virus strain EP-2 in the cells of a primary fibroblast cultures (PFC) and chorion-allantoic membrane (CAM) of chick embryos (CE). The sequential stages of viral morphogenesis and the structure of A-type inclusions were described. The parameters of viral reproduction in PFC and CE CAM were compared. The formation of crystalloid tubular structures in PFC, unusual electron dense inclusions in the cells of CE CAN, and different variants of A-type inclusions in the cells of a pock was found. The histological and ultrastructural characteristics of pocks in CE CAM are described

Allen, W.R., Mathias, S., Ford, M., 2005. Placentation in the African elephant, Loxodonta africana. IV. Growth and function of the fetal gonads
550. Reproduction. 130, 713-720.
Abstract: The gonads, both ovaries and testes, of 44 elephant fetuses weighing 0.09-112 kg (6.1-21.3 months gestation) were examined grossly and histologically. As in equids, elephant fetal gonads undergo a phase of marked growth and enlargement during the second half of gestation, which is more pronounced in ovaries than testes due to growth and antrum formation of numerous follicles in the former. Stromal cells undergo hypertrophy and transformation to form zones of interstitial cells that are associated with the enlarged follicles in the ovaries and in which the primitive seminiferous tubules are embedded in the testes. The interstitial cells have the capacity to synthesize 5alpha-dihydroprogesterone and other 5alpha-reduced progestagens from cholesterol and pregnenelone and the hypothesis is raised that these fetal gonadal progestagens may supplement significantly the progestagens secreted by the multiple large corpora lutea of pregnancy in the elephant

Andrews, J., Mecklenborg, A., Bercovitch, F.B., 2005. Milk intake and development in a newborn captive African elephant  (Loxodonta africana). Zoo Biology 24, 275-281.
Abstract: In August 2003 the San Diego Zoo's Wild Animal Park (WAP) and the Lowry Park Zoo, under the auspices of the AZA's Elephant SSP and a USFWS permit, imported 3.8 African elephants (Loxodonta africana ) from the Kingdom of Swaziland. When they were captured, transrectal ultrasound examinations revealed that one nulliparous cow was approximately 10 months pregnant.  At the time of their arrival (August 2003), all of the animals were estimated to be approximately 13 years old and were thought to be nulliparous.  Based on the ultrasound examination results and the average African elephant gestation period, parturition was predicted to be 20 February 2004. In this report, we provide the first detailed data about nursing activity around the clock and newborn calf development, describe maternal and neonatal nighttime activity budgets, and explore maternal weight changes during suckling and lactation. The newborn calf suckled significantly more at night than during the day, but suckled for only about 2 hr per 24-hr period. Regression analysis revealed that through the first 3 months of life the calf gained 0.385 kg/day while it suckled on a regular basis. We compare our findings with published information on wild elephants, and conclude that although the growth rate is reduced compared to hand-reared elephant calves, the suckling patterns are almost identical to those reported for wild calves.

Deem, S.L., Brown, J.L., Eggert, L., Wemmer, C., Htun, W., Nyunt, T., Murray, S., Leimgruber, P. Health and management of working elephants in Myanmar (Burma). Procedings American Association of Zoo Veterinarians.  228-231. 2005.
Ref Type: Conference Proceeding
Abstract: Myanmar has approximately 6,000 working elephants.  Remaining wild elephants are declining, partly because of live-capture for captivity.  Through health and reproductive assessments, genetic analyses and GPS tracking of captive and wild elephants, we are exploring linkages between the two populations and conducting studies to reduce morbidity and mortality of captive elephants. Captive elephants live and work in Myanmar's forests in close proximity and contact to the remaining wild herds. We propose that reducing morbidity and mortality in the captive elephants will decrease the need for live-capture, and the risk of disease transmission, to wild elephants.
Introduction
There are an estimated 6,000 working elephants in Myanmar - half owned by the government operated Myanmar Timber Enterprise (MTE) and half owned privately^.5 This may be one of the largest captive elephant populations in the world and its management will have a significant impact on remaining wild herds in Myanmar.^4,6,8  With mortality rates higher than birth rates, the working population is probably maintained by supplementing it with elephants captured from the wild.⁵ There is evidence that continued harvest of wild elephants may have reduced the remaining wild populations of Myanmar.  Recent surveys of wild populations in two of Myanmar's protected elephant ranges revealed extremely low dung counts, indicative of small and declining herds. Constant contact with captive elephants in Myanmar's forests may exacerbate the threat to Myanmar wild elephants by increasing the transmission of disease between these two groups. For both the above reasons, we believe that the conservation of wild elephants in Myanmar will require significant improvements in the care and management of currently existing captive populations.  
Elephants owned by MTE receive veterinary care from the Burmese veterinarians that work for the timber company and travel extensively throughout the country to sites were the elephants are located.¹ There is a dire need for veterinary supplies and laboratory capabilities in the country. Currently, veterinary practices are based on the extensive field experience of lead MTE veterinarians. However, MTE veterinarians frequently rely on older published work ^3,7 and would benefit significantly from training that incorporates new insights into elephant health and new veterinary techniques. Similarly, because of their close-up experience of elephant health problems in the forests, MTE veterinarians may be able to make important new contributions to the care and management of elephants elsewhere.     
The overall objective of our study is to work jointly with MTE veterinarians to develop long-term captive population management strategies to reduce mortality and increase births in the working timber elephants and stop the continued off-take of animals from the wild to supplement captive herds.
Methods
The health component of this study has five major objectives.  These are to:
1              Conduct a training workshop, in conjunction with MTE veterinarians, on elephant management and veterinary care. 
2              Develop protocols so that the MTE veterinarians can collect samples for reproductive, genetic, and health status assessments.
3              Analyze samples and provide data to MTE veterinarians to improve husbandry, preventive care and disease treatment of working elephants.
4              Develop a comprehensive bibliography of all published information on the health and management of Myanmar elephants.
5              Perform an epidemiologic evaluation of records available on the historic and current working elephant population.
Specific steps to achieve these objectives include: 
1              Determine causes and rates of morbidity and mortality of captive MTE elephants.
2              Determine causes of low rates of reproduction in captivity.
3              Develop a genetic profile of the captive herds.
4              Develop a protocol to assess oozies-Burmese mahout-expertise in parallel with endocrine and health assessments to determine quality of care and potentially related stress.
5              Develop small population viability models to assess how current mortality effects long-term survival of the captive population and what supplementation from the wild is needed for short- and long-term sustainability.
6              Use population viability models to demonstrate how supplementation from the wild will negatively affect that population.
7              Get baseline health parameter data on free-ranging elephants.
8              Quantify habitat/space use using GPS and satellite tracking of captive and wild elephants. 
Results and Discussion
During an initial exploratory visit in November 2004, we learned that the annual mortality rate for MTE working elephants was 2.4% (66) in 2003.  Deaths occurred in all age groups (>18 yr, n = 40; 4 - 17 yr, n = 11; <4 yr, n = 15) and included preventable diseases (i.e., poor nutrition, heat stroke, diarrhea, dystocia, infectious and parasitic agents).  Additionally, we collected samples for performing health, genetic and endocrine analyses of 22 elephants maintained in one of the working camps (results to be presented). A relationship also was established with the veterinary staff at the Yangon Zoo, including follow up donations of veterinary literature and journals to the zoo. We provided medical advice for the care of an orphaned elephant calf and other animals housed at the zoo during our brief visit. We are seeking funds for a training course to be conducted in late 2005 and hope to perform health evaluations on a larger number of zoo and working elephants during that visit.
The National Zoo already has an extensive conservation program for wild elephants in Myanmar.^4,6,8  This program has focused on assessing wild elephant populations in protected areas and satellite-tracking of four wild elephants to learn more about their conservation status and ecology in Myanmar.  Currently this work is being extended to a national elephant survey. Part of this work included collecting fecal samples for genetic and health assessments.
The Smithsonian team of researchers involved in this project includes a veterinarian, reproduction physiologist, geneticist, conservation biologist, and landscape ecologist.  All members of this multidisciplinary team have extensive experience working with elephants and together provide the necessary expertise to study and understand the numerous factors affecting Myanmar's captive elephants and the long-term survival of elephants in Myanmar.  These challenges range from human land use and elephant population fragmentation, human-elephant conflict, poor reproduction and health care of captive elephants and lack of information on the health status of the wild elephants.  A viable conservation initiative for the elephants of Myanmar requires that health issues be addressed as one component of a comprehensive program to address the anthropogenic pressures on both working and wild elephants.²
The elephants of Myanmar are an excellent example of the fine line that exists between captive and wild animals, especially as it relates to health.  Captive and wild elephants are regularly in direct and indirect contact.  The working elephants live with their oozies who may expose them to diseases, such as tuberculosis.  The working elephants in turn may encounter wild elephants at night in the forests where they forage and live during non-working hours. In fact, the majority of captive born calves are said to be sired by wild bulls.  Potentially, the use of working elephants in selectively extracting valuable timber provides new strategies for the conservation of elephants and forests. Most likely, "elephant-logging" is less damaging than machine-operated timbering projects that tend to clear-cut areas and also damage the soil and streams.  However, decreasing the negative impact of such practices (i.e., minimizing off-take of elephants from the wild, decreasing disease risks to the wild elephants) is imperative.  
LITERATURE CITED
1 Aung, T., and T. Nyunt.  2002.  The care and management of the domesticated Asian elephant in Myanmar.  In: Baker, I., and M. Kashio (eds.): Giants on our hands. Proc. Int. Workshop Domesticated Asian Elephant. Dharmasarn Co., Ltd. Bangkok, Thailand. Pp. 89 - 102.
2 Deem, S.L., W.B. Karesh, and W. Weisman.  2001.  Putting theory into practice: wildlife health in conservation.  Conserv. Biol. 5: 1224-1233.
3 Evans, G.H. 1910.  Elephants and Their Diseases.  Government Printing. Rangoon. 323 
4 Kelly, D.S. 2005.  Habitat selection in declining elephant populations of Alaungdaw Kathapa National Park. Masters Thesis.  George Mason University.
5 Lair, R.C. 1997.  Myanmar. In: Gone Astray: The Care and Management of the Asian Elephant in Domesticity. FAO Regional Office for Asia and the Pacific, Thailand.  RAP Publication. Pp. 99-131
6 Leimgruber, P., and C. Wemmer.  2004.  National elephant symposium and workshop. Report to the USFWS and the Myanmar Forest Department.
7 Pfaff, G. 1930.  Reports on Diseases of Elephants.  Government Printing. Rangoon. 91
8 Wemmer, C., P. Leimgruber and D. S. Kelly.  2005.  Managing wild elephants in Alaungdaw Kathapa National Park and Htamanthi Wildlife Sanctuary.  Report to the USFWS and the Myanmar Forest Department.

Delves, P.J., Roitt, I.M., 2005. Vaccines for the control of reproduction--status in mammals, and aspects of comparative interest
592. Dev. Biol. (Basel) 121, 265-273.
Abstract: The objective of producing vaccines which target elements of the reproductive system to control fertility has been pursued for many years. Of the many targets for such vaccines, several sperm-associated antigens have been proposed for antibody-mediated intervention before fertilization but the very abundance of antigen to be neutralized has been a barrier. Zona pellucida antigens associated with the surface of the oocyte have also been targeted and used successfully for control of 'wild' elephant populations but worries concerning immunopathologically-mediated tissue damage have been mooted. Vaccines using human chorionic gonadotropin (hCG) which is required for the implantation and maintenance of the fertilized egg, although of interest for the development of fertility control in human populations, has no relevance in the context of the present conference because external fertilization of fish eggs is independent. The pathways by which gonadotropin-releasing hormone (GnRH) secreted by the hypothalamus promote release of luteinizing (LH) and follicle-stimulating hormone (FSH) which govern the physiological maturation and maintenance of the reproductive organs, provide many targets for immunological intervention. Most consistent success has been reported using GnRH-based vaccines which are immunosterilizing in a variety of mammalian species such as pigs, rodents and white-tailed deer. The fact that the structure of the decapeptide, GnRH, has been maintained over so many years of evolution and been conserved across so many animal species, encourages the view that a strategy for control of sexual maturation in fish based upon stimulation of GnRH antibodies may well prove to be a practical proposition, provided the formulation of an appropriate highly immunogenic vaccine can be achieved

Ganswindt, A., Rasmssen, H.B., Heistermann, M., Hodges, J.K., 2005. The sexually active states of free-ranging male African elephants (Loxodonta africana): defining musth and non-musth using endocrinology, physical signals, and behavior. Horm Behav 47, 83-91.
Abstract: Musth in male African elephants, Loxodonta africana, is associated with increased aggressive behavior, continuous discharge of urine, copious secretions from the swollen temporal glands, and elevated androgen levels. During musth, bulls actively seek out and are preferred by estrous females although sexual activity is not restricted to the musth condition. The present study combines recently established methods of fecal hormone analysis with long-term observations on male-female associations as well as the presence and intensity of physical signals to provide a more detailed picture about the physical, physiological, and behavioral characteristics of different states of sexual activity in free-ranging African elephants. Based on quantitative shifts in individual bull association patterns, the presence of different physical signals, and significant differences in androgen levels, a total of three potential sub-categories for sexually active bulls could be established. The results demonstrate that elevations in androgen levels are only observed in sexually active animals showing temporal gland secretion and/or urine dribbling, but are not related to the age of the individual. Further, none of the sexually active states showed elevated glucocorticoid output indicating that musth does not represent an HPA-mediated stress condition. On the basis of these results, we suggest that the term "musth" should be exclusively used for the competitive state in sexually active male elephants and that the presence of urine dribbling should be the physical signal used for defining this state.

Ganswindt, A., Heistermann, M., Hodges, K., 2005. Physical, physiological, and behavioral correlates of musth in captive African elephants (Loxodonta africana)
594. Physiol Biochem. Zool. 78, 505-514.
Abstract: Although musth in male African elephants (Loxodonta africana) is known to be associated with increased aggressiveness, urine dribbling (UD), temporal gland secretion (TGS), and elevated androgens, the temporal relationship between these changes has not been examined. Here, we describe the pattern of musth-related characteristics in 14 captive elephant bulls by combining long-term observations of physical and behavioral changes with physiological data on testicular and adrenal function. The length of musth periods was highly variable but according to our data set not related to age. Our data also confirm that musth is associated with elevated androgens and, in this respect, show that TGS and UD are downstream effects of this elevation, with TGS responding earlier and to lower androgen levels than UD. Because the majority of musth periods were associated with a decrease in glucocorticoid levels, our data also indicate that musth does not represent a physiological stress mediated by the hypothalamic-pituitary-adrenal axis. Furthermore, we demonstrate that the occurrence of musth is associated with increased aggression and that this is presumably androgen mediated because aggressive males had higher androgen levels. Collectively, the information generated contributes to a better understanding of what characterizes and initiates musth in captive African elephants and provides a basis for further studies designed to examine in more detail the factors regulating the intensity and duration of musth

Ganswindt, A., Rasmussen, H.B., Heistermann, M., Hodges, J.K., 2005. The sexually active states of free-ranging male African elephants (Loxodonta africana): defining musth and non-musth using endocrinology, physical signals, and behavior
652. Horm. Behav. 47, 83-91.
Abstract: Musth in male African elephants, Loxodonta africana, is associated with increased aggressive behavior, continuous discharge of urine, copious secretions from the swollen temporal glands, and elevated androgen levels. During musth, bulls actively seek out and are preferred by estrous females although sexual activity is not restricted to the musth condition. The present study combines recently established methods of fecal hormone analysis with long-term observations on male-female associations as well as the presence and intensity of physical signals to provide a more detailed picture about the physical, physiological, and behavioral characteristics of different states of sexual activity in free-ranging African elephants. Based on quantitative shifts in individual bull association patterns, the presence of different physical signals, and significant differences in androgen levels, a total of three potential sub-categories for sexually active bulls could be established. The results demonstrate that elevations in androgen levels are only observed in sexually active animals showing temporal gland secretion and/or urine dribbling, but are not related to the age of the individual. Further, none of the sexually active states showed elevated glucocorticoid output indicating that musth does not represent an HPA-mediated stress condition. On the basis of these results, we suggest that the term "musth" should be exclusively used for the competitive state in sexually active male elephants and that the presence of urine dribbling should be the physical signal used for defining this state

Garstang, M., 2005. Long-distance, low-frequency elephant communication. J Comp Physiol A Neuroethol Sens Neural Behav Physiol. 191, 299.
Abstract: Erratum: J Comp Physiol A Neuroethol Sens Neural Behav Physiol. 2004; Oct;190(10):791-805. Epub 2004 Sep 2. The production, transmission, and reception of and the behavioral response to long-distance, low-frequency sound by elephants is reviewed. The structure of low-frequency calls generated by elephants is separated into the "source" and the "filter" roles played by the lungs, larynx and vocal track, the composition of the expired air and the ambient air temperature. Implications regarding the size, age, sex, sexual and physical status follow from the call structure and detection. Reception of the signal is discussed in terms of the characteristics of the elephant's ear with particular attention to the determination of the threshold of hearing and the ability to locate the source of low-frequency sounds. Factors which influence the transmission of near infrasound are related to atmospheric structure. The critical role played by the thermal stratification and vertical gradient and magnitude of the wind in determining both the range and the detection of a signal are discussed for open and closed elephant habitats. Infrasound plays a pervasive role in reproduction, resource utilization, avoidance of predation and other social interactions. Current and future technology can be expected to contribute to the detection and interpretation of elephant communication. This will aid in the understanding of behavior and in efforts to sustain the species.

Glickman, S.E., Short, R.V., Renfree, M.B., 2005. Sexual differentiation in three unconventional mammals: Spotted hyenas, elephants and tammar wallabies. Hormones and Behaviour 48, 403-417.
Abstract: The present review explores sexual differentiation in three non-conventional species: the spotted hyena, the elephant and the tammar wallaby, selected because of the natural challenges they present for contemporary understanding of sexual differentiation. According to the prevailing view of mammalian sexual differentiation, originally proposed by Alfred Jost, secretion of androgen and anti-Mullerian hormone (AMH) by the fetal testes during critical stages of development accounts for the full range of sexually dimorphic urogenital traits observed at birth. Jost's concept was subsequently expanded to encompass sexual differentiation of the brain and behavior. Although the central focus of this review involves urogenital development, we assume that the novel mechanisms described in this article have potentially significant implications for sexual differentiation of brain and behavior, a transposition with precedent in the history of this field. Contrary to the ''specific'' requirements of Jost's formulation, female spotted hyenas and elephants initially develop male-type external genitalia prior to gonadal differentiation. In addition, the administration of anti-androgens to pregnant female spotted hyenas does not prevent the formation of a scrotum, pseudoscrotum, penis or penile clitoris in the offspring of treated females, although it is not yet clear whether the creation of masculine genitalia involves other steroids or whether there is a genetic mechanism bypassing a hormonal mediator. Wallabies, where sexual differentiation occurs in the pouch after birth, provide the most conclusive evidence for direct genetic control of sexual dimorphism, with the scrotum developing only in males and the pouch and mammary glands only in females, before differentiation of the gonads. The development of the pouch and mammary gland in females and the scrotum in males is controlled by genes on the X chromosome. In keeping with the ''expanded'' version of Jost's formulation, secretion of androgens by the fetal testes provides the best current account of a broad array of sex differences in reproductive morphology and endocrinology of the spotted hyena, and androgens are essential for development of the prostate and penis of the wallaby. But the essential circulating androgen in the male wallaby is 5α androstanediol, locally converted in target tissues to DHT, while in the pregnant female hyena, androstenedione, secreted by the maternal ovary, is converted by the placenta to testosterone (and estradiol) and transferred to the developing fetus. Testicular testosterone certainly seems to be responsible for the behavioral phenomenon of musth in male elephants. Both spotted hyenas and elephants display matrilineal social organization, and, in both species, female genital morphology requires feminine cooperation for successful copulation. We conclude that not all aspects of sexual differentiation have been delegated to testicular hormones in these mammals. In addition, we suggest that research on urogenital development in these non-traditional species directs attention to processes that may well be operating during the sexual differentiation of morphology and behavior in more common laboratory mammals, albeit in less dramatic fashion.

Glickman, S.E., Short, R.V., Renfree, M.B., 2005. Sexual differentiation in three unconventional mammals: spotted hyenas, elephants and tammar wallabies
566. Horm. Behav. 48, 403-417.
Abstract: The present review explores sexual differentiation in three non-conventional species: the spotted hyena, the elephant and the tammar wallaby, selected because of the natural challenges they present for contemporary understanding of sexual differentiation. According to the prevailing view of mammalian sexual differentiation, originally proposed by Alfred Jost, secretion of androgen and anti-Mullerian hormone (AMH) by the fetal testes during critical stages of development accounts for the full range of sexually dimorphic urogenital traits observed at birth. Jost's concept was subsequently expanded to encompass sexual differentiation of the brain and behavior. Although the central focus of this review involves urogenital development, we assume that the novel mechanisms described in this article have potentially significant implications for sexual differentiation of brain and behavior, a transposition with precedent in the history of this field. Contrary to the "specific" requirements of Jost's formulation, female spotted hyenas and elephants initially develop male-type external genitalia prior to gonadal differentiation. In addition, the administration of anti-androgens to pregnant female spotted hyenas does not prevent the formation of a scrotum, pseudoscrotum, penis or penile clitoris in the offspring of treated females, although it is not yet clear whether the creation of masculine genitalia involves other steroids or whether there is a genetic mechanism bypassing a hormonal mediator. Wallabies, where sexual differentiation occurs in the pouch after birth, provide the most conclusive evidence for direct genetic control of sexual dimorphism, with the scrotum developing only in males and the pouch and mammary glands only in females, before differentiation of the gonads. The development of the pouch and mammary gland in females and the scrotum in males is controlled by genes on the X chromosome. In keeping with the "expanded" version of Jost's formulation, secretion of androgens by the fetal testes provides the best current account of a broad array of sex differences in reproductive morphology and endocrinology of the spotted hyena, and androgens are essential for development of the prostate and penis of the wallaby. But the essential circulating androgen in the male wallaby is 5alpha androstanediol, locally converted in target tissues to DHT, while in the pregnant female hyena, androstenedione, secreted by the maternal ovary, is converted by the placenta to testosterone (and estradiol) and transferred to the developing fetus. Testicular testosterone certainly seems to be responsible for the behavioral phenomenon of musth in male elephants. Both spotted hyenas and elephants display matrilineal social organization, and, in both species, female genital morphology requires feminine cooperation for successful copulation. We conclude that not all aspects of sexual differentiation have been delegated to testicular hormones in these mammals. In addition, we suggest that research on urogenital development in these non-traditional species directs attention to processes that may well be operating during the sexual differentiation of morphology and behavior in more common laboratory mammals, albeit in less dramatic fashion

Greenwood, D.R., Comeskey, D., Hunt, M.B., Rasmussen, L.E., 2005. Chemical communication: chirality in elephant pheromones
528. Nature 438, 1097-1098.
Abstract: Musth in male elephants is an annual period of heightened sexual activity and aggression that is linked to physical, sexual and social maturation. It is mediated by the release of chemical signals such as the pheromone frontalin, which exists in two chiral forms (molecular mirror images, or enantiomers). Here we show that enantiomers of frontalin are released by Asian elephants in a specific ratio that depends on the animal's age and stage of musth, and that different responses are elicited in male and female conspecifics when the ratio alters. This precise control of communication by molecular chirality offers insight into societal interactions in elephants, and may be useful in implementing new conservation protocols

Lacasse, C., Gamble, K.C., Terio, K., Farina, L.L., Travis, D.A., Miller, M. Mycobacterium szulgai osteoarthritis and pneumonia in an African elephant (Loxodonta Africana). 2005 Proceedings AAZV, AAWV, AZA Nutrition Advisory Group.  170-172. 2005.
Ref Type: Conference Proceeding
Abstract: Tuberculosis, particularly Mycobacterium bovis and M. tuberculosis, is an important health issue in zoological collections.  Zoos are a particular public health concern because of the close contact between tuberculosis-susceptible animals and humans, specifically animal handlers and visitors.¹⁶ Evidence of M. tuberculosis transmission between humans and elephants, confirmed by DNA fingerprinting, has been reported.¹³ Between 1994 and 2001, M. tuberculosis was isolated from trunk washes of captive elephants from 11 herds in the United States.¹⁷  To date, most reported cases of tuberculosis have occurred in captive Asian elephants (Elephas maximus).¹⁴ In 1997, the National Tuberculosis Working Group for Zoo and Wildlife Species partnered with the USDA to formulate the "Guidelines for the Control of Tuberculosis in Elephants." ¹⁵ This document outlines criteria for the testing, surveillance, and treatment of tuberculosis in elephants. The guidelines recommend annual monitoring of elephants by mycobacterial culture of three direct trunk washes collected over 1 wk.  Isolation of Mycobacterium avium and non-tuberculous mycobacteria from elephant trunk wash samples is common, but these organisms have not been associated with clinical disease.^14,18 This case report details clinical disease with fatal complications of an atypical mycobacterial infection in an African elephant (Loxodonta africana). In September 2003, an African elephant presented with acute, severe lameness of the left rear limb with subsequent swelling of the stifle.  Diagnostic procedures included aspiration cytology of the swelling, radiographs, and thermographic imaging.  The exact location of the injury could not be detected, but a lesion to the stifle or coxofemoral articulation was suspected.  After 13 mo of treatment, including pulse therapy with a variety of nonsteroidal anti-inflammatory drugs (NSAIDs), weekly to biweekly injections of polysulfated glycosaminoglycan, and intensive foot care efforts to treat secondary pedal lesions of both rearlimbs, the animal died acutely.  Gross necropsy revealed granulomatous osteomyelitis with necrosis/loss of the femoral head and acetabulum and pulmonary granulomas.  Both of these lesions contained acid-fast bacteria on cytology. While awaiting confirmatory culture results, quarantine procedures were established for the elephant facility and a program was established to screen all zoo personnel in close contact with the elephant or who participated in the necropsy.  All personnel were tested by the Chicago Department of Public Health without documented conversion. Mycobacterium szulgai was ultimately cultured from both coxofemoral and pulmonary lesions. Mycobacterium szulgai is an uncommon nontuberculous mycobacterium that is usually isolated from pathologic lesions in humans.²¹ This bacterial species was first identified in 1972.¹¹ The lungs are the main locality for pathologic manifestation in humans and several cases have been in patients with acquired immunodeficiency syndrome.^9,20,21 Infection due to M. szulgai most frequently produces thin-walled cavities in lungs resembling tuberculosis.⁴ Other documented sites of infection include the skin, bone, and tendon sheath (causing a carpal tunnel syndrome).^{2,9,10,12,19,20}  Intra-operative contamination from ice water has led to M. szulgai keratitis after laser-assisted ophthalmic surgeries^.6 A case of disseminated disease in a previously healthy young human has been reported.⁵  No evidence of human-to-human transmission of this organism has been documented and human cases are believed to originate from environmental sources.¹²  The natural habitat of the organism is unknown, but previous reports suggest an association of the bacteria with water of swimming pools and fish tanks.^1,21 The organism has been cultured from a snail and tropical fish.^1,3 No standard recommendation for the treatment of M. szulgai infection currently exists.  In general, triple antibiotic therapies used in standard mycobacterial treatments are reported with a low rate of relapses and sterilization of sputum cultures within a mean of 3 mo.³ Pulmonary lesions in this elephant were chronic; it was not possible to determine when initial infection occurred. Infection could have occurred in captivity or in the wild prior to captivity. Three trunk washes over the past year had been negative for mycobacterial culture. Osteomyelitis in the hip may have developed secondary to hematogenous spread from the lungs with the acute lameness resulting from a pathologic fracture associated with this infection. Alternatively, though considered less likely, a traumatic fracture of the hip could have occurred, with bacterial inoculation and secondary osteomyelitis as a result of increased blood flow to the site. The source of infection for this elephant remains unknown.  Prevalence of this organism in the natural habitat or captive environment of the elephants has not been previously documented.
LITERATURE CITED
1 Abalain-Colloc, M.L., D. Guillerm, M. Salaun, S. Gouriou, V. Vincent, and B. Picard.  2003.  Mycobacterium szulgai isolated from a patient, a tropical fish, and aquarium water.  Eur. J. Clin. Microbiol. Infect. Dis.  22: 768-769.
2.Cross, G.M., M. Guill, and J.K. Aton.  1985.  Cutaneous Mycobacterium szulgai infection. Arch. Dermatol. 121: 247-249.
3. Davidson, P.T. 1976. Mycobacterium szulgai: a new pathogen causing infection of the lung.  Chest 69: 799- 801.
4. Dylewski, J.S., H.M. Zackon, A.H. Latour, and G.R. Berry.  1987.  Mycobacterium szulgai: an unusual pathogen.  Rev. Infect. Dis.  9: 578-580.
5. Gur, H., S. Porat, H. Haas, Y. Naparstek, and M. Eliakim.  1984.  Disseminated mycobacterial disease caused by Mycobacterium szulgai. Arch. Intern. Med. 144: 1861-1863.
6.Holmes, G.P., G. Bond, R.C. Fader, and S.F. Fulcher.  2002. A cluster of cases of Mycobacterium szulgai keratitis that occurred after laser-assisted in situ keratomileusis.  Clin. Infect. Dis. 34: 1039-1046.
7.Horusitzky, A., X. Puechal, D. Dumont, T. Begue, M. Robineau, and M. Boissier.  2000.  Carpal tunnel syndrome caused by Mycobacterium szulgai. J. Rheumatol 27: 1299-1302.
8.Hurr, H., and T. Sorg.  1998.  Mycobacterium szulgai osteomyelitis.  J. Infect.  37: 191-192.
9.Luque, A.E., D. Kaminski, R. Reichman, and D. Hardy. 1998.  Mycobacterium szulgai osteomyelitis in an AIDS patient. Scand. J. Infect. Dis. 30: 88-91.
10.Maloney, J.M., C.R. Gregg, D.S. Stephens, F.A. Manian, and D. Rimland.  1987.  Infections caused by Mycobacterium szulgai in humans.  Rev. Infect. Dis.  9: 1120-1126.
11.Marks, J., P.A. Jenkins, and M. Tsukamura.  1972.  Mycobacterium szulgai: a new pathogen.  Tubercle 53: 210.
12.Merlet, C., S. Aberrane, F. Chilot, and J. Laroche.  2000.  Carpal tunnel syndrome complicating hand flexor tenosynovitis due to Mycobacterium szulgai. Joint Bone Spine 67: 247-248.
13.Michalak, K., C. Austin, S. Diesel, J.M. Bacon, P. Zimmerman, and J. N. Maslow.  1998. Mycobacterium tuberculosis infection as a zoonotic disease: transmission between humans and elephants. Emerg. Infect. Dis. 4: 283-287.
14.Mikota, S.K., R.S. Larsen, and R.J. Montali.  2000.  Tuberculosis in elephants in North America.  Zoo Biol. 19: 393-403.
15.National Tuberculosis Working Group for Zoo and Wildlife Species. 2000. Guidelines for the control of tuberculosis in elephants.  USDA Animal and Plant Health Inspection Services.
16.Oh, P., R. Granich, J. Scott, B. Sun, M. Joseph, C. Stringfield, S. Thisdell, J. Staley, D. Workman-Malcolm, L. Borenstein, E. Lehnkering, P. Ryan, J. Soukup, A. Nitta, and J. Flood.  2002.  Human exposure following  Mycobacterium tuberculosis infection of multiple animal species in a metropolitan zoo.  Emerg. Infect. Dis. 8: 1290-1293.
17.Payeur, J.B., J.L. Jarnagin, J.G. Marquardt, and D.L. Whipple.  2002.  Mycobacterial isolations in captive elephants in the United States.  Ann. N.Y. Acad. Sci. 969: 256-258.
18.Shojaei, H., J.G. Magee, R. Freeman, M. Yates, N.U. Horadagoda, and M. Goodfellow.  2000. Mycobacterium elephantis sp. nov., a rapidly growing non-chromogenic Mycobacterium isolated from an elephant.  Int. J. Syst. Evol. Microbiol.  50: 1817-1820.
19.Stratton, C.W., D.B. Phelps, and L.B. Reller.  1978.  Tuberculoid tenosynovitis and carpal tunnel syndrome caused by Mycobacterium szulgai.  Am. J. Med.  65: 349-351.
20.Tappe, D., P. Langmann, M. Zilly, H. Klinker, B. Schmausser, and M. Frosch.  2004.  Osteomyelitis and skin ulcers caused by Mycobacterium szulgai in an AIDS patient.  Scand. J. Infect. Dis. 36: 883-885.
21.Tortoli, E., G. Besozzi, C. Lacchini, V. Penati, M.T. Simonetti, and S. Emler.  1998.  Pulmonary infection due to Mycobacterium szulgai, case report and review of the literature.  Eur. Respir. J.  11: 975-977.

Leong, K.M., Burks, K., Rizkalla, C.E., Savage, A., 2005. Effects of reproductive and social context on vocal communication in captive female African elephants (Loxodonta africana). Zoo Biology 24, 331-347.
Abstract: Female African elephants advertise changes in reproductive condition to males through a variety of modalities, including an increase in low-frequency vocalizations, presumed to travel long distances.  Although males respond to these vocalizations, it has been suggested that their proximate function may be to signal to nearby females rather than to distant males. Because elephants live in a female-bonded society, it is likely  that  changes in female reproductive condition also affect close-range  interactions between high- and low-ranking females and that  vocalizations  may mediate these interactions. To examine female-female interactions related to vocal production and the ovulatory cycle, this year-long study monitored behavior, vocalizations and hormonal cycles for a group of six female captive African elephants at Disney's Animal Kingdom. Rates of several types of close-range interactions were observed to change over the phases of the estrous cycle, and rank seemed to affect whether or not low-frequency vocalizations were given in association with these interactions. Results of this study suggest that a female African  elephant's  immediate social context and rank in the social hierarchy interact with  the  hormonal cycle in the production of low-frequency vocalizations, thus  many  of these vocalizations may not function proximately as signals to  distant  males, but may be a result of the changing dynamics among females.

Mobasheri, A., Gent, T.C., Womack, M.D., Carter, S.D., Clegg, P.D., Barrett-Jolley, R., 2005. Quantitative analysis of voltage-gated potassium currents from primary equine (Equus caballus) and elephant (Loxodonta africana) articular chondrocytes
618. Am. J. Physiol Regul. Integr. Comp Physiol 289, R172-R180.
Abstract: In this comparative study, we have established in vitro models of equine and elephant articular chondrocytes, examined their basic morphology, and characterized the biophysical properties of their primary voltage-gated potassium channel (Kv) currents. Using whole cell patch-clamp electrophysiological recording from first-expansion and first-passage cells, we measured a maximum Kv conductance of 0.15 +/- 0.04 pS/pF (n = 10) in equine chondrocytes, whereas that in elephant chondrocytes was significantly larger (0.8 +/- 0.4 pS/pF, n = 4, P </= 0.05). Steady-state activation parameters of elephant chondrocytes (V = -22 +/- 6 mV, k = 11.8 +/- 3 mV, n = 4) were not significantly different from those of horse chondrocytes (V = -12.5 +/- 4.3 mV, k = 12 +/- 2, n = 10). This suggests that there would be slightly more resting Kv activation in elephant chondrocytes than in their equine counterparts. Kinetic analysis revealed that both horse and elephant chondrocyte Kv currents had similar activation and inactivation parameters. Pharmacological investigation of equine chondrocyte Kv currents showed them to be powerfully inhibited by the potassium channel blockers tetraethylammonium and 4-aminopyridine but not by dendrotoxin-I. Immunohistochemical studies using polyclonal antibodies to Kv1.1-Kv1.5 provided evidence for expression of Kv1.4 in equine chondrocytes. This is the first electrophysiological study of equine or elephant chondrocytes. The data support the notion that voltage-gated potassium channels play an important role in regulating the membrane potential of articular chondrocytes and will prove useful in future modeling of electromechanotransduction of fully differentiated articular chondrocytes in these and other species

Murwira, A., Skidmore, A.K., 2005. The response of elephants to the spatial heterogeneity of vegetation in a Southern African agricultural landscape. Landscape Ecology 20, 217-234.
Abstract: Based on the agricultural landscape of the Sebungwe in Zimbabwe, we investigated whether and how the spatial distribution of the African elephant (Loxodonta africana) responded to spatial heterogeneity of vegetation cover based on data of the early 1980s and early 1990s. We also investigated whether and how elephant distribution responded to changes in spatial heterogeneity between the early 1980s and early 1990s.  Vegetation cover was estimated from a normalised difference vegetation index (NDVI).  Spatial heterogeneity was estimated from a new approach based on the intensity (i.e., the maximum variance exhibited when a spatially distributed landscape property such as vegetation cover is measured with a successively increasing window size or scale) and dominant scale (i.e., the scale or window size at which the intensity is displayed). We used a variogram to quantify the dominant scale (i.e., range) and intensity (i.e., sill) of NDVI based congruent windows (i.e., 3.84 km x 3.84 km in a 61 km x 61 km landscape). The results indicated that elephants consistently responded to the dominant scale of spatial heterogeneity in a unimodal fashion with the peak elephant presence occurring in environments with dominant scales of spatial heterogeneity of around 457-734 m. Both the intensity and dominant scale of spatial heterogeneity predicted 65 and 68% of the variance in elephant presence in the early 1980s and in the early 1990s respectively.  Also, changes in the intensity and dominant scale of spatial heterogeneity predicted 61% of the variance in the change in elephant distribution. The results imply that management decisions must take into consideration the influence of the levels of spatial heterogeneity on elephants in order to ensure elephant persistence in agricultural landscapes.

Naz, R.K., Gupta, S.K., Gupta, J.C., Vyas, H.K., Talwar, A.G., 2005. Recent advances in contraceptive vaccine development: a mini-review
577. Hum. Reprod. 20, 3271-3283.
Abstract: Contraceptive vaccines (CV) may provide viable and valuable alternatives to the presently available methods of contraception. The molecules that are being explored for CV development either target gamete production [luteinizing hormone-releasing hormone (LHRH)/GnRH, FSH], gamete function [sperm antigens and oocyte zona pellucida (ZP)], and gamete outcome (HCG). CV targeting gamete production have shown varied degrees of efficacy; however, they either affect sex steroids causing impotency and/or show only a partial rather than a complete effect in inhibiting gametogenesis. However, vaccines based on LHRH/GnRH are being developed by several pharmaceutical companies as substitutes for castration of domestic pets, farm and wild animals, and for therapeutic anticancer purposes such as in prostatic hypertrophy and carcinoma. These vaccines may also find applications in clinical situations that require the inhibition of increased secretions of sex steroids, such as in uterine fibroids, polycystic ovary syndrome, endometriosis and precocious puberty. CV targeting molecules involved in gamete function such as sperm antigens and ZP proteins are exciting choices. Sperm constitute the most promising and exciting target for CV. Several sperm-specific antigens have been delineated in several laboratories and are being actively explored for CV development. Studies are focused on delineating appropriate sperm-specific epitopes, and increasing the immunogenicity (specifically in the local genital tract) and efficacy on the vaccines. Anti-sperm antibody (ASA)-mediated immunoinfertility provides a naturally occurring model to indicate how a vaccine might work in humans. Vaccines based on ZP proteins are quite efficacious in producing contraceptive effects, but may induce oophoritis, affecting sex steroids. They are being successfully tested to control feral populations of dogs, deer, horses and elephants, and populations of several species of zoo animals. The current research for human applicability is focused on delineating infertility-related epitopes (B-cell epitopes) from oophoritis-inducing epitopes (T-cell epitopes). Vaccines targeting gamete outcome primarily focus on the HCG molecule. The HCG vaccine is the first vaccine to undergo Phase I and II clinical trials in humans. Both efficacy and lack of immunopathology have been reasonably well demonstrated for this vaccine. At the present time, studies are focused on increasing the immunogenicity and efficacy of the birth control vaccine, and examining its clinical applications in various HCG-producing cancers. The present article will focus on the current status of the anti-sperm, anti-ZP, anti-LHRH/GnRH and anti-HCG vaccines

Ortolani, A., Leong, K., Graham, L., Savage, A., 2005. Behavioral indices of estrus in a group of captive African elephants (Loxodonta africana). Zoo Biology 24, 311-329.
Abstract: This study investigated behavioral signals of estrus by systematically monitoring the interactions of one male with four female African elephants housed in a naturalistic outdoor enclosure at Disney's Animal Kingdom over a period of 11 months. We measured changes in five spatial behaviors and 22 tactile-contact behaviors, as well as changes in serum progestagen and LH concentrations, across three ovarian cycles for each female. Two females did not cycle during the study. Three different phases of the ovarian cycle were identified: mid luteal, anovulatory follicular, ovulatory follicular.  The male followed more and carried out more genital inspections, flehmen, and trunk-to-mouth behaviors toward cycling females during their ovulatory phase. Genital inspections by the male peaked above baseline levels on  the  day of an LH surge, and up to 9 days before, in both cycling females  and,  thus, might be a useful behavioral index of estrus. The male also carried out more genital inspections, flehmen, and trunk touches to the back leg toward ovulatory cycling than noncycling females. Overall, our results  indicated that: 1) a single subadult African elephant male could  discriminate two females in the ovulatory phase of their cycle (i.e.,  during  the 3 weeks preceding ovulation) from the mid luteal phase; 2) the male  also  discriminated two cycling females in the ovulatory and anovulatory  follicular phases from two noncycling females; 3) two females in the  ovulatory phase of the cycle displayed a greater variety of  tactile-contact  behavior toward the male compared to the other cycle phases.

Perez-Barberia, F.J., Gordon, I.J., 2005. Gregariousness increases brain size in ungulates
586. Oecologia. 145, 41-52.
Abstract: The brain's main function is to organise the physiological and behavioural responses to environmental and social challenges in order to keep the organism alive. Here, we studied the effects that gregariousness (as a measurement of sociality), dietary habits, gestation length and sex have on brain size of extant ungulates. The analysis controlled for the effects of phylogeny and for random variability implicit in the data set. We tested the following groups of hypotheses: (1) Social brain hypothesis-gregarious species are more likely to have larger brains than non-gregarious species because the former are subjected to demanding and complex social interactions; (2) Ecological hypothesis-dietary habits impose challenging cognitive tasks associated with finding and manipulating food (foraging strategy); (3) Developmental hypotheses (a) energy strategy: selection for larger brains operates, primarily, on maternal metabolic turnover (i.e. gestation length) in relation to food quality because the majority of the brain's growth takes place in utero, and finally (b) sex hypothesis: females are expected to have larger brains than males, relative to body size, because of the differential growth rates of the soma and brain between the sexes. We found that, after adjusting for body mass, gregariousness and gestation length explained most of the variation in brain mass across the ungulate species studied. Larger species had larger brains; gregarious species and those with longer gestation lengths, relative to body mass, had larger brains than non-gregarious species and those with shorter gestation lengths. The effect of diet was negligible and subrogated by gestation length, and sex had no significant effect on brain size. The ultimate cause that could have triggered the co-evolution between gestation length and brain size remains unclear

Poole, J.H., Tyack, P.L., Stoeger-Horwath, A.S., Watwood, S., 2005. Animal behaviour: elephants are capable of vocal learning
623. Nature 434, 455-456.
Abstract: There are a few mammalian species that can modify their vocalizations in response to auditory experience--for example, some marine mammals use vocal imitation for reproductive advertisement, as birds sometimes do. Here we describe two examples of vocal imitation by African savannah elephants, Loxodonta africana, a terrestrial mammal that lives in a complex fission-fusion society. Our findings favour a role for vocal imitation that has already been proposed for primates, birds, bats and marine mammals: it is a useful form of acoustic communication that helps to maintain individual-specific bonds within changing social groupings

Rasmussen, L.E.L., Krishamurthy, V., Sakumar, R., 2005. Behavioural and chemical confirmation of the preovulatory pheromone, (Z)-7-dodecenyl acetate, in wild Asian elephants: its relationship to musth. Behaviour 142, 351-396.
Abstract: Mammalian breeding strategies vary depending on particular social contexts and sensory systems emphasized in various species. Among sexually dimorphic non-territorial Asian elephants,  Elephas maximus, a multiplex olfactory chemical signaling system has been implicated in ensuring effective reproduction. This study explores how, using chemosensory mechanisms, widely roaming, wild male elephants locate periovulatory females in matriarchal-led female family units and precisely assess their ovulatory status. In this species, the dual obstacles of separately living sexes and infrequent oestrus are overcome by lengthy female cycles. During an extended preovulatory period captive females release increasing concentrations of the urinary pheromone (Z)-7-dodecenyl acetate, timed to reach a maximum just before ovulation. The current field studies combined chemical identification and quantification of female urinary (Z)-7-dodecenyl acetate with behavioural observations, monitoring the frequencies of chemosensory responses and premating  behaviours by various categories of males. The results suggest the temporal extension of the preovulatory period effectively provides a synchrony between sexes for successful reproduction. Male elephants undergo a two-decade-long maturation process that involves physical, sexual, social, and physiological maturation. Males older than 30 years are generally large, sexually active, socially adept and capable of sustaining long periods of musth, during which they release secretions distinctive of adult musth.  These older adult males in musth demonstrated significantly more chemosensory responses and premating behaviours than their younger or nonmusth counterparts; they apparently are more skilled at detecting the precise ovulatory status of females. Male-male interactions are affected by size, age, and musth; the winners gain greater access to females, as indicated by the high incidence of mate guarding.  The Asian elephant shares some breeding tactics common to other mammals including some primates (e.g. orangutans) and whales, while the musth parameter adds a unique feature. Fusion-fission events are influenced by elephant reproductive strategies, as roving males join female groups while tracking preovulatory pheromone concentrations.

Raubenheimer, E.J., Ngwenya, S.P., 2005. The role of ivory in the survival of the African elephant
510. SADJ. 60, 426, 430.
Abstract: The unique chequered pattern of polished ivory has created a perverted commercial demand for elephant tusks. The morphologic basis of the pattern, which makes ivory a sought after product for the manufacturing of works of art, is discussed. Chemical analyses of ivory holds great potential in tracing the source of illegally harvested tusks and exposing poorly managed elephant sanctuaries. The impact of uncontrolled ivory hunting on the population genetics of the African elephant is briefly reviewed

Saragusty, J., Hildebrandt, T.B., Natan, Y., Hermes, R., Yavin, S., Goeritz, F., Arav, A., 2005. Effect of egg-phosphatidylcholine on the chilling sensitivity and lipid phase transition of Asian elephant (Elephas maximus) spermatozoa. Zoo Biology 24, 233-245.
Abstract: This study was conducted in an effort to improve our understanding of the response of Asian elephant (Elephas maximus, Em) spermatozoa to chilling.  Semen was collected from two elephant bulls by means of the manual rectal stimulation method. Five out of seven semen collections were deemed to be suitable for use based on motility (ranging from 20% to 60%) and membrane integrity. We evaluated the chilling sensitivity by incubating the sperm with a fluorescent dye (5-carboxyfluorescein diacetate (cFDA)) at 16 degrees C, 12 degrees C, 4 degrees C, and 22 degrees C (control). Cells with an intact membrane retained the dye and were identified as viable. The membrane lipid phase transition (LPT) temperature curve was determined with a Fourier transform infrared (FTIR) spectrometer connected to an FTIR microscope.  The LPT center, T-m, was determined by statistical analysis. The LPT and T-m were also assessed in fresh spermatozoa and spermatozoa incubated with egg yolk or egg-phosphatidylcholine (EPC) liposomes at 16 degrees C, 12 degrees C, 4 degrees C, and 26 degrees C (control). The results show that the membrane integrity of spermatozoa incubated at 16 degrees C, 12 degrees C, and 4 degrees C decreased by 39%, 62%, and 67%, respectively, compared to the control. The LPT temperatures were between room temperature (26 degrees C) and 10 degrees C, with Tm at 14-16 degrees C. The T-m for sperm incubated with liposomes or egg-yolk extender was below the measured range (2 degrees C). Chilling sensitivity was found at a wide range of temperatures and transition temperatures, suggesting the presence of a wide variety of fatty acids (FAs) in the membrane with a high ratio of saturated-to-polyunsaturated FAs. Here we show that the protection  afforded  by the presence of egg yolk or liposomes in the extender is  accomplished by  shifting the T. to below the 4 degrees C point at which chilled semen  is  maintained for transport, or the point at which fast freezing begins to  minimize cellular damage.

Steinetz, B.G., Brown, J.L., Roth, T.L., Czekala, N., 2005. Relaxin concentrations in serum and urine of endangered species: correlations with physiologic events and use as a marker of pregnancy
596. Ann. N. Y. Acad. Sci. 1041, 367-378.
Abstract: Many mammalian species are facing extinction due to problems created by human encroachment, agriculture, pollution, and willful slaughter. Among those at risk are the Asian and African elephant, Sumatran rhinoceros, and giant panda. Conservation groups try to save species in the wild by preserving habitat and limiting animal-human conflicts, often with limited success. Another alternative is to preserve the extant gene pool through captive breeding as a hedge against extinction. Measurement of circulating reproductive hormones is impractical for most wildlife species; determination of urinary or fecal hormone metabolites provides a more viable approach. To aid breeding management, one important tool is the ability to diagnose and monitor pregnancy, especially in species with long gestations (e.g., rhinos over 15 mo and elephants over 20 mo). Unfortunately, measuring progestins often is not useful diagnostically, because concentrations are similar during at least part of the pregnancy and the nonpregnant luteal phase in some species (e.g., elephants, rhinoceroses, and giant pandas). As serum relaxin reliably distinguishes between pregnancy and pseudopregnancy in bitches, relaxin measurement might also provide a method for detecting a successful pregnancy in endangered species. Appropriate immunoassay reagents have enabled the estimation of relaxin concentrations in the serum of elephants and rhinos and the determination of pregnancy establishment and the outcome. Relaxin was also detected in panda serum and urine. However, the extreme variability of the time between observed mating and parturition and the confounding factors of delayed implantation, pseudopregnancy, and frequent fetal resorptions made it impossible to use the panda relaxin data as a specific marker of pregnancy

Stetter, M., Grobler, D., Zuba, J.R., Hendrickson, D., Briggs, M., Castro, L., Neiffer, D., Terrell, S., Robbins, P.K., Stetter, K., Ament, B.S., Wheeler, L. Laprascopic reproductive sterilization as a method of population control in free-ranging African elephants (Loxodonta africana). 2005 Proceedings AAZV, AAWV, AZA Nutrition Advisory Group.  199-200. 2005.
Ref Type: Conference Proceeding

Vinogradov, I.V., Kochneva, G.V., Malkova, E.M., Shchelkunov, S.N., Riabchikova, E.I., 2005. [Intranasal infection in mice inoculated with cowpox virus strain EP-2 isolated from the elephant]
579. Vopr. Virusol. 50, 37-42.
Abstract: The specific features of reproduction of EP-2 strain of cowpox virus (CPV) were studied in intranasally infected BALC/C mice by light and electron microscopy. Virus replication was found in the ciliated, intercalary, basal, and goblet cells (the nasal respiratory area), basal and supporting cells (the nasal olfactory area), ciliated, intercalary, goblet cells (the tracheal and bronchial epithelium), and collagen-producing, Schwann's, endothelial, smooth muscle, and adventitial cells. It has been shown that the CPV strain EP-2 locally replicates in the nasal cavity, trachea, and large bronchi and that there is no generalized infection

Walker, C.L., Stewart, E.A., 2005. Uterine fibroids: the elephant in the room. Science 308, 1589-1592.
Abstract: Uterine fibroids (leiomyomas) have historically been viewed as important chiefly as the major indication for hysterectomy. As new therapies are developed, the heterogeneity of this disease becomes therapeutically relevant. An awareness of the role of genetics, the extracellular matrix, and hormones in tumor etiology is key to understanding this disease. 

Wooding, F.B., Stewart, F., Mathias, S., Allen, W.R., 2005. Placentation in the African elephant, Loxodonta africanus: III. Ultrastructural and functional features of the placenta
598. Placenta 26, 449-470.
Abstract: Successful transfer of nutrients to the elephant fetus during pregnancy relies on a variety of placental modifications. Our light and electron microscopical investigations show that the structure is endotheliochorial from implantation to term, with unicellular, never syncytial trophoblast. Light and electron microscope immunocytochemistry shows the restriction of the glucose transporter 1 isoform to the basolateral surfaces of the trophoblast, with the glucose transporter 3 restricted to the apical plasmalemma of the trophoblast. Glucose transport to the fetus therefore requires a sequential use of both isoforms. Light and electron microscope cytochemistry indicate the presence of iron deposits only in the haemophagous zones confirming their iron transport function. No trophoblast areas with high concentrations of Calcium binding protein, specialised for Calcium transport were found. In situ hybridisation demonstrated the presence of IGF-II mRNA in the trophoblast from the earliest stage, with TGFbeta1 and HGF-SF mRNA expressed subsequently but only IGF-II and HGF mRNA present in the second half of pregnancy. The results are briefly discussed in terms of placental growth and function and indicate that the elephant placenta is another example of a unique solution to the variety of problems posed by a resident fetus

 2004. Elephant Husbandry Resource Guide. International Elephant Foundation, Azle. TX.

Agnew, D.W., Munson, L., Ramsay, E.C., 2004. Cystic endometrial hyperplasia in elephants
741. Vet. Pathol. 41, 179-183.
Abstract: Most captive female elephants are nulliparous and aged and many have endometrial disease, factors that may hinder fertility. This study characterized the pathologic features and demographic distribution of endometrial lesions from 27 captive Asian (Elephas maximus) and 13 African elephants (Loxodonta africanus), 12- to 57-years of age. The principal lesion was marked cystic and polypoid endometrial hyperplasia (CEH), present in 67% of Asian and 15% of African elephants ranging from 26 to 57 years. The lower prevalence in African elephants likely reflects their younger age range in this study. Fourteen of 15 affected elephants with breeding information were nulliparous. These results suggest that CEH and polyps are common in aged nulliparous elephants, and the severity of these lesions may impair fertility. These findings will be useful in the interpretation of ultrasonographic findings during reproductive examinations of potential breeding cows. Also, breeding programs should focus on younger animals

Brown, J.L., Olson, D., Keele, M., Freeman, E.W., 2004. Survey of the reproductive cyclicity status of Asian and African elephants in North America. Zoo Biology 23, 309-321.
Abstract: The Asian and African elephant populations in North America are not self sustaining, and reproductive rates remain low. One problem identified from routine progestagen analyses is that some elephant females do not exhibit normal ovarian cycles. To better understand the extent of this problem, the Elephant TAG/SSP conducted a survey to determine the reproductive status of the captive population based on hormone and ultrasound evaluations. The survey response rates for facilities with Asian and African elephants were 81% and 71%, respectively, for the studbook populations, and nearly 100% for the SSP facilities. Of the elephants surveyed, 49% of Asian and 62% of African elephant females were being monitored for ovarian cyclicity via serum or urinary progestagen analyses on a weekly basis. Of these, 14% of Asian and 29% of African elephants either were not cycling at all or exhibited irregular cycles. For both species, ovarian inactivity was more prevalent in the older age categories (>30 years); however, acyclicity was found in all age groups of African elephants. Fewer elephant females (B30%) had been examined by transrectal ultrasound to assess reproductive-tract integrity, and corresponding hormonal data were available for about three-quarters of these females. Within this subset, most (B75%) cycling females had normal reproductive-tract morphologies, whereas at least 70% of noncycling females exhibited some type of ovarian or uterine pathology. In summary, the survey results suggest that ovarian inactivity is a significant reproductive problem for elephants held in zoos, especially African elephants. To increase the fecundity of captive elephants, females should be bred at a young age, before reproductive pathologies occur. However, a significant number of older Asian elephants are still not being reproductively monitored. More significantly, many prime reproductive-age (10-30 years) African females are not being monitored. This lack of information makes it difficult to determine what factors affect the reproductive health of elephants, and to develop mitigating treatments to reinitiate reproductive cyclicity.

Brown, J.L., Walker, S.L., Moeller, T., 2004. Comparative endocrinology of cycling and non-cycling Asian (Elephas maximus) and African (Loxodonta africana) elephants
729. Gen. Comp Endocrinol. 136, 360-370.
Abstract: Up to 14% of Asian and 29% of African elephants in captivity are not cycling normally or exhibit irregular cycles based on progestin profiles. To determine if ovarian acyclicity is related to other disruptions in endocrine activity, serum pituitary, thyroid, adrenal, and ovarian hormones in weekly samples collected for 6-25 months were compared between normal cycling (n=22 each species) and non-cycling (n=6 Asian; n=30 African) elephants. A subset of cycling females (n=4 Asian, 7 African) also were blood sampled daily during the follicular phase to characterize the peri-ovulatory period. In normal cycling females, two leutinizing hormone (LH) surges were observed 3 weeks apart during a normal follicular phase, with the second inducing ovulation (ovLH). Serum FSH concentrations were highest at the beginning of the non-luteal phase, declining to nadir concentrations within 4 days of the ovLH surge. FSH remained low until after the ovLH surge and then increased during the luteal phase. A species difference was noted in prolactin secretion. In the African elephant, prolactin was increased during the follicular phase, but in Asian elephants concentrations remained stable throughout the cycle. Patterns of thyroid hormones (thyroid-stimulating hormone, TSH; free and total thyroxine, T4; free and total triiodothyronine, T3) and cortisol secretion were not affected by estrous cycle stage or season in cycling elephants. In non-cycling elephants, there were no fluctuating patterns of LH, FSH, or prolactin secretion. Overall mean concentrations of all hormones were similar to those in cycling animals, with the exception of FSH, prolactin, and estradiol. Mean serum FSH concentrations were lower due to females not exhibiting normal cyclic increases, whereas serum estradiol was higher overall in most acyclic females. Prolactin concentrations were significantly increased in 11 of 30 non-cycling females, all of which were African elephants. In sum, while there were no consistent endocrine anomalies associated with ovarian acyclicity, hyperprolactinemia may be one cause of ovarian dysfunction. The finding of elevated estrogens in some acyclic females also deserves further investigation, especially determining how it relates to reproductive tract pathologies

Brown, J.L., Goritz, F., Pratt-Hawkes, N., Hermes, R., Galloway, M., Graham, L.H., Gray, C., Walker, S.L., Gomez, A., Moreland, R., Murray, S., Schmitt, D.L., Howard, J., Lehnhardt, J., Beck, B., Bellem, A., Montali, R., Hildebrandt, T.B., 2004. Successful artificial insemination of an Asian elephant at the National Zoological Park. Zoo Biology 23, 45-63.
Abstract: For decades, attempts to breed elephants using artificial insemination (AI) have failed despite considerable efforts and the use of various approaches. However, recent advances in equipment technology and endocrine-monitoring techniques have resulted in 12 elephants conceiving by AI within a 4-year period (19982002). The successful AT technique employs a unique endoscope-guided catheter and transrectal ultrasound to deliver semen into the anterior vagina or cervix, and uses the "double LH surge" (i.e., identifying the anovulatory LH (anLH) surge that predictably occurs 3 weeks before the ovulatory LH (ovLH) surge to time insemination. This study describes the 6-year collaboration between the National Zoological Park (NZP) and the Institute for Zoo Biology and Wildlife Research (IZW), Berlin, Germany, that led to the refinement of this AI technique and subsequent production of an Asian elephant calf. The NZP female was the first elephant to be inseminated using the new AI approach, and was the fifth to conceive. A total of six AI trials were conducted beginning in 1995, and conception occurred in 2000. Semen was collected by manual rectal stimulation from several bulls in North America. Sperm quality among the bulls was variable and was thus a limiting factor for AI. For the successful AI, semen quality was good to excellent (75-90% motile sperm), and sperm was deposited into the anterior vagina on the day before and the day of the ovLH surge. Based on transrectal ultrasound, ovulation occurred the day after the ovLH surge. Pregnancy was monitored by serum and urinary progestagen, and serum prolactin analyses in samples collected weekly. Fetal development was assessed at 12, 20, and 28 weeks of gestation using transrectal ultrasound. Elevated testosterone measured in the maternal circulation after 36 weeks of gestation reliably predicted the calf was a male. Parturition was induced by administration of 40 IU oxytocin 3 days after serum progestagens dropped to undetectable baseline levels. We conclude that AI has potential as a supplement to natural breeding, and will be invaluable for improving the genetic management of elephants, provided that problems associated with inadequate numbers of trained personnel and semen donors are resolved.

Carter, A.M., Enders, A.C., Kunzle, H., Oduor-Okelo, D., Vogel, P., 2004. Placentation in species of phylogenetic importance: The Afrotheria. Animal Reproduction Science 82-83, 35-48.
Abstract: Afrotheria, one of four mammalian superorders, comprises elephants, sea cows, hyraxes, aardvark, elephant shrews, tenrecs and golden moles. Their placentas either form an equatorial band or are discoid in shape. The interhemal region, separating fetal and maternal blood, is endotheliochorial in elephants, aardvark and possibly the sea cows, but hemochorial in the remaining orders. There is a secondary epitheliochorial placenta in elephant shrews while a similar structure in tenrecs erodes maternal tissues. Specialized hemophagous regions are a striking characteristic of some of these placentas yet absent in hyraxes, elephant shrews, and golden moles. It is possible that the common ancestor of the Afrotheria had an endotheliochorial placenta. Establishment of a hemochorial condition, as seen in rock hyraxes, elephant shrews, tenrecs, and golden moles, would be a more recent development. The elephant, manatee, and aardvark all have circumferential placentas. Thus the formation of a discoid placenta with a more or less extensive secondary placenta in elephant shrews and tenrecs would also be a derived state.

Carter, A.M., Enders, A.C., 2004. Comparative aspects of trophoblast development and placentation. Reproductive Biology and Endocrinology 2.
Abstract: Based on the number of tissues separating maternal from fetal blood, placentas are classified as epitheliochorial, endotheliochorial or hemochorial. We review the occurrence of these placental types in the various orders of eutherian mammals within the framework of the four superorders identified by the techniques of molecular phylogenetics. The superorder Afrotheria diversified in ancient Africa and its living representatives include elephants, sea cows, hyraxes, aardvark, elephant shrews and tenrecs. Xenarthra, comprising armadillos, anteaters and sloths, diversified in South America. All placentas examined from members of these two oldest superorders are either endotheliochorial or hemochorial. The superorder Euarchontoglires includes two sister groups, Glires and Euarchonta. The former comprises rodents and lagomorphs, which typically have hemochorial placentas. The most primitive members of Euarchonta, the tree shrews, have endotheliochorial placentation. Flying lemurs and all higher primates have hemochorial placentas. However, the lemurs and lorises are exceptional among primates in having epitheliochorial placentation. Laurasiatheria, the last superorder to arise, includes several orders with epitheliochorial placentation. These comprise whales, camels, pigs, ruminants, horses and pangolins. In contrast, nearly all carnivores have endotheliochorial placentation, whilst bats have endotheliochorial or hemochorial placentas. Also included in Laurasiatheria are a number of insectivores that have many conserved morphological characters; none of these has epitheliochorial placentation. Consideration of placental type in relation to the findings of molecular phylogenetics suggests that the likely path of evolution in Afrotheria was from endotheliochorial to hemochorial placentation. This is also a likely scenario for Xenarthra and the bats. We argue that a definitive epitheliochorial placenta is a secondary specialization and that it evolved twice, once in the Laurasiatheria and once in the lemurs and lorises.

Dahl, N.J., Olson, D., Schmitt, D., Blasko, D.R., Kristipati, R., Roser, J.F., 2004. Development of an enzyme-linked immunosorbent assay (ELISA) for luteinizing hormone (LH) in the elephant (Loxodonta africana and Elephas maximus). Zoo Biology 23, 65-78.
Abstract: A simple, rapid enzyme-linked immunosorbent assay (ELISA) for the measurement of LH in plasma and serum of elephants (Loxodonta africana and Elephas maximus) has been developed, validated, and used for comparative studies. Purified elephant LH (eleLH) diluted in elephant plasma was used as standards (0.78-50 ng/ml). A monoclonal antibody against the beta-subunit of bovine LH (518B(7)) was used as the capture antibody. The second antibody (a polyclonal rabbit anti-human LH antibody), conjugated to horseradish peroxidase, cleaved a substrate (tetramethyl benzidine), resulting in a color change. The total assay time was approximately 21/2 hr, with incubations at room temperature. Sensitivity was found to be 1.56 ng/ml. Cross-reactivities to elephant FSH and TSH were low: 0.9% and 0.15%, respectively. The accuracy of the assay was demonstrated by comparing the ELISA with a validated eleLH radioimmunoassay (RIA), progesterone data, and  ultrasound observations. Blood samples from 18 Asian and African elephant cows were analyzed with the ELISA and RIA, and an additional 11 cows were used to describe endocrine parameters for LH and progesterone using only RIA. No difference was found in LH peak concentrations between the ELISA and RIA. The time from the progesterone decline to the first LH peak, and the time between the two peaks were similar between species. Asian cows had higher LH peaks than African cows. Ultrasound confirmed the time of ovulation occurring with the second LH peak. Three cows were inseminated and confirmed to be pregnant using this ELISA as a timing device. Instrumentation is not always required, as LH peaks approximating 3 ng/ml can be visually observed. In conclusion, this ELISA can be used as a field test to determine time of ovulation for artificial insemination (AI) or natural breeding of both species of elephants, and thus is an important tool for the preservation of captive populations worldwide.

de Oliveira, C.A., West, G.D., Houck, R., Leblanc, M., 2004. Control of musth in an Asian elephant bull (Elephas maximus) using leuprolide acetate
716. J. Zoo. Wildl. Med. 35, 70-76.
Abstract: The results of long-term administration of leuprolide acetate (LA) depot in a 52-yr-old Asian elephant bull (Elephas maximus) for control of musth are presented. Twelve injections were administered for 6 yr during our interpretation of early musth or "premusth." Intervals between musth periods during the study varied from 2 to 34 mo. Blood samples, drawn weekly, were assayed for serum testosterone concentrations; mean levels were 11.78 +/- 1.97 nmol/L throughout the first 26 mo of the study, 7.28 +/- 1.28 nmol/L during the following 21 mo, and 0.45 +/- 0.035 nmol/L in the last 34 mo of this study. Early musth signs ceased within 3 days of drug administration after 10 of 12 injections. The mean serum testosterone concentrations were significantly decreased by the last 34 mo of the study. The results suggest leuprolide is a suitable alternative for controlling or preventing (or both) musth in captive Asian elephants, although permanent reproductive effects may occur. Zoos and wildlife conservation institutions could benefit from the use of LA in Asian elephants to increase the male availability in captivity, consequently ensuring genetic diversity and the perpetuation of the species

deOliveira, C.A., West, G.D., Houck, R., Leblanc, M., 2004. Control of musth in an Asian elephant bull (Elephas maximus) using leuprolide acetate. Journal of Zoo and Wildlife Medicine 35 , 70-76.
Abstract: The results of long-term administration of leuprolide acetate (LA) depot in a 52-yr-old Asian elephant bull (Elephas maximus) for control of musth are presented. Twelve injections were administered for 6 yr during our interpretation of early musth or "premusth". Intervals between musth periods during the study varied from 2 to 34 mo. Blood samples, drawn weekly, were assayed for serum testosterone concentrations; mean levels were 11.78 +/- 1.97 nmol/L throughout the first 26 mo of the study. 7.28 +/- 1.28 nmol/L during the following 21 mo. and 0.45 +/- 0.035 nmol/L in the last 34 mo of this study. Early musth signs ceased within 3 days of drug administration after 10 of 12 injections. The mean serum testosterone concentrations were significantly decreased by the last 34 mo of the study. The results suggest leuprolide is a suitable alternative for controlling or preventing (or both) musth in captive Asian elephants, although permanent reproductive effects may occur. Zoos and wildlife conservation institutions could benefit from the use of LA in Asian elephants to increase the male availability in captivity, consequently ensuring genetic diversity and perpetuation of the species.

Freeman, E.W., Weiss, E., Brown, J.L., 2004. Examination of the interrelationships of behavior, dominance status, and ovarian activity in and African elephants. Zoo Biology 23, 431-448.
Abstract: Ovarian inactivity has been identified in captive African (Loxodonta africana) and Asian (Elephas maximus) elephants and is thought to be mediated in part by social influences. Thus, a survey was conducted to determine how behavior and dominance status relate to each other and to ovarian cyclicity. For both Asian and African elephants, dominance status was positively correlated with relative size, age, temperament, disciplinary nature, and willingness to share novel objects. Relative size and temperament were also related to disciplinary nature toward herdmates. Behavior toward keepers was a good indicator of the willingness of elephants to follow cmmands, whereas sharing novel objects was positively correlated with object curiosity. Finally, dominance status, temperament, and disciplinary nature were all correlated with willingness to share. Comparisons of ovarian cyclicity status with behavior rankings were conducted only for African elephants because of the low number of noncycling Asian elephants surveyed. Overall, social status appeared to be the best predictor of ovarian activity in African elephants. Noncycling African elephant females ranked higher in the dominance hierarchy and gave more discipline to herdmates than cycling cohorts. It remains to be determined whether these are cause or effect relationships, but clearly it is important to understand how physical and social attributes impact physiological processes, such as reproduction. Captive management now needs to focus on optimizing social and environmental conditions to maximize reproductive potential in elephants.

Garstang, M., 2004. Long-distance, low-frequency elephant communication. J Comp Physiol A 190, 791-805.
Abstract: The production, transmission, and reception of and the behavioral response to long-distance, low-frequency sound by elephants is reviewed. The structure of low-frequency calls generated by elephants is separated into the ''source'' and the ''filter'' roles played by the lungs, larynx and vocal track, the composition of the expired air and the ambient air temperature. Implications regarding the size, age, sex, sexual and physical status follow from the call structure and detection. Reception of the signal is discussed in terms of the characteristics of the elephant's ear with particular attention to the determination of the threshold of hearing and the ability to locate the source of low-frequency sounds. Factors which influence the transmission of near infrasound are related to atmospheric structure. The critical role played by the thermal stratification and vertical gradient and magnitude of the wind in determining both the range and the detection of a signal are discussed for open and closed elephant habitats. Infrasound plays a pervasive role in reproduction, resource utilization, avoidance of predation and other social interactions. Current and future technology can be expected to contribute to the detection and interpretation of elephant communication. This will aid in the understanding of behavior and in efforts to sustain the species.

Garstang, M., 2004. Long-distance, low-frequency elephant communication
686. J. Comp Physiol A Neuroethol. Sens. Neural Behav. Physiol 190, 791-805.
Abstract: The production, transmission, and reception of and the behavioral response to long-distance, low-frequency sound by elephants is reviewed. The structure of low-frequency calls generated by elephants is separated into the "source" and the "filter" roles played by the lungs, larynx and vocal track, the composition of the expired air and the ambient air temperature. Implications regarding the size, age, sex, sexual and physical status follow from the call structure and detection. Reception of the signal is discussed in terms of the characteristics of the elephant's ear with particular attention to the determination of the threshold of hearing and the ability to locate the source of low-frequency sounds. Factors which influence the transmission of near infrasound are related to atmospheric structure. The critical role played by the thermal stratification and vertical gradient and magnitude of the wind in determining both the range and the detection of a signal are discussed for open and closed elephant habitats. Infrasound plays a pervasive role in reproduction, resource utilization, avoidance of predation and other social interactions. Current and future technology can be expected to contribute to the detection and interpretation of elephant communication. This will aid in the understanding of behavior and in efforts to sustain the species

Graham, L.H., Bando, J., Gray, C., Buhr, M.M., 2004. Liquid storage of Asian elephant (Elephas maximus) sperm at 4 degrees C
737. Anim Reprod. Sci. 80, 329-340.
Abstract: The Asian elephant (Elephas maximus) population in the wild has been in decline for several decades and breeding in captivity has not been self-sustaining. The use of artificial insemination (AI) can help overcome many of the difficulties associated with breeding elephants in captivity; however, the ability to store semen for extended periods of time is critical to the successful application of AI to elephants. The objective of the present study was to assess the effects of four different semen extenders and the presence of egg yolk on the viability and motility of Asian elephant semen stored at 4 degrees C. High quality ejaculates (n=4) were collected from two Asian elephant bulls by rectal massage. Aliquots of each ejaculate were extended in four different diluents (Beltsville thawing solution (BTS); Tris-citric acid (TCA)/fructose-based; Beltsville F5 (BF5); dextrose-supplemented phosphate-buffered saline (PBS)) with or without egg yolk then cooled and stored at 4 degrees C. The percentages of viable (viability) and motile (motility) sperm were evaluated at 8, 24 and 48 h following collection. The addition of egg yolk significantly reduced the percentage loss in viability from initial collection to 48 h compared to extenders without egg yolk (17.0 +/- 8.2 versus 32.6 +/- 8.9 decline in percent viable sperm in the population, respectively; P<0.05). Extender and egg yolk affected (P<0.005) total motility and percent progressively motile sperm at all evaluation times during incubation. TCA + egg yolk maintained higher (P<0.05) levels of progressive motility compared to other extenders supplemented with egg yolk. These results indicate that Asian elephant semen extended in TCA diluent supplemented with egg yolk can maintain at least 50% viability and motility when stored at 4 degrees C for 48 h.

Hermes, R., Hildebrandt, T.B., Goritz, F., 2004. Reproductive problems directly attributable to long-term captivity--asymmetric reproductive aging
704. Anim Reprod. Sci. 82-83, 49-60.
Abstract: Problems attributable to long-term captivity have been identified and are responsible for the difficulties in establishing successful reproduction in captive populations of wildlife, specifically, elephants and rhinoceroses. Historically, non-reproductive periods of 10-15 years in nulliparous female rhinoceroses and elephants have not been considered problematic. New evidence suggests that prolonged exposure to endogenous sex steroids and that long stretches of non-reproductive periods induce asymmetric reproductive aging in captive animals. The consequences are reduced fertility, shortened reproductive life-span and, eventually, irreversible acyclicity. Although age-related reproductive lesions have also been documented in male rhinoceroses, they continue to maintain a longer reproductive life-span than females. Since human and domestic animal models have already indicated that early pregnancy provides natural protective mechanism against asymmetric reproductive aging processes and premature senescence, it is imperative that appropriate counter measures such as assisted reproductive technologies (ART) be utilized to ensure early pregnancy in captive animals for their preservation and to ensure increased genetic diversity of the captive populations

Hermes, R., Hildebrandt, T.B., Goritz, F., 2004. Reproductive problems directly attributable to long-term captivity-asymmetric reproductive aging. Animal Reproduction Science 82-83, 49-60.
Abstract: Problems attributable to long-term captivity have been identified and are responsible for the difficulties in establishing successful reproduction in captive populations of wildlife, specifically, elephants and rhinoceroses. Historically, non-reproductive periods of 10-15 years in nulliparous female rhinoceroses and elephants have not been considered problematic. New evidence suggests that prolonged exposure to endogenous sex steroids and that long stretches of non-reproductive periods induce asymmetric reproductive aging in captive animals. The consequences are reduced fertility, shortened reproductive life-span and, eventually, irreversible acyclicity. Although age-related reproductive lesions have also been documented in male rhinoceroses, they continue to maintain a longer reproductive life-span than females. Since human and domestic animal models have already indicated that early pregnancy provides natural protective mechanism against asymmetric reproductive aging processes and premature senescence, it is imperative that appropriate counter measures such as assisted reproductive technologies (ART) be utilized to ensure early pregnancy in captive animals for their preservation and to ensure increased genetic diversity of the captive populations.

Hildebrandt, T.B., Hermes, R., Janssen, D.L., Oosterhuis, J.E., Murphy, D., Göritz, F. Reproductive evaluation in wild African elephants prior to translocation. 2004 PROCEEDINGS AAZV, AAWV, WDA JOINT CONFERENCE.  75-76. 2004.
Ref Type: Conference Proceeding
Abstract: Translocations of wild African (Loxodonta africana) elephants have increased significantly since 1993 after Clem Coetzee developed a new method to move adult elephants in Zimbabwe. Since then the technique have been optimized mainly by the staff of the Kruger National Park (KNP) and over 750 elephants in family units and almost 100 mature bulls have been translocated by the KNP capture team.1 The translocations were mainly performed for reducing the number of elephants in KNP and for stocking other reserves. Few elephants were also moved for overseas export to international zoological institutions. However, each elephant translocation is always a logistic challenge and is extremely costly. Therefore, it is very important to select the right elephants or elephant groups for the future translocation. If the main goal of a translocation is the establishment of a new breeding group, it is especially important to select infertile individuals and highly pregnant females which could have a miscarriage due to the transport stress. The IZW team developed a field applicable portable ultrasound technique which allows the reproductive ev ry Killmar (ZSSD), and Randy Rieches (ZSSD).

LITERATURE CITED
1. Hofmeyr, M. 2003. Translocation as a management tool for control of elephant populations. Managing African Elephant Populations: Act or Let Die. Beekbergen, The Netherlands, 6.-7.Nov., Pp. 38-39.
2. Hildebrandt T. B., F. Göritz, N. C. Pratt, D. L. Schmitt, S. Quandt, J. Raath and R. R. Hofmann. 1998. Reproductive assessment of male elephants (Loxodonta africana and Elephas maximus) by ultrasonography. J. Zoo Wildl. Med. 29: 114-128.

Jones, C.J., Wooding, F.B., Mathias, S.S., Allen, W.R., 2004. Fetomaternal glycosylation of early placentation events in the African elephant Loxodonta africana
740. Placenta 25, 308-320.
Abstract: During implantation in the African elephant (Loxodonta africana), fetal trophoblast displaces the surface uterine epithelium and superficially penetrates the uterine glands. This limited invasion is followed by the upgrowth of blunt fingers of endometrial stroma, covered with trophoblast and containing capillaries that subsequently vascularize the growing placenta. We have used lectin histochemistry to compare the glycosylation of maternal endothelial cells in the endometrium with those growing within the trophoblastic processes of a 2 g embryo (approximately 125 days' gestation), and also examine changes in the endometrial glands associated with trophoblastic invasion. Maternal vessels at the apices of the trophoblast-covered stromal upgrowths showed increased expression of terminal N-acetyl galactosamine, N-acetyl glucosamine oligomers, some sialic acids, and tri/tetra-antennate non-bisected complex N-linked glycan, as indicated by increased lectin staining. The areas of increased staining were also more resistant to neuraminidase digestion. Invaded glands had distended walls composed of flattened epithelial cells, some of which showed heavy lectin staining suggestive of intracellular glycan accumulation. The vascular changes suggest that new maternal capillary growth is accompanied by alterations in surface glycosylation. This may be the result of increased glycosyl transferase activity associated with cell proliferation and may also indicate the expression of significantly increased anti-adhesive molecules preventing blood stasis and egress of maternal immunocompetent cells into the fetal compartment

Meyer, J.M., Walker, S.L., Freeman, E.W., Steinetz, B.G., Brown, J.L., 2004. Species and fetal gender effects on the endocrinology of pregnancy in elephants
682. Gen. Comp Endocrinol. 138, 263-270.
Abstract: Quantitative and temporal progestin profiles vary during gestation in the elephant, sometimes making it difficult to determine if a pregnancy is progressing normally. The aim of the present study was to determine if circulating progestin variability was related to species or fetal gender effects. A similar comparison also was conducted for secretory profiles of prolactin, relaxin, and cortisol. Overall mean progestin concentrations during gestation in Asian (n = 19) and African (n = 8) elephants were similar; however, the temporal profiles differed (P < 0.001). Concentrations were higher in African elephants during the first half of pregnancy, but then declined to levels below those observed in Asian elephants (P < 0.05). There also was a fetal gender effect in Asian, but not African elephants. Progestin concentrations were higher in Asian cows carrying male calves (n = 9) as compared to those carrying females (n = 10) (P < 0.001). Overall prolactin concentrations were higher in Asian than in African elephants between 8 and 15 months of gestation ( P< 0.001). There were no species differences in the secretory patterns of relaxin. Cortisol was relatively stable until the end of gestation when significant surges were observed, mainly between 8 and 11 days before parturition, and again on the day of birth. In sum, a comparison of progestin patterns between Asian and African elephants identified notable differences related to species and fetal gender. A role for cortisol in the initiation of parturition also was inferred from these data. From a practical standpoint, understanding the factors affecting gestational hormone characteristics and recognizing what the species differences are will help ensure that data used in diagnosing and monitoring elephant pregnancies are properly interpreted

Ruedi, D. Collection and handling of semen and insemination in the African elephant (Loxodonta africana) at Basle Zoo.  2004.
Ref Type: Unpublished Work
Abstract: It is urgently necessary that we take measures towards enhancing the efficiency of captive breeding of African and Asian elephants. This might be accomplished in several ways: for instance, by an extensive loan of cows for natural mating and, last but not least, by breeding under artificial conditions. With the third possibility in mind we initiated a programme in 1977 to develop a technique for artificial insemination in the African elephant; it encompassed the collection and handling of semen, oestrus detection and insemination.

Sanchez, C.R., Murray, S., Montali, R.J., Spelman, L.H., 2004. Diagnosis and treatment of presumptive pyelonephritis in an Asian elephant (Elephas maximus). J Zoo Wildl Med 35, 397-399.
Abstract: A 37-yr-old female Asian elephant (Elephas maximus) presented with anorexia, restlessness, and dark-colored urine. Urinalyses showed hematuria, leukocyturia, isosthenuria, proteinuria, granular casts, and no calcium oxalate crystals. Bloodwork revealed azotemia. Urine culture revealed a pure growth of Streptococcus zooepidemicus resistant to sulfamethoxazole-trimethoprim but susceptible to cephalosporins. A presumptive diagnosis of pyelonephritis was made based on bloodwork, urinalysis, and urine culture. The animal was treated with intravenous ceftiofur, and intravenous and per rectum fluids were given for hydration. The elephant's attitude and appetite returned to normal, the abnormal blood parameters resolved, and urinary calcium oxalate crystals reappeared after treatment, supporting presumptive diagnosis. Follow-up ultrasonography revealed an abnormal outline of both kidneys with parenchymal hyperechogenicity and multiple uterine leiomyomas.

Stumpf, P., Welsch, U., 2004. Secretory and defensive functions of the duct system of the lactating mammary gland of the African elephant (Loxodonta africana, Proboscidea). Zoomorphology 123, 155-157.
Abstract: The duct system of the lactating mammary Gland of the African elephant (Loxodonta africana) was investigated with histochemical and immunohistochemical techniques and with the transmission electron microscope in order to detect specific cell biological differentiations in the ductal epithelia of this species, which is marked by an unusually long lactation period. General histology and electron microscopy allow to distinguish several segments in the entire duct system. The apical membranes of the epithelia have binding sites for several lectins [Canavalia ensiformis agglutinin (ConA), Ricinus communis agglutinin (RCA 1), Wisteria floribunda agglutinin (WFA), peanut agglutinin (PNA)] and also stain with alcian blue indicating the presence of a highly differentiated negatively charged glycocalyx forming an effective barrier between lumen and epithelium. Cytokeratins, actin, tubulin and vinculin show different expression intensities in the proximal and distal portion of the duct system. Lysozyme, lactoferrin, the secretory component of IgA and human beta defensin-2 are expressed in the epithelium of the entire duct system. In the distal portion of the ducts the staining intensity is stronger than in the proximal portion. We conclude that the duct system of the elephant mammary gland has specific secretory functions and that the secretory products are part of the defensive mechanisms against invading microorganisms.

Thongtip, N., Saikhun, J., Damyang, M., Mahasawangkul, S., Suthunmapinata, P., Yindee, M., Kongsila, A., Angkawanish, T., Jansittiwate, S., Wongkalasin, W., Wajjwalkul, W., Kitiyanant, Y., Pavasuthipaisit, K., Pinyopummin, A., 2004. Evaluation of post-thaw Asian elephant (Elephas maximus) spermatozoa using flow cytometry: the effects of extender and cryoprotectant
711. Theriogenology 62, 748-760.
Abstract: Although the development of semen cryopreservation in the African elephants (Loxodonta africana) has been accomplished, effective procedures for cryopreservation of Asian elephant (Elephas maximus) spermatozoa have not been established. In the present study, we investigate the freezing methods for conservation of Asian elephant spermatozoa under field conditions and identify the most suitable freezing protocols which provide acceptable post-thaw semen quality. Semen was collected from two Asian elephant bulls (EM1 and EM2, 10 ejaculates from each bull) by manual manipulation and were assessed for volume, pH, sperm cell concentration, and progressive motility. Eight out of 20 ejaculates were of acceptable quality (progressive motility >/= 60%), and were used for cryopreservation studies. Semen were frozen in TEST + glycerol, TEST + DMSO, HEPT + glycerol, or HEPT + DMSO. The post-thaw progressive sperm motilities were assessed, and sperm cells were stained with PI and FITC-PNA for membrane and acrosomal integrity assessment using flow cytometry. Post-thaw progressive motility of spermatozoa (EM1: 42.0 +/- 4.3%; EM2: 26.0 +/- 17.3%) and the percentage of membrane and acrosome intact spermatozoa (EM1: 55.5 +/- 8.1%; EM2: 46.3 +/- 6.4%) cryopreserved in TEST + glycerol were significantly higher than (P < 0.05) those frozen in the other medium investigated choices for cryopreservation of Asian elephant spermatozoa. The data support the use of TEST + glycerol as an acceptable cryopreservation media of Asian elephant semen for the establishment of sperm banks

Visscher, D.R., vanAarde, R.J., Whyte, I., 2004. Environmental and maternal correlates of foetal sex ratios in the African buffalo (Syncerus caffer) and savanna elephant (Loxodonta africana). Journal of Zoology 263 , 111-116.
Abstract:  Many species exhibit skewed sex ratios at birth. Here we investigate the relationships between environmental and maternal variables (as surrogates for maternal condition) and foetal sex in African buffalo Syncerus caffer and elephant Loxodonta africana of the Kruger National Park, South Africa. Using logistic regression no significant effect was found of year, maternal lactational status, maternal age, rainfall or density on foetal sex ratio. Using a subset of our data, it was also concluded that maternal body condition did not affect foetal sex ratio in buffalo. Our analyses failed to support hypotheses predicting that mothers will skew the sex ratios of their offspring in relation to their body condition. In this study, buffalo and elephant produced offspring with a sex ratio close to parity. Our results are discussed in light of the implications for testing such hypotheses in analyses of population level.

 2003. Healthcare, Breeding and Management of Asian Elephants. Project Elephant. Govt. of India, New Delhi.

Allen, W.R., Mathias, S., Wooding, F.B., van Aarde, R.J., 2003. Placentation in the African elephant (Loxodonta africana): II morphological changes in the uterus and placenta throughout gestation. Placenta 24, 598-617.
Abstract: The gross and microscopic development of the zonary endotheliochorial placenta in the African elephant was studied in 22 gravid uteri that ranged in
gestational stage from 0.5 to 20.6 months. The conceptus only ever occupies one horn of the uterus and is associated with 2-5 large corpora lutea that persist in the ipsilateral ovary throughout gestation. Initially, the trophoblast in the equatorial region of the conceptus completely replaces the lumenal epithelium of the endometrium to which it is apposed. Blunt upgrowths of endometrial stroma then develop, each closely invested by trophoblast, and  containing the capillaries that will vascularize this maternal component of the resulting placental band. With advancing gestation the lamellate stromal upgrowths increase markedly in length and become much thinner, thereby bringing the trophoblast into intimate contact with the endothelium of the maternal capillaries. They also become folded or pleated to increase the total area of intimate feto-maternal contact. At the lateral edges of the placental band the lamellae bend over towards the endometrium to form a blind cleft. Leakage of blood into this area creates haemophagous zones in which phenotypically specialized trophoblast cells phagocytose the blood components. The presence of large resorbing blood clots and circumferential scars in the uteri of three post parturient animals initiated the hypothesis that, when the standing elephant gives birth at term, the passage of the 120 kg fetus through the vagina may wrench the placenta off the endometrium by severing its very narrow maternal placental hilus. The resulting intrauterine haemorrhage may then play a role in preventing further conception for around 2 years.Department of Clinical Veterinary Medicine Equine Fertility Unit, University of Cambridge, Mertoun Paddocks,Woodditton Road, Suffolk CB8 9BH, Newmarket, UK. vetart@aht.org.uk

Bhattacharyya, B.K., 2003. Some reproductive traits of elephants. In: Das, D. (Ed.), Healthcare, Breeding and Management of Asian Elephants. Project Elephant. Govt. of India, New Delhi, pp. 37-41.

Czekala, N.M., MacDonald, E.A., Steinman, K., Walker, S., Garrigues, N.W., Olson, D., 2003. Estrogen and LH dynamics during the follicular phase of the estrous cycle in the Asian elephant. Zoo Biology 22, 443-454.
Abstract: Pituitary and corpus luteum hormone patterns throughout the elephant estrous cycle have been well characterized. By contrast, analysis of follicular maturation by measurement of circulating estrogens has been uninformative. This study tested the ability of a urinary estradiol-3-glucuronide radioimmunoassay to noninvasively assess follicular development during the nonluteal phase of the elephant estrous cycle, and to determine the relationship between estrogen production and the "double LH surge." Daily urine and serum samples were collected throughout seven estrous cycles from three Asian elephants, and urine was collected from an additional three females, for a total of 13 cycles. Serum was analyzed for luteinizing hormone (LH), and urine was analyzed for estrogens and progestins. Elephants exhibited a typical LH pattern, with an anovulatory LH (anLH) surge occurring approximately 21 days before the ovulatory LH (ovLH) surge. The urinary estrogen pattern indicated the presence of two follicular waves during the nonluteal phase. The first wave (anovulatory) began 5 days before the anLH surge and reached a maximum concentration the day before the peak. Thereafter, urinary estrogens declined to baseline for 2 weeks before increasing again to peak concentrations on the day of the ovLH surge. Urinary progestins were baseline throughout most of the follicular phase, increasing 2-3 days before the ovLH surge and continuing into the luteal phase. These results support previous ultrasound observations that two waves of follicular growth occur during the nonluteal phase of the elephant estrous cycle. Each wave is associated with an increase in estrogen production that stimulates an LH surge. Thus, in contrast to serum analyses, urinary estrogen monitoring appears to be a reliable method for characterizing follicular activity in the elephant.

Das, D., 2003. Breeding, reproduction and conservation strategies in Asian elephants. In: Das, D. (Ed.), Healthcare, Breeding and Management of Asian Elephants. Project Elephant. Govt. of India, New Delhi, pp. 45-57.

Davis, B.L., Dill, W.M., Hicks, A.R., Goodwin, T.E., Hollister-Smith, J., Alberts, S.C. Use of SPME and GC-MS for chemical analysis of urine from African elephants in musth. Abstracts Of Papers Of The American Chemical Society 225[614-CHED Part 1 MAR 2003]. 2003.
Ref Type: Abstract
Abstract: Addresses: Hendrix Coll, Dept Chem, Conway, AR 72032 USA ;Duke Univ, Dept Biol Sci, Durham, NC 27706 USA

Dehnhard, M., Hatt, J.M., Eulenberger, K., Ochs, A., Strauss, G., 2003. Headspace solid-phase microextraction (SPME) and gas chromatography-mass spectrometry (GC-MS) for the determination of 5alpha-androst-2-en-17-one and -17beta-ol in the female Asian elephant: application for reproductive monitoring and prediction of parturition. J Steroid Biochem Mol Biol. 84, 383-391.
Abstract: Asian elephants are not self-sustaining in captivity. The main reasons for this phenomenon are a low birth rate, an aging population, and poor calf-rearing. Therefore, it is essential that reproductive rates had to be improved and there is need for rapid quantitative measures to monitor reproductive functions focussing on estrous detection and the prediction of the period of parturition. The objective of this study was to develop a method which combines headspace solid-phase microextraction (SPME) and gas chromatography-mass spectrometry (GC-MS) for analyses of 5alpha-androst-2-en-17beta-ol and -17-one to prognose estrous and to predict the period of parturition. SPME was carried out with a CTC Combi Pal system.The course of the luteal phase-specific substance 5alpha-androst-2-en-17beta-ol and -17-one followed a cyclic pattern in which the follicular and luteal phases could be clearly distinguished (mean estrous cycle length, 15+/-1.4 weeks). Based on daily urine samples, estrous prognosis might be possibly based on the initial 5alpha-androst-2-en-17beta-o1 increase at the end of the follicular phase. Parturition prognosis was performed in three elephant cows based on the 5alpha-androst-2-en-17beta-o1 drop to baseline levels 5-4 days prior parturition. Experiments revealed that 5alpha-androst-3alpha-ol-17-one and probably 5alpha-androst-3alpha-ol-17beta-ol are generated from sulfate conjugates by a thermal process. Institute for Zoo Biology and Wildlife Research, PF 601103, D-10252 Berlin, Germany. dehnhard@izw-berlin.de

Dill, W.M., Davis, B.L., Hicks, A.R., Goodwin, T.E., Rasmussen, L.E.L., Loizi, H., Schulte, B.A. Chemical analysis of preovulatory female African elephant urine: A search for putative pheromones. Abstracts Of Papers Of The American Chemical Society 225[409-CHED Part I Mar 2003]. 2003.
Ref Type: Abstract

Gage, L., Schmitt, D. Dystocia in an African elephant (Loxodonta africana). Proc Amer Assoc Zoo Vet.  88. 2003.
Ref Type: Conference Proceeding
Abstract: A 24-yr-old African elephant (Loxodonta africana) was inseminated on 12/2/00 and again on 12/4/00.  Pregnancy was confirmed on 1/21/01 with an ultrasound evaluation, and elevated progesterone levels of over a twelve to sixteen week period supported the diagnosis. The pregnancy progressed without complication.  Ultrasounds confirmed calf growth and movement throughout the pregnancy.  On 10/5/02, 673 days post-insemination, the cow passed the mucous plug in the evening.  The chorio-allantois ruptured shortly after that.  There were a few modest contractions over a period of about three hours, which then ceased.  Fetal front feet could be palpated in the birth canal, just into the cranial pelvis. Fetal circulation was confirmed using color doppler ultrasound of the fetal extremities.  Oxytocin was administered on 10/6/02.  The first two doses resulted in a few good contractions.  Fetal feet entered further into the pelvis, and fetal viability was confirmed again with color doppler ultrasound. Oxytocin was administered three more times resulting in a few small contractions and some stretching.  One dose of 110 IU Oxytocin resulted in one large contraction, subsequent doses of oxytocin did not yield any appreciable additional efforts.  That evening while walking the elephant, the amniotic sac broke releasing several liters of fluid. Blood was drawn and serum chemistries were within normal range. On 10/7/02 the uterus did not respond to oxytocin. Color doppler ultrasound of the fetal legs revealed no discernable fetal circulation.  Antibiotics and supportive care were initiated.  For the next week the feet were easily palpated within the pelvis.  For the several days the elephant was stiff and moderately depressed. Her appetite was good and she was drinking water. The elephant was given another dose of oxytocin on 10/15/02, which resulted in a few moderate contractions, but no progress was made in delivering the calf.  The next day the elephant seemed more depressed and uncomfortable. Her appetite and water intake were markedly reduced and within two days she exhibited what appeared to be abdominal pain. The elephant was treated aggressively with antibiotics and fluids for several weeks.  Severe peritonitis was confirmed during a laparoscopic procedure and the elephant was euthanized. The post-mortem examination revealed a tear in the uterus and a normal-sized fetus in normal presentation in the birth canal. The fetus however was abnormal and had arthrogryposis affecting all four limbs to varying degrees.  Both rear limbs of the fetus were particularly affected and the joints of those limbs were bent in an abnormal way, anchoring the fetus inside the dam. The arthrogryposis of the calf was likely the cause of the dystocia, and was likely the contributing factor to the tear in the uterus.

Ganswindt, A., Palme, R., Heistermann, M., Borragan, S., Hodges, J.K., 2003. Non-invasive assessment of adrenocortical function in the male African elephant (Loxodonta africana) and its relation to musth. Gen Comp Endocrinol 134, 156-166.
Abstract: German Primate Centre, Department of Reproductive Biology, Kellnerweg 4, 37077 Gottingen, Germany. ganswindt@www.dpz.gdwg.de

Adult male elephants periodically show the phenomenon of musth, a condition associated with increased aggressiveness, restlessness, significant weight reduction and markedly elevated androgen levels. It has been suggested that musth-related behaviours are costly and that therefore musth may represent a form of physiological stress. In order to provide data on this largely unanswered question, the first aim of this study was to evaluate different assays for non-invasive assessment of adrenocortical function in the male African elephant by (i) characterizing the metabolism and excretion of [3H]cortisol (3H-C) and [14C]testosterone (14C-T) and (ii) using this information to evaluate the specificity of four antibodies for determination of excreted cortisol metabolites, particularly with respect to possible cross-reactions with androgen metabolites, and to assess their biological validity using an ACTH challenge test. Based on the methodology established, the second objective was to provide data on fecal cortisol metabolite concentrations in bulls during the musth and non-musth condition. 3H-C (1 mCi) and 14C-T (100 microCi) were injected simultaneously into a 16 year old male and all urine and feces collected for 30 and 86 h, respectively. The majority (82%) of cortisol metabolites was excreted into the urine, whereas testosterone metabolites were mainly (57%) excreted into the feces. Almost all radioactive metabolites recovered from urine were conjugated (86% 3H-C and 97% 14C-T). In contrast, 86% and >99% of the 3H-C and 14C-T metabolites recovered from feces consisted of unconjugated forms. HPLC separations indicated the presence of various metabolites of cortisol in both urine and feces, with cortisol being abundant in hydrolysed urine, but virtually absent in feces. Although all antibodies measured substantial amounts of immunoreactivity after HPLC separation of peak radioactive samples and detected an increase in glucocorticoid output following the ACTH challenge, only two (in feces against 3alpha,11-oxo-cortisol metabolites, measured by an 11-oxo-etiocholanolone-EIA and in urine against cortisol, measured by a cortisol-EIA) did not show substantial cross-reactivity with excreted 14C-T metabolites and could provide an acceptable degree of specificity for reliable assessment of glucocorticoid output from urine and feces. Based on these findings, concentrations of immunoreactive 3alpha,11-oxo-cortisol metabolites were determined in weekly fecal samples collected from four adult bulls over periods of 11-20 months to examine whether musth is associated with increased adrenal activity. Results showed that in each male levels of these cortisol metabolites were not elevated during periods of musth, suggesting that in the African elephant musth is generally not associated with marked elevations in glucocorticoid output. Given the complex nature of musth and the variety of factors that are likely to influence its manifestation, it is clear, however, that further studies, particularly on free-ranging animals, are needed before a possible relationship between musth and adrenal function can be resolved. This study also clearly illustrates the potential problems associated with cross-reacting metabolites of gonadal steroids in EIAs measuring glucocorticoid metabolites. This has to be taken into account when selecting assays and interpreting results of glucocorticoid metabolite analysis, not only for studies in the elephant but also in other species.

Hama, N., Yamada, A., Noda, A., Murata, K., Shimada, Y., Ashida, M., Ishikawa, K., Matsuo, Y., Okuno, K., 2003. Serum hormonal changes in a female Asian elephant (Elephas maximus) with stillbirth. Japanese Journal of Zoo and Wildlife Medicine 8, 109-113.
Abstract: A case of stillbirth by an 11-year-old Asian elephant on January 11, 2002 is reported. Swelling of the abdominal region as well as the nipples in 2000 and 2001 had been previously observed. Serum progesterone (P), estradiol (E2) and prolactin (PRL) were measured monthly from May 2000 to January 2002 using radioimmunoassay (RIA). And P and E2 were additionally measured weekly from September 4, 1999 to January 12, 2002 using enzyme immunoassay (EIA). On the basis of EIA, elevated P levels were observed during early April 2000, and remained high until the end of 2001. Serum P concentrations gradually began to decrease by December 2001; at 6 days before stillbirth, there was a sudden drop in the level of blood P. On the day that the stillbirth occurred, the P value decreased to the level first recorded prior to April 2000. E2, based on results of EIA, did not display the regular cyclic trend prior to the elevation of serum P for pregnancy. The recorded values of serum P by both EIA and RIA were positively correlated (Pearson's correlative coefficient: 0.763, P < 0.01). On the basis of RIA, serum E2 and PRL were almost below detectable levels (E2: 8.0 pg/ml, PRL:0.5 ng/ml). The period of pregnancy estimated from the change of serum P concentration by EIA was 640 days.

Hermes, R., Arav, A., Saragusty, J., Goeritz, F., Pettit, M., Blottner, S., Flach, E., Eshkar, G., Boardman, W., Hildebrandt, T.B. Cryopreservation of Asian elephant spermatozoa using directional freezing. Proc.Amer Assoc of Zoo Veterinarians.  264. 2003.
Ref Type: Conference Proceeding
Abstract: Male infertility and absence of males in a facility are contributing factors to the limited reproduction of Asian elephants in captivity. Subsequent transport for breeding purposes increase social stress, risks of disease transmission and management costs. Recent success in artificial insemination eliminated these obstacles only transporting the semen. However, the transport of fresh semen involves logistical<bold> </bold>difficulties: access to semen donors, consistent semen quality and preservation of the spermatozoa during transport. The use of cryo-preserved sperm for AI can partially overcome these problems and can additionally be used for the establishment of Genome Resource Banks. However, to date, attempts to cryo-preserve Asian elephant spermatozoa have failed due to its sensitivity to freezing. Aims of this study were to identify the temperature range during which spermatozoa is most sensitive to chilling injury, and to use directional freezing (DF) to reduce cell damage during the freezing process. Semen was collected from two Asian elephants by manual stimulation. DF was used for freezing sperm samples. In contrast to conventional freezing methods DF facilitated a fast cooling rate, controlled ice crystal formation and cryopreservation of large volumes. Samples extended with a variety of DMSO extenders showed post thaw motility of 30-40%. DF was able to cryo-preserve Asian Elephant spermatozoa for the first time. As DF seems to reduce cryo injury it may become of interest to optimize existing cryopreservation protocols of other endangered species, or to make cryopreservation even possible in species with cryo-sensitive spermatozoa.

Hildebrandt, T.B., Strike, T., Flach, E., Sambrook, B.S., Dodds, J., Lindsay, N., Goeritz, F., Hermes, R., McGowan, M. Fetotomy in the elephant. Proc Amer Assoc Zoo Vet.  89-92. 2003.
Ref Type: Conference Proceeding
Abstract: There were several reports about dystocia and its treatment in elephants since they were kept in captivity in western zoos and safari parks. ^{1, 2, 3, 4, 5, 6, 7, 8, 9, 10}  Especially in the last two years there has been a dramatic accumulation of reports about birth associated problems (Tab. 1). Causes for this development are probably the intensified captive elephant breeding programs and the involvement of more older nulliparous cows.

Over the last 100 years, there were more dystocia cases in captive Asian elephants than in African elephants. ⁷  However, this difference seems to be abolished with enhanced number of pregnancies in African elephants by now. The fact of an increased percentage of dystocia cases in both species can have two consequences for captive management. Firstly, older nulliparous cows will be strictly excluded from breeding in the future or secondly, the birth management will be improved.

Three (Table 1, Em 4, 5; La 3) of 10 cows died from the consequences of dystocia in western zoological institutions over the last two years. Totally, there were eleven cases of fatal dystocia cases listed in the literature since 1972 (Table 2). ^{5, 6, 7}  The seven cesarian performed (Table 2) as the ultimate intervention to treat the dystocia in elephants ended all with euthanasia or death of the females. The following presentation will described a potential new way of birth management in the elephant, the fetotomy. The authors believe that the fetotomy is prospective tool instead of the unsuccessful cesarian for saving the life of the dam with severe dystocia.  This method is recommended by the authors as an ultimate tool besides all the other important preparations in pregnant cows for an upcoming birth. Most important classical preparations are following: (I) intensive physical exercise, (ii) optimization of the body weight with a pregnancy associated diet, (iii) training for safe handling in case of necessary intervention in free or protected contact.  The fetotomy was never applied before in elephants because of the limited exploration field and the high degree of skeleton calcification in comparison to domestic and exotic hoofstock. 

In this particular case it was firstly tried to remove the dead fetus by episiotomy. ^{2, 3, 4, 7, 9, 10} However, the episiotomy attempt failed due to he inability to extract a dead oversized or malpositioned fetus.  From the two options to leave the dead fetus inside the uterus despite the surgically opened genital tract or to cut out parts of the fetus it was decided to perform a fetotomy. The initial hesitation to perform such drastic and complicated procedure led to the complication of an emphysemic fetus and progressive peritonitis in the female even before the procedure started. Fetus was successfully removed in six parts using an extra long and durable carthorse embryotom (Thygesen's type). Unfortunately, the patient died 3 days later because of the treatment resistant peritonitis.

In conclusion, it was demonstrated that fetotomy can successfully be applied in elephants. The authors suggested in order to achieve a positive outcome for the dam to decide on such ultimate intervention earlier in the dystocia process. Active elephant birth management should always be prepared for dystocia treatment ranging from (I) the administration of oxytocin, (ii) rectal massage of the caudal birth channel, (iii) episiotomy to the last option (iv) of fetotomy to safe the life of the mother.  Based on the complex preparation for an elephant birth and the knowledge about the huge range of potential complications it is advised to consult veterinary elephant specialists well in advance to ensure maximum expertise and experience.

Acknowledgements
The authors like to thank Charlie Gray who helped us to collect the relevant data from the dystocia cases and the elephant staff from Whipsnade Wild Animal Park for their active support. We also like to acknowledge professor Peter Glatzel, DVM and Andrea Krause for helping with the logistic preparation.

Literature cited

1.Lang, E.M. 1963. Geburtshilfe bei einem Indischen Elefanten. Acta Trop. 20, 87-114.
2.Merkt, H., D., Ahlers, H., Bader, H.-P., Brandt, M., Boer and L. Dittrich. 1985a. Bildbericht über den Auszug eines toten Elefantenfetus (Elephas maximus) am 645. Tag p.c. und 65 Stunden nach Geburtsbeginn via Damschnitt (vorläufige Mitteilung). Praktischer Tierarzt 5: 377-378.
3.Merkt, H., D., Ahlers, H., Bader, H.-P., Brandt, M., Boer and L. Dittrich. 1985b. Der Damschnitt, eine geburtshilfliche Interventionsmöglichkeit bei einer Elefantenkuh. Deutsche tieraerztliche Wochenschrift 92: 428-432.
4.Merkt, H., D., Ahlers, H., Bader, D., Rath, H.-P., Brandt, M., Boer and L. Dittrich. 1986. Nachbehandlung und Heilungsverlauf bei einer Elefantenkuh nach Geburtshilfe durch Damschnitt. Berliner Münchner Tierärztliche Wschrift 99: 329-333.
5.Furley, C.W. 1993. A caesarean section in an elephant (The first in europe). Help-Newsletter,15: 2931.
6. Foerner, J.J. 1998. Dystokia in the Elephant. In: Fowler, M. E. & E., Miller (eds.): Zoo & Wild Animal Medicine. 4. Ed. W. B. Saunders Company, 522-525.
7.Lange, A., T.B., Hildebrandt, G., Strauss, O., Czupalla, F., Goeritz and W. Schaftenaar. 1999. Möglichkeiten und Grenzen der Geburtshilfe bei Elefanten. Verhandlungsbericht Erkrankungen der Zootiere 39: 47-58.
8.Fluegger, M., F. Goeritz, R. Hermes, E. Isenbuegel, A. Klarenbeek, W. Schaftenaar, K. Schaller and G. Strauss. 2001. Evaluation of physiological data and veterinary medical experience in 31 Asian elephant (Elephas maximus) births in six European zoos. Verhandlungsbericht Erkrankungen der Zootiere, 40: 123-134.
9.Schaftenaar, W. 1996. Vaginal Vestibulotomy in an Asian Elephant (Elephas maximus). 1996 Proceedings American Association Zoo Veterinarians,  434-439.
10. Schaftenaar, W., T.B., Hildebrandt, M., Fluegger, F., Goeritz, D.J., Schmidt, and G. West. 2001. Guidelines for veterinary assistance during the reproductive process in female elephants. Proceedings American Association Zoo Veterinarians, 348-355.

Note: See source for Table 1: Dystocia cases in captive elephants in the last 2 yr and Table 2: Known dystocia cases with a fatal oucome for the dam

Hildebrandt, T.B., Strike, T., Flach, E., Sambrook, L., Dodds, J., Lindsay, N., Furley, C.F., Glatzel, P.S., McGowan, M., Wisser, J.ed., Hofer, H.e., Frolich, K. Fetotomy in the elephant. Erkrankungen der Zootiere.  315-318. 2003.
Ref Type: Conference Proceeding

Hunt, K.E., Wasser, S.K., 2003. Effect of long-term preservation methods on fecal glucocorticoid concentrations of grizzly bear and african elephant. Physiol Biochem Zool. 76, 918-928.
Abstract: Center for Conservation Biology, Department of Biology, University of Washington, Seattle, WA, 98195-1800, USA.

Leong, K.M., Ortolani, A., Graham, L.H., Savage, A., 2003. The use of low-frequency vocalizations in African elephant (Loxodonta africana) reproductive strategies. Horm Behav. 43, 433-443.
Abstract: Fertility-advertisement calls in females are predicted to occur in nonmonogamous species where males and females are widely separated in space. In African elephants, low-frequency vocalizations have thus been suggested as a reproductive strategy used by fertile females to attract mates. This study examined the use of low-frequency vocalizations with respect to different phases of the estrous cycle in African elephants by simultaneously monitoring vocalizations, behavior, and hormonal profiles. Subjects were one male and six female African elephants housed at Disney's Animal Kingdom. No acoustically distinct vocalizations were restricted to the ovulatory follicular phase. However, overall rate of low-frequency vocalization as well as the rate of one acoustically distinct vocalization changed over the estrous cycle, with highest rates of calling related to the first period of follicular growth, or anovulatory follicular phase. Elevated rates of vocalization thus were not restricted to behavioral estrus and occurred much earlier in the estrous cycle than in most species that produce fertility-advertisement calls. Both herd composition and elephant identity also affected rates of vocalization. Vocalizations therefore may not be reliable signals of actual fertility. However, the increase in vocalizations in advance of estrus may attract males to the herd prior to ovulation, facilitating both male-male competition and female choice. Once present in the herd, males may then switch strategies to use more reliable chemical and visual cues to detect ovulating females. Disney's Animal Kingdom, Lake Buena Vista, FL 32830, USA. Kristen.Leong@disney.com

Malhotra, A.K., Kumar, M., 2003. Management of musth Indian elephant at National Zoological Park, New Delhi. Zoos' Print Journal 18, 10.

Nath, K.C., 2003. Oestrous cycle and pregnancy in elephant. In: Das, D. (Ed.), Healthcare, Breeding and Management of Asian Elephants. Project Elephant. Govt. of India, New Delhi, pp. 42-44.

Pucher, H.E., Stremme, C., Schwarzenberger, F., 2003. Priapism in a semiwild Asian elephant (Elephas maximus) in Vietnam. Vet Rec 153, 717-718.

Rajaram, A., Krishnamurthy, V., 2003. Elephant temporal gland ultrastructure and androgen secretion during musth. Current Science 85, 1467-1471.
Abstract: We have investigated the ultrastructure of the temporal gland of the Asian elephant (Elephas maximus) in the musth condition. We find that the organelles are highly evolved for the production of the androgen, testosterone which is reported to be very high in the Asian male elephant in full musth. The mitochondria bear cristae which are profuse and tubular, and occur along with many Golgi bodies. There is hypertrophy of smooth endoplasmic reticulum. All the structures involved in the production of androgen, as in the Leydig cell or the cells of the adrenal cortex, are thus found in abundance. Cellular structures also seem singularly evolved for the secretion of androgen and its degradation products.

Rasmussen, B., 2003. Why musth elephants use pheromones? Biologist 50, 195-196.

Rasmussen, L.E., Greenwood, D.R., 2003. Frontalin: a chemical message of musth in Asian elephants (Elephas maximus). Chemical Senses 28, 433-446.
Abstract: Musth is an important male phenomenon affecting many aspects of elephant society including reproduction. During musth, the temporal gland secretions (as well as the urine and breath) of adult male Asian elephants (Elephas maximus) discharge a variety of malodorous compounds together with the bicyclic ketal, frontalin. In contrast, teenage male elephants in musth release a sweet-smelling exudate from their facial temporal gland. We recently demonstrated that the concentration of frontalin becomes increasingly evident as male elephants mature. In the present study, we demonstrate that behaviors exhibited towards frontalin are consistent and dependent on the sex, developmental stage and physiological status of the responding conspecific individual. To examine whether frontalin functions as a chemical signal, perhaps even a pheromone, we bioassayed older and younger adult males, and luteal- and follicular-phase and pregnant females for their chemosensory and behavioral responses to frontalin. Adult males were mostly indifferent to frontalin, whereas subadult males were highly reactive, often exhibiting repulsion or avoidance. Female chemosensory responses to frontalin varied with hormonal state. Females in the luteal phase demonstrated low frequencies of responses, whereas pregnant females responded significantly more frequently, with varied types of responses including those to the palatal pits. Females in the follicular phase were the most responsive and often demonstrated mating-related behaviors subsequent to high chemosensory responses to frontalin. Our evidence strongly suggests that frontalin, a well-studied pheromone in insects, also functions as a pheromone in the Asian elephant: it exhibits all of the determinants that define a pheromone and evidently conveys some of the messages underlying the phenomenon of musth. Department of Biochemistry and Molecular Biology, OGI School of Science and Engineering, Oregon Health & Science University, Beaverton, OR 97006-8921, USA. betsr@bmb.ogi.edu

Rees, P.A., 2003. The welfare and conservation of Asian elephants – a reply to Sukumar. Oryx 37, 25.
Abstract: Since my summary of the global fate of Asian elephants in zoos (this issue) was written Clubb & Mason (2002) have published a review of the welfare of zoo elephants in Europe, commissioned by the Royal Society for the Prevention of Cruelty to Animals in the UK. In an attempt to collect data on behaviour, reproduction, group composition, welfare and other aspects of husbandry, they sent questionnaires to the directors of the 18 zoos in the UK that hold elephants. Professor Sukumar doubts my contention that zoo directors lack the commitment necessary to manage the zoo elephant population as viable breeding units. Why then did none of the zoos contacted by Clubb & Mason reply?

Sarma, K.K., 2003. Managing troublesome bulls with special reference to musth in captive Asian elephants. In: Das, D. (Ed.), Healthcare, Breeding and Management of Asian Elephants. Project Elephant. Govt. of India, New Delhi, pp. 58-66.

Schmitt, D.L., 2003. Proboscidea (Elephants). In: Fowler, M.E., Miller, R.E. (Eds.), Zoo and Wild Animal Medicine. Elsevier Science USA, pp. 541-550.

Slade, B.E., Schulte, B.A., Rasmussen, L.E.L., 2003. Oestrous state dynamics in chemical communication by captive female Asian elephants. Animal Behaviour 65, 813-819.
Abstract: In many mammals, reproductive status is revealed through chemical cues in urine. The reproductive status of receivers may influence their interest in such signals. For social mammals that live in matrilineal groups, females may benefit by detecting the reproductive condition of herdmates. Responses to urine during oestrous cycles of senders and receivers are potential indicators of signal functions. We examined the chemosensory responses, first by four captive female Asian elephants, Elephas maximus, over their oestrous cycles to familiar follicular and luteal phase urine and second by 14 different female Asian elephants to unfamiliar conspecific follicular and luteal phase urine. We asked whether females could distinguish the reproductive state of another female as measured by their differential response to luteal-and follicular-phase urine. We further examined the influence of the receiver's reproductive status on response levels. Females responded more with specific tactolfactory trunk behaviours to follicular- than to luteal-phase urine, but only when the receiving female was in her follicular phase. Like their male conspecifics, Asian elephant females can detect changes in the reproductive state of conspecifics. The functional significance of this ability has yet to be determined but may be related more to the resource holding power of females in follicular phase than to a means for females to synchronize oestrous cycles. Such female-female communication may have important effects on social group dynamics.

Sleeman, J.M., Clyde, V.L., Finnegan, M.V., Ramsay, E.C., Shires, M.G., 2003. Mammary botryomycosis and mastectomy in an African elephant (Loxodonta africana). Vet Rec 152, 54-55.

Teng, M.S., Yang, X.L., Wu, D.H., 2003. Characteristics of reproductive biology of Asian elephants. Chinese Journal of Zoology 38, 86-90.
Abstract: The biological characteristics of 1 female Asian elephant that had produced 2 fetuses consecutively were studied through whole-day observation and recording of oestrus, mating, gestation, parturition, postpartum behaviour, and the behaviour of the 2 young elephants (China). Daily frequencies of the young elephant's sucking the breast of the mother reduced day by day. There were differences in the characteristics of different fetuses, including gestation omen, gestation period, defaecation of the young elephant, etc.

West, J.B., Fu, Z., Gaeth, A.P., Short, R.V., 2003. Fetal lung development in the elephant reflects the adaptations required for snorkeling in adult life. Respir Physiol Neurobiol 138, 325-333.
Abstract: The adult elephant is unique among mammals in that the pleural membranes are thickened and the pleural cavity is obliterated by connective tissue. It has been suggested that this peculiar anatomy developed because the animal can snorkel at depth, and this behavior subjects the microvessels in the parietal pleura to a very large transmural pressure. To investigate the development of the parietal pleura, the thickness of the endothoracic fascia (ET) was measured in four fetal African elephants of approximate gestational age 111-130 days, and the appearances were compared with those in human, rabbit, rat and mouse fetuses of approximately the same stage of lung organogenesis. The mean thicknesses of ET in the elephant, human, rabbit, rat and mouse were 403, 53, 29, 27 and 37 microm, respectively. This very early development of a thick parietal pleura in the elephant fetus is consistent with the hypothesis of a long history of snorkeling in the elephant's putative aquatic ancestors. Department of Medicine, University of California San Diego, 9500 Gilman Drive, La Jolla, CA 92093-0623, USA. jwest@ucsd.edu

Whitehouse, A.M., Schoeman, D.S., 2003. Ranging behaviour of elephants within a small, fenced area in Addo Elephant National Park, South Africa. African Zoology 38, 95-108.
Abstract: The elephant population (n = 324, December 2000) in Addo Elephant National Park (AENP), South Africa, is restricted to an area (103 km(2)) considerably smaller than most elephant ranges. The Addo elephants' ranging behaviour was studied in order to determine whether natural patterns of male and female ranging behaviour can be maintained within a confined area. Radio-tracking was used to facilitate measurements of hourly distances travelled, larger scale directional movements, home range sizes, centres of activity and associations. Female home ranges overlap, and interactions between family groups are frequently observed. Females and non-musth males travel similar distances and speeds on an hourly basis, but female core range sizes are larger, resumably because bigger areas are needed to satisfy the nutritional requirements of their young. contrary to expectations, Addo males do not travel further and faster on an hourly basis when they are in musth in comparison to when they are not in musth. However, the movement of males in musth is more directional, so that they cover a greater area of the park. Males associate more frequently with females when in musth than when non-musth. Unlike in other populations, non-musth 'male retirement areas' in AENP are not exclusive, as there is spatial and temporal overlap with female ranges, and hence with the ranges of males in musth. It is suggested that the resulting increased frequency of contact between musth and non-musth males may result in elevated mate competition, and this could contribute to the high levels of aggression observed between Addo bulls.

Wilson, J.D., Leihy, M.W., Shaw, G., Renfree, M.B., 2003. Androgen physiology: unsolved problems at the millennium. Molecular and Cellular Endocrinology 198, 1-5.
Abstract: Androgen physiology differs from that of other steroid hormones in two major regards. First, testosterone, the predominant circulating testicular androgen, is both an active hormone and a prohormone for the formation of a more active androgen, the 5alpha-reduced steroid dihydrotestosterone. Genetic evidence indicates that testosterone and dihydrotestosterone work via a common intracellular receptor, and studies involving in vitro reporter gene assays and intact mice in which both steroid 5alpha-reductase isoenzymes have been disrupted by homologous recombination indicate that dihydrotestosterone acts during embryonic life to amplify hormonal signals that can be mediated by testosterone at higher concentrations. However, in post-embryonic life dihydrotestosterone plays unique roles that have not been elucidated. Studies of other 5alpha-reduced steroids, including the plant hormone brassinolide, the hog pheromones androstanol and androstenol, and 5alpha-dihydroprogesterone (in horses and elephants) indicate that this reaction serves different functions in different systems. Second, during embryonic life androgen causes the formation of the male urogenital tract and hence is responsible for development of the tissues that serve as the major sites of androgen action in postnatal life. It has been generally assumed that androgens virilize the male fetus by the same mechanisms as in the adult, namely by the conversion of circulating testosterone to dihydrotestosterone in target tissues. However, in marsupial mammals there is no sexual dimorphism in the levels of testosterone or dihydrotestosterone at the time the male phenotype forms, and in the pouch young of one marsupial, the tammar wallaby, the testes secrete another 5alpha-reduced steroid, 5alpha-androstane-3alpha, 17beta-diol (5alpha-adiol), into plasma. The administration of 5alpha-adiol to female pouch young causes profound virilization of the urogenital sinus and external genitalia, but within target tissues 5alpha-adiol appears to work after oxidation to dihydrotestosterone. Thus, two separate mechanisms evolved for the formation of dihydrotestosterone in target tissues. 5alpha-adiol is the predominant androgen in neonatal testes in several placental mammals, but it is unclear whether it plays a similar role in other mammalian species.

Wingfield, J.C., Sapolsky, R.M., 2003. Reproduction and resistance to stress: When and how. Journal of Neuroendocrinology 15, 711-724.

Zuba, J.R., Stetter, M.D., Dover, S.R., Briggs, M. Development of rigid laparoscopy techniques in elephants and rhinoceros. Proc Amer Assoc Zoo Vet.  223-227. 2003.
Ref Type: Conference Proceeding
Abstract: Diagnostic and surgical laparoscopy has become a routine procedure in human and veterinary medicine and has similar potential uses in zoological medicine. Surgical telescopes and fiberoptic cables allow the veterinarian to look inside body cavities of the patient and specialized instruments provide the ability to perform a wide variety of diagnostic and surgical procedures.  Rigid laparoscopy is considered minimally invasive surgery and is associated with a more rapid post-operative recovery rate and an improved prognosis as compared to conventional surgical techniques.³  Laparoscopic surgery is commonly being utilized in horses and other zoo animals for a variety of abdominal surgical procedures including tubal ligation and ovariectomy.^7,8,12  Recent advances in technology now provide the ability to perform laparoscopy in the largest mammalian species maintained in zoological collections.¹¹

A variety of disease problems and reproductive disorders have been documented in the rhinoceros and elephant.  Due to their size and anatomy, many of the standard diagnostic tests available in human and veterinary medicine are not routine in these animals (radiography, advanced imaging techniques, liver biopsy, abdominal tap, etc.) at this time.  Although conventional abdominal surgery has been performed in rhinoceros and elephants^1,5,10 survival rates have been extremely low.  Laparoscopy can provide a variety of additional diagnostic options, and may also provide an avenue for performing surgical procedures that would otherwise be impossible in these animals.

A multi-institutional collaboration has been organized to address common goals with regard to enhancing diagnostic capabilities and improving surgical techniques in elephants and rhinoceros.  Specifically, we are aiming to: Develop laparoscopic techniques and equipment for use in rhinoceros and elephants, which can be used to significantly expand our diagnostic and treatment capabilities in these large mammals. Improve international conservation efforts in both rhinoceros and elephants by helping improve animal health and welfare of these species in captive and free ranging situations.

This collaborative effort has been organized to critically review current equipment, techniques and uses with the ultimate goal of overcoming some of the inherent difficulties with laparoscopy in these large vertebrates.  This includes further development and modification of equipment, investigation of surgical techniques, and expansion of clinical applications.

Cooperation in the development of innovative surgical equipment for the anatomic variety of our zoological species is necessary for the advancement of zoological medicine.  Karl Storz Veterinary Endoscopy of America (KSVEA, Goleta, California 93117, USA) was instrumental in the initial development of this specialized equipment at the San Diego Wild Animal Park and by generously providing research and development for our continuing investigations.  The availability of appropriately sized equipment has limited the application of laparoscopic techniques to animals of a size consistent with the intended species of its production.  The largest laparoscopy equipment commercially available is marketed for use in equine medicine (57 cm telescope and accessories).  This system has been used with limited success in rhinoceros¹¹ and elephants.  In many cases, to adequately visualize and manipulate visceral organs, a longer telescope and associated instrumentation is required.  We continue to work closely with KSVEA on the production of specialized equipment (Table 1).  Standard light source, fiberoptic light cable and electronic insufflator units manufactured for use in domestic large animals have been used successfully in megavertebrates.  Due to its compact size and versatility, the authors suggest the use of a portable, battery operated laparoscopy kit (Techno Pack, KSVEA) (monitor, light source, camera and digital recording device) especially under field conditions.

To date we have evaluated the utility of specially designed laparoscopic equipment in approximately seven elephants and four rhinoceros.  Experiences from these limited cases have identified several technical and procedural challenges, which need to be overcome if megavertebrate laparoscopy is to be successfully performed.¹¹  Some of these challenges are summarized below:

Technical challenges:
Equine laparoscopic equipment too fragile and/or short for certain surgical applications in rhinoceros and elephant
Size and disposition of megavertebrates (> 1000 kg)
Thick, non-pliable, pachydermatous skin puts unusual pressure (and risk of damage) on equipment
Insufflation and illumination of large abdominal cavity
Great depth to visualize and physically reach/manipulate large organs
Thick, redundant, fibro-elastic peritoneum which is difficult to puncture during surgical attempts to enter the abdominal cavity
Influence of patient positioning:  unable to use conventional laparoscopic positioning techniques in these species
Both rhinoceros and elephants are hindgut fermentors and have very large and extensive lower intestinal tracts.  Gas dilation of these bowel loops combined with limited positioning options, can make laparoscopic visualization of certain organs a problem

Procedural challenges:
Cost of developing new and specialized laparoscopic equipment is high; duplicate equipment is not available at this time
Charismatic nature of megavertebrates makes it difficult to perform surgery on such important and high exposure species 
Limited clinical cases for testing equipment due to the relatively low number of megavertebrates in zoological facilities
Use of laparoscopy in zoo mammals may be out of the comfort zone for many veterinarians and curators due to lack of experience with this instrumentation
Lack of published surgical procedures in these species
Inability to perform laparoscopic procedure in "surgical suite" as with most other species
Risks associated with megavertebrate anesthesia and sedation including difficulty in providing safe anesthetic procedures, proper restraint, and safety of personnel

A multidisciplinary, systematic approach has been initiated to critically review current instrumentation and procedures with the objective of overcoming these technical difficulties. 

The initial phase of our project has been to develop laparoscopic techniques and equipment (Table 1) for use in rhinoceros and elephants by utilizing individuals that may have died of natural causes, or live animals with medical conditions that warrant abdominal surgery.  We have already had the opportunity to perform laparoscopic surgery on both live and deceased white rhinoceros, black rhinoceros, African and Asian elephants.  From our initial studies, it is apparent that laparoscopy in these megavertebrates is possible and may greatly enhance our ability to care for and manage these animals in captive and free ranging situations.  The authors have received some funding and institutional support that allows us to travel to institutions which may benefit from the use of this equipment and/or our experience.  We are also interested in those zoological institutions which may find themselves dealing with a terminal case in an elephant or rhinoceros, to please contact us directly and to consider allowing laparoscopy to be conducted on the animal prior to a post-mortem examination.

Ultimately, we envision the results of our studies on captive animals to be applicable to the management and conservation of elephants and rhinoceros in the wild.  A variety of medical disorders are commonly reported in black and white rhinoceros.  Many of these medical conditions are difficult to diagnose, monitor and treat.  Furthermore, there is a paucity of information on the incidence of these disease conditions in free ranging populations.  The use of minimally invasive laparoscopic techniques will greatly enhance our diagnostic abilities in this species and would be extremely valuable to the understanding of medical conditions of captive and free ranging rhinoceros and to conservation efforts overall.

Throughout many parts of Africa, wildlife professionals are seriously concerned about the negative effects large elephant herds are having on the native flora and fauna within parks and reserves.^2,6,9  The historical rangelands of the elephant have become interrupted by national borders and artificial barriers.  Habitats surrounding wildlife parks are increasingly being converted to agricultural lands. The encroachment of human populations has caused a dramatic increase in the number and severity of human-elephant conflicts.^2,9  These conflicts are commonplace in many parts of East and Southern Africa.  Although there have been a variety of plans to reduce human-elephant and elephant environment impacts, little overall success has been achieved in most countries.⁹

Historically elephant population control has primarily been limited to culling and translocation of small groups.^2,6  Immunocontraception has been attempted with a small population of elephants but is not currently realistic in many situations.^4,6  In wildlife parks where large herds of elephants exist there is currently no effective, humane method of population control.

One of our long-range goals is to develop laparoscopic techniques, such as ovariectomy and tubal ligation, in free ranging African elephants that can be used to sterilize reproductive females.  Once these techniques have been developed, it is our intention to train local wildlife veterinarians and health professionals to perform laparoscopic sterilization of elephants in the field and thus provide local wildlife officials with a tool to help manage elephant populations.  It is our hope this will improve conservation efforts across Africa by reducing human-elephant conflicts and helping to save critical ecosystems.

Acknowledgements
This ongoing project is possible due to the generous contributions of the following individuals:  Hans Lunneman, Christopher Chambliss, Michele McCutcheon, Lynn Richardson, Dean Hendrickson, Robin Radcliffe, Rolf Radcliffe, Laurie Gage, Larry Galuppo, Bill Lindsay, John Olsen, Genny Dumonceaux, and the veterinary staffs at the San Diego Wild Animal Park and Disney's Animal Programs.

LITERATURE CITED
1.Byron, H., J. Olsen, M. Schmidt, J. Copeland, and L. Byron. 1985. Abdominal surgery in three adult male Asian elephants. J Am Vet Med Assoc. 187:1236-1237.
2.Chalfota J. and Owen-Smith N. 1996. Options for the management of elephants in northern Botswana. Pachyderm. 22:67-73.
3.Cook, R.A., and D.R. Stoloff. 1999. The application of minimally invasive surgery for the diagnosis and treatment of captive wildlife. In: Fowler, M., Miller, E. (Eds): Zoo and Wild Animal Medicine: Current Therapy 4. W.B. Saunders, Philadelphia, PA. Pp 30-40.
4.Delsink, A.K., van Altena, J.J., Kirkpatrick, J. Grobler, D., and Fayrer-Hosken, R.A. 2002. Field applications of immunocontraception in African elephants (Loxodonta africana).  J Soc.Reprod. Fert. 60: 117-124.
5.Fowler, M.E., and R. Hart. 1973. Castration of an Asian elephant using etorphine anesthesia. J Am Vet Med Assoc. 163: 539-543.
6.Garai, M.E.  2001.  Managing elephants on private reserves in South Africa.  Proceedings of the International Elephant and Rhino Research Symposium. Vienna, Austria. 259-261.
7.Hendrickson, D.A. 2002. New techniques for performing equine laparoscopic ovariectomy. DVM Best Practices Magazine. Oct. 2002.
8.Hendrickson, D.A., and D.G. Wilson. 1996. Instrumentation and techniques for laparoscopic and thoracoscopic surgery in the horse. Vet. Clin. N.A. Equine Pract.12; 2: 235.
9.Hoare, R., Update on the study and management of human-elephant conflict in Africa. Pachyderm. 33: 91-92.
10.Olsen, J., and H. Byron. 1993. Castration of the elephant. In: Fowler, M. (Ed): Zoo and Wild Animal Medicine: Current Therapy 3, 3^rd ed. W.B. Saunders, Philadelphia, PA. Pp. 441-444.
11.Radcliffe R.M., D.A. Hendrickson, G.L. Richardson., J.R. Zuba, and R.W. Radcliffe. 2000. Standing laparoscopicguided uterine biopsy in a southern white rhinoceros (Ceratotherium simum simum). J. Zoo Wildl. Med. 31:201207.
12.Rogerson, D., M. Brown, B. Watt, C. Keoughan, and M. Hanrath. 2002. Hand-assisted laparoscopic technique for removal of ovarian tumors in standing mares. J. Am. Vet. Med. Assoc. 220(10):1503-1507.

See source for Table 1.  Current laparoscopy equipment specifically manufactured for use in megavertebrates.^a
a All equipment was specially manufactured by Karl Storz Veterinary Endoscopy of America (KSVEA, Goleta, California, USA) and is not commercially available at this time.

Journal of Indian Veterinary Assocaition Kerala. Journal of Indian Veterinary Association Kerala 7[3], 1-64. 2002.
Ref Type: Journal (Full)

Alex, P.C., 2002. The Musth, the vicious and the rogue elephants - a review. Journal of Indian Veterinary Association Kerala 7, 26-27.

Allen, W.R., Mathias, S.S., Wooding, F.B., Skidmore, J.A., van Aarde, R.J., 2002. Placentation in the African elephant, Loxodonta africana. I. Endocrinological aspects. Reprod Suppl 60, 105-116.
Abstract: Placental and fetal tissues were recovered from the uteri of 59 pregnant elephant that ranged in estimated age from day 18 to month 21 of gestation. Incubation of placenta and fetal gonad, alone or in combination, with tritium-labelled cholesterol, pregnenolone and androstenedione failed to yield any labelled progestagens or oestrogens from placenta, but did produce small amounts of labelled progesterone and 5alpha-dihydroprogesterone from fetal gonad. Immunochemical staining of tissues with four antisera specific for enzymes involved in the steroidogenic pathway revealed no staining in sections of placenta but positive labelling for P450 side chain cleavage enzyme (SCC450) and 3beta-hydroxysteroid dehydrogenase (3beta-HSD) of the interstitial cells that comprise the bulk of the enlarged fetal gonads during the second half of gestation. Saline extracts of placental tissue showed no activity in three different gonadotrophin assays. In view of this endocrinological inactivity in the zonary elephant placenta and the probable reliance on maternal luteal sources of progestagens for maintenance of the pregnant state, the argument is advanced that uncomplicated abortion would probably follow a single administration of a PGF analogue given at any stage of pregnancy. If so, the treatment might constitute an efficacious method for controlling population increases in elephants maintained in enclosed game parks in Africa.

Chandrasekharan, K., 2002. Elephant - an overview. Journal of Indian Veterinary Association Kerala 7, 8-11.

Cheeran, J.V., Radhakrishnan, K., Chandrasekharan, K., 2002. Musth. Journal of Indian Veterinary Association Kerala 7, 28-30.

Cheeran, J.V., Chandrasekharan, K., Radhakrishnan, K., 2002. Tranquilization and translocation of elephants. Journal of Indian Veterinary Association Kerala 7, 42-46.

Cheeran, J.V., 2002. Elephant facts. Journal of Indian Veterinary Association Kerala 7, 12-14.

Davis, S.A., Pech, R.P., 2002. Dependence of population response to fertility control on the survival of sterile animals and their role in regulation. Reprod Suppl 60, 89-103.
Abstract: The species for which fertility control is presently used, or for which it is being developed, range from small mammal pests, such as the house mouse (Mus domesticus), to large mammals, such as the African elephant (Loxodonta africana). However, the possibility of a population response other than a reduction in abundance proportional to the fraction of animals rendered infertile has been shown in field trials. For example, when intermediate levels of sterility were imposed on wild populations of European rabbits (Oryctolagus cuniculus), there was an increase in their abundance, on an annual basis, due to enhanced survival of juveniles and adult females. In this article, we relate intraspecific regulatory processes to the response of populations to fertility control using a set of density-dependent structured-population models. In each of the models, the population is exposed periodically to a fertility control agent that renders a fraction of fertile females sterile.  Although our intention is not to predict the population response of any one particular species, the results of the models are illustrated using parameter values that are representative of populations of the European fox (Vulpes vulpes) in south-eastern Australia. When populations were regulated by density-dependent mechanisms in which sterile females did not participate, such as competition for resources among young animals or competition among fertile females for breeding sites or territories, then populations could increase in abundance for low and intermediate levels of imposed sterility. For other intraspecific regulatory mechanisms, such as competition for resources between all individuals, all levels of sterility were observed to reduce abundance. The population response was sensitive to (i) whether the survival of sterile adults was higher than that of fertile adults, (ii) whether animals could be sterilized before sexual maturity, and (iii) whether density dependence was modelled as a threshold process.

Delsink, A.K., van Altena, J.J., Kirkpatrick, J., Grobler, D., Fayrer-Hosken, R.A., 2002. Field applications of immunocontraception in African elephants (Loxodonta africana). Reprod Suppl 60, 117-124.
Abstract: The primary aim of the Makalali elephant immunocontraception programme is to test the efficacy of porcine zona pellucida (PZP) vaccine for practical population control of elephants in small, enclosed reserves, with the goal of stabilizing the current growth rate and reducing it to the 5-10% per annum displayed currently in the Kruger National Park. A secondary aim is to test the hypothesis that PZP treatment does not affect patterns of elephant social behaviour. Eighteen sexually mature cows (age > 12 years) were vaccinated in May 2000 using remote darts. Behavioural observations before, during and after vaccination included noting the activity of individual animals every minute for 15 min. No changes in general behaviour patterns have been noted to date although the animals' spatial use of the reserve was erratic during the period of vaccination, indicating irregular or disturbed patterns associated with vaccination. Normality was resumed on completion of the vaccinations. No aggressive or indifferent behaviour related to nursing, calf proximity or bull-cow interactions have been noted. Ten of the females were in various stages of pregnancy when treated. Subsequently, seven of them gave birth to healthy calves and the other three females are expected to calve shortly. It is too early in the study to draw conclusions about stabilization of growth rates.

Duer, C., Carden, M., Schmitt, D., Tomasi, T., 2002. Utility of maternal serum total testosterone analysis for fetal gender determination in Asian elephants (Elephas maximus). Anim Reprod Sci 69, 47-52.
Abstract: It has been shown in some species that fetal testes produce testosterone early in gestation. This study investigated the possibility that fetal testosterone may be reflected in maternal serum levels in the Asian elephant (Elephas maximus). Weekly serum samples were collected from seventeen pregnant captive Asian elephants and analyzed via radioimmunoassay (RIA) for total testosterone levels. Nine of the cows carried male fetuses and eight carried female fetuses. A non-random pattern over time (P<0.01) was observed in cows carrying either a male or female fetus. Mean maternal serum total testosterone was significantly higher in cows carrying male versus female fetuses (P<0.01). Mean trimester values indicate that first trimester values are not significantly different among male versus female groups. The second and third trimester values of cows carrying male fetuses were higher than cows carrying female fetuses, (P<0.01 and <0.05, respectively). The results of this study show that it is possible via RIA of maternal serum for total testosterone to determine the gender of calves during gestation.

Forsyth, I.A., Wallis, M., 2002. Growth hormone and prolactin--molecular and functional evolution. J Mammary Gland Biol Neoplasia 7, 291-312.
Abstract: Growth hormone, prolactin, the fish hormone, somatolactin, and related mammalian placental hormones, including placental lactogen, form a family of polypeptide hormones that share a common tertiary structure. They produce their biological effects by interacting with and dimerizing specific single transmembrane-domain receptors. The receptors belong to a superfamily of cytokine receptors with no intrinsic tyrosine kinase, which use the Jak-Stat cascade as a major signalling pathway. Hormones and receptors are thought to have arisen as a result of gene duplication and subsequent divergence early in vertebrate evolution. Mammalian growth hormone and prolactin show a slow basal evolutionary rate of change, but with episodes of accelerated evolution. These occurred for growth hormone during the evolution of the primates and artiodactyls and for prolactin in lineages leading to rodents, elephants, ruminants, and man. Placental lactogen has probably evolved independently on three occasions, from prolactin in rodents and ruminants and from growth hormone in man. Receptor sequences also show variable rates of evolution, corresponding partly, but not completely, with changes in the ligand. A principal biological role of growth hormone, the control of postnatal growth, has remained quite consistent throughout vertebrate evolution and is largely mediated by insulin-like growth factors. Prolactin has many and diverse roles. In relation to lactation, the relative roles of growth hormone and prolactin vary between species. Correlation between the molecular and functional evolution of these hormones is very incomplete, and it is likely that many important functional adaptations involved changes in regulatory elements, for example, altering tissue of origin or posttranscriptional processing, rather than change of the structures of the proteins themselves. The Babraham Institute, Babraham, Cambridge, United Kingdom. isabel.forsyth@bbsrc.ac.uk

Graham, L.H., Bolling, J., Miller, G., Pratt-Hawkes, N., Joseph, S., 2002. Enzyme-immunoassay for the measurement of luteinizing hormone in the serum of African elephants (Loxodonta africana). Zoo Biology 21, 403-408.
Abstract: Circulating patterns of progesterone and luteinizing hormone (LH) in the elephant have been well characterized, and routine monitoring of these hormones is now viewed as a valuable tool for making informed decisions about the reproductive management of elephants in captivity. Currently LH monitoring in elephants is done with radio-immunoassays (RIAs); unfortunately, the use of radioactive materials in RIAs limits their application to institutions with laboratory facilities equipped for the storage and disposal of radioactive waste. Enzyme-immunoassays (EIAs)offer an inexpensive and more zoo-friendly alternative to RIA. This work reports on an EIA capable of quantifying circulating LH in African elephants. The EIA employs a biotin label and microtiter plates coated with goat anti-mouse gamma globulin. LH surges in African elephants (n=3) increased fivefold over baseline concentrations (1.00±0.1 ng/ml vs. 0.2±0.1 ng/ml) and occurred 19.3±0.2 days apart. Ovulatory LH surges were associated with an increase in serum progestogens from 4.8±0.4 ng/ml to 11.7±0.4 ng/ml. The ability to quantify reproductive hormones in elephants via EIA is an important step in the process of making endocrine monitoring more accessible to zoos housing these species.

Grandy, J.W., Rutberg, A.T., 2002. An animal welfare view of wildlife contraception. Reprod Suppl 60, 1-7.
Abstract: Although there is some dissent, the animal protection community generally supports the concept of wildlife contraception. However, some contraceptive agents, delivery mechanisms and specific applications will be opposed by animal welfare advocates on environmental, humane or other ethical grounds, and some animal rights advocates may oppose wildlife contraception entirely. The Humane Society of the United States (HSUS) has supported and conducted wildlife contraception studies for more than 10 years. In general, we have invested in contraceptive agents (such as porcine zona pellucida) that we believe will prove environmentally, physiologically and behaviourally benign, and in delivery mechanisms that are narrowly targeted. As we consider contraception to be a major intervention into natural processes, we believe that wildlife contraception should be applied judiciously, locally and in a manner that is sensitive to the needs of animals, humans and ecosystem function.

Khawnual, P., Clarke, B., 2002. General care and reproductive management of pregnant and infant elephants at the Ayutthaya Elephant Camp. In: Baker, I., Kashio, M. (Eds.), Giants on Our Hands: Proceedings of the International Workshop on the Domesticated Asian Elephant, Bangkok, Thailand, 5-10 February 2001. FAO Regional Office for Asia and the Pacific (RAPA), Bangkok; Thailand, pp. 249-256.
Abstract: The management practices used at the Ayutthaya Elephant Camp (Thailand), where animals are kept for ecotourism purposes, are described. The aspects covered include feeding, provision of water, waste management and veterinary care (including health monitoring and disease control). Detailed descriptions are given of the procedures for management of pregnant females. The topics covered include mating, pregnancy diagnosis, parturition and postpartum management of the female and her offspring. These practices had resulted in 4 successful births in 2000.

Kumar, G.A., Ghosh, K.N.A., Sreekumaran, T., Chandrasekharan, K., 2002. Reproduction in elephants. Journal of Indian Veterinary Association Kerala 7, 38-40, 48-48.

Rasmussen, L.E., Riddle, H.S., Krishnamurthy, V., 2002. Mellifluous matures to malodorous in musth. Nature 415, 975-976.
Abstract: Male Asian elephants in musth--an annual period of heightened sexual activity and intensified aggression--broadcast odoriferous, behaviourally influential messages from secretions of the temporal gland. From our observations in the wild, together with instantaneous chemical sampling and captive-elephant playback experiments, we have discovered that young, socially immature males in musth signal their naivety by releasing honey-like odors to avoid conflict with adult males, whereas older musth males broadcast malodorous combinations to deter young males, facilitating the smooth functioning of male society. As elephant--human conflicts can upset this equilibrium, chemically modulating male behaviour may be one way to help the conservation of wild elephants.

Rasmussen, L.E., Wittemyer, G., 2002. Chemosignalling of musth by individual wild African elephants (Loxodonta africana): implications for conservation and management. Proc R Soc Lond B Biol Sci 269, 853-860.
Abstract: Elephants have extraordinary olfactory receptive equipment, yet this sensory system has been only minimally investigated in wild elephants. We present an in-depth study of urinary chemical signals emitted by individual, behaviourally characterized, wild male African elephants, investigating whether these compounds were the same, accentuated, or diminished in comparison with captive individuals. Remarkably, most emitted chemicals were similar in captive and wild elephants with an exception traced to drought-induced dietary cyanates among wild males. We observed developmental changes predominated by the transition from acids and esters emitted by young males to alcohols and ketones released by older males. We determined that the ketones (2-butanone, acetone and 2-pentanone, and 2-nonanone) were considerably elevated during early musth, musth and late musth, respectively, suggesting that males communicate their condition via these compounds. The similarity to compounds released during musth by Asian male elephants that evoke conspecific bioresponses suggests the existence of species-free 'musth' signals. Our innovative techniques, which allow the recognition of precise sexual and musth states of individual elephants, can be helpful to managers of both wild and captive elephants. Such sampling may allow the more accurate categorization of the social and reproductive status of individual male elephants.

Rasmussen, L.E.L., Riddle, H.S., Krishnamurthy, V., 2002. Mellifluous matures to malodorous in musth; Mood-altering secretions by excited male elephants smooth out social interactions. Nature 415, 975-976.

Sanchez, C.R., Murray, S.Z., Montali, R.J., Spelman, L.H. Medical Management of Acute Pylelonephritis in an Asian Elephant. Baer, C. K. American Association of Zoo Veterinarians Annual Conference.  162-164. 2002. 2002.
Ref Type: Conference Proceeding

Stokke, S., duToit, J.G., 2002. Sexual segregation in habitat use by elephants in Chobe National Park, Botswana. African Journal of Ecology 40, 360-371.
Abstract: We report on a study conducted on free-ranging African elephants in the woodlands of northern Botswana.We compared bull groups and family units with regard to (1) their patterns of habitat use and (2) their ranging distances from perennial water sources. During the dry season, adult males frequented more habitat types than family units, whereas family units used a wider diversity of habitats than bulls during the wet season. Bulls roamed widely (>10 km) from perennial drinking water in the dry season, when family units congregated within 3.5 km of the rivers. During the wet season,when ephemeral pans were abundant, all elephant groups were found at intermediate distances (5 km) from the rivers.The spacing of elephants in the dry season is consistent with sexual segregation but we reject the hypothesis that this is an outcome of indirect competition for food, because our concurrent studies on elephant feeding ecology found no evidence for intraspecific competition. Instead, we propose that most adult male elephants space themselves to avoid conflict with musth bulls and roam widely in the dry season between discretely distributed feeding 'hotspots'.The small proportion of males that are in musth remain close to family units to maximize mating opportunities, and family units are unable to range far from water in the dry season. This is due to (1) comparatively high rates of water turn-over among juveniles and lactating cows and (2) the reduced mobility of neonates.

Suedmeyer, W.K. Transabdominal ultrasonic Gestational monitoring in an African elephant (Loxodonta africana). Baer, C. K. American Association of Zoo Veterinarians Annual Conference.  219-220. 2002. 2002.
Ref Type: Conference Proceeding

Welsch, U., Unterberger, P., Hofter, E., Cuttitta, F., Martinez, A., 2002. Adrenomedullin in mammalian and human skin glands including the mammary gland. Acta Histochem 104, 65-72.
Abstract: Adrenomedullin is a peptide that has been ascribed numerous functions. In the present paper, adrenomedullin has been localized immunhistochemically in a variety of skin glands of humans, elephants and impalas: apocrine scent glands, eccrine sweat glands, holocrine glands and mammary glands. In the apocrine glands expression of adrenomedullin varied with respect to staining intensity and intracellular localization. In general, glands which appeared to be actively secreting were more strongly stained than quiescent glands. However, within a single glandular tubule, individual cells differed considerably in the staining intensity of adrenomedullin. Adrenomedullin was present in both non-lactating and lactating mammary secretory epithelia, both ducts and alveoli reacted positively. In human mammary glands displaying apocrine metaplasia, the apical protrusions were strongly positive. Furthermore, positive immunostaining was found in endothelium and often in smooth muscle cells of small arteries and veins and in mast cells as well. Many of the adrenomedullin-positive epithelial cells were most strongly stained in the area of the Golgi apparatus, the cellular apex and particularly close to the basal side of the cell membrane.This pattern suggests packaging of adrenomedullin into secretory granules and secretion both at the apex of cells and at their basis. The first form of secretion suggests exocrine secretion, the latter form endocrine secretion of adrenomedullin. A possible hormonal function is in line with basally located electron dense small secretory granules, which have been found by electron microscopy in the glandular epithelia studied.

Barber, M.R., Lee, S.M., Steffens, W.L., Ard, M., Fayrer-Hosken, R.A., 2001. Immunolocalization of zona pellucida antigens in the ovarian follicle of dogs, cats, horses and elephants. Theriogenology 55, 1705-1717.
Abstract: A comparative evaluation of the location of immunoreactive porcine zona pellucida (pZP) glycoproteins was performed with polyclonal rabbit anti-pZP antibodies on ovarian sections of the dog, cat, horse and elephant. For this, formalin (light microscopy) and glutaraldehyde (transmission electron microscopy [TEM]) fixed ovarian sections were incubated with antibodies raised against highly purified pZP. Staining patterns were determined with diaminobenzidine (DAB) at the light level. The dog ZP had a distinct staining distribution that is characterized by intense staining around the periphery of the ZP and the oolemma and less dense staining throughout the width of the ZP. In dog follicles that contained multiple oocytes, there were oocytes of identical and dissimilar stages. Cat ovarian sections showed uniform staining of the ZP. Horse results showed uniform staining of ZP and ooplasm, and granulosa cells (GC). Elephant sections showed staining of the ZP with dense staining at the oolemma, as well as staining of the ooplasm. In all species the staining of the ZP was not evident until GC differentiation. In all cases there was no staining of ovarian tissue with control normal rabbit serum. Specific staining patterns of ZP were evaluated by TEM and immunogold staining. The immunogold-linked anti-pZP antibodies stained the ZP matrix in all species. There was staining of ooplasm organelles suggesting that ZP secretion originates from the oocyte of the dog and cat. In addition, follicular and ZP measurements were taken that allowed accurate characterization of follicle stage. These findings suggest that in all 4 species the ZP is recognized by anti-pZP antibodies and there is also evidence to suggest the possible origins of ZP glycoproteins.

Campbell, M.M., 2001. Pachyderms, primates, plants and population. Reprod Fertil Dev 13, 697-703.
Abstract: In the past, growth in human population has often been associated with species loss. Current rates of population growth, both globally (1 million more births than deaths every 103 hours) and regionally, pose a threat of additional ecological damage. There is a well-documented unmet demand for family planning in nearly all high-fertility countries. Improved family planning and safe abortion services will improve the health of women and their families, accelerate fertility decline, and help preserve the environment. Many ecologically vulnerable areas are especially poorly served by family planning services. Examples are given here of improving family planning services through private health providers near the Kakamega Forest in western Kenya, and of adding family planning choices to a reforestation project run by the Jane Goodall Institute near the Gombe National Park, Tanzania. Wildlife biologists can play a critical role in identifying local professionals and institutions with the potential to improve family planning.

Dastig, B. Birth and Reproduction Rate in a Herd of Captive Asian Elephants at the Pinnawela Elephant Orphanage. A Research Update on Elephants and Rhinos; Proceedings of the International Elephant and Rhino Research Symposium, Vienna, June 7-11, 2001.  19-23. 2001. Vienna, Austria, Schuling Verlag. 2001.
Ref Type: Conference Proceeding
Abstract: The majority of Asian elephants currently or previously held in zoos and circuses in Europe and North America came into captivity as young animals between two and five years of age. This was possible due to the low transportation costs. These young animals, either captured or born to working elephants, were merely a by-product and thus sold at a low price. Today, Asian elephants destined for zoos come from working elephants or from jungle camps. These animals are all orphans and have never lived in a natural social structure. Circuses and zoos usually keep only female elephants, which are then deprived of a social structure including an alpha cow, aunts, sisters and their young. This results in the animals' rarely giving birth in captivity. This is why gathering data and conducting observation in this field is particularly difficult. It is for this reason that the Pinnawela Elephant Orphanage offers an ideal location for learning more about reproduction and social structure in Asian elephants. The orphanage is located in Sri Lanka halfway between the capitol, Colombo, and Kandy, the ancient royal city. The orphanage represents the largest herd of Asian elephants in human care. In 1997 the herd consisted of 56 elephants; in 1998 the population had grown to include 63 elephants.

Dehnhard, M., Hildebrand, T., Rohleder, M., Strauss, G., Meyer, H.H.D., Goritz, F., 2001. Application of an enzyme-immunoassay (EIA) for rapid screening of 5alpha-pregnane-3,20-dione (DHP) in blood plasma of the Asian elephant, Elephas maximus. Berliner und Munchener Tierarztliche Wochenschrift. 114, 161-165.
Abstract: Populations of African (Loxodonta africana) and Asian elephants (Elephas maximus) in zoos and safari parks are at risk due to their low reproductive success. To extend the limited knowledge of their reproductive physiology, easy and practical methods for the analysis of relevant reproductive hormones must be developed to support assisted reproduction, for instance. Blood samples from 2 nonpregnant and 2 pregnant Asian elephants were used in the following study. For the measurement of 5alpha-pregnane-3,20-dione (DHP), the predominant ovarian gestagen in both species, an enzyme-immunoassay (EIA) based on commercial reagents was applied. Advantages of this EIA are the small volume of plasma needed for evaluation (5 µl) and the possibility of direct processing without an extraction stage. The lower limit of detection was 0.16 ng/ml, mean recovery was 101% and the mean coefficients of variation were 7.3 (intra-assay) and 9.9% (inter-assay). In the Asian elephants, DHP levels reached 15 ng/ml during the luteal phase and 21 ng/ml during pregnancy. Oestrous cycle lengths based on the lowest DHP concentrations varied from 12 to 20 weeks (mean of 15.4±2.3). In two Asian elephant cows, a calf was stillborn. Thereafter, ovarian activity in the animals resumed after approximately 8 and 13 weeks, respectively. In one animal, estradiol implants for hormonal contraception caused a down regulation of ovarian function as demonstrated by an irregular pattern of DHP secretion over a period of 48 weeks. It is proposed that the direct DHP-EIA is a suitable method for reproductive monitoring in elephants, as it can be easily established in laboratories.

Dehnhard, M., Hesitermann, M., Goritz, F., Hermes, R., Hildebrand, T., Haber, H., 2001. Demonstration of 2-unsaturated C19-steroids in the urine of female Asian elephants, Elephas maximus, and their dependence on ovarian activity. Reproduction-Cambridge 121, 475-484.
Abstract: An oestrous-related pheromone of the female Asian elephant (Elephas maximus) is known to induce behavioural responses in elephant bulls. Additional data revealed that timing of oestrus in females with close social relationships tends to be synchronized. Therefore, urine from female Asian elephants might be expected to contain luteal phase-dependent volatile substances, which may function as additional chemical signals in this species. The aim of the present study was to identify such compounds and to investigate their pattern of excretion throughout the ovarian cycle. Urine samples were collected 3 times a week during the follicular phase and 1 to 3 times a week during the luteal phase from 5 adult female Asian elephants from a total of 13 non-conception cycles and one conception cycle, including the first 72 weeks of pregnancy. A simple headspace solid-phase microextraction method has been developed for quantification of urinary volatile substances and analysis was performed by gas chromatography. The comparison of urine collected during the follicular and the luteal phase indicated the presence of two luteal phase-dependent substances. Mass spectrometry was used to identify one substance as 5alpha-androst-2-en-17-one and a second substance as the corresponding alcoholic compound 5alpha-androst-2-en-17beta-ol. The 5alpha-androst-2-en-17beta-ol and -17-one profiles reflected cyclic ovarian activity with clear (10-20-fold) luteal phase increases. Furthermore, measurements of both compounds were correlated positively with the concentration of urinary pregnanetriol and indicated cycle duration (15.1±1.2 weeks) similar to that obtained from pregnanetriol measurements (15.2±1.6 weeks). The results demonstrated the presence of 2 luteal phase-specific steroidal volatile compounds in elephant urine. One of the substances, 5alpha-androst-2-en-17-one, has been demonstrated in human axillary bacterial isolates. The measurement of both volatile substances in elephant urine can be used for rapid detection of the stage of the ovarian cycle, as the analysis can be completed within 2 h.

Delsink, A.K., van Altena, J.J., Kirkpatrick, J., Grobler, D., Fayrer-Hosken, R. Field applications of immunocontraception in African elephants (Loxodonta africana). Proceedings of the Fifth International Symposium on Fertility Control in Wildlife.  2001.  Society for Reproduction and Fertility; Cambridge; UK. 2001.
Ref Type: Conference Proceeding
Abstract: The primary aim of the Makalali elephant immunocontraception programme is to test the efficacy of porcine zona pellucida (PZP) vaccine for practical population control of elephants in small, enclosed reserves, with the goal of stabilizing the current growth rate and reducing it to the 5-10% per annum displayed currently in the Kruger National Park. A secondary aim is to test the hypothesis that PZP treatment does not affect patterns of elephant social behaviour. Eighteen sexually mature cows (age > 12 years) were vaccinated in May 2000 using remote darts. Behavioural observations before, during and after vaccination included noting the activity of individual animals every minute for 15 min. No changes in general behaviour patterns have been noted to date although the animals' spatial use of the reserve was erratic during the period of vaccination, indicating irregular or disturbed patterns associated with vaccination. Normality was resumed on completion of the vaccinations. No aggressive or indifferent behaviour related to nursing, calf proximity or bull-cow interactions have been noted. Ten of the females were in various stages of pregnancy when treated. Subsequently, seven of them gave birth to healthy calves and the other three females are expected to calve shortly. It is too early in the study to draw conclusions about stabilization of growth rates.

Doi, O., Yamada, T., Terazono, M., Wada, S., 2001. Macroscopic changes in the mucous membrane of vaginal vestibule during the estrous cycle of female Asian elephant. Japanese Journal of Zoo and Wildlife Medicine 6, 55-59.

Flugger, M., Goritz, F., Hermes, E., Isenbugel, A., Klarenbeek, W., Schaftenaar, W., Schaller, K., Strauss, G. Evaluation of physiological data and veterinary medical experiences in 31 Asian elephant (Elephas maximus) births in six European zoos. Verh ber Erkrg Zootiere (Proc. 40th Intl Symp  Zoo and Wild Anim  Med).  123-133. 2001. Rotterdam, Netherlands. 2001.
Ref Type: Conference Proceeding

Foley, C.A.H., Papageorge, S., Wasser, S.K., 2001. Noninvasive stress and reproductive measures of social and ecological pressures in free-ranging African elephants. Conserv Biol 15, 1134-1142.

Fritsch, G., Hermes, R., Maltzan, J. New Aspects of Sexual Maturation in Male Elephants. A Research Update on Elephants and Rhinos; Proceedings of the International Elephant and Rhino Research Symposium, Vienna, June 7-11, 2001.  25. 2001. Vienna, Austria, Schuling Verlag. 2001.
Ref Type: Conference Proceeding

Ganslober, U. Behavioural Ecology, Social Relationships, Life History and Evolutionary Constraints in Megaherbivores. A Research Update on Elephants and Rhinos; Proceedings of the International Elephant and Rhino Research Symposium, Vienna, June 7-11, 2001.  26-31. 2001. Vienna, Austria, Schuling Verlag. 2001.
Ref Type: Conference Proceeding

Ganswindt, A., Heistermann, M., Borragan, S., Hodges, J.K., 2001. Assessment of testicular endocrine function in captive African elephants by measurement of urinary and fecal androgens. Zoo Biology 21,  27-36.

Ganswindt, A., Heistermann, M., Hodges, J.K. Faecal Glucocorticoid and Androgen Metabolite Excretion in Male African Elephants (Loxodonta africana). A Research Update on Elephants and Rhinos; Proceedings of the International Elephant and Rhino Research Symposium, Vienna, June 7-11, 2001.  258. 2001. Vienna, Austria, Schuling Verlag. 2001.
Ref Type: Conference Proceeding

Goeritz, F., Hildebrandt, T.B., Hermes, R., Quandt, S., Jewgenow, K., Hofmann, R.R., Hofer, H., Meyer, H.H.D. Results of Hormonal Contraception in Free-Ranging African Elephants. A Research Update on Elephants and Rhinos; Proceedings of the International Elephant and Rhino Research Symposium, Vienna, June 7-11, 2001.  262. 2001. Vienna, Austria, Schuling Verlag. 2001.
Ref Type: Conference Proceeding

Graham, L., Schwarzenberger, F., Möstl, E., Galama, W., Savage, A., 2001. A versatile ezyme immunoassay for the determination of progestogens in feces and serum. Zoo Biology 20, 227-236.
Abstract: The ability of zoos to monitor the reproductive status of their animals can vastly improve the effectiveness of husbandry/management practices, and noninvasive methods such as fecal steroid analysis are the easiest to apply in a zoo setting. Furthermore, enzyme immunoassay (EIA) is preferred to radioimmunoassay (RIA) as the method of quantifying hormones because EIAs do not involve the use, storage, and disposal of radioactive materials. However, progesterone is excreted in the feces as predominantly unconjugated metabolites (progestogens) and, until recently, antibodies able to cross-react with a variety of progestogens were used primarily in RIAs. An EIA using a broad-spectrum progestogen antibody is described and applied to serum and/or fecal samples from female African elephants, black rhinoceros, white rhinoceros, okapi, and hippopotami. The clear progestogen profiles generated in these species suggest that the described EIA would be as versatile as the RIA using the same antibody and could be a practical and economical alternative to RIAs for monitoring gonadal function via progestogen analysis in zoo species.

Hanks, J., 2001. Conservation strategies for Africa's large mammals. Reprod Fertil Dev 13, 459-468.
Abstract: Africa's large mammals are conserved for their aesthetic, scientific and economic values. Many of these species face a gloomy future precipitated by a combination of factors directly and indirectly influenced by the activities of man, including habitat loss, overexploitation, poor management of designated protected areas, and the vulnerability of small isolated populations. Africa's designated protected areas and biodiversity hotspots are also under threat, highlighting the importance of embracing community participation to address accelerating poverty and malnutrition. Innovative strategies are required for the conservation of Africa's mammals, such as the integration of a wide range of species in the production landscape, including the farming community. Transfrontier conservation areas (TFCAs) have been established with the combined objectives of conserving biodiversity, creating new jobs in the tourism and wildlife industry, and promoting a culture of peace. These areas extend far beyond traditional national parks, providing opportunities for integrating large mammals into sustainable land-use practices, at the same time as addressing some of the continent's more pressing socioeconomic needs. Research on African mammals will inevitably have to change direction to accommodate the growing threats and changed circumstances. Priorities will include the identification of corridors associated with TFCA establishment, the determination of the economic value of certain species in consumptive use programmes, research on contraception as a management option in restricted areas, and further work on the indirect use value of species. There will also be worthwhile opportunities to be pursued with ex situ conservation programmes, but these need to be focussed more efficiently.

Hildebrandt, T.B., Hermes, R., Pratt, N.C., Brown, J.L., Schwammer, H., Schmitt, D., Jewgenow, K., Olson, D., Lehnhardt, J.L., Goritz, F. Results of Artificial Insemination Programmes in Asian and African Elephants Kept Under Different Management Systems. A Research Update on Elephants and Rhinos; Proceedings of the International Elephant and Rhino Research Symposium, Vienna, June 7-11, 2001.  52. 2001. Vienna, Austria, Schuling Verlag. 2001.
Ref Type: Conference Proceeding

Kirkman, S., ., Wallace, E.D., van Aarde, R.J., Potgieter, H.C., 2001. Steroidogenic correlates of pregnancy in the rock hyrax (Procavia capensis). Life Sciences 68, 2061-2072.
Abstract: In pregnant rock hyraxes (P. capensis) isolated leucocytes metabolize both [3H]pregnenolone and [3H]progesterone while whole blood, erythrocytes and an erythrocyte/leucocyte mixture only metabolized [3H]progesterone. Plasma displayed no tendency to metabolically convert any one of these two steroids. In whole blood, [3H]progesterone appears to be converted to 5alpha-pregnane-3,20-dione and a compound with chromatographic properties similar to that of 5alpha-pregnan-3alpha-ol-20-one. 5alpha-Pregnane-3,20-dione exhibited a high relative binding affinity for the uterine progesterone receptor (94%), but 5alpha-pregnan-3alpha-ol-20-one displayed very little affinity for the same receptor (0.4%). 5alpha-Pregnane-3,20-dione may therefore aid in the maintenance of pregnancy. Corpora lutea metabolized progesterone to 17alpha-hydroxyprogesterone, a compound exhibiting no progestational function because of its low relative binding affinity for the uterine progesterone receptor (2%). Progesterone appears to be the main product of the corpus luteum. However, 5alpha-pregnane-3,20-dione circulated at concentrations approximately 8.5 times higher than progesterone, probably due to the metabolic conversion of progesterone to 5alpha-pregnane-3,20-dione by the blood. We conclude that in the hyrax, progesterone, produced by the corpora lutea, enters the circulation, where it is reduced to 5alpha-pregnanes. 5alpha-Pregane-3,20-dione may then be transported to the uterus where it binds to the progesterone receptor to assist in the maintenance of pregnancy. This mechanism appears to be analogous to that of the African elephant (Loxodonta africana) which is phylogenetically related to the hyrax, except that in the elephant the 5alpha-reduced metabolites are produced by luteal tissue and not the blood.

Kurt, F., Touma, C. Musth in Wild - Living and Captive Asian Elephants in Sri Lanka. A Research Update on Elephants and Rhinos; Proceedings of the International Elephant and Rhino Research Symposium, Vienna, June 7-11, 2001.  64-69. 2001. Vienna, Austria, Schuling Verlag. 2001.
Ref Type: Conference Proceeding

Lehnhardt, J., Bolling, J., Pratt, N., Joseph, S., Miller, G., Graham, L., Miller, M., Neiffer, D., Hildebrandt, T., Goeritz, F. Elephant Artificial Insemination (AI) in Protected Contact. A Research Update on Elephants and Rhinos; Proceedings of the International Elephant and Rhino Research Symposium, Vienna, June 7-11, 2001.  70. 2001. Vienna, Austria, Schuling Verlag. 2001.
Ref Type: Conference Proceeding

Miller, D.L., Dougherty, M.M., Decker, S.J., Bossart, G.D., 2001. Ultrastructure of the spermatozoa from a Florida manatee (Trichechus manatus latirostris). Anat Histol Embryol 30, 253-256.
Abstract: Semen was opportunistically collected from a free-ranging, 10-year-old, 275 cm (total length) Florida manatee (Trichechus manatus latirostris) during rehabilitation treatments. Ultrastructure of the spermatozoa was examined by scanning and transmission electron microscopy and differed slightly from that described for other mammals. Comparisons to the manatee's closest phylogenetic relatives, the elephant and hyrax, were made. The manatee spermatozoa had a similar acrosome but a distinct annulus and lacked the dense bodies observed in the neck of the elephant spermatozoa. Additionally, manatee spermatozoa lacked the lateral vacuoles observed in the nuclear chromatin from of the hyrax spermatozoa. These data add to our understanding of manatees and allow for comparative studies with other species that may be useful in phylogenetic and reproductive studies.

Moss, C.J., 2001. The demography of an African elephant (Loxodonta africana) population in Amboseli, Kenya. J. Zool. , Lond. 255, 145-156.
Abstract: This paper presents basic demographic parameters of African elephants (Loxodonta africana) living in and around Amboseli National Park, Kenya. The study was conducted from 1972 to the present and results are based on the histories of 1778 individually known elephants. From 1972 to 1978, the Amboseli elephant population declined and then increased steadily from 1979 to the present. Births occurred throughout the year but over 80% occurred between November and May. Birth rate varied from year to year with a pattern of peaks and troughs at 4- to 5-year intervals. The birth sex ratio did not differ significantly from 1:1. Mean age at first birth was 14.1 years, determined from a sample of 546 known-age females. Mean birth interval (n = 732) was 4.5 years for 255 females. Fecundity and calf survival varied by age of the females. Mortality fluctuated from year to year. Sex-specific mortality rates were consistently higher for males than females at all ages.

Oerke, A., Heistermann, M., Hodges, K. Reproductive Characteristics of the European Elephant Population: Long-Term Cycle and Pregnancy Data Based on Non-Invasive Methodology. A Research Update on Elephants and Rhinos; Proceedings of the International Elephant and Rhino Research Symposium, Vienna, June 7-11, 2001.  103. 2001. Vienna, Austria, Schuling Verlag. 2001.
Ref Type: Conference Proceeding

Pimm, S.L., van Aarde, R., 2001. African elephants and contraception. Nature London 411, 766.

Rasmussen, L.E.L., Riddle, H.S. Musth in Teenage Male Asian Elephants (Elephas maximus): The What & the Why of their Chemical Signals. A Research Update on Elephants and Rhinos; Proceedings of the International Elephant and Rhino Research Symposium, Vienna, June 7-11, 2001.  110. 2001. Vienna, Austria, Schuling Verlag. 2001.
Ref Type: Conference Proceeding

Rasmussen, L.E.L., 2001. Source and cyclic release pattern of (Z)-7-dodecenyl acetate, the pre-ovulatory pheromone of the female Asian elephant. Chemical Senses 26, 611-623.
Abstract: Female Asian elephants (Elephas maximus) release a pre-ovulatory urinary pheromone, (Z)-7-dodecenyl acetate (Z7-12:Ac), to signal males of their readiness to mate. Z7-12:Ac is quantitatively elevated during the follicular stage of oestrus, reaching maximum concentrations just prior to ovulation, as demonstrated by two complementary headspace techniques: (1) evacuated canister capture followed by cryogenic trapping and (2) solid phase microextraction (SPME) used prior to gas chromatography/mass spectrometry (GC/MS). These patterns were coincident with observed male behaviours and were consistent with biochemical and binding properties of the active ligand, including optimal binding pH. To release maximum amounts of Z7-12:Ac for quantitation, serum and urine samples from three mature female Asian elephants in their luteal and follicular stages of several oestrous cycles were subjected to heat and pH changes and were then treated with protease prior to SPME-GC/MS analyses. When the post-luteal serum progesterone concentrations declined to baseline levels, Z7-12:Ac became detectable in the female urine. Throughout the follicular stage, pheromone concentrations increased linearly with no apparent relationship to the two serum luteinizing hormone peaks. Pre-ovulatory urine also contained related compounds, including (Z)-7-12-dodecenol. The relative amount of this alcohol increased relative to acetate during long-term storage, with a proportional reduction in bioactivity. Z7-12:Ac was not detected in mucus samples from the urogenital tract. A potential precursor of Z7-12:Ac was identified in liver homogenates from female elephants in the follicular stage.  Erratum in: Chem Senses 2001 Sep;26(7):935

Sarma, K.K., 2001. Musth in Asian Elephant. Central Zoo Authority, New Delhi, India.

Schaftenaar, W., Hildebrandt, T.B., Flugger, M., Goritz, F., Schmitt, D., West, G. Guidelines for veterinary assistance during the reproduction process in female elephants. Proceedings American Association of Zoo Veterinarians, American Association of Wildlife Veterinarians, Association of Reptilian and Amphibian Veterinarians, and the National Association of Zoo and Wildlife Veterinarians Joint Conference.  348-355. 2001. USA.
Ref Type: Conference Proceeding
Abstract: In February 2000, a group of European zoo veterinarians met at Tierpark Hagenbeck, Hamburg to evaluate a questionnaire about 31 parturitions in Asian elephants.  The results were presented at the 40th International Symposium on Diseases of Zoo and Wild Animals.  The results were combined with the experiences of some North-American zoo veterinarians, which resulted in the protocol presented in this paper.  The protocol may serve as a guideline for institutions that wish to breed elephants. The proper application of the recommendations given in these guidelines should increase the reproductive success in elephants.  It is the moral obligation of everyone who is responsible for the  management and breeding of elephants to consider utilizing the guidelines as they may apply to their situation and to collect data that may help increase our knowledge. The breeding process in elephants requires monitoring of several parameters in both males and females. The most crucial parameters are the determination of the estrous cycle through progesterone and, perhaps, LH assay, evaluation of the genital tract in both sexes, determination of the number of fetuses and finally, parturition.  The first part of the paper will mention briefly the tools that can be used in female elephants to achieve these goals.  The second part describes a protocol for veterinary intervention in elephant parturition.

Schmitt, D. Riddles's Elephant amd Wildlife Sanctuary Elephant Birth Protocol.  2001.
Ref Type: Internet Communication

Schmitt, D., Krywko, R., Reichard, T.A., Shellabarger, W., Bailey, K., Short, J. Surgical approach to artificial insemination in elephants. Proceedings American Association of Zoo Veterinarians, American Association of Wildlife Veterinarians, Association of Reptilian and Amphibian Veterinarians, and the National Association of Zoo and Wildlife Veterinarians Joint Conference.  338. 2001. USA. 1.
Ref Type: Conference Proceeding
Abstract: Artificial insemination is a recent development for assisted reproduction in elephants.  Non-surgical insemination requires a cooperative elephant,   well-habituated to the various procedures.  In addition, a well-trained and equipped insemination team is needed to successfully complete the procedure.  A surgical approach for artificial insemination in elephants reduces both the technology needed for success and the level of cooperation needed from the elephant to be inseminated.  The first successful, surgical, artificial insemination was accomplished by making a 3-cm incision into the urogenital canal just below the anus.  The vestibulotomy incision was guided by the placement of 8-cm PVC tube with a 3-cm opening at the upper end, up through the vulva to a level just below the anus.  The opening in the PVC tube was used as a guide by palpating the opening through the skin.  The incision was made following injection of a local anesthetic above the proposed incision site.  The PVC guide prevents incision into the opposite wall of the urogenital tract.  After the incision is complete a sterile disposable vaginal speculum is introduced into the urogenital canal.  The intact hymen or cervix can be visualized directly with a flashlight or, for documentation of the procedure, a short endoscope can be utilized.  Placement of semen into the vagina can be accomplished with little difficulty using sterile disposable horse insemination pipettes.  Multiple inseminations are possible through the incision for the 2-3 days of estrus.  Following the last insemination, a local anesthetic is administered and the edges of the incision are freshened and four to six simple interrupted sutures are placed to close the incision.  Healing of the incision requires 4 to 6 weeks with good aftercare.

Schmitt, D.L., Krywko, R., Reichard, T.A., Shellabarger, W. Surgical approach to artificial insemination in elephants. Kirk Baer, C. and Wilmette, M. W. Proceedings American Association of Zoo Veterinarians, American Association of Wildlife Veterinarians, Association of Reptilian and Amphibian Veterinarians and the National Association of Zoo and Wildlife Veterinarians Joint Conference  2001.  338. 2001.  American Association of Zoo Veterinarians. 9-18-2001.
Ref Type: Conference Proceeding

Schmitt, D.L., Krywko, R.L., Reichhardt, T., Shellabarger, R.W., Bailey, K.M., Short, J.N. Surgical approach to artificial insemination in elephants. A Research Update on Elephants and Rhinos; Proceedings of the International Elephant and Rhino Research Symposium, Vienna, June 7-11, 2001.  129-131. 2001. Vienna, Austria, Schuling Verlag. 2001.
Ref Type: Conference Proceeding
Abstract: Artificial insemination in elephants is a recent development for assisted reproduction in elephants. Non-surgical insemination requires both a cooperative and well-habituated elephant to the various procedures. In addition a well-trained and equipped insemination team is needed to successfully complete the procedure. A surgical approach for artificial insemination reduces the technology needed for success and the level of cooperation needed from the elephant to be inseminated. The first successful surgical artificial insemination was accomplished by making a 3cm incision into the urogenital canal just below the anus. The vestibulotomy incision was guided by placement of an 8 cm diameter PCV tube, with a 3cm opening near the upper end, through the vulva up to the level just below the anus. The opening in the PCV tube was used as a guide by palpating above the proposed incision site. This guide prevents incision into the opposite wall of the urogenital tract. After the incision is complete a sterile disposable vaginal speculum is introduced into the urogenital canal. The intact hymen or cervix can be visualized directly with a flashlight or, for documentation of the procedure, a short endoscope can be utilized. Placement of semen into the vagina or hymen can be accomplished with little difficulty through the incision for the two to three days of estrus. Following the last insemination, a local anesthetic is administered and the edges of the incision are freshened and four to six simple interrupted sutures are placed to close the incision. Healing of the incision requires four to six weeks with good aftercare.

Schulte, B.A. Examining Ideas on the Evolution of Musth. A Research Update on Elephants and Rhinos; Proceedings of the International Elephant and Rhino Research Symposium, Vienna, June 7-11, 2001.  287. 2001. Vienna, Austria, Schuling Verlag. 2001.
Ref Type: Conference Proceeding

Schulte, B.A., Slade, B.E., Rasmussen, L.E.L. The Trunk and Tail of Elephant Communication: Studies on Captive Asian Elephants. A Research Update on Elephants and Rhinos; Proceedings of the International Elephant and Rhino Research Symposium, Vienna, June 7-11, 2001.  286. 2001. Vienna, Austria, Schuling Verlag. 2001.
Ref Type: Conference Proceeding

Schwammer, H. From Artificial Insemination to Birth. A Case Study on African Elephants (Loxodonta africana). A Research Update on Elephants and Rhinos; Proceedings of the International Elephant and Rhino Research Symposium, Vienna, June 7-11, 2001.  289-292. 2001. Vienna, Austria, Schuling Verlag. 2001.
Ref Type: Conference Proceeding

Suedmeyer, W.K. Serum hydrocortisone levels in a manually restrained African elephant (Loxodonta africana)  pre- and post- semen collection. Kirk Baer, C. and Wilmette, M. W. Proceedings American Association of Zoo Veterinarians, American Association of Wildlife Veterinarians, Association of Reptilian and Amphibian Veterinarians and the National Association of Zoo and Wildlife Veterinarians Joint Conference.  388-389. 2001.   American Association of Zoo Veterinarians. 9-18-2001.
Ref Type: Conference Proceeding

Wingate, L., Lasley, B. Is Musth a Reproductive Event: An Examination of Arguments For and Against this View. A Research Update on Elephants and Rhinos; Proceedings of the International Elephant and Rhino Research Symposium, Vienna, June 7-11, 2001.  150-156. 2001. Vienna, Austria, Schuling Verlag. 2001.
Ref Type: Conference Proceeding

Wisser, J., Pilaski, J., Strauss, G., Meyer, H., Burck, G., Truyen, U., Rudolph, M., Frolich, K., 2001. Cowpox virus infection causing stillbirth in an Asian elephant (Elephas maximus). Veterinary Record 149, 244-246.

Wyatt, J. Elephant breeding soundness examination forms and database. Kirk Baer, C. and Wilmette, M. W. Proceedings American Association of Zoo Veterinarians, American Association of Wildlife Veterinarians, Association of Reptilian and Amphibian Veterinarians, and the National Association of Zoo and Wildlife Veterinarians Joint Conference 2001.  396-400. 2001. USA, AAZV. 1.
Ref Type: Conference Proceeding
Abstract: Routine use of standardized soundness examination forms for male and female elephants provides data for an inter-relational database.  Through queries we may answer population-based reproductive questions essential for promoting self-sustaining populations. This poster presentation demonstrates breeding soundness examination forms and accompanying database used in a pilot project to evaluate 25 elephants.

Agnew, D.W., Munson, L., Gage, L.J., Fowler, M.E., Ramsay, E. Cystic Endometrial Hyperplasia in Nulliparous Asian Elephants. 2000 Proceedings AAZV and IAAAM Joint Conference.  442. 2000. 2000.
Ref Type: Conference Proceeding
Abstract: Maintaining self-sustaining populations of elephants through captive breeding is a new goal of the Elephant Species Survival Plan. Most elephants available for breeding in U.S. zoos are nulliparous and aged, and their fertility is unknown. Endometrial hyperplasia has been noted in aged elephants, and this condition may affect their fertility. The purpose of this study was to better characterize the gross and histopathologic features of these lesions and assess the demographic distribution. Clinical histories, necropsy reports, and endometrial samples from Asian elephants (Elephas maximus) at least 20 yr of age that died from 1985 to 1999 were reviewed. Gross pathologic findings in all cases were similar and consisted of a multifocal to diffuse distribution of 1-2 cm diameter cysts in the endometrium. Pedunculated edematous endometrial polyps up to 10 cm in length also were present, projecting from the endometrial surface or free within the lumen. Some polyps were necrotic. Histologically, the endometrium was characterized by varying sized cystic endometrial glands lined by cuboidal to tall columnar epithelium. Other glands were present in small clusters and lined by hyperplastic endometrium. The endometrial polyps consisted of a core of edematous stroma containing clusters of cystic glands. Tissues expelled from the urogenital tract of another aged, nulliparous cow were also reviewed. These fragments consisted of necrotic tissue with ghost-like remnants of glands similar to endometrial glands. These fragments may represent expelled pedunculated endometrial polyps, which had become necrotic and sloughed. These results indicate that aged nulliparous Asian elephants commonly develop cystic endometrial hyperplasia and that the pedunculated polyps may represent a more advanced form of this disease. Sloughing of these pedunculated polyps may be noted clinically and may offer information about the condition of a cow's endometrium. The effect of endometrial hyperplasia on fertility in elephants is unknown, but in other species large numbers of cysts can interfere with implantation. The prevalence of these lesions in aged elephants suggests that younger animals would be better candidates for breeding and that efforts should be made to clinically evaluate potential breeding cows for endometrial health.

Ananth, D., 2000. Musth in elephants. Zoos' Print Journal 15, 259-262.

Barber, M.R., Fayrer-Hosken, R.A., 2000. Possible mechanisms of mammalian immunocontraception. J Reprod Immunol 46, 103-124.
Abstract: Ecological and conservation programs in ecosystems around the world have experienced varied success in population management. One of the greatest problems is that human expansion has led to the shrinking of wildlife habitat and, as a result, the overpopulation of many different species has occurred. The pressures exerted by the increased number of animals has caused environmental damage. The humane and practical control of these populations has solicited the scientific community to arrive at a safe, effective, and cost-efficient means of population control. Immunocontraception using zona pellucida antigens, specifically porcine zona pellucida (pZP), has become one of the most promising population control tools in the world today, with notable successes in horses and elephants. A conundrum has risen where pZP, a single vaccine, successfully induces an immunocontraceptive effect in multiple species of mammals. This review describes the most current data pertaining to the mammalian zona pellucida and immunocontraception, and from these studies, we suggest several potential mechanisms of immunocontraception.

Brown, J.L. Zoo Biology. Special Issue on elephant biology 19[5], 1-184. 2000.
Ref Type: Journal (Full)
Abstract: This issue focuses on elephant biology and includes the following topics: ultrasonography of the urogenital tract in elephants Loxodonta africana and Elephas maximus (an important tool for assessing female and male reproductive function); reproductive endocrine monitoring of elephants (an essential tool for assisting captive management); ultrasonography of the oestrous cycle in female African elephants; review of a newly recognized disease of elephants caused by endotheliotropic herpesviruses; tuberculosis in elephants in North America; how chemical signals integrate Asian elephant society; elephant communication; social structure and helping behaviour in captive elephants; a postcryogenic comparison of membrane fatty acids of elephant spermatozoa; and first disclosure and preliminary investigation of a liquid released from the ears of African elephants.

Brown, J.L., 2000. Reproductive endocrine monitoring of elephants: an essential tool for assisting captive management. Zoo Biology 19, 347-367.
Abstract: Considerable information now is available about the basic reproductive biology of elephants, especially females. However, as important as this knowledge is, it no longer is enough to simply compile it into a database. The potential exists for using endocrine monitoring techniques to solve real problems. This review summarizes our current knowledge of elephant endocrinology and offers suggestions on how to use the technology to maximize reproductive potential. The oestrous cycle can be monitored through the analysis of serum progestogens, primarily 5alpha-reduced compounds, and consists of an 8- to 12-week luteal phase and a 4- to 6-week inter-luteal period. Proof of ovarian cyclicity currently is mandatory before Species Survival Plan breeding recommendations are approved. However, because many adult females are not cycling normally, the reproductive monitoring of all cows throughout their life span is now encouraged. Complete endocrine evaluations in conjunction with ultrasound examinations and behavioral assessments are needed to identify causes of reproductive failure and develop mitigating treatments. Progestogen analyses also are effective for monitoring pregnancy, but only if longitudinal samples are collected. Alternatively, pregnancy can be diagnosed in occasional samples using serum prolactin or possibly relaxin measurements after 20 weeks of gestation. Parturition can be predicted on the basis of the rapid decrease in progestogens that occurs about 2-5 days before birth. An updated model of ovarian dynamics during the oestrous cycle suggests that two waves of follicular development occur 3 weeks apart during the non-luteal phase, possibly under the control of follicle-stimulating hormone. Each follicular wave culminates in a luteinizing hormone (LH) surge, with the second surge inducing ovulation and corpus luteum formation. The functional significance of the first, anovulatory LH surge is under investigation, but from a practical perspective it can be used to schedule breeding (by artificial insemination or natural mating) to coincide with the ovulatory LH surge. Less is known about the reproductive biology of bulls, aside from the fact that musth is associated with dramatic changes in androgen secretion. Studies are needed to determine whether poor libido and inadequate semen quality observed in some mature elephants are due to testicular steroidogenic dysfunction. When blood samples cannot be collected for routine hormone analysis, gonadal activity can be monitored non-invasively through the measurement of excreted steroid metabolites (males: androgens; females: estrogens, progestogens) in urine and faeces. Lastly, suggestions for future research priorities are provided.

Doi, O., Komatsumoto, M., Terazono, M., Wada, S., 2000. Exfoliative cytology in vaginal vestibule of female Asian elephants: relation to circulating progesterone concentrations. Zoological Science 17, 1303-1309.
Abstract: The oestrous cycle of the Asian elephant (Elephas maximus) was monitored by analysis of exfoliative cytology in the vaginal vestibule and serum progesterone concentrations. Appearance frequency of each 5 exfoliative cells; parabasal, intermediate, superficial anuclear and nuclear cells and leukocytes; on the smear collected from two elephants was calculated, and serum progesterone concentrations were measured by radioimmunoassay. Serum progesterone concentrations changed regularly with the cycle between 14 and 17 weeks. Using spectrum analysis (Yule-Walker method) to appearance frequency of exfoliative cells, it was found that the time when a superficial cell markedly appeared in vaginal vestibule corresponded to the time when serum progesterone concentration was almost negligible. It is suggested that the time when numbers of two kinds of superficial (anuclear and nuclear) cells and parabasal and intermediate cells increase to the smear of the elephant, means the period from pro-oestrus to oestrus and from metoestrus to dioestrus, respectively.

Emanuelson, K.A., Kinzley, C.E. Salmonellosis and subsequent abortion in two African elephants. Proc. AAZV and IAAAM Joint Conf.  269-274. 2000.
Ref Type: Conference Proceeding

Fayrer-Hosken, R.A., Grobler, D., van Altena, J.J., Bertschinger, H., Kirkpatrick, J.F., 2000. Immunocontraception of African elephants. Nature London 407, 6801.
Abstract: Sum: Based on a South African trial of 41 adult females, it is argued that pZP immunocontraception is a humane method to control elephant populations without behavioural side effects.

Fritsch, G., Göritz, F., Hermes, R., Jewgenow, K., Maltzan, J., Hildebrandt, T.B., 2000. Physiology of sexual maturity in male elephants. Reprod Dom Anim 35, 26.

Hermes, R., Olson, D., Goritz, F., Brown, J.L., Schmitt, D.L., Hagan, D., Peterson, J.S., Fritsch, G., Hildebrandt, T.B., 2000. Ultrasonography of the estrous cycle in female African elephants (Loxodonta africana). Zoo Biology 19, 369-382.
Abstract: The endocrinology of the elephant oestrous cycle has been well characterized, but little emphasis has been placed on evaluating corresponding changes in the reproductive tract. Ultrasound was used to document changes in reproductive tract morphology throughout the oestrous cycle in four cycling female African elephants. During a 7-month period, frequent ultrasound examinations (n=190) during the luteal and non-luteal phase were compared with serum progesterone and luteinizing hormone (LH) concentrations during a 7-month period. Ultrasonographic images documented vaginal and cervical oedema and changes in mucus consistency during the non-luteal phase. The cross-sectional diameter of the endometrium showed a dramatic increase during the non-luteal phase and followed cyclic changes. A different pattern of follicular development on the ovary was associated with the two LH surges. Follicle growth associated with the first, anovulatory LH surge was characterized by the formation of multiple small follicles, in contrast to the maturation of a single large follicle at the second, ovulatory LH (ovLH) surge. Ovulation and the subsequent formation of a corpus luteum (CL) were observed only after the ovLH surge. Ultrasound data in combination with endocrine assessments suggest that the African elephant is non-ovulatory, although multiple non-ovulatory luteal structures developed during the late non-luteal phase of each cycle. Both ovulatory CL and non-ovulatory luteal structures were present only through one cycle and regressed at the end of the luteal phase in conjunction with the drop in serum progesterone. We conclude that periodic reproductive-tract ultrasound assessments in association with continued endocrine monitoring of the oestrous cycle should be incorporated into the routine reproductive health assessment of elephants. This information is necessary for determining reproductive fitness before making breeding recommendations. It also has proven to be an invaluable tool for use with assisted reproductive techniques and has enormous potential for evaluating the efficiency of hormonal therapies used to treat reproductive dysfunction.

Hildebrandt, T.B., Goritz, F., Pratt, N.C., Brown, J.L., Montali, R.J., Schmitt, D.L., Fritsch, G., Hermes, R., 2000. Ultrasonography of the urogenital tract in elephants (Loxodonta africana and Elephas maximus): an important tool for assessing female reproductive function. Zoo Biology 19, 321-332.
Abstract: At present, the worldwide captive elephant population is not self-sustaining.  The major reason for low reproductive rates is the heretofore undiagnosed reproductive disorders of nulliparous females of prime breeding age.  Recent advances in ultrasound technology have facilitated the detection of these disorders in non-sedated animals.  Approximately 2000 ultrasonographic examinations were performed in more than 280 captive and wild African and captive Asian female elephants.  The entire urogenital tract was scanned, measured and documented to provide a reference for ultrasound specialists involved in elephant breeding programs. The primary pathological lesions that influenced reproductive rates in these females were uterine tumors and endometrial cysts, and ovarian cysts that resulted in acyclicity.  The detection of these disorders and their stage of development can be used by elephant managers to make decisions approximately which animals to include in breeding programs.  Ultrasonography can be used as an effective tool for assessing the reproductive fitness of female breeding candidates in both African and Asian elephants.

Hildebrandt, T.B., Hermes, R., Pratt, N.C., Fritsch, G., Blottner, S., Schmitt, D.L., Ratanakorn, P., Brown, J.L., Rietschel, W., Goritz, F., 2000. Ultrasonography of the urogenital tract in elephants (Loxodonta africana and Elephas maximus): an important tool for assessing male reproductive function. Zoo Biology 19, 333-345.
Abstract: The success rate of captive elephant breeding programs worldwide is poor. Along with undiagnosed reproductive disorders in females and fatal diseases such as the newly discovered herpesvirus infection, male infertility now is considered a major contributing factor in the failure to maintain self-sustaining captive populations. To address questions related to male reproductive dysfunction, approximately 309 ultrasonographic assessments combined with semen collection were performed in captive (n=10) and wild (n=4) African (Loxodonta africana) and captive (n=61) Asian (Elephas maximus) elephants. Bulls ranged from 4 to 50 years of age and were examined at 9 institutions in North America, 13 in Europe, 2 in Africa, and 7 in Asia. About half of the reproductive assessments were performed in protected contact situations with elephants handled in a restraint device, and half involved assessments of trained Asian bulls managed in free contact. Four wild African and two Asian elephant bulls were evaluated after receiving general anaesthesia. Transrectal ultrasound was used to characterize the morphology and functionality of the entire urogenital tract, including the testes and accessory sex organs. Bulls were categorized on the basis of breeding status (breeders vs. non-breeders) and social history (i.e., type of interaction with conspecifics and keepers). Most of the bulls were non-breeders (designated Types I-V). Type I (n=3 African, 6 Asian) and Type V (n=1 Asian) were immature and castrate, respectively. On the basis of keeper evaluations, Type II bulls (n=2, 4) were subordinate to older cows and keepers, whereas Type III bulls (n=4, 28) were dominated by other bulls. Type IV (n=1, 8) were older bulls of unknown history that exhibited numerous testicular pathologies resulting in poor semen quality. Non-breeding bulls included those that were exposed to females, but failed to breed, as well as those that had no opportunities to breed. Type VI individuals (n=4, 14) were proven breeders. The percentage of observable reproductive tract pathology in adult males was remarkably low (14%), even in older bulls. However, apparent infertility of non-organic cause (i.e., not due to specific anatomical abnormalities) in these otherwise healthy bulls was high (32%). Semen quality varied markedly in ejaculates collected from the same bull, as well as from different bulls. In conclusion, although many of these bulls could serve as semen donors for natural mating or artificial insemination, the inconsistent production of good-quality ejaculates raises questions as to the reliability of these individuals to participate in breeding programs. The apparent inhibitory effect of suppressive social interactions on reproductive potential also needs to be investigated. Ultrasound examinations combined with semen collection should be conducted periodically to estimate the reproductive value of each bull and determine whether altered management strategies are needed to enhance captive breeding.

Kahl, M.P., Armstrong, B.D. Visual displays in wild musth males of the Arican Elephant. Proceedings of the Fifth International Elephant Research Symposium.  2000.
Ref Type: Conference Proceeding

Kitiyanant, Y., Schmidt, M.J., Pavasuthipaisit, K., 2000. Evaluation of sperm acrosome reaction in the Asiatic elephant. Theriogenology 53, 887-896.
Abstract: This study focuses on the effect of chemicals on acrosome reaction in elephant spermatozoa. Semen was collected at the Washington Park Zoo in Portland, Oregon, from an 11-yr-old Asian elephant by artificial vagina (7 ejaculates) and transported to Mahidol University in Bangkok in extender at 4 to 5 degrees C within 24 to 28 h. A total of 500 x 10(6) sperm/mL was used for the control and for each of the 4 treatment groups: 1) cAMP (0.1 mM); 2) caffeine (0.1 mM); 3) Penicillamine hypotaurine and epinephrine, PHE (penicillamine 2 mM, hypotaurine 1 mM, epinephrine 1 mM); and 4) heparin (10 microg/mL) at 39 degrees C for 2 h. Aliquots were removed and the sperm viability, abnormal morphology, and acrosome status were evaluated by triple stain technique. Transmission electron microscopy (TEM) was used to observe changes of the sperm head membrane in all treatment groups. Trypan blue reliably stained dead spermatozoa, while rose Bengal stained only the spermatozoa with intact acrosomes. The concentration of dead sperm cells was similar in the 4 groups. The percentages of live acrosome-reacted spermatozoa in the control and in groups treated with caffeine, PHE, cAMP and heparin were 19.5 +/- 4.3, 38.1 +/- 4.0, 34.8 +/- 3.7, 29.8 +/- 0.8 and 28.0 +/- 4.2, respectively. The acrosome reaction rate was higher in the treatment groups than in the control (P<0.05). Caffeine and PHE caused significantly higher acrosome reaction of the sperm head than cAMP or heparin (P<0.05). The electron micrographs showed that the acrosome reaction occurred by the presence of apical vesiculation. The results indicated that 1) the triple stain technique allowed for evaluation of both viability and acrosome reaction simultaneously in elephant spermatozoa,2) acrosome reaction occurred at a high rate in all 3 treatment groups. 3) the effects of caffeine and PHE were significantly higher (P<0.05) than of cAMP and heparin, and 4) the data obtained from the triple stain technique corresponded to those from TEM.

Lazar, J., Prestwich, G.D., Rasmussen, L.E.L., 2000. Urinary and trunk mucus protein carriers of (Z)-dodecenyl acetate, the sex pheromone of the Asian elephant. Chemical Senses 25, 603.

Milewski, A., 2000. Iodine as a Possible Controlling Nutrient for Elephant Populations. Pachyderm 28, 78-90.
Abstract: The geography and physiology of iodine deficiency in humans and domestic ungulates suggests that the nutritional content of ground water may hold a key to humane and efficient management of population sizes of elephants. Artificial bore water in dry climates in southern Africa appear to be, on average, a good supplement of this easily leached element, and may have inadvertently boosted the reproductive rates of elephants in several conservation areas. The largest land mammals are likely to be limited by deficiency of iodine, inasmuch as their plant foods are deficient in this element relative to the hormonal requirements associated with exceptional brain size and relatively great thyroid size. Extrapolation from domestic ecosystems suggests that elephants exceed medium-sized wild herbivores in the sensitivity of their reproductive rates to subclinical deficiency of iodine, partly because the rate of loss of iodine from the body is likely to be hyperallometric to those of energy, protein, and water, with increasing body size. Elephants pass food and water rapidly for their body size, but this allows maximal intakes of iodine, which can potentially be further supplemented by absorption through the skin. The great variation in concentrations of iodine between adjacent aquifers suggests a versatile approach to population control. Closure of iodine-rich bore holes in overpopulated areas may reduce rates of sexual maturation, conception, birth, and weaning, with minimum artificial distress to adults or surviving juvenile elephants. Conversely, selection of the bore waters richest in iodine may help to promote population growth in areas recently restocked with elephants. All proboscideans became extinct in the Americas and temperate Eurasia at the end of the Pleistocene, when glacial melting had profoundly depleted iodine, and humans had the means to monopolize the remaining sources of supplementary iodine. The maximal intelligence and fecundity of those megaherbivores which have survived the era of domestication may have made these species depend on supplementation of iodine.

Olson, D., Weise, R.J., 2000. State of the North American African elephant population and projections for the future. Zoo Biology 19, 311-320.
Abstract: The African elephant has historically received less attention in the captive community than the Asian elephant.  One manifestation of this lack of attention is that only 25 African elephant calves had been born in captivity in North America as of 01 January 1999.  With the recent attention to both elephant species, it is imperative to evaluate the African elephant's potential to maintain a self-sustaining population in North America.  Review of the raw data indicates that African elephants have reproduced poorly and experienced low juvenile survival in North America.  However, using realistic life table models the future of the North American African elephant population can be predicted.  The current population is relatively young compared to the captive Asian elephant population and has a much greater potential to become self-sustaining with increased focus and efforts toward reproduction.  Unlike the Asian elephant population, the African elephant population may be able to become self-sustaining without further importation, if reproduction and juvenile survivorship increase significantly in the next ten years.

Prasad, A., Dinesh, M.T., Hareesh, P.S., Biju, S., Harikumar, S., Saseendran, P.C., 2000. Analysis of musth episodes in captive Asian elephants (Elephas maximus). Zoos' Print Journal 15, 322-327.
Abstract: Analysis of data on musth episodes of 29 Asian elephants (Elephas maximus) collected from Punnathoor Kotta of Guruvayoor (1990-2000) revealed that the duration of musth was 99±36 days, increasing as age advances. Hours of bright sunshine had pronounced influence on the duration of musth. The number of elephants coming to musth showed peaks during January and August, which are the post monsoon periods in Kerala. Musth first occurred at 23 years of age (range 16 to 35).

Rasmussen, L.E.L., Krishnamurthy, V., 2000. How chemical signals integrate Asian elephant society: the known and the unknown. Zoo Biology 19, 405-423.
Abstract: The importance of chemical senses to elephants was recognized in anecdotal observations by ancient humans. Modern scientific tools, such as molecular biological techniques, highly sensitive gas chromatographic/mass spectrometric instrumentation, and statistically valid ethological methods, have allowed the study of real events of chemical communication between elephants. Such communication encompasses long- and short-range navigation, relationship recognition, and inter- and intra-sexual exchange of reproductive condition, metabolic state, and social status. Asian elephants emit large amounts of complex chemical mixtures in breath and urine, and in secretions from the temporal gland, inter-digital glands, and ears. Some emitted chemicals originate in blood and may be metabolic products; others are secretory products, at times apparently under hormonal control. The wide variety of emitted compounds includes hormones, proteins, and volatile compounds; selected volatile ketones and an acetate ((Z)-7-dodecenyl acetate) function as chemical signals and a pheromone, respectively. Some of these specific chemicals identified in emissions from Asian elephants dwelling in the United States have been found to be present in the exudates from elephants in India. This similarity is demonstrable for three metabolic conditions: pregnancy in females and pre- and post-musth males. Future chemical communication studies on male elephants should focus on musth and its relevance to reproduction and male social structures. Such investigations should include hormones, metabolites, brain chemistry, and possible primer pheromones. For females, the factors influencing possible estrous synchrony, what role primer pheromones play in female reproduction, how chemical signals influence social behavior, and whether luteinizing hormone influences pheromone production are among remaining fundamental questions.

Schmitt, D.L., Hildebrandt, T.B., 2000. Corrigendum to "Manual collection and characterization of semen from Asian elephants". Anim Reprod Sci 59, 119.

Schulte, B.A., Feldman, E., Lambert, R., Oliver, R., Oliver, R., 2000. Temporary ovarian inactivity in elephants: relationship to status and time outside. Physiol Behav 71, 123-131.
Abstract: The captive elephant population in North America is in reproductive decline and,without importation from the wild, may cease to be viable within the next several decades. The estrous cycle of three captive, reproductive-age African elephants was monitored for 3 years by measuring serum progesterone concentrations. Each elephant experienced one or more episodes of extended low progesterone (>12 weeks), analogous to supposed terminal cessation of estrous cyclicity or 'flatlining' that has been described in some captive Asian and
African elephants. Other studies have reported lengthy non-luteal (follicular)phases that indicate extended episodes of ovarian inactivity; however, this phenomenon has not been examined in detail. In this study, total duration of temporary ovarian inactivity or acyclicity followed a social rank pattern, with the most subordinate female having the longest and the dominant female the shortest duration. During periods of acyclicity, the number of hours the elephants spent outside was significantly less than during non-luteal or luteal phases of the cycle. Except in one instance, behavioral data recorded by elephant keepers during their interactions with the elephants showed no change in handling during periods of ovarian inactivity. Further study is necessary to distinguish the causative agent for temporary cessation of estrous cyclicity. Understanding this phenomenon is imperative for the future reproductive viability of captive elephant populations.

Smith, B., Hutchins, M., 2000. The value of captive breeding programmes to field conservation:elephants as an example. Pachyderm 28, 101-109.
Abstract: Wildlife conservation is among the highest priorities of professionally managed zoological parks and aquariums.  Even if reintroduction is not the goal of a captive breeding programme, zoos and aquariums can contribute to wildlife and habitat conservation in a number of ways, including public education,scientific research, development of relevant technologies, professional training and technology transfer, ecotourism, political action and involvement in field conservation.  Here, we use elephants as an example of how such efforts support conservation activities.

Swain, J.E., Miller, R.R., Jr., 2000. A postcryogenic comparison of membrane fatty acids of elephant spermatozoa. Zoo Biology 19, 461-473.
Abstract: Cryogenic protocols have been successful in storing spermatozoa collected from African elephants (Loxodonta africana). However, these same protocols and modifications of these protocols have failed to preserve spermatozoa collected from Asian elephants (Elephas maximus), Because the success or failure of cryogenic freezing may rely on differences in membrane composition, a postcryogenic comparison of the membrane fatty acid composition of spermatozoa isolated from African and Asian elephants was studied. The spermatozoa of African elephants possessed significantly higher levels of docosahexaenoic acid (22 : 6, n-3) and docosapentaenoic acid (22 : 5, n-6) as compared to the spermatozoa of Asian elephants. Meanwhile, the spermatozoa of Asian elephants had higher levels of myristic acid (14 : 0), arachidonic acid (20 : 4, n-6), and docosatetraenoic acid (22 : 4, n-6) as compared to the spermatozoa of African elephants. The most abundant membrane fatty acid was docosahexaenoic acid (22 : 6, n-3). The percentage of membrane docosahexaenoic acid (22 : 6, n-3) in the spermatozoa of African elephants was 68.13±0.52 as compared to 42.88±0.87 in the spermatozoa of Asian elephants (t=31.48, P_0.0001).

Wallis, M., 2000. Episodic evolution of protein hormones: molecular evolution of pituitary prolactin. J Mol Evol 50, 465-473.
Abstract: Previous studies have shown that pituitary growth hormone displays an episodic pattern of evolution, with a slow underlying evolutionary rate and occasional sustained bursts of rapid change. The present study establishes that pituitary prolactin shows a similar pattern. During much of tetrapod evolution the sequence of prolactin has been strongly conserved, showing a slow basal rate of change (approx 0.27x10(9) substitutions/amino acid site/year). This rate has increased substantially ( approximately 12- to 38-fold) on at least four occasions during eutherian evolution, during the evolution of primates, artiodactyles, rodents, and elephants. That these increases are real and not a consequence of inadvertent comparison of paralogous genes is shown (for at least the first three groups) by the fact that they are confined to mature protein coding sequence and not apparent in sequences coding for signal peptides or when synonymous substitutions are examined. Sequences of teleost prolactins differ markedly from those of tetrapods and lungfish, but during the course of teleost evolution the rate of change of prolactin has been less variable than that of growth hormone. It is concluded that the evolutionary pattern seen for prolactin shows long periods of near-stasis interrupted by occasional bursts of rapid change, resembling the pattern seen for growth hormone in general but not in detail. The most likely basis for these bursts appears to be adaptive evolution though the biological changes involved are relatively small.

Wasser, S.K., Hunt, K.E., Brown, J.L., Cooper, K., Crockett, C.M., Bechert, U., Millspaugh, J.J., Larson, S., Monfort, S.L., 2000. A generalized fecal glucocorticoid assay for use in a diverse array of nondomestic mammalian and avian species. Gen Comp Endocrinol 120, 260-275.
Abstract: Noninvasive fecal glucocorticoid analysis has tremendous potential as a means of assessing stress associated with environmental disturbance in wildlife. However, interspecific variation in excreted glucocorticoid metabolites requires careful selection of the antibody used in their quantification. We compared four antibodies for detecting the major fecal cortisol metabolites in yellow baboons following (3)H cortisol administration, ACTH challenge, and HPLC separation of fecal glucocorticoid metabolites. The most effective antibody (ICN corticosterone RIA; Cat. No. 07-120102) demonstrated relatively high cross-reactivities to the major cortisol metabolites present in feces during peak excretion, following both radiolabel infusion and ACTH challenge. This same antibody also detected increased fecal glucocorticoid metabolites after ACTH administration in the African elephant, black rhinoceros, Roosevelt elk, gerenuk, scimitar-horned oryx, Alaskan sea otter, Malayan sun bear, cheetah, clouded leopard, longtailed macaque, and northern spotted owl. Results suggest that (1) fecal glucocorticoid assays reliably detect endogenous changes in
adrenal activity of a diverse array of species and (2) where comparisons were made, the ICN corticosterone antibody generally was superior to other antibodies for measuring glucocorticoid metabolites in feces.

Wiese, R.J., 2000. Asian elephants are not self-sustaining in North America. Zoo Biology 19, 299-309.
Abstract: Demographic analysis of the captive Asian elephants in North America indicates that the population is not self-sustaining.  First year mortality is nearly 30%, but perhaps more importantly the fecundity is extremely low (M_x = 0.01-0.02) throughout the prime reproductive years.  Without continued importation or a drastic increase in birth rates the Asian elephant population in North America will drop to approximately ten elephants in 50 years and be demographically extinct.  Model mortality and fecundity curves needed to establish a self-sustaining Asian elephant population in North America show that fecundity must increase 4-8 times the historical rates.  Emerging techniques such as artificial insemination may assist in making the goal of a self-sustaining population more realizable by allowing reproduction by the numerous females that do not have access to a male, but other obstacles exist as well.  A self-sustaining population will present challenges such as maintaining the significant number of male offspring that will be produced.  Importation of young females from documented self-sustaining populations overseas is one option that would alleviate the need for a self-sustaining Asian elephant population in North America and the number of imports per year would be minimal.

Wingate, L., Lasley, B. The Significance of Musth in Bull Elephants: Is It a Reproductive Event? Elephants: Cultural, Behavioral, and Ecological Perspectives; Program and Abstracts of the Workshop.  24. 2000. Davis, CA. 2000.
Ref Type: Conference Proceeding

 1999. Equine Medicine and Surgery. Mosby, St. Louis MO USA.

Bechert, U.S., Swanson, L., Wasser, S.K., Hess, D.L., Stormshak, F., 1999. Serum prolactin concentrations in the captive female African elephant (Loxodonta africana): potential effects of season and steroid hormone interactions. Gen Comp Endocrinol 114, 269-278.
Abstract: Research was conducted to determine whether seasonal changes in prolactin secretion occur in nonpregnant female African elephants and to examine potential functional interrelationships between secretion of prolactin, cortisol, and progesterone. Weekly blood samples were taken for 18 months from four female African elephants and the sera were analyzed by RIA for progesterone, cortisol, and prolactin concentrations. There was no significant effect of season on serum concentrations of prolactin. Estrous cycles averaged 14 weeks in length and were composed of a 9-week luteal phase and a 5-week follicular phase (based on progesterone concentrations consistently >200 and <200 pg/ml, respectively). Estrous cycle synchronicity was evident between pairs of elephants. Serum concentrations of prolactin (3.91 +/- 0.69 ng/ml; range: 0.84-15.8 ng/ml) were significantly lower during the luteal, compared with the follicular, phase (P < 0.0001; t test) and were positively correlated with serum concentrations of cortisol (r = 0.14; P < 0.05). Mean (+/-SE) serum concentration of cortisol was 5.7 +/- 1.3 ng/ml (range: 1.4-19.3 ng/ml), and concentrations of this adrenal steroid were negatively correlated with progesterone concentrations (r = -0.15; P < 0.01). Increased serum concentrations of prolactin detected during the follicular phase suggest that this hormone may be regulated by ovarian estrogens and may play a role in modulating ovarian function in the elephant.

Brown, J.L., Hildebrandt, T.B., Theison, W., Neiffer, D.L., 1999. Endocrine and ultrasound evaluation of a non-cycling African elephant: identification of an ovarian follicular cyst. Zoo Biology 18, 223-232.
Abstract: The reproductive rate of captive African elephants is low because of logistical difficulties associated with transporting animals for breeding, the danger of maintaining bulls and medical or physiological problems. There also is growing evidence that a significant number of mature female elephants are not experiencing normal estrous cycles. The case described in this report involves the diagnosis and attempted treatment of an ovarian follicular cyst in an African elephant at the Pittsburgh Zoo. On the basis of serum progesterone analysis, the female exhibited regular ovarian cycles from July 1993 through March 1994, but from November 1994 to the present has not shown any evidence of reproductive cyclicity. In April 1996, a large follicular structure was identified on the right ovary using transrectal ultrasound. In an attempt to luteinize the cyst, 500 ug gonadotropin-releasing hormone (GnRH) was administered intravenously in October 1996, which stimulated a modest increase in serum luteinizing hormone (LH) (approximately twofold over baseline), but no resumption of ovarian activity. The elephant was treated again 5 months later with a higher dose of GnRH (5 mg, i.v.) with the same results. An ultrasound evaluation in July 1997 indicated the structure was still present. In October 1997, the female was given human chorionic gonadotropin (hCG; 10,000 IU, i.m.), which induced estrus and breeding, but no ovulation or luteinizing of the cyst, and she remains acyclic. These results suggest that conventional methods developed in other species for treating ovarian follicular cysts may not necessarily be effective in the elephant. It is also important that reproductive age females be monitored via continuous progesterone analysis and occasional reproductive tract ultrasound evaluations to understand better the etiology of ovarian dysfunction so that effective treatments can be developed to induce consistent ovarian activity.

Brown, J.L., 1999. Difficulties Associated with Diagnosis and Treatment of Ovarian Dysfunction in Elephants - The Flatliner Problem. Journal of the Elephant Managers Association 10, 55-61.

Brown, J.L., Schmitt, D.L., Bellem, A., Graham, L.H., Lehnhardt, J., 1999. Hormone secretion in the Asian elephant (Elephas maximus): characterization of ovulatory and anovulatory luteinizing hormone surges. Biology of Reproduction 61, 1294-1299.
Abstract: In the elephant, 2 distinct LH surges occur 3 weeks apart during the non-luteal phase of the oestrous cycle: the first is an anovulatory surge (anLH) while the second surge (ovLH) induces ovulation. To understand how the timing of these 2 surges is regulated, serum concentrations of LH, FSH, progesterone, inhibin, oestradiol and prolactin were measured throughout the oestrous cycle. Long-term dynamics of hormone secretion were examined from weekly blood samples taken from 3 Asian elephants for up to 3 years. To determine whether differences exist in the secretory patterns between the anLH and ovLH surges, daily blood samples were analysed from 21 non-luteal-phase periods from 7 Asian elephants. During the non-luteal phase, serum LH was elevated for 1-2 days during the anLH and ovLH surges; there was, however, no difference in peak concentration between the 2 surges. The anLH surge occurred 19.9±1.2 days after the end of the luteal phase and was followed by the ovLH surge 20.8±0.5 days later. Serum FSH concentrations were highest at the beginning of the non-luteal phase and gradually declined to nadir concentrations within 4 days of the ovLH surge. FSH remained low until after the ovLH surge and then increased during the luteal phase. Inhibin concentration was negatively correlated with FSH during the non-luteal phase (-0.53). Oestradiol and prolactin concentrations fluctuated throughout the oestrous cycle, without a discernible pattern. The results did not identify clear differences in hormone patterns between the anLH and ovLH surges. It is suggested that elevated FSH at the beginning of the non-luteal phase may be important for follicle recruitment, and that the anLH surge acts so as to complete the follicle selection process before ovulation.

Doyle, C., York, B., Whitely, A., 1999. A survey of Asian elephant births from 1962-1998. J Elephant Managers Assoc 10, 146-148.

Fayrer-Hosken, R.A., Bertschinger, H.J., Kirkpatrick, J.F., Grobler, D., Lamberski, N., Honneyman, G., Ulrich, T., 1999. Contraceptive potential of the porcine zona pellucida vaccine in the African elephant (Loxodonta africana). Theriogenology 52, 835-846.
Abstract: Immunocontraception has been successful in controlling free-roaming equids; however, what is the potential for the immunocontraceptive control of the African elephant (Loxodonta africana)? The porcine zona pellucida (pZP) glycoproteins share antigenic domains with the African elephant zona pellucida (elZP) glycoproteins, and anti-zona pellucida serum antibodies have been successfully stimulated. To determine the cross-reactivity of the pZP and elZP, immunocytochemistry was evaluated by light and electron microscopy. Specifically, the binding of polyclonal antibodies against total heat-solubilized-porcine zona pellucida to fixed elephant ovary sections was evaluated. The elZP of primary, secondary and tertiary follicles was recognized by the rabbit-anti-pZP serum, but there was no apparent recognition of the primordial follicles. The ability of anti-pZP antibodies to recognize the elZP demonstrates that there is molecular homology between the pZP and elZP glycoproteins. This homology makes the African elephant a candidate for pZP immunocontraception. Three captive elephants were vaccinated with 400 micrograms pZP with a synthetic trehalose dicorynomycolate (S-TDCM) adjuvant. The elephants received 2 boosters of 600 micrograms pZP at 4 wk and 10 m.o. after the primary vaccination. The vaccinated female elephants developed significant (P < 0.05) titers to pZP over prevaccination levels. These levels persisted for 12 to 14 m.o. after the third vaccination. This preliminary evidence shows that the female elephant can develop significant serum antibody levels to pZP. These levels of antibodies are comparable to those required in horses for successful immunocontraception. Thus, porcine zona pellucida immunocontraception might be used to control elephant populations.

Fies, M., Heistermann, M., Hodges, J.K., 1999. Patterns of urinary and fecal steroid excretion during the ovarian cycle and pregnancy in the African elephant (Loxodonta africana). Gen Comp Endocrinol 115, 76-89.
Abstract: The aims of the present study were to (I) determine the relative abundance of the 5alpha-reduced progestins 5alpha-pregnane-3-ol-20-one (5alpha-P-3OH) and 5alpha-dihydroprogesterone (5alpha-DHP) and progesterone (P4) in African elephant feces and to establish improved fecal progestin assays for monitoring ovarian function; and (ii) describe longitudinal profiles of urinary and fecal progestin and estrogen metabolites during pregnancy. Matched urine and fecal samples were collected weekly from six adult females throughout 18 nonfertile cycles and two complete pregnancies (89 and 93 weeks duration). Fecal samples were lyophilized and extracted with 80% methanol in water and immunoreactive 5alpha-P-3OH, 5alpha-DHP, and P4 and (for pregnant females only) estrone (E1) and estradiol (E2) determined by enzyme immunoassay. Urine samples were hydrolyzed, ether-extracted, and assayed for 5alpha-P-3OH, E1, and E2. HPLC cochromatography of fecal extracts with various radioactive progestin tracers confirmed the presence of large amounts of both 5-reduced progestins (5alpha-P-3OH > 5alpha-DHP) but not of P4. 5-Reduced progestins (but not P4) were excreted in a cyclic pattern and levels were significantly correlated with urinary 5alpha-P-3OH. Fecal 5alpha-P-3OH showed the more pronounced and consistent luteal-phase elevation and a better correspondence to urine with respect to timing of the luteal-phase rise. Fecal and urinary 5-reduced progestins increased gradually during early pregnancy to maximum values around week 40-45. Levels gradually declined during the second half of pregnancy, reaching baseline values 2 days before parturition. Urinary estrogens did not show any cyclic pattern during the preconception period and levels remained low during the first 30 weeks of gestation. Thereafter, there was a rapid 10- to 20-fold increase to maximum values at mid-pregnancy, followed by a gradual decline to birth. There was no mid-pregnancy elevation in fecal estrogens, but there was a modest increase in E1 during the second half of gestation.

Foerner, J.J., 1999. Dystocia in the elephant. In: Fowler, M.E., Miller, R.E. (Eds.), Zoo and Wild Animal Medicine: Current Therapy 4. W.B. Saunders, Philadelphia; USA, pp. 522-525.

Fowler, M.E., Miller, R.E., 1999. Zoo and Wild Animal Medicine Current Therapy 4. W.B. Saunders, Philadelphia.

Gaeth, A.P., Short, R.V., Renfree, M.B., 1999. The developing renal, reproductive, and respiratory systems of the African elephant suggest an aquatic ancestry. Proc Natl Acad Sci U S A 96, 5555-5558.
Abstract: The early embryology of the elephant has never been studied before. We have obtained a rare series of African elephant (Loxodonta africana) embryos and fetuses ranging in weight from 0.04 to 18.5 g, estimated gestational ages 58-166 days (duration of gestation is approximately 660 days). Nephrostomes, a feature of aquatic vertebrates, were found in the mesonephric kidneys at all stages of development whereas they have never been recorded in the mesonephric kidneys of other viviparous mammals. The trunk was well developed even in the earliest fetus. The testes were intra-abdominal, and there was no evidence of a gubernaculum, pampiniform plexus, processus vaginalis, or a scrotum, confirming that the elephant, like the dugong, is one of the few primary testicond mammals. The paleontological evidence suggests that the elephant's ancestors were aquatic, and recent immunological and molecular evidence shows an extremely close affinity between present-day elephants and the aquatic Sirenia (dugong and manatees). The evidence from our embryological study of the elephant also suggests that it evolved from an aquatic mammal.

Göritz, F., Hildebrandt, T.B., Hermes, R., Quandt, S., Grobler, D., Jewgenow, K., Rohleder, M., Meyer, H.H.D., Hofmann, R.R. Results of hormonal contraception program in free-ranging African elephants. Verh ber Erkrg Zootiere.  39-40. 1999.
Ref Type: Conference Proceeding

Gual-Sill, F., Pickard, A.R., Holt, W.V., Green, D. Preliminary Results of Non-Invasive Monitoring of the Estrous Cycle in Female Asian Elephants (Elephas maximus) through Fecal Steroid Analysis. 1999 Proceedings American Association of Zoo Veterinarians.  87-92. 1999. 1999.
Ref Type: Conference Proceeding
Abstract: For a number of years, estrous cycle monitoring and pregnancy detection in the Asian elephant has been performed using urinary steroid hormone metabolite analysis; this technique presents some practical problems. Monitoring the reproductive status through fecal steroid analysis is possible in this and many other species. The steroid metabolite profiles of female Asian elephants were monitored by enzyme-linked immunosorbent assay (ELISA), to provide detailed information about the estrous cycle and pregnancy in this species, and to investigate causes of reproductive failure. Fecal and matched urine samples were non-invasively collected regularly for 6 mo from captive female Asian elephants. (n = 4 cyclic; n = 1 acyclic). The samples were frozen at -20ºC. Gas Chromatography and Mass Spectroscopy (GC-MS) procedures were used to investigate the steroid hormone metabolite profile and to identify the major excretory metabolites; no steroid metabolites were found in the concentrated extracted feces of this species using the currently available methodology. The fecal pregnanetriol profile observed in three of the cyclic females showed a clear relation with their matched urinary pregnanetriol profile and a cyclic pattern was demonstrated. Fecal pregnanetriol values increased from an overall mean of 94.67 ng/g of dry feces (+/- 13.24, range 31.5 - 219.12 ng/g) during the inter-luteal period to a luteal phase mean of 334.61 ng/g dry feces (+/- 43.48, range 34.35 - 1035.1 ng/g). All the data collected from the fecal and urinary analysis of pregnanetriol in all five individuals investigated demonstrated a significant relationship between urinary and fecal pregnanetriol. The acyclic individual showed a mean fecal pregnanetriol concentration of 84.91 ng/g (+/- 13.06) and values ranged from 33.17 ng/g to 211.42 ng/g. Fecal steroid hormone metabolite analysis for monitoring estrous cycles in Asian elephants may be used in the future to monitor free-roaming, wild or semi-wild individuals as well as those in captivity to assist reproductive and conservation programs of this highly endangered species.

Hildebrandt, T.B., Fritsch, G., Hermes, R., Jewgenow, K., Rudolph, M., Maltzan, J., Wiesner, H., Pratt, N.C., Schmitt, D.L., Goritz, F. Ultrasound Monitoring of the Sexual Maturation in the Male Elephant. 1999 Proceedings of the American Association of Zoo Veterinarians.  203-204. 1999. 1999.
Ref Type: Conference Proceeding
Abstract: In general, the reproductive rate of elephants in captivity is low. This is partly because of logistic difficulties associated with transporting these large animals for breeding purposes and there may be physiologic problems which also contribute to this low reproductive rate. In context with a reproductive assessment of potential breeding bulls it appears that many adult bulls of both species (Loxodonta africana and Elephas maximus) are not producing viable sperms and/or sufficient ejaculate. Our current understanding of incomplete sexual maturation or temporary infertility in male elephants is at best fragmentary. The following study was performed for characterizing the physiologic sexual maturation process in young male elephants. Two adolescent individuals of both species have been examined in order to investigate the time of their sexual maturity. The examination utilized transrectal ultrasonography of the urogenital tract, rectal stimulation for the collection of ejaculates as well as blood samples for plasma testosterone determination. The development of the testes, the accessory glands (especially the ampullae), the concentration of the testosterone, the body-height and the success of ejaculation after manual stimulation was documented and evaluated over a 3-yr period. The results were compared with data from other bull elephants which had ultrasonographic examinations or post mortem investigations. The findings of this study led to important conclusions about the characterization of the reproductive status of male elephants by means of ultrasonographic examinations. We established criteria for reproductive soundness in connection with the recruitment of potential semen donors for future artificial insemination projects. The ultrasonographic examination combined with the semen collection were appropriate methods for characterizing the exact state of sexual maturity or for identifying potential reproductive disorders in male elephants.

Hildebrandt, T.B., Göritz, F., Schnorrenberg, A., Hermes, R., Schmitt, D.L., Hagan, D., Peterson, J.S., Brown, J.L., Loskutoff, N., Pratt, N.C., Lehnhardt, J., Miller, G., Montali, R.J., Olson, D. Successful artificial insemination of African nulliparous elephants at the Indianapolis Zoo. Verh Ber Erkrg Zootiere.  41-46. 1999.
Ref Type: Conference Proceeding

Hildebrandt, T.B., Goritz, F., Hermes, R., Schmitt, D., Brown, J.L. Artifical insemination of African (Loxodonta africana) and Asian (Elephas maximus) elephants. Proceedings of the American Association of Zoo Veterinarians.  83-86. 1999. 10-9-0099.
Ref Type: Conference Proceeding

Hodges, J.K., Fieb, M., Heistermann, M. Non-Invasive Reproductive Assessment in the African Elephant: Applications to Studies in Captivity and in the Wild. Roth, T. L., Swanson, W. F., and Blattman, L. K. Seventh World Conference on Breeding Endangered Species: Linking Zoo and Field Research to Advance Conservation.  1999. Cincinnati, OH. 1999.
Ref Type: Conference Proceeding

Holt, W.V., Pickard, A.R., 1999. Role of reproductive technologies and genetic resource banks in animal conservation. Rev Reprod 4, 143-150.
Abstract: In combination with modem reproductive technologies, there is potential to use frozen and stored germplasm (genetic resource banks) to support conservation measures for the maintenance of genetic diversity in threatened species. However, turning this idea into reality is a complex process, requiring interdisciplinary collaboration and clearly defined goals. As the number of species deserving the attention of conservation scientists is overwhelmingly large, yet detailed knowledge of reproductive physiology is restricted to relatively few of them, choosing which species to conserve is one of the most difficult issues to be tackled. Besides the direct application of technologically advanced reproductive procedures, modern approaches to non-invasive endocrine monitoring play an important role in optimizing the success of natural breeding programmes. Through the analysis of urine and faecal samples, this type of technology provides invaluable management information about the reproductive status of diverse species. For example, it is possible to diagnose pregnancy and monitor oestrous cycles in elephants and rhinos without causing stress through restraint for sample collection. In this review, we identify the potential contribution of reproductive biology and genetic resource banks to animal conservation, but also highlight the complexity of issues determining the extent to which this potential can be achieved.

Lange, A., Hildebrandt, T.B., Strauss, G., Czupalla, O., Göritz, F., Schaftenaar, W., Schmitt, D.L. Feasibilities and limits of obstetrics in elephants. Verh ber Erkg Zootiere.  47-57. 1999.
Ref Type: Conference Proceeding

Mikota, S.K., 1999. Diseases of the Elephant: A Review. Verh. ber. Erkrg. Zootiere 39, 1-15.

Poole, J.H., 1999. Signals and assessment in African elephants: evidence from playback experiments. Animal Behaviour 58, 185-193.
Abstract: A series of playback experiments using two elephant vocalizations, the 'musth rumble' and the 'oestrous call', was carried out in Amboseli National Park to examine signaling and assessment in African elephants, Loxodonta africana. In response to the musth rumble of a high-ranking male other musth males approached the speaker aggressively, whereas nonmusth males walked away from the stimulus. The call of an oestrous female, too, attracted musth males who approached the speaker rapidly, while nonmusth males listened and then walked away. Females listened and often showed considerable interest in the musth rumbles of males, approaching the speaker and sometimes responding by vocalizing and or secreting from the temporal glands. The experiments bear out earlier observational data and game theory predictions which suggest that by being in or out of musth a male may be conveying information about the relative value he places on contesting his dominance rank and his access to oestrous females. When not visibly in musth, a male may be indicating his intention not to contest access to oestrous females.

Rasmussen, L.E.L., 1999. Evolution of chemical signals in the Asian elephant, Elephas maximus: behavioural and ecological influences. Journal of Biosciences 24, 241-251.
Abstract: In antiquity, the Asian elephant, Elephas maximus, gradually spread southward and eastward to become a successfully surviving, ecologically dominant megaherbivore in the tropical environment of south-east Asia. The changing physical environment forced dynamic fluxes in its social structure and altered its metabolism. Such events shaped the production and ultimately the stability of certain chemicals released by body effluvia. Some of these chemicals took on significance as chemical signals and/or pheromones. This article demonstrates by experimental and observational evidence, and hypothesizes based on speculative reasoning, how and why specific chemical signals evolved in the modern Asian elephant. Evidence, including the functional criteria required by elephant social structure and ecology, is presented for the hypothesis that the recently identified female-emitted, male-received sex pheromone, (Z)-7-dodecenyl acetate evolved first as a chemical signal. Subsequently, the cohesiveness and harmony of small, matriarchally-led female groups were strengthened by a female-to-female chemical signal, recently defined behaviourally. The looser societal structure of freer, roaming males also became bounded by chemical signals; for the males, breath and temporal gland emissions, as well as urinary ones function in chemical signaling. Basic knowledge about elephant chemical signals is now linking chemical information to behaviour and beginning to demonstrate how these signals affect elephant social structure and enable the species to cope with environmental changes.

Rasmussen, L.E.L., Schulte, B.A., 1999. Ecological and biochemical constraints on pheromonal signaling systems in Asian elephants and their evolutionary implications. In: Johnston, R.E., Muller-Schwarze, D., Sorenson, P.W. (Eds.), Advances in Chemical Communication in Vertebrates 8. Kluwer/Academic/ Plenum Press, pp. 49-62.
Abstract: The Asian elephant is an unusual example of how intraspecies chemical communication helps maintain societal cohesiveness within familial and herd units. The amount of multi-directional chemical communication is surprising, because long-lived elephants have a highly organized society, are capable of trans-generational passage of information, possess a sophisticated vocalization system, and are capable of complex learning and tool use. This paper discusses the ecological, behavioral, and biochemical aspects of chemical signals in elephants from an evolutionary perspective. Diverse bodily emissions are utilized as intraspecies chemical signals (including pheromones), often with imposed biochemical constraints. In this chapter, chemosignals released from the temporal gland secretions and breath of male Asian elephants in musth and a urinary female-to-male preovulatory pheromone are utilized as examples of these concepts. Furthermore, specific behavioral and biochemical studies with (Z)-7-dodecenyl acetate (a female-to-male urinary sex pheromone) demonstrate that social context significantly influences responsivity (demonstrated by field studies in Myanmar) and that additional biochemical requirements, perhaps lipocalin-like proteins, may be required for full bioactivity. The remarkable convergent evolution of (Z)-7-dodecenyl acetate, both structurally and functionally, in elephants and Lepidoptera, allows the use in elephant studies of effective biochemical tools developed for insect investigations. This convergence of chemical signaling systems of elephants and insects has several interesting implications.

Rasmussen, L.E.L., Perrin, T.E., 1999. Physiological correlates of musth: lipid metabolites and chemical composition of exudates. Physiology and Behavior 67, 539-549.
Abstract: Physiological changes related to lipid metabolism, behaviour and chemicals released in body exudates were studied during musth in the Asian elephant (Elephas maximus) as a case study. During musth, changes in serum testosterone and triglyceride concentrations followed similar patterns, with the former increasing sooner than the latter. Deviant behaviour increased during changing androgen levels. The observed high concentrations of testosterone were positively and significantly correlated with increased triglycerides. Lipase activity elevated significantly immediately before and after musth. Blood pH increased significantly in alkalinity. Urine and temporal gland secretions released variable amounts of compounds, some of which may be chemical signals. During musth, temporal gland and urinary exudates demonstrated increased acetone and other ketones indicative of lipid metabolic alterations. Large quantities of nonmethane hydrocarbons, especially 2-butanone, were released from the seemingly dry orifice of the temporal gland before the start of over musth and before maximum blood elevations were observed; isoprene release was similar. However, maximal acetone levels occurred simultaneously in blood, temporal gland secretions, and urine. Metabolically, musth is a series of interwoven, changing stages of increasing and decreasing hormones and lipid-related constituents. Released chemicals can be quantitatively related to these internal physiological events; some observed behaviours appear to result from altered chemical signals.

Schmidt, M.J., 1999. Calving elephants (Normal). In: Fowler, M.E., Miller, R.E. (Eds.), Zoo and wild animal medicine: current therapy 4. W.B. Saunders, Philadelphia, PA,  USA, pp. 521-522.

Schulte, B.A., Rasmussen, L.E.L., 1999. Musth, sexual selection, testosterone and  metabolites. In: Johnston, R.E., Muller-Schwarze, D., Sorenson, P.W. (Eds.), Advances in Chemical Communication in Vertebrates 8. Kluwer Academic/Plenum Press, pp. 383-397.
Abstract: Musth is an annual, yet asynchronous, rut-like condition that is experienced by many adult African and Asian male elephants. Behaviorally, musth is characterized by heightened aggression, decreased feeding, urine dribbling, temporal gland secretion and enhanced sexual activity. Musth improves the access of a male to reproductively active females through increased mobility and a higher dominance ranking (intrasexual competition). Whether females prefer males in musth as mates is as yet uncertain (intersexual choice). Females can distinguish among the odors of males in musth and nonmusth. Although behavioral musth has been associated with greatly elevated plasma testosterone levels, a recent study in Sri Lanka shows that intensified aggressiveness follows maximal testosterone secretion and proposes that behavioral musth is a consequence of declining androgen levels. Our data from an Asian male elephant in North America suggest that either declining or rising serum testosterone may be related to "musth behaviors." Our report demonstrates that certain aspects of body physiology are greatly altered during musth. Rather than a single state, our data suggest that musth is an ever-changing condition with some typical stages. Specific chemical compounds released at different stages of musth may serve individually or in combination as honest signals of male condition.

Schulte, B.A., Rasmussen, L.E., 1999. Signal-receiver interplay in the communication of male condition by Asian elephants. Animal Behaviour 57, 1265-1274.
Abstract: Signal design and meaning are dependent on the condition of the sender and receiver as well as the response of the receiver. This study examined (1) whether female Asian elephants, Elephas maximus, can distinguish between a conspecific male in musth and nonmusth states using urinary signals, (2) how the oestrous condition of the female affects discrimination, and (3) correlation of female responses with the testosterone level of the male. Musth is a rut-like state displayed by healthy adult male elephants. Males in musth dominate nonmusth males and may be preferred by females as mates. Urine was collected from two captive male Asian elephants during nonmusth periods and from one of these males during times of musth. Samples of musth and nonmusth urine and control liquids were placed in an elephant enclosure weekly for 16 weeks, the length of a female oestrous cycle. Primary response behaviours were approach and four trunk-tip motions, namely sniff, check, place and flehmen. Musth urine consistently elicited greater responses than nonmusth and control samples. Females were more responsive during their follicular (sexually  receptive) than luteal (unreceptive) stages of oestrus. Furthermore, females appeared to be sensitive to the degree of musth as responses increased with rising serum testosterone levels of the male donor. Chemical signals from males are a likely source of honest signals related to status and reproductive condition. Female elephants appear capable of detecting differences in a male based upon urinary chemosignals.

Zecchini, A., 1999. Life and death of species. Reconstituted animals? Courrier de la Nature 177, 22-27.
Abstract: This article gives an account of a breeding project, started in 1921, to reconstitute the aurochs, which became extinct in 1627, from existing breeds of cattle, and of a project to reconstitute the quagga (Equus quagga), which became extinct in 1921, from crosses among zebra species. The possibility of reconstituting the mammoth by obtaining semen from a mammoth preserved in the permafrost of Siberia and using it to fertilize elephant ova, followed by repeated backcrossing of hybrids to the mammoth, using mammoth semen, is discussed.

Ball, R.L. Carbon monoxide and carboxyhemoglobin in captive Asian (Elephas maximus) and African (Loxodonta africana) elephants. 1998 Proceedings AAZV and AAWV Joint Conference.  506-507. 1998. 1998.
Ref Type: Conference Proceeding

Butler, V., 1998. Elephants: trimming the herd. In South Africa, biologists are experimenting with new birth control methods designed to reduce elephant populations humanely. BioScience 48, 76-81.

Carden, M., Schmitt, D., Tomasi, T., Bradford, J., Moll, D., Brown, J., 1998. Utility of serum progesterone and prolactin analysis for assessing reproductive status in the Asian elephant (Elephas maximus). Animal Reproduction Science 53, 133-142.
Abstract: Concentrations of serum progesterone and prolactin were measured in blood samples collected during the perioestrous period and throughout gestation in captive Asian elephants. In >95% of perioestrous periods (n = 35), behavioural oestrus (as determined by bull interest, mounting and/or mating) coincided with the onset of increased serum progesterone concentrations at the beginning of the luteal phase and continued through day 7 (day 1 = first significant serum progesterone rise). A transient decrease (P<0.05) in serum progesterone, lasting 1-2 days, occurred between days 2 and 9. No sexual behaviour was observed in any female after this transient fall in progesterone. Prolactin concentrations fluctuated randomly throughout the perioestrous period, with no clear pattern. During the study, 4 females conceived (1 conceived twice), and delivered 3 viable offspring. Serum progesterone concentration was above baseline throughout gestation, and declined rapidly 2-3 days before parturition. Serum prolactin concentrations were higher than baseline values (P<0.05) after 5-6 months of gestation and remained high until after parturition. It is suggested that progesterone and prolactin concentrations could be used to detect oestrus, pregnancy and impending parturition.

Gilmore, J.A., McGann, L.E., Ashworth, E., Acker, J.P., Raath, J.P., Bush, M., Critser, J.K., 1998. Fundamental cryobiology of selected African mammalian spermatozoa and its role in biodiversity preservation through the development of genome resource banking. Anim Reprod Sci 53, 277-197.
Abstract: Fundamental cryobiological characteristics of spermatozoa from threatened or endangered species must be determined for successful cryopreservation techniques to be established. In this study, spermatozoa from four diverse species, impala (Aepyceros melampus), wart hog (Phacochoerus aethiopicus), elephant (Loxodonta africana), and lion (Panthera leo), were collected by electroejaculation or epididymal aspiration. Spermatozoal plasma membrane permeability to water (hydraulic conductivity, Lp) and the osmotically inactive fraction of the sperm cell (Vb) were determined from each species. Changes in cell volume were measured over time using an electronic particle counter. A Kedem-Katchalsky membrane transport model was used to theoretically characterize the data to determine Lp and Vb for each species. In addition to determining plasma membrane characteristics, spermatozoa were also studied to determine their sensitivity to low temperatures and to permeating cryoprotectant solutes. Cells maintained at room temperature (20-22 degrees C) were slowly or rapidly exposed to cold temperatures (1-4 degrees C), and percent motility was estimated to determine the sensitivity of the cells to cooling. Spermatozoa were also in media containing 1 M glycerol, dimethyl sulfoxide or ethylene glycol, and percent motility was measured at 15, 30 and 60 min intervals to determine the sensitivity of the cells to the cryoprotectant agent over time. Results indicate that sperm motility is significantly effected by decreased temperatures and the presence of cryoprotectant agents.

Greyling, M.D., Ford, M., Potgieter, H.C., van Aarde, R.J., 1998. Influence of gestation on uterine endometrial steroid receptor concentrations in the African elephant, Loxodonta africana. Biol Reprod 58, 60-64.
Abstract: The modulatory effects of gestational age and circulating concentrations of progesterone, 5alpha-pregnane-3,20-dione, and estradiol-17beta on the uterine sex steroid hormone receptor levels of the African elephant were investigated. Uterine tissue biopsies and blood samples were obtained from animals culled in the Kruger National Park. Estrogen and progesterone receptor concentrations were determined in uterine biopsies from subadult, lactating, early-, mid-, and late-pregnant elephants, by equilibrium binding assays. Circulating estradiol-17beta and progesterone concentrations were measured by means of RIAs, while plasma concentrations of 5alpha-pregnane-3,20-dione were determined with an amplified ELISA. Significant inverse correlations of the concentrations of estrogen and progesterone receptors with the gestational stage of the elephants were observed. Pregnant uterine horns of individual animals contained lower levels of estrogen and progesterone receptors than the nonpregnant horns of the same animals. A strong positive correlation existed between uterine estrogen and progesterone receptors levels. Circulating concentrations of 5alpha-pregnane-3,20-dione and progesterone decreased with an increase in the concentrations of progesterone receptors as well as with fetal age. We conclude that the progesterone receptor concentrations are down-regulated with progressing gestation in the African elephant. This down-regulation appears to be linked to an increase in circulatory 5alpha-pregnane-3,20-dione concentration in the plasma of pregnant animals.

Gunasena, K.T., Lakey, J.R., Villines, P.M., Bush, M., Raath, C., Critser, E.S., McGann, L.E., Critser, J.K., 1998. Antral follicles develop in xenografted cryopreserved African elephant (Loxodonta africana) ovarian tissue. Anim Reprod Sci 53, 265-275.
Abstract: The preservation of germ plasm from endangered species could augment captive breeding programs aimed at maintaining genetic diversity. Mammalian female germ plasm (oocytes) is extremely difficult to collect and cryopreserve; however, a promising alternative is the cryopreservation of ovarian tissue. In the present study, athymic nude (nu/nu) Balb/C mice were used to evaluate in vivo viability of cryopreserved ovarian tissue from Institute of Cancer Research genotype (ICR) mice or elephants. Female mice were ovariectomized prior to transplant of cryopreserved-thawed ovarian tissue from ICR mice (n=4) or elephants (n=6). Control mice were sham operated (n=4) or ovariectomized (n=5). Transplants were in the ovarian bursa, enabling in vivo ovulation and pregnancies from allografts. Vaginal cytology was monitored daily, and the intervals between andduration of epithelial cells present in smears were evaluated. Appearance of epithelial cells in sham-operated and allografted mice were at intervals of 4.3+/-0.6 and 3.3+/-0.5 days, lasting for 1.4+/-0.1 and 1.6+/-0.2 days, respectively. Sporadic incidence of epithelial cells in ovariectomized animals occurred at longer intervals (8.6+/-3.8 days). Females with xenografted elephant ovarian tissue demonstrated epithelial cells in vaginal smears at intervals of 4.5+/-1.0 days, for 2.5+/-0.5 days duration, which was significantly longer than the other groups (P < 0.05). Histological evaluation of tissues at the time of epithelial cells in smears demonstrated well-developed antral follicles, although oocytes were of poor morphological appearance or only cumulus-like complexes were seen. The nude mouse model is effective for assessing cryopreserved ovarian tissue xenograft function which can support the development of antral follicles.

Hildebrandt, T., Goritz, F., Pratt, N.C., Schmitt, D., Quandt, S., Raath, J.P., Hofmann, R.R., 1998. Reproductive assessment of male elephants (Loxodonta africana and Elephas maximus) by ultrasonography. Journal of Zoo and Wildlife Medicine 29, 114-128.
Abstract: Transrectal ultrasonography was performed on five wild and two captive male African elephants (Loxodonta africana) and four captive male Asian elephants (Elephas maximus) to develop standards for assessment of reproductive health and status.  The entire internal urogenital tract was visualized ultrasonographically by using a 3.5 MHz or a 7.5 MHz transducer in combination with a probe extension adapted for elephant anatomy.  The findings were verified by postmortem ex situ ultrasound examinations in several individuals of each species.  Each part of the internal urogenital tract was sonographically detectable except for the bulbourethral glands and the cranial portion of the ureters and ductus deferentes, which were only observed in situ in the neonate.  Each structure visualized was measured and described.  The size and morphology of the urogenital structures, especially the accessory glands, were indicative of breeding status, if known.  There was a notable difference between African and Asian males in the size and morphology of the prostate gland and a slight difference in the shape of the ampullae.  No other structure showed significant species differences.  The detection of the location and description of the testes may provide information for modifying present castration procedures.  Furthermore, ultrasound examination of the male accessory glands may aid in the identification of potential semen donors for assisted reproduction programs in captive elephants.

Hodges, J.K., 1998. Endocrinology of the ovarian cycle and pregnancy in the Asian (Elephas maximus) and African (Loxodonta africana) elephant. Animal Reproduction Science 53, 3-18.
Abstract: In reviewing the literature, this paper assesses the current level of understanding of the hormonal control of the ovarian cycle and pregnancy in the
elephant. Data are compiled from two genera, Elephas maximus and Loxodonta africana, recognizing differences where known to exist. Measurements of various steroid and peptide hormones, but most notably immunoreactive progesterone (iP), indicate an overall cycle length of 15-16 weeks, comprising an 8-11-week luteal phase and a shorter inter-luteal (follicular) period of 4-6 weeks. Oestrogen related phenomena suggest the occurrence of two (possibly more) waves of follicular development with a 3-week periodicity, although oestrogen levels are low and fluctuate without clear pattern. The inter-luteal period is characterized by two discrete LH peaks also 3 weeks apart, but only the second is associated with an iP rise. Ovulation rate is variable and additional luteal structures lacking ovulation points are probably formed each cycle. The post-ovulatory period is associated with elevated progestogen levels, mainly 5alpha-reduced compounds, while progesterone itself is a minor secretory product. A revised model for the elephant ovarian cycle is presented. Detailed information on the endocrinology of pregnancy is lacking. Elevated progestins beyond the normal luteal phase indicate that conception extends the functional lifespan of the CL, with maximum steroidogenic activity between 3-15 months. The nature of the luteotrophic support is not known and placental gonadotrophins have not been conclusively demonstrated. Progestins fluctuate at or slightly above luteal phase values throughout pregnancy. There is a marked increase in prolactin levels after 16-24 weeks and a relationship with oestrogen secretion may exist since circulating and urinary levels show a progressive increase from 6-8 months. A placental contribution to progestin secretion is likely, although direct evidence is lacking. Considerable enlargement of foetal gonads during the second half of pregnancy in African elephants suggests a possible steroidogenic function, but none has been defined. The endocrine events leading to parturition are unknown. In the Asian elephant, a fall in iP precedes parturition; oestrogen levels decline and cortisol increases at the time of birth. The paper concludes with a brief selection of priorities for future research.

Jewgenow, K., Meyer, H.H., 1998. Comparative binding affinity study of progestins to the cytosol progestin receptor of endometrium in different mammals. Gen Comp Endocrinol 110, 118-124.
Abstract: The relative binding affinity of 5 alpha-reduced progestins and a newly synthesized antiprogestin J912 (progesterone 100%) was determined in a competitive receptor binding assay using [3H]ORG-2058 as radiolabeled ligand for the progestin receptor. Uteri obtained from 12 different species of four mammalian orders were examined. The relative binding affinity of 75-100% and a blood prevalence of 5 alpha-pregnane-3,20-dione in horses and African elephants suggest a biological role of this particular 5 alpha-reduced progesterone. For pigs the binding affinity of 5 alpha-pregnane-3,20-dione was about 50% of progesterone, but blood levels are unknown. In all other cases the low binding affinity of investigated progestins precludes possible biological role. For 5 alpha-pregnane-3 alpha-ol-20-one, 5 alpha-pregnane-20 alpha-ol-3-one, and 5 alpha-pregnane-3 beta,20 alpha-diol the relative binding affinity was less than 1%. A rather low binding (< 15%) was observed in 5 alpha-pregnane-3,20-dione in all ruminant species investigated. The antiprogestin J912 was found to be highly efficient in displacing progesterone from its endometrial binding sites in carnivores and might therefore be used for pregnancy interruption during diapause in certain species, e.g., in captive bears.

Keele, M.N., 1998. North American Asian elephant birth statistics: What are the numbers telling us?  Journal of the Elephant Managers' Association 7, 29-32.

Kirkpatrick, J.F., Fayrer-Hosken, R., Grobler, D., Raath, C., Bertschinger, H., Turner, J.W., Liu, I.K.M. Immunocontraception of Free-Ranging African Elephants in Kruger National Park, South Africa. 1998 Proceedings AAZV and AAWV Joint Conference.  434-435. 1998. 1998.
Ref Type: Conference Proceeding
Abstract: In order to seek a more publicly acceptable alternative to the management of African elephants by culling, a test was conducted to determine if a porcine zona pellucida (PZP) vaccination could effectively contracept females of this species. Initially, ovaries were recovered from culled animals and slices were incubated with immunogold-labeled rabbit antibodies against PZP. Significant staining of the elephant zona suggested that PZP would be an effective contraceptive vaccine. In a second experiment, three captive female zoo elephants were inoculated with the PZP vaccine (400 ug PZP + 300 mg RIBI). These non-breeding animals were tractable and blood samples were recovered and assayed for anti-PZP antibodies. Antibody titers (1:500 dilution) peaked (0.75 - 1.3 OD) at 1-2 mo following the initial inoculation, declined to 0.1 - 0.34 at 6 mo - 1 yr, and peaked again following a third inoculation (0.8 - 2.3).  These data indicated that African elephants would mount a significant antibody response to the PZP vaccine and together with the histochemical study, suggested the vaccine would be a successful immunocontraceptive in this species.

Niemuller, C., Brown, J.L., Hodges, J.K., 1998. Reproduction in elephants. In: Knobil, E., Neill, J. (Eds.), Encyclopedia of reproduction, Vol. 1. Academic Press, New York, NY. USA, pp. 1018-1029.

Niemuller, C.A., Gray, C., Cummings, E., Liptrap, R.M., 1998. Plasma concentrations of immunoreactive relaxin activity and progesterone in the pregnant Asian elephant (Elephas maximus). Animal Reproduction Science 53, 119-131.
Abstract: Immunoreactive relaxin activity and progesterone concentrations in weekly plasma samples were measured throughout 6 pregnancies in 5 Asian elephants maintained at the African Lion Safari in Ontario. One animal aborted, one delivered a stillborn calf at term and the remaining pregnancies produced living calves. Duration of pregnancy ranged from 89 to 95 weeks. Immunoreactive relaxin activity increased by week 20 of pregnancy to reach a mean peak concentration of >6 ng/ml in the 2nd trimester. This was followed by a slow decline beginning approximately 30 weeks before parturition. A smaller, secondary relaxin peak was observed during the final 8 weeks before parturition and circulating concentrations remained above the prepregnancy basal value for 72 h after birth. Progesterone concentration increased immediately after mating and rose to a maximum of >2 ng/ml by midgestation. As with relaxin, progesterone concentrations began to decline gradually approximately 30 weeks before birth. More frequently collected samples before, during and after parturition revealed a decline to the prepregnancy level between 2 and 5 days before parturition, in contrast to an elevated relaxin level (1 ng/ml) during this time. The aborted pregnancy at week 35 was accompanied by a sudden decline in immunoreactive relaxin and progesterone concentrations 1 week before the visible termination of the pregnancy. Delivery of the stillborn calf occurred 5 weeks after immunoreactive relaxin and progesterone concentrations had declined to the prepregnancy value.

Rasmussen, B., 1998. The Chemical Identification of a Preovulatory Pheromone: A Reproductive Chemosignal from Female to Male Asian Elephants. Journal of the Elephant Managers Association 7, 52-56.

Rasmussen, L.E.L., Schulte, B.A., 1998. Chemical signals in the reproduction of Asian and African elephants. Animal Reproduction Science 53, 34.

Rasmussen, L.E.L., Lazar, J., Greenwood, D., Feng, L., Prestwich, G.D., 1998. Initial characterizations of secreted proteins from Asian elephants that bind the sex pheromone, (Z)-7- dodecenyl acetate. Chemical Senses 23, 591.

Rasmussen, L.E.L., 1998. Chemical communication: An integral part of functional Asian elephant (Elephas maximus) society. Ecoscience 5, 410-426.
Abstract: The matriarchally organized Asian elephant society is characterized by long-term stability and continuity. Flux within this society results from changing ecological conditions and the dynamics of its population. Its structure is influenced by age composition and physiological states within the female herd and by impinging influences of the peripheral males, especially during reproductive times. Recent behavioral studies of captive populations have substantiated older field studies and have demonstrated that chemical signals play a significant role in elephant society. Chemical investigations, based on previously substantiated behavioral interactions, have identified specific compounds or combinations of compounds in elephant emissions (especially urine, temporal gland secretions and breath) that retain bioactivity throughout chemical extractions and playback experiments, based on behavioral and/or chemosensory responses. Chemosensory neuroreceptive systems in Asian elephants are reviewed, as well as behavioral and chemosensory effects of whole exudate chemical signals on lifestyles, especially related to mating. Several discrete and composite chemical signals have been deciphered in elephants, one of which is a preovulatory female-to-male pheromone, (Z)-7-dodecen-1-yl acetate. This pheromone and other recently described or potential chemical signals are compared to compounds in insect pheromone blends. Such knowledge of the chemical ecology of the Asian elephant has potentially important implications for conservation.

Rasmussen, L.E.L., Schulte, B.A., 1998. Chemical signals in the reproduction of Asian (Elephas maximus) and African (Loxodonta africana) elephants. Animal Reproduction Science 53, 19-34.
Abstract: Asian (Elephas maximus) and African (Loxodonta africana) elephants exhibit polygynous mating that involves female choice of mates and male-male competition for access to females. Chemical signals mediate intersexual and intrasexual interactions associated with reproduction. The need for reliable and honest signals is accentuated by the markedly different social structure of adult males and females. Adult female elephants live in matriarchal herds consisting of a dominant female and several generations of offspring. Adult males are solitary or travel with other males except during breeding periods. Because females have a long 16-week oestrous cycle with a brief 1-week receptive period and a 4-5 year interval between births, a sexually active female is a limited resource. Asian elephant females advertise a forthcoming ovulation by releasing (Z)-7-dodecen-1-yl acetate in their urine during the preovulatory period. African elephants probably produce a sex pheromone as well. Females regularly contact the ano-genital region of other females and show heightened chemosensory responsiveness to urine during the follicular phase. The physiological impacts of this ability to detect reproductive condition (e.g. possible synchronizing or suppressing of oestrus) are uncertain. Males experience an annual period of heightened aggressiveness and highly elevated testosterone concentrations known as musth. Males secrete fluid copiously from their temporal gland and dribble strongly odoriferous urine during musth. Females appear to prefer musth males as mates, and captive Asian females exhibit greater chemosensory responses to urine from males in musth than not. Males in musth are competitively dominant to all other males, even those larger than themselves. Nonmusth males avoid males in musth, and captive Asian bulls show greater interest in musth than nonmusth urine. In captivity subordinate Asian females back away from musth secretions, and females with calves sometimes display protective behaviour. Clearly, chemical signals play an important role in communication by elephants between and within the sexes. Further work is needed to identify more of these chemical messengers and to understand their complete function in mediating reproductive interactions in the elephant social system.

Sai, D.J., Chen, Z.B., Zhang, J.Z., 1998. Study on the courtship-mating behavior of Asian elephant. Chinese Journal of Zoology 33, 28-31.

Schmitt, D.L. Report of a successful artificial insemination in an Asian elephant. Proc 3^rd International Elephant Research Symposium.  1998.
Ref Type: Conference Proceeding

Schmitt, D.L., Hildebrandt, T.B., 1998. Manual collection and characterization of semen from Asian elephants (Elephas maximus). Animal Reproduction Science 53, 309-314.
Abstract: The implications of collecting semen from elephants for use in artificial insemination programs are profound in the context of propagating captive elephants. Using a manual manipulation technique, semen was collected and characterized from five adult Asian elephants (Elephas maximus) and ejaculate fluid was obtained from one castrated elephant bull. The penis was stimulated to protrusion and erection by rectal massage of the pelvic portion of the urethra. During an ejaculatory response, massage was also directed onto the area of the ampulla of the ductus deferens. Sperm rich ejaculates were usually collected as a result of each ejaculatory contraction. Ejaculates were evaluated for spermatozoal concentration and pH (when possible) and sperm rich fractions combined for determination of total volume. Mean total volume of each collection was 27.5+/-4.4 ml. Mean concentration of the first and second ejaculatory responses from a collection was 2.05+/-0.17 x 10(9) and 1.34+/-0.19 x 10(9) sperm/ml, respectively. Measurement of seminal pH revealed no significant differences between the fractions. Mean pH of the first and second ejaculatory responses were 7.05+/-0.07 and 7.04+/-0.13. This method of collecting elephant sperm can be utilized for semen evaluation of bulls of unknown reproductive status in conjunction with other evaluation techniques (i.e. ultrasonographic, endocrinologic). It also has the potential for providing valuable genetic material for genome resource banks and for use with assisted reproductive techniques like artificial insemination.

Taylor, V.J., Poole, T.B., 1998. Captive breeding and infant mortality in Asian elephants:  a comparison between twenty Western zoos and three Eastern elephant centers. Zoo Biology 17, 311-332.
Abstract: A questionnaire was designed to assess the importance of reproductive behaviour and husbandry factors on breeding success in captive Asian elephants (Elephas maximus). This was circulated to zoos in Europe and North America in 1996. Data from 20 zoos were analysed. Data were also obtained from 3 elephant centres in Asia (Pinnawela Elephant Orphanage in Sri Lanka, Myanma Timber Enterprise in Myanmar and the Tamilnadu Forest Department in India). The aims were to compare Asian elephant breeding success, establish possible causes for any differences, and make recommendations for improving the welfare and breeding success of the animals. Breeding success in most of the zoos was notably lower and the percentages of stillbirths and infant mortality were relatively higher when compared with those of the centres in Asia. Female elephants in zoos appeared to reach sexual maturity and reproduce earlier than those in the Asian establishments. However, zoo elephants produced fewer young per female. The different facilities and husbandry methods used are described. Recommendations are made for both short- and long-term changes that could be used to modify existing practices to improve the welfare and breeding success of captive Asian elephants.

Welsch, U., Feuerhake, F., van Aarde, R., Buchheim, W., Patton, S., 1998. Histo- and cytophysiology of the lactating mammary gland of the African elephant(Loxodonta africana). Cell Tissue Res 294, 485-501.
Abstract: The lactating mammary gland of the African elephant (Loxodonta africana) has been studied with a panel of morphological techniques focusing on (1) the functional changes during the secretory process, (2) proliferative process [by application of proliferating cell nuclear antigen (PCNA) immunohistochemistry] and apoptotic phenomena [by use of the TUNEL technique] in the individual lobules, and (3) components of milk and milk-fat-globule membrane. In the lactating gland, the lobules are variably differentiated; within a lobule, however, the alveoli are usually similarly differentiated. The morphology of their alveoli suggests a classification of the lobules into types 1-3. Lobules of type 1 are composed of immature tubular alveoli with mitotic figures and numerous PCNA-positive nuclei; advanced type 1 alveoli contain abundant glycogen and specific secretory granules. Lobules of type 2 are further subdivided. In type 2a lobules, the epithelial cells of the alveoli form tall apical protrusions, which in part are occupied by small lipid droplets and which are pinched off in an apocrine fashion. The number of lysosomes varies considerably. Type 2b is the most common type, with striking basal membrane foldings, abundant rough endoplasmic reticulum cisterns, large Golgi apparatus, numerous mitochondria, lipid droplets, and protein vesicles with 30- to 90-nm-wide casein micelles. The lipid droplets are pinched off with minimal amounts of cytoplasm. Type 2c is composed of alveoli with a cuboidal epithelium and few signs of secretory activity. Increasing expression of peanut-agglutinin-binding sites parallels the maturation and differentiation of the glandular cells. Type 3 lobules are marked by numerous TUNEL-positive nuclei and large lipid droplets and are apparently degenerating structures. Cytokeratin (CK) 14 is usually present in the myoepithelial cells; CK 19 and CK 7 mark ductal and immature alveolar epithelia. Milk protein content varies between 2.6% and 6.3%, and casein micelles range from 35 to 90 nm in diameter. The diameter of intra-alveolar milk fat globules ranges from 5 to 25 micrometer and the membranes bear a filamentous surface coat composed of membrane-anchored mucins; gel-electrophoretic analysis of these mucins from different individuals demonstrates the presence of mucin MUC 1, which is expressed with considerable genetic heterogeneity.

Whyte, I., van Aarde, R., Pimm, S.L., 1998. Managing the elephants of Kruger National Park. Animal Conservation 1, 77-83.
Abstract: The elephant population in Kruger National Park, Republic of South Africa, is growing rapidly. To prevent damage to the Park's ecosystems, the management has culled about 7% of the population annually. Such culls are very controversial. At first glance, contraceptives seem an attractive alternative means of control. We examine contraception as a management option, review the relevant aspects of elephant reproduction, physiology and demography and conclude that this optimism is probably misplaced. First, contraceptives have a wide range of physiological and behavioural side-effects that may prove to be damaging to the individual female and those around her. Second, the elephants in the Park have near-maximal growth rates with inter-calving intervals of less than four years. To achieve zero population growth, about three-quarters of the adult female elephants would need to be on contraceptives. There are no simple alternatives. The smallest numerical target for controlling population numbers is to kill or sterilize females about to become pregnant for the first time. Such a solution is unlikely to appease those who consider killing elephants to be unethical. It may, however, be the one closest to the natural patterns of elephant mortality.

Brown, J.L., Lehnhardt, J., 1997. Secretory patterns of serum prolactin in Asian (Elephas maximus) and African (Loxodonta africana) elephants during different reproductive states:  comparison with concentrations in a noncycling African elephant.  Zoo Biology 16, 149-159.
Abstract: Serum prolactin was quantified in adult female Asian (Elephas maximus) and African (Loxodonta africana) elephants during various reproductive states and the profiles compared to that in a noncycling African elephant.  In reproductively normal elephants, there was no effect of season, estrous cycle stage, or lactational status on quantitative or qualitative prolactin secretion (P > 0.05), nor were there any differences (P > 0.05) in overall prolactin concentrations between species.  In pregnant elephants, prolactin concentrations remained at baseline for the first 4-6 months of gestation. Thereafter, concentrations during early pregnancy averaged ~four-fold higher than those during the estrous cycling, increasing to ~100-fold over baseline during mid- to late gestation in both species.  In contrast to cycling elephants, prolactin concentrations in an African elephant exhibiting chronic anovulation (on the basis of an acyclic serum progesterone profile) and mild galactorrhea were consistently about five-fold higher (P < 0.05) suggesting she is hyperprolatinemic.  Other endocrinological assessments confirmed the hypogonadal state of this female.  Serum estradiol concentrations were consistently at or below dectectable levels.  Additionally, no preovulatory leutinizing hormone (LH) surges occured in daily serum samples analyzed over a 12-month period.  the pituitary was not totally, refractory, however, and responded with a several-fold increase in serum LH concentration (peak, 3.07 ng/ml) over baseline (0.75 ng/ml) after i.v. injection of gonadotropin-releasing hormone.  this study describes normal baseline serum prolactin values for Asian and African elephants and is the first to identify hyperprolactinemia as a possible cause of reproductive acyclicity and galactorrhea in and African elephant.

Dickerman, R.D., Zachariah, N.Y., Fouraker, M., McConathy, W.J., 1997. Neuroendocrine-associated behavioral patterns in the male Asian elephant (Elephas maximus). Physiology and Behavior 61, 771-773.
Abstract: Steroid-responsive behaviors have been reported in various species; however, the reports thus far on the male Asian elephant (bull) during musth are few in number and most have been conducted on single captive animals for short time periods. The purpose of this investigation was to perform a longitudinal study on steroid-responsive behavior in 3 male Asian elephants from a captive herd of 11 male Asian elephants in Nepal. Male Asian elephants were 18, 25, and 43 years old. The animals had serum collected for 11 months and were observed on a daily basis for aggressive behavior according to the Species Survival Plan (SSP) collection protocol on SSP data sheets. Testosterone (T) and dihydrotestosterone (DHT) were measured in each animal by radioimmunoassay. Testosterone levels rose during musth 26-fold compared to nonmusth, and DHT was elevated 12-fold in musth. Maximal aggressive behavior episodes occurred during peak elevations of T and DHT, with correlation coefficients of 0.82 and 0.89, respectively. Therefore, we suggest that the aggressive episodes are dependent on elevated circulating androgens acting on androgen-responsive neural tissues.

Fayrer-Hosken, R.A., Brooks, P., Bertschinger, H.J., Kirkpatrick, J.F., Turner, J.W., Liu, I.K.M., 1997. Management of African elephant populations by immunocontraception. Wildlife Society Bulletin 25,  18-21.

Goyal, A.K., Rastogi, S.C., Nayak, A.K., Jain, V.K., 1997. Herbal oral contraceptives: retrospects and prospects. Advances in Plant Sciences 10, 141-143.
Abstract: The potential of herbs and animal matter for use as alternative oral contraceptives in India is discussed. Some non-conventional herbal contraceptives are identified, together with Lawsonia inermis, Butea monosperma and elephant fecal matter, which have recently been tested for their potency and require further chemical and biological analysis

Greyling, M.D., vanAarde, R.J., Potgieter, H.C., 1997. Ligand specificity of uterine oestrogen and progesterone receptors in the subadult African elephant, Loxodonta africana. Journal of Reproduction and Fertility 109, 199-204.
Abstract: The ligand specificity of progesterone and oestrogen receptors in the uteri of four nonpregnant, nonlactating African elephants, killed during routine culling in the Kruger National Park, were determined. The mean (+/-SEM) Kd values of the oestrogen (0.18 +/- 0.019 x 10(-9) mol l-1, n = 12) and progesterone (0.22 +/-0.025 x 10(-9) mol l-1, n = 12) receptors were essentially similar when [3H]promegestone was used as radioligand in the progesterone receptor assays. However, when [3H]progesterone was used as radioligand, the progesterone receptor exhibited a significantly higher Kd value (1.03 +/- 0.132 x 10(-9) mol l-1, n = 12) than that of the oestrogen receptor. The use of the different radioligands did not significantly affect the quantitative values obtained for the progesterone receptor. Both the oestrogen and the progesterone receptors displayed a high ligand specificity. The 5 alpha-reduced metabolites of progesterone exhibited a high relative binding affinity for the progesterone receptor (5 alpha-pregnane-3,20-dione: relative binding affinity = 43%; 5 alpha-pregnane-3 alpha-ol-20-one: relative binding affinity = 20%) but the synthetic antiprogestin RU 486 did not compete successfully with progesterone in competitive binding studies. However, norethindrone (relative binding affinity = 293%) competed successfully for binding to the progesterone receptor, and may have some potential in the future development of a technique to control reproductive output in the African elephant.

Heisterman, M., Trohorsch, B., Hodges, J.K., 1997. Assessment of ovarian function in the African elephant (Loxodonta africana) by measurement of 5-reduced progesterone metabolites in serum and urine.  Zoo Biology 16, 273-284.
Abstract: We have previously shown that 5-pregnane-3,20-dione (5-DHP) and 5-pregnane-3-ol-20-one (5-OH) are the major luteal and circulating progestins in the African elephant. Therefore, the aim of the present study was to determine 1) circulating levels and patterns of secretion of 5-DHP in relation to progesterone (P4) throughout the ovarian cycle, 2) the presence and relative abundance of 5-reduced progestins in urine and 3) whether their measurement in urine would provide a non-invasive method for monitoring luteal function.  Urine samples were collected weekly throughout a total of 13 ovarian cycles from 5 females.  In addition, matched blood samples were collected during 6 cycles from 2 of the 5 animals.  All hormone measurements were carried out by enzymeimmunoassay following extraction.  Urine was hydrolized prior to extraction.  Profiles of P4 and 5-DHP in serum followed a similar cyclic pattern and both measurements were significantly correlated (r = 0.78-0.98, mean 0.89, P < 0.001).  Concentrations of 5-DHP were, however 10-20-fold higher than those if P4.  In addition, 5-DHP measurements showed a more pronounced luteal phase increase than that of immunoreactive P4.  HPLC co-chromatography confirmed the presence of large amounts of 5-P-3-OH in urine as a single immunoreactive peak, whereas 5-DHP was present in very low levels and measurable only as one of several immunoreactive substances.  Measurements of urinary 5-P-3-OH were significantly correlated to serum 5 -DHP measurements in each of the 6 cycles (r = 0.72-0.93, mean 0.81, P < 0.001), whereas correlation coefficients between urinary and serum 5-DHP values were generally lower (r = 0.34-0.83, mean 0.69) and significant in only 4 of 6 cycles.  Accordingly, only urinary excretion of 5-P-3-OH, but not of 5-DHP, exhibited a clear cyclic pattern, with consistently low levels of 0.15-.020 g/mg Cr in the follicular phase and 10-fold elevated levels (1.8-2.2 g/mg Cr) in the luteal phase.  Based on the intervals between successive luteal phase increases in urinary 5-P-3-OH, a mean cycle length of 14.1+/- 1.8 weeks, comprising a follicular phase of 5.0 +/- 0.9 weeks and a luteal phase of 9.10+/-01.4 weeks was determined for the 13 cycles studied.  The results indicate that measurements of 5-P-3-OH in urine provide a reliable non-invasive method for monitoring luteal function in the African elephant.

Hildebrandt, T.B., Göritz, F., Pratt, N.C., Schmitt, D.L., Lehnhardt, J., Hermes, R., Quandt, S., Raath, J., West, G., Montali, R.J. Assessment of health and reproductive status in African elephants by transrectal ultrasonography. Proc: Am Assoc Zoo Vet Ann Conf.  207-211. 1997.
Ref Type: Conference Proceeding

Hodges, J.K., Heisterman, M., Beard, A., van Aarde, R.J., 1997. Concentrations of progesterone and the 5-reduced progestins, 5-pregnane-3,20-dione, and 3--hydroxy-5-pregnan-20-one, in luteal tissue and circulating blood and their relationship to luteal function in the African elephant (Loxodonta africana). Biology of Reproduction 56, 640-646.
Abstract: The 5 alpha-reduced metabolites 5 alpha-pregnane-3,20-dione (5 alpha-DHP) and 3 alpha-hydroxy-5 alpha-pregnan-20-one (5 alpha-P-3 alpha-OH) are the principal progestins biosynthesized by the African elephant corpus luteum. The aim of the present study was to determine luteal and circulating concentrations of these 5 alpha-reduced progestins in relation to progesterone (P4) and to examine whether their measurement reflects corpus luteum function. Ovarian (luteal) tissue (30 corpora lutea and 3 corpora rubra from 8 animals) and plasma samples (30 animals) were collected from pregnant and nonpregnant adult elephants shot in the Kruger National Park. Specific immunological measurement for both 5 alpha-reduced progestins and P4 was achieved by enzymeimmunoassay of tissue and plasma extracts following purification by HPLC. Mean (+/- SEM) luteal concentrations of 5 alpha-DHP and 5 alpha-P-3 alpha-OH were 79.5 +/- 9.4 micrograms/g and 196.5 +/- 24.8 micrograms/g, respectively, approximately 2-3 orders of magnitude higher than those of P4 (mean +/- SEM, 0.16 +/- 0.01 microgram/g). Whereas 5 alpha-reduced progestin concentrations tended to be lower in corpora lutea from late pregnancy compared with earlier stages and were lowest in corpora rubra, P4 levels were similar in all tissues/stages examined. The 5 alpha-reduced progestins also predominated over P4 in plasma (mean 5 alpha-DHP:P4 and 5 alpha-P-3 alpha-OH:P4 ratios 20.3 and 13.4, respectively). Similar to results for luteal tissue, plasma concentrations of 5 alpha-reduced progestins, but not of P4, were lower in late pregnancy than in earlier gestation stages and in nonpregnant animals. Moreover, plasma levels of both 5 alpha-reduced metabolites were negatively correlated with gestation age, whereas those of P4 were not. Levels of 5 alpha-reduced metabolites (without chromatography) were also measured in weekly blood samples throughout two complete ovarian cycles in one captive female. Both measurements showed a cyclic profile (similar to that of P4) with a luteal-phase elevation of 10- to 15-fold. The results indicate that 5 alpha-reduced compounds are the predominant progestins contained within and secreted by the corpus luteum of the African elephant, both during the ovarian cycle and throughout pregnancy. They also provide preliminary evidence to suggest that measurements of 5 alpha-reduced metabolites may reflect corpus luteum function more closely than those of P4.

Kurt, F. Captive breeding of the Asian elephant (Elephas maximus) in Asian and in western zoos - A comparison of different managment systems. International Seminar on the Conservation of Asian Elephants ( Myanmar).  1997.
Ref Type: Conference Proceeding

Mar, D.K. Reproductive parameters of Myanmar cow elephants based on 500 calvings. VII International Seminar on Conservation of Asian Elephants.  1997.
Ref Type: Conference Proceeding

Meyer, H.H.D., Jewgenow, K., Hodges, J.K., 1997. Binding activity of 5-reduced gestagens to the progestin receptor from African elephant (Loxodonta africana). General and Comparative Endocrinology 105, 164-167.
Abstract: Recent findings in the African elephant (Loxodonta africana) indicate that the major progestins contained within and biosynthesized by corpora lutea are 5alpha-reduced metabolites and that progesterone is quantitatively of minor importance. The specific gestagenic action within the reproductive tract of elephants was determined by measurement of relative binding affinity of the respective progestins to the gestagen receptor extracted from elephant endometrium. The cytosol was incubated with 40 nmol/liter [3H]ORG-2058 and increasing concentrations of the tested progestin. Progesterone (P4), 5alpha-pregnane-3,20-dione (DHP), and 5alpha-pregnane-3alpha-ol-20-one (5alpha-P-3OH) were used. The competition for binding sites on the progestin receptor was shown by decreasing counts measured after extraction with scintillation fluid. The progestin concentration which induced a 50% reduction of measured counts was estimated (C50) and relative binding affinity of progesterone to other progestins was calculated (RBA = C50progestin/C50p4). The relative binding affinity of DHP to P4 at the gestagen receptor of elephant endometrium was equivalent. The other 5alpha-reduced progestin (5alpha-P-3OH) showed no competition to the [3H]ORG-2058 receptor binding. We conclude that the biological significance of P4 and DHP at the receptor level is very similar. The higher quantitative levels of DHP in corpus luteum and serum support the hypothesis that this progestin is the major gestagen in the elephants, whereas 5alpha-P-3OH is an inactive metabolite.

Montali, R.J., Hildebrandt, T., Goritz, F., Hermes, R., Ippen, R., Ramsay, E.C., 1997. Ultrasonography and pathology of genital tract leiomyomas in captive Asian elephants: implications for reproductive soundness.  Verh. ber. Erkrg. Zootiere 38, 199-204.

Niemuller, C., Shaw, H.J., Hodges, J.K., 1997. Pregnancy determination in the Asian elephant (Elephas maximus):  A change in the plasma progesterone to 17 hydroxyprogesterone ratio. Zoo Biology 16, 415-426.
Abstract: The measurement of circulating progesterone (P) is widely used to confirm and monitor pregnancy in the captive Asian elephant.  Consistently elevated progesterone (P) concentrations for a minimum of 12 weeks can be used as a positive  indication of pregnancy, although two instances of prolonged P secretion during the cycle have been recorded [Rubel, 1987; Olsen et al, 1994].  Previously, we demonstrated that pregnanetriol was the major urinary gestagen metabolite enabling the noninvasive monitoring of the reproductive cycle of the Asian elephant [Niemuller et al., 1993] as well as pregnancy [unpublished data].  the importance of this unusual urinary metabolite triggered an investigation into the secretion of circulating concentrations of 17 hydroxyprogesterone (17 OHP) during pregnancy and reproductive cycles, as this steroid is the only steroid precursor of pregnanetriol.  Comparison of the profiles between 17 OHP and P during early pregnancy (n = 5) and nonconceptive cycles (n = 15) demonstrated a decline in 17 OHP , but not P, as early as week 3 postmating (designated as week 1) and lasting up to week 13.  Otherwise, secretions of 17 OHP mimicked P concentrations throughout pregnancy and in nonconceptive cycles.  Examination of the mean ratio values of 17 OHP to P demonstrated a significant drop in the ratio during weeks 2-7 of early pregnancy from >/= 0.7 to < 0.7 compared with the same time period in a nonreceptive cycles (p < 0.05m N = 5),  A 2x2 table analysis of the 17 OHP:P ratio during weeks 2-7 indicated that the possibility of a false positive or false negative result was 3.4 and 6.5%, respectively, based on the sensitivity and specificity of the test.  Overall, gestation lengths of the pregnancies completed during this study (N = 4) ranged from 91 to 98 weeks, with a mean of 93+/-2.9 (SEM) weeks.  A birth-to-conception interval of 47 weeks was noted in one animal.  The results described in this report provide additional data on the reproductive endocrinology of the pregnant Asian elephant and also present the earliest means to date of determining pregnancy by analysis of the 17 OHP:P ratio.

Poole, T.B., Taylor, V.J., Fernando, S.B.U., Ratnasooriya, W.D., Ratnayeke, A., Lincoln, G.A., Manatunga, A.M.V.R., Mcneilly, A.S., 1997. Social behaviour and breeding physiology of a group of captive Asian elephants. International Zoo Yearbook 35, 297-310.
Abstract: In 1986 at the Pinnawala Elephant Orphanage, Sri Lanka, a study of 4.4 Asian elephants Elephas maximus was carried out to establish the physical and behavioural changes associated with the female oestrous cycle and the male musth cycle. At Pinnawala the elephants are maintained in mixed-sex groups which has resulted in successful breeding. This paper describes the management of elephants at the orphanage and details their social and reproductive behaviour. It is suggested that successive matings throughout the oestrous cycle and on multiple cycles may be required to ensure successful breeding.

Raju, R., Rao, B.S.G., Khadri, S.M., Asha, D., 1997. Chemical manipulation of delayed parturition in captive Asiatic elephant at Mysore Zoo. Indian Forester 123, 910-916.

Rasmussen, L.E., Lee, T.D., Zhang, A., Roelofs, W.L., Daves, G.D.Jr., 1997. Purification, identification, concentration and bioactivity of (Z)-7-dodecen-1-yl acetate: sex pheromone of the female Asian elephant, Elephas maximus. Chemical Senses 22, 417-437.
Abstract: In their natural ecosystems, adult male and female Asian elephants, Elephas maximus, live separately. For several weeks prior to ovulation, female elephants release a substance in their urine which elicits a high frequency of non-habituating chemosensory responses, especially flehmen responses, from male elephants. These responses occur prior to, and are an integral part of, mating. Using bioassay-guided fractionation, quantitatively dependent on these chemosensory responses, a specific sex pheromone was isolated and purified by an alternating series of organic and/or aqueous extractions, column chromatography, gas chromatography and high-performance liquid chromatography. Using primarily 1H-proton nuclear magnetic resonance (NMR) spectrometry and gas chromatography-mass spectrometry (GC-MS) of the urine-derived pheromone and its dimethyl disulfide derivative, we determined the structure of the active compound to be (Z)-7-dodecen-1-yl acetate (Z7-12:Ac). Concentrations of Z7-12:Ac in the female urine increased from non-detectable during the luteal phase to 0.48 microgram/ml (0.002 mM) early in the follicular phase and to 33.0 micrograms/ml (0.146 mM) just prior to ovulation. Bioassays with commercially available authentic synthetic Z7-12:Ac, using 10 Asian male elephants at several locations in the US, demonstrated quantitatively elevated chemosensory responses that were robust during successive tests, and several mating-associated behaviors. Bioassays with Z7-12:Ac with adult male elephants dwelling in more natural social situations in forest camps in Myanmar revealed some differing contextual pre-mating behavioral components. The remarkable convergent evolution of this compound suggests that compounds identified in mammalian exudates that are also present in pheromone blends of insects should be re-evaluated as potential mammalian chemosignals.

Rasmussen, L.E.L., Gunawardena, R.A., Rasmussen, R.A., 1997. Do Asian elephants, especially males in musth, chemically signal via volatiles in breath? Chemical Senses 22, 775.

Sarma, K.K., Dutta, B., 1997. Preputial diverticulum in an Asian elephant (Elephas maximus) - a case report. Indian Veterinary Journal 74, 59-60.

Schwarzenberger, F., Strauss, G., Hoppen, H.-O., Schaftenaar, W., Dieleman, S.J., Zenker, W., Pagan, O., 1997. Evaluation of progesterone and 20-oxo-progestins in the plasma of Asian (Elephas maximus) and African (Loxodonta africana) elephants. Zoo Biology 16, 403-413.
Abstract: The corpus luteum of African elephants produces high amounts of 5-reduced progesterone metabolites (5-pregnane-3,20-dione and 5--pregnane-3-ol-20-one), whereas progesterone itself is quantitatively less important, and plasma levels of progesterone during the estrous cycle in elephants are considerably lower than those of other mammals.  The objective of this study was to compare the concentration of progesterone in plasma of Asian and African elephants as determined by specific progesterone assays with those of total immunoreactive progestagens containing a 20-oxo-group (20-oxo-P).  These metabolites were determined by an enzyme immunoassay using an antibody against 5--pregnane-3-ol-20-one, 3HS:BSA.  Plasma of non-pregnant Asian (n = 4) and African (n = 4) elephants was collected at weekly intervals for periods of 8-15 months  and at random intervals during pregnancy in one Asian elephant.  High-performance liquid chromatography separation of plama samples of both species demonstrated that in the 20-oxo-P assay, 5-pregnane-3,20-dione makes up ~60% of the total immunoreactive material. The progesterone and 20-oxo-P values during the estrous cycle showed a parallel pattern and were significantly correlated (P < 0.001; Asian:  r = 0.80; y = 3.76 x -0.10; African: r = 0.75; y = 2.66 x -0.08.  Progesterone and 20-oxo-P values in Asian and African elephants were < 15 ng/mL during the follicular phase (weeks -4 to 0) of the estrous cycle; progesterone values during the luteal phase (weeks 2-9) were 0.60+/-0.03 and 053+/-0.03 ng/mL, and the 20-oxo-P values were 2.19+/-0.16 and 1.48+/-0.12 ng/mL, respectively.  The 20-oxo-P values of the pregnant animal, although slightly higher, were comparable to those of non-pregnant elephants during the luteal phase.  Total immunoreactive 20-oxo-P values are about three times higher than those of progesterone during the luteal phase, and 5-pregnane-3,20-dione is the major immunoreactive 20-oxo-P in the plasma of Asian and African elephants.

Sukumar, R., Krishnamurthy, K.V., Wemmer, C., Rodden, M., 1997. Demography of captive Asian elephants (Elephas maximus) in southern India. Zoo Biology 16, 263-272.
Abstract: Historically, the Asian elephant has never bred well in captivity.  We have carried out demographic analyses of elephants captured in the wild or born in captivity and kept in forest timber camps in southern India during the past century.  The average fecundity during this period was 0.095/adult female/year.  During 1969-89, however, the fecundity was higher at 0.155/adult female/year, which compares favorably with wild populations. there was a seasonality in births with a peak in January.  The sex reation of 129 male to 109 female calves born is not significantly different from equality, although the excess of male calves born mainly to mothers 20-40 years old may have biological significance. Mortality rates were higher in females than in males up to age 10, but much lower in females than in males above age 10 years.  The population growth rate, based on fecundity during 1969-89, was 1.8% per year.  The analyses thus showed that timber camp elephants in southern India could potentially maintain a stationary or increasing population without resorting to captures from the wild.  Breeding efforts for elephants in zoos can thus profitably learn from the experience of traditional management systems in parts of Asia.

Tiedemann, R., 1997. Sexual selection in Asian elephants. Science 278, 1550-1551.

Allen, W.R., Mathias, S.S., Skidmore, J.A., Wooding, F.B.P., van Aarde, R.J. Fetoplacental function the African elephant. Stone, G. and Evans, G. Proceedings of the 13th International Congress on Animal Reproduction.  9-10. 1996.  Sydney, Australia.
Ref Type: Conference Proceeding

Chandrasekharan, K., Cheeran, J.V., 1996. Use of antiandrogen in controlling musth in captive elephants. Zoos' Print Journal XI, 25.

Hearn, J.P., 1996. Mechanisms regulating the reproduction and fertility of some mammalian species in their natural environments. Journal of Public Health Policy 17, 152-158.
Abstract: This paper discusses examples, taken from the author's research programmes, of physiological mechanisms that link fertility regulation to environmental constraints. The species considered are the giant panda, black rhinoceros, African elephant, tamar wallaby, marmoset, stumptailed macaque and man.

Hildebrandt, T., Goritz, F., Pratt, N.C., Quandt, S., Lehnhardt, J., Montali, R.J., Pitra, C., 1996. Ultrasonography as a tool to evaluate the reproductive tract in female Asian elephants ultrasound in elephants. J. Ultras. Med 15, 59.

Kaimal, R., 1996. Musth:observations based on studies on 140 elephants in Kerala over 10 years. Zoos' Print Journal XI, 26-27.

Kapustin, N., Critser, J.K., Olson, D., Malven, P.V., 1996. Nonluteal estrous cycles of 3-week duration are initiated by anovulatory lutienizing hormone peaks in African elephants. Biology of Reproduction 55, 1147-1154.
Abstract: Previous attempts to characterize the estrous cycle of elephants have yielded conflicting estimates of cycle length and LH profiles. In order to establish artificial breeding programs in this species, resolution of these issues is needed. Therefore, four female African elephants housed at the Indianapolis Zoo were studied for approximately 6 mo beginning in December 1994. Blood was collected weekly, and the serum was immediately analyzed for progesterone (P4). Whenever the weekly concentration of P4 was found to be low, blood was collected one or four times per day. All serum samples were assayed for LH, and the daily samples were assayed for P4 and estradiol. Transient increases of serum LH (designated as peaks) were observed four times in each of the four females. Of these 16 LH peaks, 8 were classified as ovulatory LH (ovLH) peaks and 8 were classified as anovulatory LH (anLH) peaks. Peaks designated ovLH averaged 3.60 +/- 0.67 ng/ml (mean +/- SEM); serum P4 measured during these peaks began to increase 2-3 days before each ovLH peak and continued to increase for several weeks thereafter, reaching a peak of 675 +/- 35 pg/ml. The eight other LH peaks, designated anLH peaks, were of similar (p > 0.05) magnitude averaging 3.07 +/-0.72 ng/ml, but the serum concentration of P4 remained very low (< 80 pg/ml) during and for several weeks after these peaks. Six peaks designated anLH occurred an average of 12.2 +/- 1.4 days after serum P4 had declined below 80 pg/ml. In each elephant, there was a regular sequence in which each ovLH peak was followed by a luteal-active period lasting about 60 days and then about 12 days later by one anLH peak. Each anLH peak was followed 19-22 days later by one ovLH peak, but serum P4 remained at nonluteal levels throughout this interval between peaks. The authors propose to designate this interval after the anLH peak and before the next ovLH peak as a nonluteal (i.e., low P4) estrous cycle of only 3-wk duration. Following each short nonluteal estrous cycle, there was a single ovLH peak that initiated one luteal-active estrous cycle lasting 10-11 wk until terminated by the next anLH peak. The present results demonstrate that nonpregnant African elephants, housed in the absence of males, alternate between short nonluteal estrous cycles and long luteal-active estrous cycles. Daily measurements of serum P4 can be used to distinguish between the two types of estrous cycles and thereby provide a clinical prediction about the optimum time for artificial insemination.

Kirkpatrick, J.F., Turner, J.W., Jr., Liu, I.K., Fayrer-Hosken, R., 1996. Applications of pig zona pellucida immunocontraception to wildlife fertility
control. J Reprod Fertil Suppl 50, 183-189.
Abstract: A unique application of pig zona pellucida (PZP) immunocontraception is the control of wildlife populations. A native PZP vaccine has been successfully applied to wild horse and donkey populations. A single annual booster inoculation was capable of maintaining contraception. Seven consecutive years of PZP treatment in wild mares resulted in no detectable debilitating side effects, and reversibility of contraception has been documented among mares treated for up to 4 consecutive years. Long-term treatment (5-7 years) is associated with some ovulation failure and depressed urinary oestrogen concentrations. Complex social behaviours in horses were unaffected by treatment. PZP immunocontraception has also been successfully applied to white-tailed deer, with no detectable changes in ovarian histology after 2 years of treatment. Seventy-four species of captive zoo animals have been treated with the PZP vaccine, with documented success in 27 species, including members of the orders
Perissodactyla (Equidae), Artiodactyla (Cervidae, Capridae, Giraffidae, Bovidae), and Carnivora (Ursidae, Mustelidae, Felidae).  Immunocytochemistry studies have demonstrated a high degree of crossreactivity between anti-PZP antibodies and African elephant zona pellucida. The need for a one-inoculation form of the vaccine has led to the incorporation of PZP into lactide-glycolide microspheres, which cause a delayed release of the PZP. PZP immunocontraception of wildlife has potential because of (1) > 90% effectiveness, (2) the ability for remote delivery, via darts, (3) reversibility after short-term use, (4) a wide breadth of effectiveness across many species, (5) a lack of debilitating side-effects even after long-term treatment, and (6) minimal effects upon social behaviours.

Kurt, F., Mar, D.K., 1996. Neonate mortality in captive Asian elephants (Elephas maximus). International Journal of Mammalian Biology 61, 155-164.
Abstract: One third of Asian elephants born in European zoos and circuses are stillborn (16.0%) or killed or refused by their mothers (15.7%). Stillbirths and infanticides are rare in extensively kept and wild-living elephants. Infanticide could be related to life history of the mothers: Females which had grown up in the company of an older, motherly female adopted their offsprings without complications. Those having lacked such affection, tended to kill or at least not to adopt their neonates. Stillborn calves show higher neonate weights (124.6 +/- 20.8 kg) than surviving calves (92.0 +/-27.6 kg). Positive correlations were found between gestation period and neonate weight as well as between neonate weight and relative weight (body weight/shoulder height) of the mother. As female elephants in modern zoos and circuses are relatively heavier than those living in Asian camps, they produce calves after longer gestation periods (644.4 +/- 19.5 days) with larger neonate weights (105.6 +/- 26.6 kg) than extensively kept females in Asia (598.1 +/- 51.6 days; 74.0 +/- 21.6 kg). Chances to survive parturition are negatively correlated with length of gestation and neonate weight.

Lincoln, G.A., Ratnasooriya, W.D., 1996. Testosterone secretion, musth behaviour and social dominance in captive male Asian elephants living near the equator. Journal of Reproduction and Fertility 108,  107-113.
Abstract: Blood samples collected weekly over a 5-year period from 6 adult (19- to 40-year old) male Asian elephants (Elephas maximus maximus) living in captivity in Sri Lanka (7°N). Testosterone profiles were very variable within and between animals. Long-term phasic changes in blood concentrations of testosterone, associated with periods of musth (temporal gland secretion, drip urination and aggressive behaviour), occurred in 3 of the 6 elephants, the most pronounced cyclicity occurring in the oldest animal. Musth occurred annually after periods of high androgen secretion and the duration of musth was positively correlated with the mean concentration of testosterone during the previous 2 months. The time of musth, while consistent for an individual, varied between animals. In 4 bulls living in 1 social group, social rank was positively correlated with the mean concentration of testosterone over the 5-year period, and only the dominant animal showed periodic musth. Short-term changes in testosterone concentration occurred in blood samples collected every 15 min for 7 h, and after the injection of 20 µg GnRH, consistent with regulation through the pulsatile secretion of LH. The results support the view that fully mature male Asian elephants living near the equator express an asynchronous, cyclical, circannual pattern of gonadal activity. The periodic increase in testosterone secretion during the gonadal cycle induces the development of musth; however, androgen withdrawal following a period of hypersecretion may be the cause of some aspects of musth behaviour (aggression, unpredictability, disobedience) which make bull elephants very difficult to manage in captivity.

Mar, K.U., 1996. Captive Breeding of Asian Elephants in Myanmar; An analysis of the reproductive parameters of domestic female elephants of the Union of Myanmar. Tiger Paper 23, 6-13.
Abstract: Three hundred and forty calvings (males = 172, females = 168) born during the fiscal year 1991/92 to 1995/96 from 322 working cow elephants which were raised from two different birth types (cows captured from the wild, n = 166; and cows born in captivity, n = 174), were analyzed in order to identify the mean age at first calving and the age at the prime reproductive performance, with special reference to fertility rates, sex ratios at birth, interbirth/calving interval, and correlation of age and birth type of dam to calving potential. Two cows produced twin births and 16 cows (age = 12.6 - 43 yr) gave birth twice during the study period of 5 years, with a mean standard deviation of interbirth interval of 3.4 +/- 0.57 years (range 3.1 - 4.9 years). The average annual calving rate was 68 calves per year. Sixty-six percent of the total calvings (255 vs 340) were born as first, second and third calves. Fewer calvings should be expected from the older working cows or after third calvings. The prime reproductive age for reproduction in female working elephants in Myanmar was in the age group of 21-25 years, with 70 calvings. Infanticides (n = 1) and stillbirths (n = 10) were rare in working elephants, which seemed to be common in heifers and young inexperienced nulliparous cows.

Mosley, J. Hand-Rearing a Captive-Born Asian Elephant (Elephas maximus): (I) A Study of Physical Development as a Response to the Rearing Regime, and (ii) Social Interactions. Spooner, N. G. and Sharp, K. The Ninth UK Elephant Workshop.  36-65. 1996. England, The North of England Zoological Society. 1996.
Ref Type: Conference Proceeding

Murray, S., Bush, M., Tell, L.A., 1996. Medical management of postpartum problems in an Asian elephant (Elephas maximus) cow and calf. Journal of Zoo and Wildlife Medicine 27, 255-258.
Abstract: An 18-yr old female Asian elephant (Elephas maximus) gave birth to a 120-kg female calf following 22 mo of gestation.  Immediately after parturition, the cow became agitated and aggressive towards the calf.  Before the keepers were able to safely intervene and remove the calf, the cow stepped on the calf's head and right front leg.  Within 30 min, the cow calmed down, allowing the calf's safe reintroduction under close keeper supervision and control.  The cow had a retained placenta, poor mammary development, and low milk production.  The calf's injuries, in combination with the cow's low milk production, impeded the calf's ability to nurse and gain weight.  Within 10 days, the calf lost 10% of its weight.  Serum protein electrophoresis indicated failure of passive transfer of maternal immunoglobulin.  On day 10, the calf received a transfusion of concentrated immunoglobulin extracted and concentrated from the cow's previously banked plasma.  On day 13, the calf developed a urinary tract infection, as diagnosed by white blood cells and bacteria in the urine.  Following immunoglobulin administration and antibiotic therapy, clinical signs slowly resolved and the calf gained weight.  The cow passed the fetal membranes during parturition, but the placenta was retained.  Despite prophylactic systemic antibiotics and vaginal flushing, the cow became depressed and developed a leukocytosis and anemia.  A mucopurulent vaginal discharge and ventral edema were noted on day 3, and milk production was minimal.  Because decreased milk production has been reported as a common sequel to retained placenta, efforts were focused on removing the placenta.  Intermittent oxytocin therapy on days 2-14 did not result in expulsion of the placenta and produced only transient abdominal contractions and minimal increases in milk letdown.  On day 15, 10 mg estradiol cypionate was administered i.m. followed by 200 IU oxytocin i.v.  An additional 10 IU oxytocin was administered i.v. on day 16.  The friable placenta was palpable within the vaginal vault on day 17.  The remaining placenta was removed by gentle traction applied by a modified weighted pressure cuff.  Once the placenta was removed, the cow's clinical problems slowly resolved and the calf continued to gain weight.

Perrin, T.E., Rasmussen, L.E.L., Gunawardena, R., Rasmussen, R.A., 1996. A method for collection, long-term storage, and bioassay of labile volatile chemosignals. J. Chemical Ecology 21, 207-221.
Abstract: A procedure for headspace sampling and long-term storage of organic volatiles coupled with gas chromatographic-mass spectrometric (GC-MS) analysis was used to study the volatile chemosignals in a biological secretion prior to bioassay. The approach involved collecting the volatiles in evacuated canisters from an apparatus in which 1 ml of secretion was dispersed for headspace sampling. These canisters, stainless steel, 850 ml, and 100% internally eletropolished, have been demonstrated to store volatile compounds, in chemically stable form, for several weeks. The GC-MS analyses provided the quantitation and identification of compounds from C₃ through C₁₄ at concentrations as low as 0.10 parts per billion volume. The approach was used to study chemosignals of musth temporal gland secretions (TGS) from a male Asian elephant (Elephas maximus). Fresh TGS material loses its biological activity within 1 hr. TGS material stored at -20ºC usually loses its activity within 30 days. The usefulness of this method for long-term storage of the volatile chemosignals was demonstrated by the retention of biologically active TGS headspace compounds, as determined through bioassays, stored in these canisters for one year.

Rasmussen, L.E.L., Lee, T.D., Roelofs, W.L., Zhang, A., Daves, G.D., 1996. Insect pheromone in elephants. Nature 379, 1.

Rasmussen, L.E.L., Hall-Martin, A.J., Hess, D.L., 1996. Chemical profiles of African bull elephants, Loxodonta africana: physiological and ecological implications. Journal of Mammalogy 77, 422-439.
Abstract: This study reports concentrations of testosterone and dihydrotestosterone in both serum and temporal-gland secretion of male African elephant (Loxodonta africana), including radiocollared elephants, and identifies a spectrum of volatile components in the temporal-gland secretions. Androgens in the serum (testosterone and dihydrotestosterone) were measured in 111 adult male African elephants, ages 21-40 years, from two national parks in South Africa during several  years and seasons.  About one-fifth (18.6%) of these mature, male, African elephants exhibited dramatically increased concentrations of testosterone in serum characteristic of male Asian elephants during musth.  In Krueger National Park, six radiocollared male African elephants, ages 25-35 years, were tracked and serially samples for both serum and temporal-gland secretions during a 5-year period. Concentrations of testosterone in serum and temporal gland secretions were elevated cyclically at times when typical musth behaviors, including aggression, were observed.  This study reports the first chemical characterization of the volatile compounds of the temporal gland secretions from male African elephants in musth. It reveals many similarities between the chemical constituents of the temporal-gland secretions of these male African elephants and the compounds identified in male Asian elephants.  In addition, several compounds, not previously identified in temporal-gland secretions of African elephants, are described.  Such chemical data support the behavioral observations by ourselves and other researchers that male African elephants experience musth. Especially convincing are the concurrent hormonal and chemical data from the radiocollared males during episodic periods of behavioral musth. Implications of the incidence of musth in the past and present ecology of African elephants are discussed in view of the increasing compression within national parks.

Sarma, K.K., Dutta, B., 1996. Musth and its management in Asian elephant: a discussion based on four clinical cases. Zoos' Print Journal April, 21-22.

Schaftenaar, W. Vaginal vestibulotomy in an Asian elephant (Elephas maximus). Proceedings American Association of Zoo Veterinarians.  434-439. 1996.
Ref Type: Conference Proceeding
Abstract: Due to its dimensions, dystocia in elephants presents a difficult problem.  This paper describes the delivery of a dead calf by surgical intervention.  A vestibulotomy was performed under local anesthesia.  Complications in wound healing resulted in a permanent fistula of the vestibulum.  The difficulties in decision making and the interpretation of clinical signs are discussed.

Schmid, J., Mar, D.K., 1996. Reproductive performance of captive Asian elephants in Myanmar.  Gajah 16, 23-42.

Schwarzenberger, F., Mostl, E., Palme, R., Bamburg, E., 1996. Faecal steroid analysis for non-invasive monitoring of reproductive status in farm, wild, and zoo animals. Animal Reproduction Science 42, 515-526.

Thakuria, D.B., Barthakur, T., 1996. Management of musth in a male African elephant by chemical sedatives in the Assam state zoo, Guwahati. Indian Veterinary Journal 73, 339-340.

Wasser, S.K., Papageorge, S., Foley, C., Brown, J.L., 1996. Excretory fate of estradiol and progesterone in the African elephant (Loxodonta africana) and patterns of fecal steroid concentrations. General and Comparative Endocrinology 102, 255-262.
Abstract: We developed and validated a noninvasive method to quantify fecal estrogens and progestins as a tool for monitoring long-term ovarian activity in free-ranging African elephants. The lag times between iv injection of [(3)H]estradiol and [(14)C]progesterone and peak excretion of radioactivity in urine and feces were approximately 4 hr and 48 hr, respectively. The majority of progesterone metabolites recovered was excreted in feces (55%) versus urine (45%), whereas comparatively little of the recovered estradiol metabolites were excreted in feces (5%) compared to urine (95%). Intrasample variation in fecal hormone concentrations was extremely high but could be substantially reduced by extracting well-mixed fecal powder from freeze-dried samples, taken from the central or premixed portion of the wet sample. This method resulted in a close correspondence between matched serum and fecal progestins (mean correlation =0.81, range 0.61-0.94) collected from five nonpregnant adult females over a 7-month period. Fecal estrogen profiles were more ambiguous, tending to overlap with those of fecal progestins. We conclude that analyses of fecal progestins can provide an effective, noninvasive means of characterizing ovarian activity in free-ranging African elephants.

 1995. A Week with Elephants; Proceedings of the International Seminar on Asian Elephants. Bombay Natural History Society; Oxford University Press, Bombay, India.

Brown, J.L., Lehnhardt, J., 1995. Serum and urinary hormones during pregnancy and the peri- and postpartum period in an Asian elephant (Elephas maximus). Zoo Biology 14, 555-564.

Desai, A.A., Johnsingh, A.J.T. Social Organization and Reproductive Strategy of the Male Asian Elephant (Elephas maximus).  532. 1995. Bombay, India, Bombay Natural History Society; Oxford University Press.
Ref Type: Abstract

Hildebrandt, T., Goritz, F., 1995. Sonographischer nachweis von leiomyomen im genitaltrakt weiblicher elefanten. verh. ber Erkrankg. Zootiere 37, 287-294.

Hildebrandt, T.B., Goritz, F. Transrectal ultrasonography for ovary and pregnancy in Indian elephant. Verh ber Erkrg Zootiere.  261-268. 1995.
Ref Type: Conference Proceeding

Joshua, J., Johnsingh, A.J.T., 1995. Ranging Patterns of Elephants in Rajaji National Park: Implications for Reserve Design. In: Daniel, J.C. (Ed.), A Week with Elephants; Proceedings of the International Seminar on Asian Elephants. Bombay Natural History Society; Oxford University Press, Bombay, India, pp. 256-260.

Krishnamurthy, V., 1995. Reproductive Pattern in Captive Elephants in the Tamil Nadu Forest Department: India. In: Daniel, J.C. (Ed.), A Week with Elephants; Proceedings of the International Seminar on Asian Elephants. Bombay Natural History Society; Oxford University Press, Bombay, India, pp. 450-455.
Abstract: The Forest Department of the State of Tamil Nadu (formerly the Madras Presidency) in India has been capturing and maintaining elephants for more than 130 years. These elephants which are mainly utilised for timber extraction work are stationed in forest camps. The elephants are maintained as mixed herds, and able to socialize both when they are in camp or when they are let out for foraging in the forests. Records were maintained on the various aspects of elephant management which included the breeding records in captivity of all elephants, varying over periods of time. From these records the birth of 210 elephant calves over a period of 104 years could be collected and the data analysed. The average fertility of the captive population particularly during the last two decades compares favourably with wild population both in Asia and Africa. A peak in births was observed during the early dry season i.e. in the months of January and February. The sex ratio at birth is not statistically significantly different from 1:1. The active reproductive phase in cow elephant extended over 40 years. During earlier periods the mortality rate among captive born calves was high, but by better management practices the mortality rate has been considerably reduced, particularly during the last two decades.

Magunna, C. Oestrous cycle and pregnancy in Asian elephants (Elephas maximus). Zyklus und Trachtigkeit beim asiatischen Elefanten (Elephas maximus).  1-117. 1995. Tierarztliche Hochschule Hannover.
Ref Type: Thesis/Dissertation
Abstract: Blood samples were collected at weekly intervals over a period of 18 months from 10 females at a zoo in Germany. Increases in blood progesterone to >0.4 ng/ml indicated the onset of the oestrous cycle; the concentration averaged 0.72±0.14 and 0.2±0.07 ng/ml in the luteal and follicular phases respectively. The average duration of 45 oestrous cycles of 8 cycling females was 13.93±0.59 weeks, the luteal phase averaging 8.11 weeks and the follicular phase 5.86 weeks. Blood LH concentration increased at the end of the follicular phase. Of 6 females mated on the basis of their blood progesterone curve, 5 conceived. Data on the cycles of individual females are tabulated.

Mar, K.U., Maung, M., Thein, M., Khaing, A.T., Tun, W., Nyunt, T., 1995. Electroejaculation and Semen Characteristics in Myanmar Timber Elephants. In: Daniel, J.C. (Ed.), A Week with Elephants; Proceedings of the International Seminar on Asian Elephants. Bombay Natural History Society; Oxford University Press, Bombay, India, pp. 473-482.
Abstract: Six bull elephants between 16 and 31 years of age (mean = 21 years) were used in this study. Semen quality was evaluated in field condition using light microscope immediately after ejaculation (EEJ). The sperm morphology was studied in wet and preparations made from formol-saline fixed samples under phase-contrast microscope and in stained (Giemsa) smear preparations using light microscope. Morphological abnormalities were recorded as a percentage of the total number of counted spermatozoa. The number of sperm per millilitre of semen was determined by Neubauer Haemocytometer and its range was 1020-2000 x 10⁵. Morphological characteristics used in this study were abnormal detached heads, abnormal acrosomes, proximal and distal cytoplasmic droplets, pouch formation, abnormal midpiece and abnormal tails (simple bent and double folded). Although this study failed to clarify the statistically significant standard norms of semen characteristics for Myanmar elephants due to the limited number of bull elephants successfully ejaculated by EEJ, the normal and abnormal sperm morphology as well as data regarding EEJ were observed and recorded for the first time in Myanmar. Further investigations are needed to clarify the standard norms of semen characteristics, to determine the acceptable values of sperm abnormalities, and to indicate the differences between individuals. These findings emphasize the importance of selecting the best sires for successful natural and artificial breeding in Myanmar elephants.

Munson, L., Karesh, W.B., Shin, S., Balke, J.M.E., Calle, P., Cambre, R.C., Cranfield, M., Citino, S., Junge, R.E., 1995. Lymphoid follicular vulvitis in African (Loxodonta africana) and Asian (Elephas maximus) elephants. Journal of Zoo and Wildlife Medicine 26, 353-358.
Abstract: Hyperemic nodules and plaques in the distal urogenital canal of African (Loxodonta africana) and Asian (Elephas maximus) elephants were investigated to determine if they represented a potentially transmissible venereal disease.  The distal urogenital canals of 29 captive Asian, 19 captive African, and 30 free-ranging African elephants were examined.  Biopsies were obtained from 10 captive Asian, four captive African, and 28 free-ranging African elephants.  Biopsies from four elephants (three Asian, one African) were examined ultrastructurally.  Bacteriologic cultures of the distal urogenital canal were performed on 15 captive elephants (nine African, six Asian), nine with lesions and six without lesions.  Hyperemic nodules and plaques were identified in the distal urogenital canals of 62% of captive Asian, 89% of captive African, and 90% of free-ranging African elephants examined, including 10 of 11 pregnant free-ranging elephants.  These lesions were characterized histopathologically by aggregates of coalescing reactive lymphoid follicles.  No viral agents were identified, and no specific bacteria were consistently associated with lesions.  These highly prevalent lesions appear to be reactions of mucosa-associated lymphoid tissues to non-specific antigenic challenges in the distal urogenital canal and appear to have no clinical significance.

Palm, M., 1995. Oestrus interval in Indian elephants. Svensk Veterinartidning 47, 519-521.

Papageorge, S., Wasser, S.K., Foley, C., Brown, J. Fecal steroid analysis: validation of extraction and radioimmunoassay for estradiol and progestagens in African elephants (Loxodonta africana) and analysis of fecal samples utilizing a validated method. Joint Conf AAZV/WDA/AAWV.  447. 1995.
Ref Type: Conference Proceeding

Foerner, J.J., Houck, R., Copeland, J.F.Jr., Schmidt, M.J., Byron, H.T., Olsen, J.H., 1994. Surgical castration of the elephant (Elephas maximus and  Loxodonta africana). Journal of Zoo and Wildlife Medicine 25, 355-359.
Abstract: The purpose of this project was to develop a reliable, safe, and efficient technique for surgical castration of elephant (Elephas maximus and Loxodonta africana). To achieve this, there have been several modifications in the surgical technique. Initially, sterilization by injecting caustic agents into the testicles via laparotomy was attempted, but results were unpredictable and had serious side effects. Castration of young males under 5 years of age was relatively easy using a standard equine chain ecraseur through a single laparotomy incision. For larger males, most cases required two laparotomy incisions with several variations in techniques for removal of the testicles. Initially, self-locking stainless steel bands were placed on the cord as ligatures, and the testicles were removed with an obstetrical wire saw. Because of technical difficulties, this method was abandoned, and an alternative technique was developed. The testicle was removed with an obstetrical wire saw, and then the artery was isolated by digital palpation. A Kelly forceps was secured on the vessel. The forceps was passed through a loop of an equine chain ecraseur and the chain positioned over the artery. The ecraseur was then closed, crushing the vessel. The most promising technique is the development of a large chain ecraseur that will allow removal of both testicles through a single laparotomy approach.

Formenty, P., Domenech, J., Lauginie, F., Ouattara, M., Diawara, S., Raath, J.P., Grobler, D., Leforban, Y., Angba, A., 1994. Epidemiological study of bluetongue in sheep, cattle and various wild animal species in the Cote d'Ivoire. Revue Scientifique et Technique Office International des Epizooties 13, 737-751.
Abstract: Between 1992 and 1993, serum samples from 623 sheep, 215 cattle and 211 other ruminants from Cote d'Ivoire were tested for bluetongue virus antibodies using the agar gel immunodiffusion test. Seroprevalence was 52±4% in sheep, 95±3% in cattle and 56±7% in wild herbivores. Bluetongue antibodies were detected in kob (Kobus kob), common waterbuck (Kobus ellipsiprymnus), roan antelope (Hippotragus equinus), buffalo (Syncerus caffer), hartebeest (Alcelaphus buselaphus) and elephant (Loxodonta africana). A significant geographical variation was observed in presence of bluetongue in sheep. Antibody prevalence increased significantly with age in sheep and wild herbivores, and seroprevalence was higher in dams with a history of abortion. It is concluded that bluetongue is enzootic in Cote d'Ivoire.

Gross, M.E., Clifford, C.A., Hardy, D.A., 1994. Excitement in an elephant after intravenous administration of atropine. Journal of the American Veterinary Medical Association 205, 1437-1438.
Abstract: A 28-year-old Asian elephant (Elephas maximus) was anaesthetized for cesarean section to remove a dead calf. The elephant was sedated with azaperone, and atropine was administered i.v. 90 minutes later in preparation for induction of anaesthesia with etorphine HCl. Within a minute of the injection of atropine the elephant began swaying kicking and moving in an agitated manner around the stall. It was concluded that there is considerable variation among species in the toxicity of atropine, although development of toxicosis usually is associated with overdosage.

Hattingh, J., Knox, C.M., Raath, J.P., 1994. Arterial blood pressure of the African elephant (Loxodonta africana) under etorphine anaesthesia and after remobilisation with diprenorphine. Veterinary Record 135, 458-459.
Abstract: Six adult, male elephants (bodyweight approximately 5000 kg) were immobilized, with 8 mg etorphine (M99) for semen collection by electroejaculation. Before electrostimulation (about 10 minutes after the elephants initially became recumbent) their mean arterial pressure was 186 ± 25 mm Hg. During the electrostimulation procedure to which each elephant was subjected intermittently over a period of about 20 minutes using a rectal probe, the mean was 263 ± 30 mm Hg. After 10 to 15 minutes stabilization, 26 mg diprenorphine (M50/50) was administered i.v. The elephants adopted a rocking motion in an attempt to stand up. This motion was accompanied by wide fluctuations in arterial pressure which peaked at 245 ± 19 mm Hg immediately before they rose. Arterial pressure subsequently decreased to a mean of 200 ± 28 mgHg once they were standing. Since these values were higher than that previously observed in standing, conscious elephants (145 ± 3 mmHg) it appears the standing, remobilized elephants in this study were hypertensive. Possible reasons for this are discussed. It is suggested that in view of the observed and possible detrimental increase in arterial pressure during electrostimulation simultaneous blood pressure monitoring should be carried out when this procedure is employed.

Hildebrand, T., Göritz, F., 1994. Einsatz der transrektalen Sonographie zur Beurteilung des Genitaltraktes weiblicher Elefanten. Imaging 61(suppl 2), 98.

Hodges, J.K., van Aarde, R.J., Heisterman, M., Hoppen, H.-O., 1994. Progestin content and biosynthetic potential of the corpus luteum of the African elephant (Loxodonta africana). Journal of Reproduction and Fertility 102, 163-168.
Abstract: The aim of this study was to examine the progestin content and biosynthetic potential of the corpus luteum of the African elephant (Loxodonta africana). Luteal tissue was collected from nonpregnant and early, mid- and late pregnant elephants (n = 2 per group) shot in the Kruger National Park. Pieces of individual corpora lutea (2-3 per animal; 23 in total) were stored directly in ethanol before hormone analysis. Matching tissue pieces were incubated for 2 h with [3H]pregnenolone (2 x 10(5) c.p.m.), after which tissue plus medium were also stored in ethanol. Progesterone and 17 alpha-hydroxyprogesterone immunoreactivity in tissue extracts were determined by enzymeimmunoassay and radioimmunoassay, respectively, before and after reverse phase HPLC. Progesterone immunoreactivity predominated over that of 17 alpha-hydroxyprogesterone in all corpora lutea examined but concentrations of both hormones were very low (73-374 ng g-1 and 3-93 ng g-1, respectively after HPLC). There were no obvious differences in hormone concentrations in corpora lutea from animals at different reproductive stages. Progesterone and 17 alpha-hydroxyprogesterone immunoreactivity assayed before HPLC was 10-30 times higher than that measured after chromatographic separation. HPLC consistently revealed two large immunoreactive peaks associated with relatively nonpolar compounds, which together accounted for most (at least 75%) of all progesterone immunoreactivity measured. Large amounts of radioactivity with the same retention times as these peaks were also detected after HPLC in samples incubated with [3H]pregnenolone. Analysis of conversion products from four corpus luteum incubations indicated that between 52% and 84% of [3H]pregnenolone had been converted; 19-33% was accounted for by progesterone, and 12-50% by the two substances represented by the unidentified peaks. Subsequent GCMS analysis identified the two immunoreactive peaks as 5 alpha-pregnane-3 alpha-ol-20-one and 5 alpha-pregnane-3,20-dione (5 alpha-dihydroprogesterone). These results indicate that the major progestins contained within and biosynthesized by corpora lutea of African elephants are 5 alpha-reduced metabolites, and that progesterone and 17 alpha-hydroxyprogesterone are quantitatively of minor importance.

Jayewardene, J., 1994. The Elephant in Sri Lanka. WHT Publications Ltd., Colombo, Sri Lanka.

Olsen, J.H., Chen, C.L., Boules, M.M., Morris, L.S., Coville, B.R., 1994. Determination of reproductive cyclicity and pregnancy in Asian elephants (Elephas maximus) by rapid radioimmunoassay of serum progesterone. Journal of Zoo and Wildlife Medicine 25, 349-354.
Abstract: The stages of the reproductive cycle and pregnancy of 15 Asian elephants at Busch Gardens in Tampa, Florida were determined by measuring serum progesterone concentrations using a commercial radioimmunoassay (RIA) kit with a sensitivity of 40 pg/ml of serum. Blood samples were collected and analyzed weekly for a 45-mo period. Serum progesterone concentrations ranged from nondetectable (<40 pg/ml) in nonpregnant elephants to 2110 pg/ml in pregnant elephants. The highest concentration of progesterone during the oestrous cycle was 1490 pg/ml. The length of oestrous cycle ranged from 10 to 23 wk, with an average of 15.1±0.3 wk for 103 oestrous cycles. The duration of the nonluteal phase was 4.6±0.2 wk, and the duration of the luteal phase was 10.5±0.2 wk. The serum progesterone concentration during the oestrous cycle was 214.0±5.2 pg/ml. The individual elephant average cycle length was used to estimate the time of oestrus and ovulation. During the nonluteal phase, elephant cows were placed with a bull for mating. Two weeks after oestrus began (based on rising progesterone at the end of the nonluteal phase), the cow was separated from the bull. Ovulation was predicated to occur during the first week after oestrus began. Eight elephants became pregnant, with a serum progesterone concentration of 554.6±16.4 pg/ml and a range of <40-2110 pg/ml. These results confirm that weekly measurement of serum progesterone by RIA can be used to characterize the luteal and nonluteal phases of the Asian elephant oestrous cycle. Using this information, cows can be managed and placed with a bull at the appropriate time for breeding. Serum concentrations of progesterone can also be used to confirm pregnancy and to monitor and evaluate potential problems during pregnancy. The approximate date of parturition can then be predicted from average gestation length data, thus allowing staff to prepare facilities and be ready to assist with delivery.

Perrin, T.E., Rasmussen, L.E.L., 1994. Chemosensory responses of female Asian elephants (Elephas maximus) to cyclohexanone. Journal of Chemical Ecology 20, 2577-2586.
Abstract: Cyclohexanone, a naturally occurring component of male Asian elephant temporal gland secretion, was tested as a candidate elicitor of bioresponses from female Asian elephants (Elephas maximus). Four female Asian elephants were presented with synthetic cyclohexanone samples during a standardized bioassay. Four types of bioresponses, some or all of which may be important in intersexual communication, were monitored: flehmen, palatal pit area contact, scrub, and check responses. Cyclohexanone evoked persistent responses of all bioresponse types by two females, moderate response by a third female, and very few responses by a relatively unresponsive female. The results suggest that cyclohexanone may provide chemical information to females about male elephants, particularly regarding their state of musth.

Plouzeau, E., daCunha, S., Shaw, H.J., 1994. The ovarian cycle in Asian and African elephants (Elephas maximus and Loxodonta africana). Techniques for the monitoring of female fertility in captivity. Revue de Medecine Veterinaire 145, 905-911.
Abstract: A discussion. During the oestrous cycle, circulating blood progesterone in elephants alternates between high and low concentrations over a cycle of 16±2 weeks. Some of the data have suggested a 3-week cycle and other data a 15- to 16-week cycle. Captive females show no signs of oestrus, although males show a Flehmen-like response to female urine, which is inversely related to plasma progesterone concentration. Pregnancy is associated with a sustained increase in circulating progesterone concentration and with an increase in total oestrogen, prolactin and oestrogen concentrations during the second half of pregnancy. The most reliable method of monitoring the oestrous cycle and pregnancy is by weekly analysis of plasma progesterone in both species, or of its urinary metabolite (pregnanetriol) in Asian elephants.

Rasmussen, L.E.L., Perrin, T.E., Rasmussen, R.A., Gunawardena, R., 1994. Isolation of potential musth-alerting signals from temporal gland secretions of male Asian elephants (Elephas maximus). Chemical Senses 19, 540.

Wallace, C., Byron, T.H., Foerner, J.J., Weston, H., Kilpatrick, J., Jastremski, M.S. Clinical case report: the medical management and treatment of a 36 year old premiparturient Asian elephant cow with a dystocia and following a Caesarian section.  1994.
Ref Type: Unpublished Work
Abstract: The medical history and management of a 36 year old premiparturient Asian elephant (Elephas maximus indicus) with a dystocia requiring a caesarian section are discussed.  The examination and complete medical evaluation to determine the health of the cow and viability and position of the calf are described.  The medical management of the post-operative complications and the changes in complete blood counts, differential, serum biochemistry values and urinalysis prior to the elephant's death are described.  Complications included peritonitis with systemic sepsis, renal failure, and hepatic failure.  Pertinent necropsy findings on the cow included severe diffuse subacute peritonitis, uterine transmural necrosis, diffuse renal tubular nephrosis, and hepatic centrolobular degeneration.

Armbrusters, P., Lande, R., 1993. A population viability analysis for African elephant (Loxodonta africana): How big should reserves be? Conservation Biology 7, 602-610.
Abstract: We present an age-structured, density-dependent model of elephant population dynamics in a fluctuating environment drawing primarily upon the life history parameters obtained from studies in semi-arid land at Tsavo National Park, Kenya. Density regulation occurs by changes in the age of first reproduction and calving interval. We model environmental stochasticity with drought events affecting sex- and age-specific survivorships. Results indicate a maximum population growth rate of 3% per year and an equilibrium elephant density of 3.1/square mile. Analysis of the demographic results and their sensitivity to changes in juvenile survivorship and drought frequencies, supported by genetic considerations, suggests that in semi-arid regions a minimum reserve size of 1000 square mile is necessary to attain a 99% probability of population persistence for 1000 years. The effect of age-independent culling on population viability is also analyzed.

Brown, J.L., Bush, M., Wildt, D.E., Raath, J.P., de Vos, V., Howard, J.G., 1993. Effects of GnRH analogues on pituitary-testicular function in free-ranging African elephants (Loxodonta africana). Journal of Reproduction and Fertility 99, 626-634.
Abstract: In the first of 2 experiments, 6 free-living adult male elephants were given 4 or 12 mg GnRH antagonist (Detirelix) or saline intramuscularly on day 0. Animals were then recaptured about 48 h later and given 300 µg GnRH intravenously to assess the ability of the antagonist to block pituitary activity. Detirelix reduced (P<0.05) basal concentrations of serum LH and testosterone on day 2 compared with day 0, with no effect of dose. Similarly, LH and testosterone release induced by GnRH were also reduced (P<0.05) in the Detirelix-treated bulls (50-70% reduction in peak concentration). In the 2nd experiment, elephants were given 30 mg of a structurally similar GnRH antagonist (103-201-40; n = 6), 22.5 mg of a long-acting GnRH agonist (Lupron Depot; n = 4) or D-mannitol carrier (n = 4) intramuscularly on day 0. All bulls were recaptured and given GnRH on day 2 (103-201-40 treatment group) or on days 2 and 20 (Lupron Depot group) after the initial injection. In contrast to Detirelix, the antagonist 103-201-40 did not inhibit basal or GnRH-induced LH or testosterone secretion. Pituitary-testicular responses to Lupron Depot were initially stimulatory, as evidence by increased (P<0.05) LH and testosterone secretion on days 0 and 2. By day 20, basal LH concentrations had returned to baseline values and the response to GnRH was markedly reduced (P<0.05), indicating that the pituitary was at least partially desensitized. Basal testosterone concentrations had also returned to baseline values by day 20 after Lupron Depot treatment. However, despite the attenuated LH response to GnRH, subsequent testosterone secretion was increased (P<0.05) compared with controls, suggesting that the testes of agonist-treated bulls had become hyper-responsive to small increases in LH secretion. It is suggested that GnRH analogues can suppress the pituitary-gonadal axis in African elephants. However, longer treatment periods, more frequent injection intervals or higher doses are probably needed to completely inhibit testosterone secretion and, thus, musth.

Christensen, C.L., Schmidt, M.J., Hess, D.L., Alak, B.M. Oocyte nuclear maturation in vitro in the African (Loxodonta africana) and Asian elephant (Elephas maximus). Biology of Reproduction 48[supp.1], 80. 1993.
Ref Type: Abstract
Abstract: Assisted reproductive technologies may contribute to the long term survival of threatened and endangered species.  In this study one pair of ovaries were collected at late follicular phase and early luteal phase of the estrus cyle from an African and an Asian elephant, respectively.  Both elephants were considered fertile, but were euthanized due to chronic arthritis.  The ovaries from the African elephant were transported from the Brookfield Zoo, Chicago, IL.  The left ovary (approximate size 10cm x 7cm x 3cm) contained predominantly corpora albicantia and oocyte recovery was not attempted.  The right ovary (approximate size 10cm x 7cm x 3.5cm) had a dominant follicle (>30 mm) and multiple preovulatory follicles (5-12mm).  Eight oocytes in tight cumulus were recovered from the preovulatory follicles and cultured immediately in simple medium (TALP) containing 20% fetal calf serum for 96 hours.  The oocyte from the dominant follicle was lost.  Oocyte size with zona ranged from 195-300 micrometers in diameter.  Zona thickness ranged from 7.5-19 micrometers.  An additional 12 oocytes were either atretic and discarded or were lost due to the extreme stickiness of the cumulus complexes, follicular fluid and stromal tissues.  An intact germinal vesicle (GV) was observed in several oocytes at the beginning of the culture (12 h lapse in time between ovariectomy and onset of culture).  GV breakdown (GVBD) was observed after 24 h of culture and two oocytes subsequently extruded a polar body within 40 h of culture.  Five metaphase I oocytes were inseminated with fresh sperm (final concentration 750,000 sperm/ml) from an Asian elephant, but there was no indication of fertilization or activation.  Three inseminated oocytes were acetolacmoid stained, no evidence of fertilizatio was observed.  Ovaries of the Asian elephant obtained from the Washington Park Zoo were characterized by several fresh corpora lutea in one ovary (ovary size 9cm x 6.5cm x 2.5cm), and several dominant and/or cystic follicles on the other (ovary size 8cm x 6cm x 2.5cm).  Two naked GV oocytes with cracked zona pellucida were recovered from one ovary and were cultured immediately. Oocyte size with zona ranged from 130-160 micrometers.  After 24 hours in culture one oocyte degenerated, one underwent GV breakdown and extruded either a cytoplasmic fragment or a polar body through the cracked zona.  Both oocytes were degenerated within 96 h.  This study suggests that it is possible to salvage the genetic contribution of fertile African and Asian elephants when they are not capable of natural reproduction.  Partially supported by the: Washington Park Zoo; Brookfield Zoo; H.D. 18185; and RR-00163.

Diephuis, E.P., 1993. Oestrus and pregnancy detection by flehmen-like responses of Asian bull elephants to urine samples of Asian female elephants. Zoologische Garten 63, 235-245.
Abstract: Urine samples were taken twice a week over a period of 130 days from 12 female Asian elephants of various reproductive status at 4 Dutch zoos. Pools, each of 12 urine samples from 6 cows, were offered to 4 male Asian elephants at 3 Dutch zoos. Flehmen responses by the bulls to the individual urine samples were recorded to detect oestrus and pregnancy in the cows. Serum progesterone data were used as a reference. During the tests, one of the 4 bulls showed hardly any flehmen responses; the others showed flehmen responses, with considerable variation between the 3 bulls in number of responses per sample (0.68±0.90, 0.93±1.24 and 1.14±1.52 per sample resp.). Correlations of flehmen responses between bulls were very low or moderate and negative. The repeatability of the response to urine samples averaged 64%. In the present study oestrus could not be detected by recording flehmen responses, and the information obtained about pregnancy was not conclusive. Several peaks of flehmen responses scattered over the 130-day period per cow were found, not exclusively during periods of low serum progesterone level (i.e. oestrous periods). Peaks for individual bulls did not usually match those of the other bulls. Urine samples from 2 cows evoked relatively few flehmen responses, which might indicate pregnancy. One of these 2 cows gave birth to a full-term calf on 1 June 1992. Pregnancy of the 2nd cow could not be confirmed. The present study showed that bulls display flehmen responses to urine samples from cows regardless of whether the cow is present or is familiar to the bull. Probably musth did not affect the interest of bulls in urine samples during flehmen tests. However, the maturity of the bull and the degree of sexual activity the bull displays may influence the interest of bulls in urine samples.

Dubiel, A., Gucwinski, A., Bielas, W., Birger, M., Nizanski, W., Bakaj, W., 1993. Treatment of vaginal prolapse in an elephant (Elaphas maximus). Zycie-Weterynaryjne 68, 138-139.

Fowler, M.E., 1993. Zoo and Wild Animal Medicine Current Therapy 3. W.B. Saunders, Philadelphia.

Niemuller, C.A., Shaw, H.J., Hodges, J.K., 1993. Non-invasive monitoring of ovarian function in Asian elephants (Elephas maximus) by measurement of urinary 5beta-pregnanetriol. Journal of Reproduction and Fertility 99, 617-625.
Abstract: An enzymeimmunoassay for 5beta-pregnanetriol is described. Immunoreactive pregnanetriol concentrations were significantly correlated with the concentrations of progesterone (0.98, n = 269, P<0.01) and 17alpha-hydroxyprogesterone (0.95, n = 205, P<0.01), the precursor of pregnanetriol. The duration of cycles as determined by measurements of plasma progesterone, plasma 17alpha-hydroxyprogesterone and urinary pregnanetriol averaged 15.54±1.5 weeks (23 cycles), 15.21±1.7 weeks (15 cycles) and 15.45±0.94 weeks (20 cycles) respectively. The results demonstrated that it is possible to monitor ovarian function in Asian elephants by measuring urinary pregnanetriol concentration.

Olsen, J.H., Byron, H.T., Jr., 1993. Castration of the elephant. In: Fowler, M.E. (Ed.), Zoo and Wild Animal Medicine Current Therapy 3. W.B. Saunders Company, Philadelphia, PA, USA, pp. 441-444.

Rasmussen, L.E.L., Lee, T.D., Daves, G.D., Schmidt, M.J., 1993. Identification of indolo [2,1-b] quinazoline-6,12-dione in the pre-ovulatory, estrous urine of Elephas maximus. Journal of Chemical Ecology 19, 2115-2128.

Rasmussen, L.E.L., Lee, T.D., Daves, G.D.Jr., Schmidt, M.J., 1993. Female-to-male sex pheromones of low volatility in the Asian elephant, Elephas maximus. Journal of Chemical Ecology 19, 2115-2128.
Abstract: In their natural ecosystems, the sexes of Asian elephant, Elephas maximus, live separately.  For several weeks prior to ovulation, the urine and cervical mucus of female Asian elephants contain extractable chemical agents of low volatility that elicit a high frequency of flehmen responses from bull elephants as an integral part of mating.  Subsequent to flehmen responses, male sexual arousal occurs and, if the female is available, mating results.  During the course of our project to determine the agent(s) and describe the responses associated with female to male sexual communication, we have identified an unusual compound.  This compound, apparently the sole component of the active fraction, was identified by mass, proton nuclear magnetic resonance, ultraviolet/visible, and infrared spectrometries as indolo-[2,1-b]quinazoline-6,12-dione (tryptanthrine).  Exhaustive and repetitive bioassays established that pure authentic (synthetic) typtanthrine was not the compound responsible for the bioresponse.   Rather a coeluting minor component, of low volatility, elicited the male bioresponse.

Rietkerk, F.E., Hiddingh, H., Van Dijk, S., 1993. Hand-rearing an Asian elephant Elephas maximus at the Noorder Zoo, Emmen. Iowa State University Veterinarian 32, 244-252.

Schmidt, M.J., 1993. Breeding elephants in captivity. In: Fowler, M.E. (Ed.), Zoo and wild animal medicine. W.B. Saunders Company, Philadelphia, PA,  USA, pp. 445-448.

Sukumar, R., Santiapillai, C., 1993. Asian elephant in Sumatra Population and Habitat Viability Analysis. Gajah 11, 59-63.

Taya, K., 1993. The reproductive physiology of the elephant. Journal of Reproduction and Development 39, 77-91.
Abstract: A discussion. In the females the vagina opens ventrally, ovarian cycles average 15-17 weeks in length, the gestation period is 22 months, and the ovary has a number of corpora lutea, although elephants are monovular. In males, there are permanent intra-abdominal testes, there is no distinct epididymis, but instead there is an extremely tortuous and convoluted duct which connects the testes to the openings of the seminal vesicles, the accessory organs are extremely well developed, especially the seminal vesicles and the bulbo-urethral glands. During musth, adult bulls become disobedient, aggressive and extremely dangerous, often attempting to kill their mahouts, or anybody else who comes within range. Bulls in musth have high testosterone levels and show urine dribbling and swollen temporal glands.

Turczynski, C.J. The endocrinology of musth in the male Asiatic elephant (Elephas maximus): Serum estradiol, serum LH and serum, fecal and urinary testosterone.  1993. College Station, TX. USA, Texas A&M University.
Ref Type: Thesis/Dissertation

 1992. Elephantine contraception. The Lancet 340, 583-584.

Acharjya, L.N., Tripathy, S.B., Rao, A.T., Mohanty, B.N., 1992. Chronic metritis in a captive Indian elephant: a case report. Pashudhan 7, 5.

Chandrasekharan, K., Radhakrishnan, K., Nair, K.N.M., Prabhakaran, T., 1992. Some observations on musth in captive elephants in Kerala (India). In: Silas, E.G., Nair, M.K., Nirmalan, G. (Eds.), The Asian Elephant: Ecology, Biology, Diseases, Conservation and Management (Proceedings of the National Symposium on the Asian Elephant held at the Kerala Agricultural University, Trichur, India, January 1989). Kerala Agricultural University, Trichur, India, pp. 71-74.

Czekala, N.M., Roocroft, A., Bates, M., Allen, J., Lasley, B.L., 1992. Estrogen metabolism in the Asian elephant (Elephas maximus). Zoo Biology 11, 75-80.
Abstract: Estradiol-17B metabolism was studied in two female Asian elephants (Elephas maximus).  In an initial study, 500 microCi of tritiated estradiol-17B was injected iv into a single animal, and 0, 30, and 60 min serum samples were collected as well as all excreted urine and feces for 24hr.  In a second study, 1.5 mg unlabeled estradiol-17B was injected iv into a second animal and 0, 5, 15, 30 and 60 min serum samples and a 30 min urine sample were collected postinjection. Analyses of samples from both studies demonstrated a rapid conversion of free estradiol to conjugated forms in the serum. The first (5 min) serum sample following the injection of unlabelled estradiol contained unconjugated estradiol:conjugated estradiol: conjugated estrone at a ratio of 60:29:10, respectively, and at 30 min a ratio of 33:43:24.  The urinary estrogen metabolites were in the conjugated form with an estradiol:estrone ratio of 60:40.  No radiolabelled estrogen was found in the fecal samples during the 24 hr following administration of the radiolabelled estradiol.  These data indicate a rapid clearance of circulating free estradiol in the elephant, with a major metabolite in the serum and urine being estradiol conjugate.

George, P.O., 1992. Some common surgical conditions encountered in elephants. In: Silas, E.G., Nair, M.K., Nirmalan, G. (Eds.), The Asian Elephant: Ecology, Biology, Diseases, Conservation and Management (Proceedings of the National Symposium on the Asian Elephant held at the Kerala Agricultural University, Trichur, India, January 1989). Kerala Agricultural University, Trichur, India, pp. 173.

Hagenbeck, D. Attempts to monitor oestrus and pregnancy in Indian elephants by urine testing. Diagnostische Zyklusansprache und Trachtigkeitsbestimmung beim asiatischen Elefanten (Elephas maximus).  1-66. 1992. Hannover, Germany, Tierarztliche Hochschule Hannover.
Ref Type: Thesis/Dissertation

Mar, K.U., 1992. Development of artificial insemination in Myanmar elephants. Captive Breeding Specialist Group News 3, 32.
Abstract: So far, no elephants have been reproduced successfully by artificial insemination (AI).  Researchers have determined the estrous cycle in cow elephants, which come into heat regularly about every 13-16 weeks. After tracing the hormonal pattern of progesterone by radioimmunoassay, zoo veterinarians were able to identify when the cows would ovulate in order to attempt AI. Four main problems need to be solved: 1) understanding the estrous cycle of female elephants in order to correctly time AI; 2) developing a safe and reliable method of collecting semen from bulls; 3) developing a way to preserve elephant semen for storage and transportation and ; 4) devising a method for insemination through the cow's twisting 3-4 foot long urogenital canal.  Timber production, which is the second largest source of Myanmar's export earnings (about $100 million U.S. per year), is still dependent to a large extend on elephant logging.  The ultimate goal of AI for elephants is not only to maintain the numbers of domestic working elephants but also to establish a reserve stock of new-generation elephants by preventing or reducing inbreeding.

Mar, K.U., Khaing, U.A.T., Tun, U.W., Nyunt, U.T., 1992. Electroejaculation and semen characteristics in Myanmar timber elephants. Captive Breeding Specialist Group News 3, 32.
Abstract: Six bull elephants between 16 and 31 years of age (mean = 21 years) were used in this study.  Semen quality was evaluated in field conditions using a light microscope immediately after ejaculation (EEJ). The sperm morphology was studied in wet preparations made from formal-saline fixed samples under phase-contrast microscope and in Giemsa-stained smear preparations using a light microscope.  Morphological abnormalities were recorded as a percentage of the total number of counted spermatozoa.  Morphological categories used in this study were: abnormal detached (loose or tailless) heads, abnormal acrosomes, proximal and distal cytoplasmic droplets, pouch formation, abnormal midpiece, and abnormal tails (simple bend, coiled and double-folded).  Detached (loose or tailless) heads were found in 20.5% of the samples while acrosome defects were found in 245 [sic] of the bulls.  Pouch formation, distal cytoplasmic droplets, coiled tails, and double-folded tails were found in 0.5% of the samples.  Although this study failed to clarify the statistically-significant standard norms of semen characteristics for Myanmar elephants due to the limited number of bull elephants successfully ejaculated by EEJ, the total incidences of normal and abnormal morphology as well as the data regarding EEJ were recorded and observed for the first time in the Union of Myanmar.  In the future, further investigations are needed to clarify the standard norms of semen characteristics in order to determine the acceptable values of sperm abnormalities and to indicate the differences between individual.  Serious attention should be paid to record as much detailed information as possible for every adult bull elephant by veterinarians and authorities concerned of the Ministry of Forestry.  These findings emphasized the importance of selecting the best bull sires for successful natural and artificial breeding in Myanmar elephants.  These investigations were carried out in Ngalaik Reserved Forest in February, 1992 with the collaboration of Mrs. and Dr. Michael J. Schmidt, Washington Park Zoo, Oregon, U.S.A. to indicate the acceptable values of sperm abnormalities and to indicate the differences.

Nair, P.G., Radhakrishnan, K., Chandrasekharan, K., 1992. Mating behaviour of the Asian elephant in captivity. In: Silas, E.G., Nair, M.K., Nirmalan, G. (Eds.), The Asian Elephant: Ecology, Biology, Diseases, Conservation and Management (Proceedings of the National Symposium on the Asian Elephant held at the Kerala Agricultural University, Trichur, India, January 1989). Kerala Agricultural University, Trichur, India, pp. 38-40.

Owen-Smith, R., 1992. Megaherbivores. The influence of very large body size on ecology. Cambridge University Press, Cambridge; UK.

Rasmussen, B., Davies, G.D., Lee, T.D. An unusual compound and further characterization of a preovulatory pheromone of Asian elephants, Elephas maximus. Chemical Senses 17, 687. 1992.
Ref Type: Abstract
Abstract: Abstract.  Full text.  A compound isolated from pre-ovulatory urine of Asian elephants as apparently a single entity (as assessed by a single band on TLC, a single peak by HPLC and a single dominant mass by field desorption mass spectrometry [FDMS]), was consistently active during bioassay and exhibited a reproducible dose-response curve.  Once pure (apparently), this compound was rapidly identified using a combination of spectral and mass spectral techniques. The principal component of the active fraction exhibited a molecular ion (m/z) at 248.  An exact mass measurement on the molecular ion was obtained by electron ionization (EI) mass spectrometry (MS) analysis.  From the mass of 248.056, the composition C15H8N2O2 was established. The isotope distribution of the molecular ion calculated from this composition was consistent with that observed in the mass spectrum.  UV spectral data indicated an extended, complex chromophore, probably a nitrogen heterocyclic.  Fragmentation information by collision-activated, EIMS demonstrated ions at 220 and 192.  The fragment ions in the EI spectrum (m/z 220 and 192) were consistent with the sequential loss of carbonyl groups; Fourier transform infrared (FTIR) spectra also indicated carbonyls.  Definitive NMR data indicated eight aromatic hydrogens, assigned on the basis of their coupling characteristics observed in the 2-D spectrum that were individually assigned to two different benzene rings.  All of these spectral data and comparison with an authentic sample established unambiguously the structure as indolol[2,1-b]-quinazoline-6,12-dione (tryptanthrine). Subsequent bioassays of the synthetic, authentic compound exhibited an initial high, novel substance response, followed by a sustained low-level response which gradually diminished to zero during a 6th month test period.  Several hundred bioassays of wide ranges of concentrations and conditions were conducted such that we are reasonably sure that tryptanthrine is not the active pheromone.  Re-evaluation of the active elephant preparation by HPLC and UV spectrometry demonstrated an earlier eluting, UV distinctive peak that, when isolated and bioassayed by itself, was active.  Preliminary data on this compound are reported.  Supported by NIH grant HD  19219-06.

Ratnasooriya, W.D., Fernando, S.B.U., Manatunga, A.N.V.R., 1992. Serum testosterone levels of Sri Lankan female elephants (Elephas maximus maximus). Med. Sci. Res. 20, 79-80.

Turczynski, C.J., Pernikoff, D., Garcia, R., Gross, T.S., Kraemer, D.C., 1992. Ovarian cycle influence in the Asian elephant (Elephas maximus). Biol Reprod (suppl 1) 46, 95.

Wallace, C., Doyle, C. The labor, birth, and post delivery management of an Asian elephant and her calf. Proc. Joint Meeting AAZV/AAWV.  104-109. 1992.
Ref Type: Conference Proceeding

Brown, J.L., Citino, S.B., Bush, M., Lehnhardt, J., Phillips, L.G., 1991. Cyclic patterns of luteinizing hormone, follicle-stimulating hormone, inhibin and progesterone secretion in the Asian elephant (Elephas maximus). Journal of Zoo and Wildlife Medicine 22, 49-57.
Abstract: Serum samples were collected one to three times weekly from four unanesthetized Asian elephants (Elephas maximus) for 6-18 consecutive months.  Based on circulating progesterone profiles, 14 complete ovarian cycles were observed.  The estrous cycle averaged 13.2 + 0.7 wk in length, with an active luteal phase of 9.8 + 0.7 wk.  Increases in serum luteinizing hormone (LH) were observed immediately before or during the progesterone rise in 11 of 14 cycles.  In eight cycles, a second LH surge was detected 11-19 days later. Radioimmunoassays for follicle-stimulating hormone (FSH) and inhibin were validated for elephant serum.  Fluctuations in FSH and inhibin secretion were observed at 12-14 wk intervals, although their temporal profiles differed from each other and from that of progesterone.  Follicle-stimulating hormone concentrations were lowest during the late follicular and early luteal phases and then increased to peak levels during the later part of the luteal phase.  In contrast, serum inhibin concentrations were inversely related to FSH levels throughout the estrous cycle (r = -0.78, P < 0.01).  In summary, progesterone analyses confirm that the luteal phase in the Asian elephant is approximately 10 weeks long.  Furthermore, the 12-14-wk oscillations in serum FSH and inhibin secretion provide additional evidence that the ovarian cycle of this species is several months in duration.  The inverse relationship between serum FSH and inhibin suggests that inhibit may regulate FSH secretion, as is described for other species.  Elevated FSH secretion throughout the mid-and late luteal phase may stimulate waves of follicular growth that are responsible for the short "follicular cycles" described in earlier reports.

Glatston, A., 1991. Unique observations on the birth of an elephant. Dieren 7, 164-167.

John, M.C., Suramanian, R., 1991. The elephant. Zoos' Print Journal 1-4.

Kramer, B., Teixeira, M., Hattingh, J., 1991. The histology of the adrenal gland of the African elephant, Loxodonta africana. South African Journal of Zoology 26, 193-198.
Abstract: The histology, particularly the ultrastructural cytology, of the adrenal gland of the African elephant, Loxodonta africana, is virtually unknown. Tissue from 14 adult male and female elephants was processed for light and transmission electron microscopy. The gland is surrounded by a thick capsule composed of an outer layer of dense connective tissue and an inner layer in which smooth muscle fibres predominate. Below the layer of smooth muscle, a continuous layer of relatively undifferentiated "capsular" cells occur. Where the capsular cells abut on the zona glomerulosa, they appear to be differentiating into glomerulosa cells, as small lipid droplets are present in their cytoplasm. The cortex is divided into three zones as is found in the adrenal glands of other mammals. Large amounts of collagenous and reticular tissue support the secretory cells, which have a marked lipid content. With electron microscopy, the cortical cells show features typical of steroid-producing cells. The medulla is characterized by an outer region of pale-staining chromaffin-positive (adrenaline) cells and an inner region of intensely staining chromaffin-positive (noradrenaline) cells. The latter cells contain granules of different sizes and structure.

Lang, E.M., Eggenberger, U., 1991. Protocol of a parturition in the elephant. Zoologische Garten 61, 5-7.

Niemuller, C., Liptrap, R.M., 1991. Altered androstenedione to testosterone ratios and LH concentrations during musth in the captive male Asian elephant (Elephas maximus). Journal of Reproduction and Fertility 91, 139-146.
Abstract: Greater concentrations of androstenedione than testosterone were usually present during periods of non-musth in plasma collected weekly for periods up to 2 years in 8 male Asian elephants (4-35 years of age).  For the 6 males that exhibited musth the androstenedione/testosterone ratio shifted greatly in favour of testosterone.  The severity of musth was assessed weekly using a scale of 1 to 5 for each of 8 behavioural traits including urine dribbling, temporal gland secretion and aggression.  Brief shifts in the ratio of the two androgens when testosterone predominated (n=106) were seen during the non-musth period in 3 of the males studied continuously for 2 years.  In 82% of these instances, stimuli of a sexual or aggressive nature had occurred in the preceding 48 h (x², p < 0.01).  A heterologous bovine assay was used to measure LH values in plasma collected every 15 minutes for 12h.  Increases in testosterone concentrations followed pulsatile increases in plasma LH concentrations during 7 non-musth periods in 4 animals.  Apart from pulse frequency, increases in the variables describing pulsatile LH secretion were seen in 2 strong musth and 2 mild musth episodes compared to non-musth values.  A strong musth, however, was characterized by a much greater increase in pulsatile testosterone secretion than was a mild musth and which may be a function of the duration of musth.

Rasmussen, L.E.L., Lee, T.D. Purification and initial characterization of a pre-ovulatory pheromone from female Asian elephants (Elephas maximus). Chemical Senses 16, 569. 1991.
Ref Type: Abstract
Abstract: Abstract.  Full text.  Urine from female Asian elephants in the pre-ovulatory phase of the ovarian cycle elicits a high frequency of flehmen responses from Asian bulls in a non-habituating manner.  These flehmen responses are an integral part of the mating sequence and suggest the presence of a sex pheromone.  Extraction and partial purification of components with retention of high biological activity was accomplished several years ago [Rasmussen et al. (1982) Science, 217, 159-162].  Subsequently, standard isolation techniques and molecular weight characterization by conventional mass spectrometric methods proved ineffective.  The pheromone was not identifiable by gas chromatography/ mass spectrometry (both electron impact and chemical ionization) nor by solid probe inlet electron impact mass spectrometry.  The pheromone appeared to be a compound of low volatility, of low molecular weight (200-500) and to be thermally labile.  It was not a peptide. The purification was hampered by close association of high concentrations of inactive components, often aromatics, which possessed similar chromatographic properties.  Purified by an empirically determined series of low pressure and high performance liquid chromatography fractionation sequences, guided at each step and in each preparation by high frequency flehmen responses from Asian bull elephants, the active sex pheromone is apparently a single entity.  Recent developments in field desorption mass spectrometric techniques allow molecular weight determinations on several micrograms of thermally labile substances; by this technique the protonated molecular weight was determined to be 249 and a tentative molecular weight of 248 is assigned.  Further information is presented on the physical and chemical properties of the elephant pheromone including its ultraviolet absorption maximum and it nuclear magnetic resonance spectrum.  Supported by NIH grant HD-19219-06.

Ratnasooriya, W.D., Fernando, S.B.U., Manatunga, A.N.V.R., 1991. Pregnancy duration of Sri Lankan elephant (Elephas maximus maximus) in captivity. Med. Sci. Res. 19, 623-624.

Taya, K., Komura, H., Kondoh, M., Ogawa, Y., Nakada, K., Watanabe, G., Sasamoto, S., Tanabe, K., Saito, K., Tajima, H., Narushima, E., 1991. Concentrations of progesterone, testosterone and estradiol-17B in the serum during the estrous cycle of Asian elephants (Elephas maximus). Zoo Biology 10, 299-307.

 1990. The story of Babe, the Asian elephant. Veterinary Viewpoints 2.

Cooper, K.A., Harder, J.D., Clawson, D.H., Fredrick, D.L., Lodge, G.A., Peachey, H.C., Spellmire, T.J., Winstel, D.P., 1990. Serum testosterone and musth in captive male African and Asian elephants. Zoo Biology 9, 297-306.
Abstract: Testosterone concentrations in serum samples collected weekly over a 5-year period from a young adult male Asian elephant (Elephas maximus) and a young adult male African forest elephant (Loxodonta africana cyclotis) were measured by radioimmunoassay.  Testosterone profiles during this maturational period were compared between the two species and related to the occurrence of musth, a recurring physiological and behavioral condition exhibited by most mature Asian, and some African, bull elephants.  Musth is characterized by secretion from the bull's temporal glands, dribbling urine, and increased aggression.  Serum testosterone concentrations in the Asian bull were elevated substantially between April and September each year, coincident with the presence of temporal gland secretion, urine dribbling, and aggressive behavior. Testosterone levels from April through September averaged (± SEM) 41.2 ± 2.8 ng/ml, compared to 7.6 ± 1.0 ng/ml during the rest of the year.  In contrast, the testosterone profile of the African bull showed greater variability and lower levels overall, the only pattern being a tendency for levels to be lowest from November to February (avg. 6.8 ± 1.5 vs. 10.3 ± 0.8 ng/ml during the rest of the year).  Temporal gland secretion and other signs of musth were first observed in this bull in 1988, at age 17.  While his testosterone values did not show a pattern comparable to that in the Asian bull, average testosterone values were significantly greater in 1988 compared to previous years.  The Asian bull showed sexual attention to preovulatory (estrous) cows whether in musth or not, and exposure to estrous cows did not appear to alter the highly consistent, annual pattern of musth as evidenced by temporal gland flow.

Dale, S. An elephant they'll never forget. Chicago Tribune October 8. 1990.
Ref Type: Newspaper

Diaz-Samayoa-de-Aguirre, L. Sex hormones in blood plasma, urine and faeces from female and male Indian elephants. Vergleichende Hormonuntersuchungen in Blutplasma, Harn und Kot beim weiblichen und mannlichen asiatischen Elefanten (Elephas maximus).  1-145. 1990. Hannover, Germany, Tierarztliche Hochschule.
Ref Type: Thesis/Dissertation

Foerner, J.J. Caesarian Section in the Elephant. 11th International Elephant Workshop Proceeding, Oct 24-27, 1990, Milwaukee County Zoo.  65-71. 1990. 1990.
Ref Type: Conference Proceeding

Mainka, S.A., Lothrop, C.D., Jr., 1990. Reproductive and hormonal changes during the estrous cycle and pregnancy in Asian elephants. Zoo Biology 9, 411-419.
Abstract: Serum progesterone and urinary total estrogen concentrations were determined weekly to bi-weekly in 2 female Asian elephants for 96 weeks.  The mean estrous interval was approximately 16 weeks in the nonpregnant animal.  A total of 5 cycles were observed in the 96 weeks study period.  The serum progesterone concentration ranged from 150 pg/ml to greater than 350 pg/ml during the luteal phase of the estrous cycle. The serum progesterone was elevated for 8-12 weeks of the 16 week estrous cycle.  The urinary total estrogen concentration ranged from less than 10 to greater than 300 pg/microgram creatinine.  The second animal was pregnant at the beginning of the study period.  The serum progesterone concentration was elevated (> 100 pg/ml) in the pregnant animal until parturition.  The urinary total estrogens increased from approximately 50 pg/microgram creatinine to greater than 400 pg/microgram creatinine during the first year of pregnancy and remained elevated until parturition.  Estrous cycling had not resumed by 3 months post partum.

McFarlane, J.R., Cabrera, C.M., Oosthuizen, M.M.J., Papkoff, H., 1990. Elephant pituitary gonadotropins. General and Comparative Endocrinology 79, 193-200.
Abstract: We describe for the first time the purification and some properties of luteinizing hormone (LH) and follicle-stimulating hormone (FSH) isolated from anterior pituitary tissue of the African Elephant (Loxodonta africana). Methodology previously applied to equine and donkey pituitaries was used to obtain purified preparations of elephant LH and FSH in yields of 8.8 and 0.48 mg, respectively, per 10 g pituitary powder.  The preparations were characterized by HPLC gel filtration and amino acid analysis, both of which showed the elephant LH and FSH to be very similar to ovine LH and FSH.  The preparations were also characterized by radioimmunoassays and bioassays for LH and FSH and a radioreceptor assay for FSH. Results showed virtually no cross-contamination of hormonal activities in the elephant LH and FSH preparations.  Elephant LH potencies ranged from 50 to 66% of highly purified ovine LH and elephant FSH potencies ranged from 21 to 52% of highly purified ovine FSH in the various assays employed.  No evidence was found for any demonstrable intrinsic FSH activity in elephant LH. The assays employed suggest possible usage for making physiological measurements of gonadotropins in elephants.

Niemuller, C., Gentry, P.A., Liptrap, R.M., 1990. Longitudinal study of haematological and biochemical constituents in blood of the Asian elephant (Elephas maximus). Comparative Biochemistry and Physiology [A] 96, 131-134.
Abstract: 1. Haematological parameters and biochemical analyses were determined in four elephants over a period of one year. 2. The haematological profile remained constant over time and was similar between animals. 3. Values for biochemical analyses were stable except for alkaline phosphatase, gamma glutamyl transferase and creatinine which rose during musth in male elephants. 4. The association of elevated enzyme levels and increased testosterone concentration is discussed.

Oosterhuis, J.E., 1990. The performance of a caesarian section on an Asian elephant (Elephas maximus indicus). Proceedings American Association of Zoo Veterinarians 157-158.

Rasmussen, L.E.L., Hultgren, B., 1990. Gross and microscopic anatomy of the vomeronasal organ in the Asian elephant. In: McDonald, D.W., Muller-Schwarze, D., Natynczuk, S.E. (Eds.), Chemical signals in vertebrates 5. Oxford University Press, New York, pp. 154-161.

Rasmussen, L.E.L., Hess, D.L., Haight, J.D., 1990. Chemical analysis of temporal gland secretions collected from an Asian bull elephant during a four-month musth episode. Journal of Chemical Ecology 16, 2167-2181.
Abstract: The temporal glands, modified facial apocrine sweat glands unique to elephants, release collectable secretions during an unusual physiological state termed "musth" in the Asian bull elephant (Elephas maximus).  Recently we began the characterization of the chemical components of musth, especially in the temporal gland secretions (TGS), and the examination of the role of such secretions as agents for chemical communication among elephants.  The presents study focuses on possible correlations between testosterone levels and the serum and temporal gland secretions.  We were especially interested in possible qualitative and/or quantitative changes in volatile compounds as the testosterone levels varied during a discrete musth period.  Ouantitative changes in TGS and serum testosterone were determined by radioimmunoassay.  Qualitative and semiquantitative changes occurring in volatile composition were studied by high-resolution gas chromatography (fused silica capillary column, on column injection).  Compound identification was by nuclear magnetic resonance, gas chromatography-mass spectrometr, and gas chromatography internal standards. Twenty-three major compounds and a number of minor components were identified.  Androgen concentrations were correlated with TGS-specific volatiles including benzoic acid, 2-nonanone, 5-nonanol, tetradecanoic acid and decanoic acid.  The latter two compounds and (E)-farnesol, a major component of African TGS, demonstrated an inverse relationship to T levels.

Rasmussen, L.E.L., Hess, D.L., Hall-Martin, A. Chemical profiles of temporal gland secretions from captive Asian bull elephants during musth and from African bull elephants living in wild but crowded conditions. Chemical Senses 15, 628. 1990.
Ref Type: Abstract
Abstract: Full-text: This study compares the volatile components of the temporal gland secretions of captive Asian bull elephants in musth and a distinctive group of wild African bull elephants, confined to a national park.  The captive Asian population has been well studied (Rasmussen et al., 1984; Rasmussen, 1988). Serum testosterone was elevated at specific times; aggressive behaviors occurred concomitantly with temporal gland secretions, although aggression and elevated serum testosterone were not always related.  Selected volatiles among the 23 compounds identified demonstrated concentration changes during the progression of musth, at times simultaneously with alterations in testosterone levels (Rasmussen et al., in press).  The African bull elephants have been monitored, behaviorally and physiologically, by radiocontrolled tracking and monthly sampling during the past 5 years.  Aggressive behaviors similar to those of Asian bull elephants have been documented; serum and temporal gland testosterone were elevated concomitantly in a cyclical fashion similar to musth in Asian elephants.  Chemical characterization of the volatiles of the temporal gland secretions from these bulls revealed several similarities to the compounds described in Asian bulls, including several compounds not previously described in African temporal gland secretions.  It is suggested that these chemicals, or other, more ephemeral compounds, may chemically inform other bulls and cows of the musth-like state of these bulls.

Roach, R., Briggs, M., Fithian, C. Determining the estrous cycle in a group of African elephants by evaluating serum progesterone levels. AAZPA Reg.Conf.Proc.  185-188. 1990.
Ref Type: Conference Proceeding

Brannian, J.D., Terranova, P.F., Griffin, F. Long and short estrous cycles in the African elephant: an endocrine profile. AAZPA Reg.Conf.Proc.  16-18. 1989.
Ref Type: Conference Proceeding

Brannian, J.D., Griffin, F., Terranova, P.F., 1989. Urinary androstenedione and luteinizing hormone concentrations during musth in a mature African elephant. Zoo Biology 8, 165-170.
Abstract: Musth has not been well documented in captive African elephants.  A 37-year-old African bull elephant in the Kansas City Zoological Park was observed during periods of behavioral musth and non-musth.  Androstenedione and luteinizing hormone (LH) concentrations in urine were measured by radioimmunoassay. Urinary androstenedione and LH levels were significantly higher in musth urine than in non-musth samples.  A positive correlation (P> 0.001) existed between urinary LH and androstenedione concentrations.  These results indicate that musth can occur in a zoo-maintained African elephant and that urinary androgen levels are elevated during musth, possibly as a result of LH stimulation of testicular steroidogenesis.

Chen, C.L., Olsen, J.H., Morris, L.S. Determination of reproductive cycle and pregnancy by radioimmunoassay (RIA) of serum progesterone in 13 Asian elephants. Proc.Am.Assoc.Zoo Vet.  203. 1989.
Ref Type: Conference Proceeding

Cooper, K.A. Reproductive endocrinology of male and female Asian and African elephants at the Columbus Zoo.  1989. Columbus, Ohio, USA,  Ohio State Unversity.
Ref Type: Thesis/Dissertation

Czekala, N.M., Roocroft, A., Bates, M. Estrogen metabolism in the Asian elephant (Elephas maximus). Biology of Reproduction 40[suppl.1], 119. 1989.
Ref Type: Abstract
Abstract: Understanding follicular development in elephants has been confounded by an inability to detect serum estradiol (E₂) in cyclic patterns.  Serum levels remain < 20 pg/ml throughout the cycle.  To understand E₂ dynamics in the elephant, two metabolic studies were initiated. First 500 microCi tritiated E₂ was injected (iv).  All urine and feces voided during the following 24 hrs. and two blood samples (30 min and 2 hrs) were collected.  No radiolabel appeared in feces during the 24 hrs.  Urinary radiolabel analyzed by HPLC appeared 100% conjugated, 52% E₂-3-conjugate (E₂C) and 48% estrone conjugate (E₁C). Peak levels appeared in urine 30 min post-injection, decreased rapidly during the next 2 hrs, then gradually to 16.5 hrs when levels stabilized.  Serum radiolabel at 30 min appeared as conjugated and free (66:34). In the second study, unlabelled E₂ (1.5 mg) was injected (iv). Blood was taken at 0, 5, 15, 30, and 60 min and urine at 30 min. In serum, peak estrogen levels appeared at 5 min (E₂:E₂C:E₁C, 38:39:22 or 61:38 conjugated:free).  Half-life of E₂ is ca.10 min and 60 min for E₂C.  E₁C peaked at 15 min and declined by 20% in 45 min.  Urine yielded similar results as the label study (E₂C:E₁C, 60:40).  Daily urine E₂C and weekly serum E₂C and progesterone (P) were measured by RIA in five mature female elephants.  Preliminary results indicated elevated levels of E₂C during the luteal phase similar to serum P profile.  The follicular phase contains a biphasic profile of E₂C.  These results suggest that E₂ is rapidly converted to E₂C and excreted in urine.

Dahl, K.D., Czekala, N.M., Hsueh, A.J.W. Measurement of urinary bioactive follicle-stimulating hormone (FSH) levels during reproductive cycles in diverse mammalian species. Biology of Reproduction 40[suppl.1], 119. 1989.
Ref Type: Abstract
Abstract: Recently an in vitro granulosa cell aromatase bioassay (GAB) was used to measure FSH in serum and urine samples (JCEM, March, 1987).  We now adapted the GAB assay to measure urinary bio-FSH levels in conjunction with the determination of urinary immunoreactive pregnanediol-3-glucoronide (PdG) and/or estrone conjugates (EC). Daily urine samples were collected from 2 monkey species, lion-tailed macaque (Macaca silenus) and golden monkey (Rhinopithecus roxellana), and 3 ungulates, giraffe (Giraffa camelopardalis), okapi (Okapi johnstoni) and Asian elephant (Elephas maximus).  The monkeys displayed a follicular phase rise in EC followed by a midcycle bio-FSH surge which declined as PdG concentrations increased during the luteal phase. Although both monkey species had similar cycle lengths (30-35 days), the golden monkey's follicular phase was much longer and displayed 2 bio-FSH peaks compared to 1 peak for the macaque. Although EC was not detectable in the ungulates, a midcycle FSH surge was followed by a luteal phase increase in PdG.  The closely related giraffe and okapi had similar cycle lengths (16-17 days), follicular phase lengths (8-9 days), and only 1 follicular phase FSH peak.  Conversely, the elephants had cycles lasting 16-19 weeks, and multiple FSH peaks were observed during the 5-6 week follicular phase.  Thus, 1) in contrast to the single follicular phase increase and preovulatory surge of bio-FSH seen in the macaque, giraffe and okapi, the finding of multiple follicular phase peaks in the golden monkey and elephants suggests the involvement of complex regulatory mechanisms; 2) the GAB assay provides a noninvasive and practical method for monitoring reproductive cycles in endangered species for future breeding programs.

de Villiers, D.J., Skinner, J.D., Hall-Martin, A.J., 1989. Circulating progesterone concentrations and ovarian functional anatomy in the African elephant (Loxodonta africana). Journal of Reproduction and Fertility 86, 195-201.
Abstract: Mean plasma progesterone concentrations measured in pregnant and non-pregnant elephants did not differ significantly from each other because of considerable variation, particularly for stage of pregnancy.  Maximum progesterone values were recorded during early pregnancy (5-8 months) and declined towards term (22 months).  The numbers of corpora lutea or total luteal tissue volume were not critical in maintaining progesterone secretion.  An increase in plasma progesterone concentrations with the luteal phase of the ovarian cycle was evident.  A possible role of the placenta in the second half of gestation is indicated by an increase in fetal progesterone concentrations towards term.

Harris, C.R. In search of a cervix. Proc.Ann.Elephant Workshop 10.  43-50. 1989.
Ref Type: Conference Proceeding

Li, C.H., Oosthuizen, M.M.J., Chung, D., 1989. Primary structure of elephant pituitary prolactin. International Journal of Peptide and Protein Research 33, 67-69.
Abstract: Tryptic digests of elephant pituitary prolactin (ele PRL) were separated by reverse-phase HPLC and paper electrophoresis. From the amino acid composition, the amino acid sequencing of selected peptides and from their alignment with expected tryptic peptides from ovine prolactin (based on the assumption that protein hormones with similar origins and bioactivities show significant degrees of homogenity), the primary sequence of ele PRL was proposed.

Payne, K., 1989. Elephant talk. National Geographic 176, 264-277.

Poole, J., 1989. Mate guarding, reproductive success and female choice in African elephants. Animal Behavior 37, 842-849.
Abstract: Male guarding of females, male mating success and female choice were studied for 8 years among a population of African elephants, Loxodonta africana.  Males were not able to compete successfully for access to oestrous females until approximately 25 years of age.  Males between 25 and 35 years of age obtained mating during early and late oestrous, but rarely in mid-oestrus.  Larger, older males ranked above the younger, smaller males and the number of females guarded by males increased rapidly late in life.  Body size and longevity are considered important factors in determining the lifetime reproductive success of male elephants.  Oestrous females exercised choice by soliciting guarding behavior from musth, but not non-musth males.  Females in mid-oestrus gave loud, very low frequency calls that may attract distant males and incite male-male competition.  The behavior of oestrous females resulted in their mating with males who were old, vigorous and healthy.

Poole, J.H., Moss, C.J. Elephant mate searching: group dynamics and vocal and olfactory communication. Symposia of the Zoological Society of London.  111-125. 1989.
Ref Type: Conference Proceeding

Poole, J.H., 1989. Announcing intent: the aggressive state of musth in African elephants. Animal Behavior 37, 140-152.
Abstract: Predictions derived from game theory suggest that animals should not signal their intentions during conflict situations.  However, during the period of musth, male elephants, Loxodonta africana, announce a state of heightened aggression with signals that are unbluffable.  Since smaller musth males in poor condition are able to dominate larger, normally higher-ranking, non-musth males in good condition, musth provides a useful system with which to examine the possibility of honest signaling of motivation, rather than of fighting ability.  Despite the highly aggressive state of males in musth, escalated contests are extremely rare.  The behaviour of musth and non-musth males suggests that opponents are able to estimate their often rapidly changing roles in the asymmetries with relative accuracy.  Since, unlike most other rutting animals, elephants have asynchronous sexually active periods, resource value varies both with age and the fluctuating sexual state of a particular individual.  It is suggested that musth may be a case where information about resource value is conveyed.

Ramsay, E.C., Leach, M.W. Postmortem reproductive findings in a female Asian elephant. Proc.Am.Assoc.Zoo Vet.  55. 1989.
Ref Type: Conference Proceeding

Schmidt, M.J. The role of the veterinarian in a captive elephant breeding program. Proc.Am.Assoc.Zoo Vet.  44-53. 1989.
Ref Type: Conference Proceeding
Abstract: Renewed interest in breeding elephants has led to the creation of new captive elephant breeding programs.  The veterinarian has a key role to play in the elephant breeding program.  The role of the veterinarian in an elephant breeding program is outlined; followed by brief discussion of specific elephant breeding problems which the veterinarian will be called upon to manage.

Schmidt, M.J., 1989. The fine art of elephant breeding. Animal Kingdom Sept / Oct, 45-51.

Turczynski, C.J., Jensen, J., Clarke, S., Kraemer, D.C. Immobilization, electroejaculation and semen characteristics of a male Asiatic elephant. Proc.Ann.Elephant Workshop 10.  51-56. 1989.
Ref Type: Conference Proceeding

Ullrey, D.E. Is vitamin E really the key to sexual satisfaction? Proc.8th Ann.Scholl Conf.Nutrition of Captive Wild Animals.  49-57. 1989.
Ref Type: Conference Proceeding

van der Horst, G., Kitchin, R.M., Curry, P.T., Atherton, R.W., 1989. Use of membrane filters and osmium tetroxide etching in the preparation of sperm for scanning electron microscopy. J. Electron Microsc. Tech. 12, 65-70.
Abstract: A new method was developed which is suitable for the preparation of mammalian sperm for scanning electron microscopy under either laboratory or field conditions. Samples of ejaculates from humans, two ferret species, and epididymal sperm from the African elephant were diluted in Millonig phosphate buffer and then fixed in glutaraldehyde solution. A small sample of the fixed sperm suspension was diluted in the same buffer, withdrawn with a syringe, and injected very slowly onto either a cellulose acetate or a polycarbonate membrane filter. This step was essential to concentrate the dilute sperm samples. During the various dilution steps most of the granular prostatic secretions were lost. However, a protein-like sheath, which remained attached to most sperm, obscured the surface features and had to be removed for SEM studies. It was removed by prolonged fixation/etching in 1% osmium tetroxide. Membrane filters containing sperm on their surfaces then were dehydrated, dried by the critical point drying method, and sputter coated with gold. Polycarbonate filters were superior to cellulose acetate filters in producing a flat and homogeneous background

Balke, J.M.E., Barker, I.K., Hackenberger, M.K., McManamon, R., Boever, W.J., 1988. Reproductive anatomy of three nulliparous Asian elephants: the development of artificial breeding techniques. Zoo Biology 7, 99-113.
Abstract: Detailed gross examinations of the reproductive tracts of three mature female nulliparous Asian elephants were conducted to develop artificial insemination (AI) techniques. Of primary concern was the determination of length characteristics and the size and configuration of the foramina between segments of the tract.  The elephants were 13, 28, and 40 years of age and had been maintained in captivity for most of their lives.  One elephant died naturally and two were euthanized for health reasons.  The reproductive tracts of two of the elephants were manually palpated in situ via the urogenital canal.  A fibreoptoscope was used to visualize the internal structures of the terminal reproductive tract of one elephant and to deposit dye into the vagina.  The reproductive organs were removed from the body cavity, dissected, measured, and photographed.  The major anatomical obstacles to overcome for standard AI procedures (the passage of an AI pipette into the reproductive tract) were the length of the urogenital canal (85-97 cm), the constriction at the urogenital-vaginal junction, and the tight cervix.  The reproductive anatomy was compared to that of previous dissections reported in the literature.

Balke, J.M.E., Boever, W.J., Ellersieck, M.R., Seal, U.S., Smith, D.A., 1988. Anatomy of the reproductive tract of the female African elephant (Loxodonta africana) with reference to development of techniques for artificial breeding. Journal of Reproduction and Fertility 84, 485-492.
Abstract: Complete reproductive tracts of 30 female African elephants (5-53 years), obtained during a population reduction procedure, were examined.  The reproductive tracts were palpated in situ via the urogenital canal.  A plastic speculum (1.3 X 170 cm) was introduced into the canal and dye was injected to simulate the procedure for artificial insemination.  The lengths of the reproductive tracts (from the vulva to the ovary) ranged from 120 to 358 cm.  The length increased with the size and age of the animal.   There was a membranous constriction (hymen) with an orifice, <2cm in diameter, between the urogenital canal and the vagina, in 4 primigravid and in all 13 nulliparous elephants.  The vaginal orifice of 13 multiparous elephants consisted of ragged folds of mucous membrane surrounding a single opening, 5-19cm in diameter.  The ages at first conception of 4 pregnant elephants with intact hymenal membranes were 10, 12, 13 and 14 years.  The hymen was not penetrated as a result of intromission and therefore the site of ejaculation would have been in the urogenital canal of the 4 primigravid elephants.

Brannian, J.D., Griffin, F., Papkoff, H., Terranova, P.F., 1988. Short and long phases of progesterone secretion during the oestrous cycle of the African elephant (Loxodonta africana). Journal of Reproduction and Fertility 84, 357-365.
Abstract: Serum samples were collected from 3 mature female African elephants once each week for 15-18 months.  Circulating concentrations of progesterone, oestradiol and LH were determined by radioimmunoassay (RIA).  The LH RIA was validated by demonstrating parallel cross-reaction with partly purified elephant LH pituitary fractions.  Changing serum progesterone concentrations indicated an oestrous cycle length of 13.3 ± 1.3 weeks (n=11).  The presumed luteal phase, characterized by elevated serum progesterone values, was 9.1 ± 1.1 weeks (n=11).  Two abbreviated phases of progesterone in serum lasting 2-3 weeks were observed in two elephants, indicating short luteal phases.  Oestradiol concentrations in serum were variable, with no clear pattern of secretion.  More frequent blood samples were collected during periovulatory periods and 9 distinct LH peaks were detected; all followed by rises in serum progesterone concentrations.  Periovulatory changes in progesterone and LH in sera correlated with external signs of oestrus and mating.

Hromadka, J. Observations made on an Asian elephant (Elephas maximus) during gestation. Proc.Ann.Elephant Workshop 9.  93-105. 1988.
Ref Type: Conference Proceeding

Koch, E., Elsaesser, F., Boer, M., Dittrich, L., Ellendorff, F., 1988. Pregnancy diagnosis in Asian and African elephants. Deutsche Tierarztliche Wochenschrift 95, 111-114.
Abstract: Serum progesterone concentrations were regularly measured by radioimmunoassay in non-pregnant African and Asiatic elephants after collection of blood from an ear vein. This revealed a cyclic pattern which did not occur in a pregnant Asiatic cow elephant. Mid-stream, morning urine was collected from spontaneously urinating animals. After cooling, both serum and urine were stored at -25°C and analyzed within 2 months. The urine of the pregnant elephant, as well as that of 3 other females which had previously been mated, was examined with human chorionic gonadotropin (HCG) cross-reacting substances, with the help of a commercial haemagglutination-inhibition test. The first female's pregnancy followed mating in May, 1983. A dead calf was born at term in February, 1985. In mid-November, 1983 the progesterone reading was 512 pg/ml and the HCG test was positive. At approximately monthly intervals for the following 3 months, the progesterone/HCG readings were, respectively 875 pg/ml positive, 265 pg/ml positive, and 439 pg/ml negative. In the 3 other females which had previously been mated, immunological cross-reactions with HCG were also detected. These results suggest that both methods, individually or in combination, might be used for pregnancy diagnosis in elephants.

Niemuller-Hare, C., Gray, C., Liptrap, R. A preliminary report on musth in male Asian elephants. Proc.Ann.Elephant Workshop 9.  106-111. 1988.
Ref Type: Conference Proceeding

Plotka, E.D., Seal, U.S., Zarembka, F.R., Simmons, L.G., Teare, A., Phillips, L.G., Hinshaw, K.C., Wood, D.G., 1988. Ovarian function in the elephant: luteinizing hormone and progesterone cycles in African and Asian elephants. Biology of Reproduction 38, 309-314.
Abstract: Serum samples were collected weekly for 3 yr from two female African elephants, for 18 mo from two other female African elephants, and for 2 yr from two female Asian elephants. Animals were not sedated at the time of blood collection. Ovarian cycles, characterized by changes in progesterone and immunoreactive luteinizing hormone (ILH) concentrations, averaged 15.9 + 0.6 wk (n=25) for African females and 14.7 + 0.5 wk for Asian females (n=10).  The length of the active luteal phase averaged 10.0 + 0.3 wk for African elephants (range 8-14 wk) and 10.6 + 0.6 wk for Asian females (range 9-13 wk).  One African female (Maliaca) had two extended interluteal phases, both occurring between the months of February and May. Excluding these two periods, there were no differences in the length of the luteal phase between species of elephant.  Serum progesterone in both species ranged from less than 50 pg/ml to 933 pg/ml.  Average progesterone concentrations during the luteal phase were significantly lower in African elephants compared with Asian elephants (328 + 13, n = 30 cycles vs. 456 + 23, n = 14 cycles; p<0.001). ILH ranged from nondetectable to 11.6 ng/ml.  These data suggest that the length of the ovarian cycle in the African elephant is about 16 wk and confirm that the length of the ovarian cycle in the Asian elephant is about 15 wk.

Rasmussen, L.E.L., 1988. Chemosensory responses in two species of elephants to constituents of temporal gland secretion and musth urine. Journal of Chemical Ecology 14, 1687-1711.
Abstract: This report discusses three areas of investigation: (1) The chemical components in the temporal gland secretion (TGS) of Asian (Elephas maximus) and African (Loxodonta africana) elephants were characterized by radioimmunoassay (RIA) for testosterone (T) and dihydrotestosterone(DHT) levels and by on-column capillary column gas chromatographic analysis of volatiles.  An inverse relationship between TGS testosterone levels and (E)-farnesol levels was observed. (2) African elephants responded preferentially toward a particular constituent of African elephant (TGS). (3) Urine from Asian bull elephants in musth was partially fractionated by high-performance liquid chromatography.  Specific chromatographic regions elicited dramatic avoidance responses from female African elephants.  These results support the suggestion that the TGS plays multiple chemocommunicative roles.

Teubner, V., Wells, S. Serum progesterone and cortisol levels in female Asian elephants (Elephas maximus).  1988.
Ref Type: Unpublished Work
Abstract: Poster presented at Symposium on Vertebrate Models in Endocrinology. National Institutes of Health, Bethesda, MD. 1988. Abstract. In the past zoological gardens and parks have not been at the forefront of endocrine research. However, zoos and wild animal parks represent a relatively untapped source of vertebrates for use as endocrine models.  The collaboration of academic institutions with zoo personnel on various projects will not only enhance the management of exotic species but will also add to the data base in endocrine research.  This is of critical importance to the propagation of endanged species.  The developement of radioimmunoassay techniques had made it possible to detect minute amounts of hormones in the blood or excreta of various species. This serves as a valuable tool in assessing the reproductive status of a given animal.  In species that do not exhibit the classic behavioral signs of estrous, measurement of hormonal activity can be the only link to evaluating fertility. The female Asian elephant (Elephas maximus) has a covert estrous cycle that only an adult bull elephant can detect.  Due to the problems associated with the maintenance of bull elephants in captivity the measurement of the hormonal activity of the female is a feasible alternative.  The goal of our study is to measure sex hormones (i.e. progesterone,and estradiol) and cortisol levels in two adult, female Asian elephants housed at the Audubon Zoo and to determine their estrous cycles.  Blood is sampled weekly from the ear vein of the each elephant.  Whole blood is then centrifuged and serum is stored at -70C for hormone measurement via radioimmunoassay.  Attempts to measure estradiol were unsuccessful as Asian elephants secrete a combination of estrone and estradiol.  Therefore, we chose to limit our analysis to progesterone, as the indicator of ovulation and to the glucocorticoid cortisol.  We determined that the elephants' estrous cycle has a duration of approximately 12 weeks with a 3.2 week follicular phase followed by a luteal phase of approximately 9 weeks.  Elevation of serum progesterone of 50 pg/ml above baseline and remaining elevated for 2 weeks was used as an indicator of luteal activity. Cortisol levels (ug/dl) were also measured and expressed as a percentage.  Serum cortisol was elevated either prior to a cycle or during a cycle.  This suggests that cortisol may also be a useful indicator of fertility. The hormonal data from the Asian elephant indicate that this species has a predictable estrous cycle that can be readily and accurately assessed using radioimmunoassay.  THe methodology used for this species may serve as a valuable research model to aid in captive breeding programs and to further our understanding of the endocrine systems of exotic species.

Balke, J.M.E., Read, B.W., Boever, W.J., Gibson, D., Miller, R.E., Junge, R.E., Seal, U.S., Plotka, E.D. Artificial insemination in elephants. AAZPA Reg.Conf.Proc.  652-658. 1987.
Ref Type: Conference Proceeding

Fischer, M.S., Trautmann, U., 1987. Fetuses of African elephants (Loxodonta africana) in photographs. Elephant 2, 40-45.
Abstract: Noticing that almost nothing has been published on the early ontogenetic development in elephants, we want to start to fill this gap by presenting pictures of elephant fetuses. All fetuses are African elephants (Loxodonta africana). Unfortunately, we do not know the age of the fetuses except for that largest one whichis about 8 months old.  All specimens were fixated in 4% formalin.  The legends will point to the peculiarities in the external morphology of each fetus.

Hall-Martin, A.J., 1987. The role of musth in the reproductive strategy of the African elephant (Loxodonta africana). South African Journal of Science 83, 616-620.
Abstract: Behavioral and endocrinological data on African elephants ( Loxodonta africana) are integrated to provide a hypothesis of the adaptations reflected in the phenomenon of musth in bulls.  Occupation of home ranges, movements, male dominance hierarchies and intra-specific agonistic behavior are reviewed.  Bulls in musth leave their home range, travel far and fast, imitate more contacts with distant breeding herds, show aggression which overrides normal social male hierarchies, probably mate more frequently than non-musth bulls and then return to their home range.  This behaviour is associated with elevated levels of serum testosterone and dihydrotestosterone. Elephants normally show a high degree of fidelity to sexually segregated adjoining home ranges, which results in regular contact between the same bulls and cows.  This breeding strategy is applicable to older, dominant bulls within the locally resident hierarchy.  The musth adaptation is a second strategy, whereby younger, lower ranking bulls (25-35 years) can ensure more contacts with cows and maximize their chances of breeding. Because musth bulls mate far from their normal ranges the strategy promotes gene flow and ensures outbreeding.  In English with Afrikaans summary.

Henneous, R.L., Schmidt, M.J., Haight, J.D. Deadly dilemmas of captive elephant breeding. AAZPA Ann.Conf.Proc.  1-5. 1987.
Ref Type: Conference Proceeding

Hodges, J.K., Mcneilly, A.S., Hess, D.L., 1987. Circulating hormones during pregnancy in the Asian and African elephants, Elephas maximus and Loxodonta africana: a diagnostic test based on the measurement of prolactin. International Zoo Yearbook 26, 285-289.

Li, C.H., Chung, D., Bewley, T.A., Cabrera, C.M., 1987. Elephant prolactin: Isolation and characterization. International Journal of Peptide and Protein Research 29, 472-477.
Abstract: Prolactin was isolated from anterior pituitary lobes of elephant pituitary glands.  It consisted of 199 amino acids with three disulfide bridges and two tryptophan residues as found in prolactin from other species.  The sequence of the NH₂ terminal 28 amino acids was determined and shown homologous with the ovine hormone.  In comparison with ovine prolactin, a marked difference was seen in the methionine content; the elephant hormone possessed only 18-34% lactogenic potency.  The conformation of elephant prolactin was examined by zero order, second order and circular dichroism spectroscopy. The alpha helical content was estimated to be about 60%.  In comparison with prolactins from other species, the second order spectra of elephant prolactin suggest [sic] that the local microenvironment for one or both tryptophan residues is somewhat different.

Marion, F. Update: Artificial insemination in the Asian elephant. Proc.Ann.Elephant Workshop. 8, 44-45. 1987.
Ref Type: Conference Proceeding

Matteri, R.L., Roser, J.F., Baldwin, D.M., Lipovetsky, V., Papkoff, H., 1987. Characterization of a monoclonial antibody which detects luteinizing hormone from diverse mammalian species. Domestic Animal Endocrinology 4, 157-165.
Abstract: The present study describes the development and characterization of a monoclonal antibody (518B7) generated against bovine LH (bLH). Although 518B7 was extremely specific for LH, very low species specificity was observed. A RIA using this antibody and radioiodinated equine LH (eLH) showed good sensitivity for all mammalian LH preparations tested, with the exception of human LH (15% relative to the eLH reference standard). Activities of most mammalian LH's ranged between approximately 50-200%. Much less activity was dectected with reptilian LH (<1.5%). Amphibian and avian LH fractions were essentially inactive. The reactivities of LH alpha and beta subunits from a variety of mammals clearly showed that the antibody reacts with the beta subunit. Sensitive RIAs were also developed utilizing 125I-bovine and 125I-rat LH. Interestingly, all hormone preparations which showed sufficient reactivity for statistical analysis within the dose ranges used in the present study (0.01-1000 ng/tube) produced a displacement curve parallel to the reference standard. We have also validated the use of 518B7 in detecting LH in serum. Parallel dilution curves relative to purified LH reference standards were observed with equine and bovine serum samples and equine pituitary extract. High (average 94%) recoveries were also seen with bovine serum with known amounts of exogenously added bLH and 518B7 and a previously described polyclonal antibody-based RIA in bovine serum samples during estrus. Thus, a monoclonal antibody for LH has been produced which can be used to develop sensitive and specific RIAs in many different mammalian species. This antibody can be readily produced in amounts sufficient to provide a stable source of a high quality LH antibody, which may find wide applications in endocrinological research dealing with both domestic and laboratory animals.

Merkt, H., Bader, H., Rath, D., Dittrich, L., 1987. An attempt to deep-freeze elephant semen. Deutsche Tierarztliche Wochenschrift 94, 488-489.

Poole, J., 1987. Elephants in musth, lust. Natural History 96, 46-55.

Poole, J., 1987. Raging bulls. Animal Kingdom 90, 18-25.

Poole, J.H., 1987. Rutting behavior in African elephants: the phenomenon of musth. Behavior 102, 283-316.

Rubel, A. Physiological and pathological conditions associated with reproduction of female Asian elephants at the Zurich zoo. Proc.1st.Intl.Conf.Zool.Avian Med.  379. 1987.
Ref Type: Conference Proceeding

Smith, D.A., Nadaraja, R., Beck, B., Honhold, N., Hale, D., Knottenbelt, D.C., Hill, F.W.G., 1987. Serum testosterone levels in male African elephants, Loxodonta africana, in Hwange National Park, Zimbabwe. Zimbabwe Veterinary Journal 18, 58-63.
Abstract: Age and serum testosterone levels were determined for 44 male African elephants (Loxodonta africana) from Hwange National Park.  Testicular weight was measured in 26 animals. Age and testicular weight were found to be highly correlated (r=0.94), while serum testosterone levels were correlated to both age (r=0.54) and testicular weight (r=0.43).  Although the range of serum testosterone levels and maximum value increased with age, the minimum value did not.

Balke, J.M.E., Read, B.W., Boever, W.J., Gibson, D., Miller, R.E., Seal, U., Plotka, E. Artificial insemination. Proc.Ann.Elephant Workshop  7.  18-21. 1986.
Ref Type: Conference Proceeding

Gorman, M.L., 1986. The secretion of the temporal gland of the African elephant, Loxodonta africana as an elephant repellant. Journal of Tropical Ecology 2, 187-190.

Hodges, J.K., 1986. Monitoring changes in reproductive status. International Zoo Yearbook 24/25, 126-130.

Howard, J.G., Bush, M., de Vos, V., Schiewe, M.C., Pursel, V.G., Wildt, D.E., 1986. Influence of cryoprotective diluent on post-thaw viability and acrosomal integrity of spermatozoa of the African elephant (Loxodonta africana). Journal of Reproduction and Fertility 78, 295-306.
Abstract: Electroejaculates from free-ranging African elephants were frozen to test various seminal diluents, freezing methods and thawing media on post-thaw sperm viability and structural integrity.  In Study I, each ejaculate was tested in each of 7 cryoprotective diluents.  After cooling to 5C and equilibration on ice (4C) for 120min, each aliquot was pellet frozen on solid CO₂, stored in liquid nitrogen and thawed (37C) in saline or tissue culture solution.  Amongst the diluents, post-thaw sperm motility, motility duration in vitro (37C) and acrosomal integrity were greatest when diluent BF5F was used.  Thawing medium had no effect on results.  In Study II, the optimal diluent from Study I (BF5F) was compared with the diluent SGI. Results were not affected by a 90- or a 150-min cooling-equilibrium interval in an electronic cooler (5C); however, post-thaw sperm motility rating and duration of motility in vitro were grater with the pellet than the straw container freezing method.  When the pelleting method was used, diluents BF5F and SGI provided comparable cryoprotection. Duration of post-thaw motility was enhanced 2-fold and up to 12h my maintaining thawed semen at 21 rather than 37C.  All diluents provided some protection on acrosomal integrity, but the overall proportion of intact acrosomes after thawing was markedly less in Study II, apparently as a result of the slower initial cooling rate (approx 1.5C/min) compared to that of Study I (approx 6.5C/min).  This study demonstrates the feasibility of cryopreserving semen from free-ranging African elephants and indicates that spermatozoa most effectively survive freezing when the BF5F or SGI diluent is used in conjunction with the pelleting method.

Kahl, V.A.L. The study of the reproductive cycle of the African elephant (Loxodonta africana) through correlation of behavior and cytology with serum and urinary hormonal patterns.  1986. Nebraska, Omaha, USA, University of Nebraska.
Ref Type: Thesis/Dissertation

Mainka, S.A. Monitoring reproductive cycles in two Sri Lankan elephants. Proc.Ann.Elephant Workshop. 7, 4-9. 1986.
Ref Type: Conference Proceeding

Merkt, H., Ahlers, D., Bader, H., Rath, D., Brandt, H.P., Boer, M., Dittrich, L., 1986. Aftercare and recovery of a female Indian elephant after delivery of a dead fetus by episiotomy. Berl. Munch. Tierarztl. Wochenschr. 99, 329-333.

Munson, L., Heuschele, W., O'Banion, M.K., Sundberg, J.P., Oosterhuis, J.E., 1986. Polyp in the urogenital canal of an African elephant. Journal of the American Veterinary Medical Association 189, 1190-1191.

Price, P., Bradford, J., Schmitt, D. Collection and semen analysis in Asian elephant. AAZPA Ann.Conf.Proc.  310-311. 1986.
Ref Type: Conference Proceeding

Ryder, O.A., 1986. Genetic investigations: tools for supportings breeding programme goals. International Zoo Yearbook 24/25, 157-162.

Byron, H.T., Olsen, J., Schmidt, M.J., Copeland, J.F.Jr., Byron, L., 1985. Abdominal surgery in three adult male Asian elephants. Journal of the American Veterinary Medical Association 187, 1236-1237.

Howard, J.G., Bush, M., Schiewe, M.C., de Vos, V., Wildt, D.E., 1985. Further developments in comparative semen freezing in free-ranging African elephants. Proceedings American Association of Zoo Veterinarians 31-32.

Jones, D.K., 1985. Horizons: Kenya: Aberdares Salient. Swara 8, 20.

Jones, D.K., 1985. New life at the Ark. Swara 8, 29-31.

Merkt, H., Ahlers, D., Bader, H., Brandt, H.P., Boer, M., Dittrich, L., 1985. Episiotomy, a new obstetrical intervention in elephant-cows. Deutsche Tierarztliche Wochenschrift 92, 428-432.

 1984. Some recorded elephant births. Loris Dec., 308-310.

Craig, G.C., 1984. Foetal mass and date of conception in African elephants: a revised formula. South African Journal of Science 80, 512-516.
Abstract: Existing information on South-Central African elephants is used to show that there is an error in the published formula for calculating date of conception from foetal mass.  A revised formula, t= 106w^1/3 + 138, is proposed, where t is the age of the foetus and w is foetal mass, which implies a faster foetal growth-rate following a longer early phase of slow growth than previously assumed.  The revised formula results in a clearer illustration of the seasonality of elephant breeding, though caution is recommended in the timing of sampling and the application of the formula to small foetuses.

Hall-Martin, A.J., van der Walt, L.A., 1984. Plasma testosterone levels in relation to musth in the male African elephant. Koedoe 27, 147-149.

Howard, J., Bush, M., de Vos, V., Wildt, D.E., 1984. Electroejaculation, semen characteristics and serum testosterone concentrations of free-ranging African elephants (Loxodonta africana). Journal of Reproduction and Fertility 72, 187-195.
Abstract: A regimented electroejaculation protocol (120 electrical stimulations; 10-30 V) was used to collect semen and characterize ejaculate quality from 9 adult, free-ranging African elephants under anesthesia.  Eight of the 9 ejaculates contained high concentrations of progressively motile spermatozoa.  The overall mean ejaculate volume, sperm concentration/ml ejaculate, sperm motility, sperm status and ejaculate pH were 93.3 ml, 2408.6 x 10(6) spermatozoa/ml, 70%, 3.9 and 7.4, respectively.  A high percentage (mean 77.5%) of spermatozoa whin each ejaculate was morphologically normal.  Of the aberrant spermatozoa, 72% had a cytoplasmic defect.  When sperm viability was tested in vitro at 37 C, sperm motility rating declined by at least half of the initial assessment within 3.5 h of semem collection.  Generally, spermatozoa maintained motility in vitro for < 6 h.  Serum testosterone ranged from 1.4 to 8.2 ng/ml in 4 males evaluated in the morning (07:30 - 08:00 h).  In 4 of the 5 bulls assessed in the afternoon (15:00 - 18:00 h), testosterone levels were < 0.9 ng/ml.  The remaining bull, evaluated at 16:00 h, had exceptionally high testosterone concentrations (peak 25.6 ng/ml) and a preputial discharge potentially indicative of "musth." The present study demonstrates that high quality semen call be collected consistently from the African elephant and that striking differences exist in serum testosterone amongst free-ranging males which may be due, in part, to a diurnal rhythm.

Kock, N., Kock, M., Arif, A., Wahid, M.N.S.A. Immobilization techniques and complications associated with a bull Indian elephant (Elephas maximus indicus) during musth. Proc.Am.Assoc.Zoo Vet.  68-74. 1984.
Ref Type: Conference Proceeding

Kock, N., Kock, M. Management of two Indian elephants (Elephas maximus indicus) in a middle eastern zoo. Proc. Amer. Assoc. Zoo Vet.  75-81. 1984.
Ref Type: Conference Proceeding

Parker, J., 1984. Preparations for artificial insemination in two Asian elephant cows. Animal Keepers' Forum 11, 420-423.

Poole, J.H., Kasman, L.H., Ramsay, E.C., Lasley, B.L., 1984. Musth and urinary testosterone concentrations in the African elephant (Loxodonta africana). Journal of Reproduction and Fertility 70, 255-260.
Abstract: Urine samples were obtained from free-ranging African elephants that were considered to be in and out of musth. Testosterone concentrations, measured by radioimmunoassay were significantly greater in males that were in or around the time of behavioral musth.  This study supports a correlation between the observed behavioral characteristics of musth and urinary testosterone levels.

Ramachandran, K.K., 1984. Observations on unusual sexual behaviour in elephants. Journal of the Bombay Natural History Society 81(3), 687.

Rasmussen, L.E., Buss, I.O., Hess, D.L., Schmidt, M.J., 1984. Testosterone and dihydrotestosterone concentrations in elephant serum and temporal gland secretions. Biology of Reproduction 30, 352-362.
Abstract: Serum and termporal gland secretions (TGS) were obtained from mature wild African (Loxodonta africana) and captive Asian (Elephas maximus).  Samples were obtained from five cows and eight bulls culled for management purposes in Kruger National Park, South Africa, and from four females and two males residing at the Washington Park Zoo, Portland, Oregon.  Our purpose was to describe the levels of androgens, testosterone, and dihydrotestosterone, and to correlate these observations with sex, species, and behavioral status.  Male-female differences in serum T were pronounced in the Asian species, whereas male and female concentrations overlapped in the African elephant serum. Serum T concentrations in African females were > than in Asian females.  Serum DHT reflected T levels, except that the striking elevation of testosterone in Asian bulls during musth was not paralleled by = increases in DHT.  A species difference observed among males was higher serum T levels in nonmusth Asian bulls (1.84-5.35ng/ml) compared to levels in African bulls (0.38-0.68ng/ml), except for one dominant African bull (6.64ng/ml).  This single African value was still considerably lower than the serum T values of the Asian males during musth. These musth values were the highest serum androgen concentrations: T was between 19 and 40ng/ml (average 26.1 ng/ml).  The TSG values of T and DHT were much higher than serum levels except in the Asian female.  T/DHT ratios in TGS were more similar than in serum.  One dominant African bull had a T TGS value of 78ng/ml, which was much higher than the rest of the African males or females, but considerably lower than an Asian bull in musth (547ng/ml).  It seems apparent that a change in androgen status as reflected in serum and TGS levels of T and DHT precedes or is concomitant with overt alteration in behavior in the Asian male.  The temporal gland appears to actively concentrate androgens in both African males and females, but in the Asian male the gland secretes only during musth when the greatest concentration of both T and DHT were observed.  The apparent difference in the degree of temporal gland secretory activity between the 2 species suggests a more specific communicative function within the Asian male.

Rowlands, I.W., Weir, B.J., 1984. Mammals: non-primate eutherians. In: Lamming, G.E. (Ed.), Marshall's physiology of reproduction. Churchill Livingstone, New York, pp. 455-658.

Short, R.V., 1984. Oestrous and menstrual cycles. In: Austin, C.R., Short, R.V. (Eds.), Hormonal control of reproduction. Cambridge University Press, New York, pp. 115-152.

Siegel, R.K., 1984. LSD-induced effects in elephants: comparisons with musth behavior. Bulletin of the Psychonomic Society 22, 53-56.
Abstract: Musth is a condition observed in male Asiatic elephants and is characterized by aggression and temporal gland secretion.  A classic and controversial 1962 study attempted to induce a musth syndrome in an elephant via treatment with LSD. Two elephants in the present study survived dosages of LSD (.003 -.10 mg/kg) and exhibited changes in the frequency or duration of several behaviors as scored according to a quantitative observational system.  LSD increased aggression and inappropriate behaviors such as ataxia.  Results are discussed in terms of musth and drug-induced perceptual-motor dysfunction.

De Alwis, L. Captive breeding of the Sri Lankan elephant (Elephas maximus maximus) -- a new approach. Elephant Symposium,Washington Park Zoo,September 1983.  1983.
Ref Type: Conference Proceeding
Abstract: .

Dublin, H., 1983. Cooperation and reproductive competition among female African elephants. In: Wasser, S.K. (Ed.), Social behavior of female vertebrates. Academic Press, New York, pp. 291-313.

Flanagan, H.O., Flanagan, F.O., 1983. Castration of African elephant Loxodonta africana africana. Zimbabwe Veterinary Journal 13, 50-51.
Abstract: The successful castration of an African bull elephant, Loxodonta africana africana, is described, with a resultant increase in docility. It is possible that, with castration, more use could be made of baby bulls captured during culling operations.

Heath, E., Jeyendran, R.S., Graham, E.F., 1983. Ultrastructure of spermatozoa of the Asiatic elephant (Elephas maximus). Zbl. Vet. Med. C. Anat. Histol. Embryol. 12, 245-252.
Abstract: A scanning and transmission electron microscopic study of semen collected with the aid of an artificial vagina was carried out.  The ultrastructural characteristics of Asiatic bull elephants is compared to that of other mammalian species.

Hess, D.L., Schmidt, M.J., Schmidt, A.M., 1983. Reproductive cycle of the Asian elephant (Elephas maximus) in captivity. Biology of Reproduction 28, 767-773.
Abstract: Serum samples were obtained either weekly or daily from six mature elephant cows for periods of 7 to 30 months from April 1979 to November 1981, to rigorously describe the hormonal and behavioral components of the reproductive cycle in the Asian elephant.  The estrous state of each elephant was concurrently monitored through quantification of the frequency of male urine testing behavior (Flehmen-like response) under controlled conditions.  Concentrations of progesterone and estradiol were determined in 1.0 ml of serum by RIA after diethylether extraction and purification on Sephadex LH-20 columns.  the presence of gonadotropic activity was assessed with a rat LH RIA and by an in vitro mouse Leydig cell bioassay.  Fifteen ovarian cycles averaging 16.3+0.4 weeks in length were observed in six females.  The luteal phase was 10.5+0.3 weeks, and 5.1+0.4 weeks separated subsequent luteal periods. Estradiol concentrations were extremely variable, and no distinct preovulatory E2 surge was associated with the onset of P4 secretion.  However, the onset of P4 release occurred during a single 24h period and reached maximum levels within 2 to 3 weeks.  Peaks of immunoreactive LH were observed, although more than 50% of such "surges" were unrelated to increments in P4 secretion, and no evidence of bioactivity was observed in any serum sample.  Pregnancy was indicated by continued secretion of P4 after the 12-week luteal phase, and elevation levels (400-1200 pg/ml) were seen throughout the first year and during the last 6 months of gestation.  Serum concentrations of P4 fell dramatically before parturition, consistent with a role for this steroid in the regulation of gestational length. Male urine testing behavior was clearly cyclical, and maximal interest during the late interluteal period was followed by a rapid decrease with the onset of P4 secretion.  These data provide the first substantive description of the hormonal milieu of the reproductive cycle, concurrent male behavior, and the serum hormonal patterns at the onset and termination of gestation in this endangered species.

Hodges, J.K., Henderson, C., Mcneilly, A.S., 1983. Circulating oestrogen concentrations during pregnancy in the African elephant (Loxodonta africana). Journal of Reproduction and Fertility 67, 121-127.
Abstract: Oestrone, Oestradiol-17B and oestriol were measured in plasma samples from non-pregnant and pregnant African elephants shot in the wild.  Enzymatic hydrolysis of plasma showed that approximately 90 and 96% of the total (i.e. conjugated plus unconjugated) concentrations of oestrone and oestradiol-17B, respectively were represented by conjugated hormones.  Unconjugated oestrogens remained low (<50pg/ml) in all samples, with no distinction between non-pregnant and pregnant animals.  Levels of total oestrone during pregnancy varied between 160 and 594 pg/ml but were not significantly different from non-pregnant values.  Total oestradiol-17B concentrations were significantly elevated during pregnancy (P < 0.01) and, despite considerable individual variation (193-1428pg/ml), were consistently higher than non-pregnant values after 6 months gestation.  The elevated levels of oestradiol-17B resulted in a reversal of the total oestradiol-17B: oestrone concentration ratio at about 6 months of pregnancy.  Concentrations of total oestriol did not exceed 103 pg/ml.  An indirect method of measurement indicated that oestradiol-17B sulfate was probably the most abundant circulating oestrogen during pregnancy in the African elephant.

Jacob, V., Cheeran, K., Chandrasekharan, K., Radhakrishnan, K. Immobilization of elephant in musth using xylazine hydrochloride. 7th Annual Symposium of the Indian Society of Veterinary Surgeons.  62. 1983. Kerala, India.
Ref Type: Conference Proceeding

Leach, E. Vaginal virus in a mixed elephant herd. Proc.Ann.Elephant Workshop 4.  79-80. 1983.
Ref Type: Conference Proceeding

Mcneilly, A.S., Martin, R.D., Hodges, J.K., Smuts, G.L., 1983. Blood concentrations of gonadotropins, prolactin and gonadal steroids in males and in non-pregnant and pregnant female African elephants (Loxodonta africana). Journal of Reproduction and Fertility 67, 113-120.
Abstract: No seasonal variation in any of the hormones measured was apparent in males or females. Testosterone levels in males increased around puberty (10-11 years) and remained significantly higher in adults than prepubertal males. This was not accompanied by any significant change in levels of LH, FSH or prolactin.  In non-pregnant females there was no apparent difference in levels of LH, FSH or prolactin with age.There was a significant increase in progesterone around puberty (12 years) but there was considerable overlap in values between prepubertal and adult females. During pregnancy, progesterone levels were significantly higher than in non-pregnant females with maximum levels occurring at mid-pregnancy (9-12 months).  However, there was considerable overlap in values between non-pregnancy and pregnancy. Concentrations of LH and FSH decreased significantly during mid-pregnancy while prolactin levels increased dramatically during pregnancy; after 7 months gestation until term levels were always at least 8ng/ml greater than in any non-pregnant female.  It is suggested that this consistent increase in plasma/serum levels of prolactin can be used to diagnose pregnancy in the elephant.

Moss, C.J., 1983. Oestrous behavior and female choice in the African elephant. Behavior 86, 167-196.

Ruedi, D., Kupfer, U., Girard, J., Gutzwiller, A., 1983. Untersuchungen zur fortflanzungsphysiologie biem Afikanischen elefanten (Loxodonta africana): Samengewihhung bei wildbullen, weitere schritte hinsichtlich kunstlicher besamung. Erkrankungen der Zootiere 341-381.

Abeyratne, A.S., 1982. Elephant breeding -- some scientific facts. Loris 16, 91-93.

Campbell, S., 1982. A matter of survival: what is the gestation period for a pregnant elephant? Zoonooz 6, 10-11.

Gehring, H., Schroder, H.D., 1982. Castration of an elephant Elephas maximus. Zoologische Garten 52, 365-368.

Hromadka, J., 1982. Birth and rearing of Elephas maximus. Animal Keepers' Forum 9, 294-299.

Rasmussen, L.E., Schmidt, M.J., Henneous, R., Groves, D., Daves, G.D.Jr., 1982. Asian bull elephants: flehmen-like responses to extractable components in female elephant estrous urine. Science 217, 159-162.
Abstract: Flehmen-like responses (urine tests) are one of the characteristic behavioral reactions of male Asian elephants (Elephas maximus) to cow elephants in estrus.  Components of the urine of estrous cow elephants were extracted with organic solvents and partially purified by chromatography and shown to evoke Flehmen-like responses when they were presented to adult bulls.

Schmidt, M.J., 1982. Studies on Asian elephant reproduction at the Washington Park Zoo. Zoo Biology 1, 141-147.
Abstract: Studies of Asian elephant reproduction at the Washington Park Zoo in the areas of estrous cycle, semen collection, bull elephant management and health care, pregnancy, and pheromones have yielded sufficient information to attempt repeated artificial insemination (AI) at the time of ovulation. While no pregnancy has been achieved to date, with the information now at hand AI can be expected to become a practical technique for breeding elephants in captivity.

Shoshani, J., Alder, R., Andrews, K., Baccala, M.J., Barbish, A., Barry, S., Battiata, R., Bedore, M.P., Berbenchuk, S.A., Bielaczyc, R., Booth, G., Bozarth, N., Bulgarelli, M.A., Church, I., Cosgriff, J.W.Jr., Crowe, H., DeFauw, S.L., Denes, L., Efthyvoulidis, E., Ekstrom, M., Engelhard, J.G., English, P., Fairchild, D.Jr., Fisher, C., Frahm, K., Frederick, D., Fried, J., Gaskins, T., Gatt, J., Gentles, W., Goshgarian, H.G., Grabowski, S., Haase, D., Hajj, K., Hall, G., Hawkins, D., Heberer, C., Helinski, A., Henry, S.R., Heyka, C., Hurt, M., Kemppainen, M., Kendra, C., Koenig, J., Konarske, P., Konwinski, S., Kopacz, S., Lakits, V.T., Jr., Lash, S.S., Laughlin, D.C., Meyers, S., Mizeres, N.J., Morehead, K.M., Muraski, A., Murphy, S., Niebala, J., Overbeck, G., Powitz, R., Rafols, J.A., Raymer, S.L., Rezzonica, L., Rossmoore, H.W., Sabo, D., Schwikert, P.J., Shy, E., Skoney, J., Smith, D., Spodarek, K.L., Sujdak, P.J., Tarrant, T., Thielman, R., Tisch, F., Wolowicz, L., Williams, J., Yehiel, D., 1982. On the dissection of a female Asian elephant (Elephas maximus maximus Linnaeus, 1758) and data from other elephants. Elephant 2, 3-93.
Abstract: A 46-year-old female Asian elephant (Elephas maximus maximus Linnaeus, 1758), named "Iki", died on July 8, 1980, at the Ringling Brothers and Barnum & Bailey Circus, Circus World, Haines City, Florida, USA.  She was transported to Detroit and was dissected by the Elephant Interest Group (EIG) and friends, Department of Biological Sciences, Wayne State University.  The purpose of this continuing study has been to collect data supplemental to that of previous workers, and to enrich knowledge of elephant anatomy, particularly in areas not thoroughly investigated in the past.  Some of these findings were compared to those observed in other elephants:  "Shirley", "Tulsa", and "Toose" and to the organs of "Ole Diamond" and "Hazel"  (see Appendix II).

Styles, T.E., 1982. Birth and early development of an African elephant Loxodonta africana at the Metro Toronto Zoo, Canada. International Zoo Yearbook 22, 215-217.

Wheeler, J.W., Rasmussen, L.E., Ayorinde, F., Buss, I.O., Smuts, G.L., 1982. Chemical constituents of temporal gland secretion of the African elephant, Loxodonta africana. Journal of Chemical Ecology 8, 821-835.
Abstract: Temporal gland secretion (TGS), obtained from 15 different mature African elephants in Kruger National Park was analyzed for volatile constituents.  Only five volatile components were present. p-Cresol was present in all samples, but phenol was found as an appreciable component of only one sample and as trace amounts in six others.  Three sesquiterpenes were identified, the latter two being new natural products: E-farnesol, farnesol hydrate (3,7,11-trimethyl-2,10-dodecadien-1,7 diol), and farnesol dihydrate (3,7,11-trimethyl-2-dodecen-1,7,11-triol).  These sesquiterpenes represent the first isolated from mammals.  Ten samples of TGS, serum, and saliva were assayed for cholesterol, urea, and proteins including several enzymes.

 1981. Annual Projects Report:  Amboseli elephant research project. Wildlife News 16, 9.

Jones, R.C., Brosnan, M.F., 1981. Studies of the deferent ducts from the testis of the African elephant, Loxodonta africana. I. Structural differentiation. Journal of Anatomy 132, 371-386.

Jones, R.C., Holt, W.V., 1981. Studies of the deferent ducts from the testis of the African elephant, Loxodonta africana. III. Ultrastructure and cytochemistry of the ductuli efferentes. J Anat 133 (Pt 3), 247-255.
Abstract: The epithelium of the ductuli efferentes is composed of ciliated, principal, halo and basal cells. The supranuclear cytoplasm of ciliated cells is penetrated by particularly long cilial rootlets which are surrounded by numerous elongate mitochondria. Microtubules are arranged along the longitudinal axis of the cells. The spaces between the microvilli of principal cells form canaliculi which penetrate the apical cytoplasm and appear to be involved in endocytotic activity. The supranuclear cytoplasm contains oval mitochondria and numerous vacuoles. Both ciliated and principal cells contain poorly developed Golgi and endoplasmic reticulum, but numerous supranuclear dense bodies are usually present. Supranuclear and basal accumulations of dense bodies were identified as lipofuscin; they were the source of brown pigmentation in the proximal two thirds of the ductuli efferentes. The halo cells were probably macrophages. They occurred quite frequently and contained crescent shaped nuclei and large
accumulations of lipofuscin material.

Mann, P.C., Bush, M., Jones, D.M., Griner, L.A., Kuehn, G.R., Montali, R.J. Leiomyomas of the genital tract in large zoo mammals. Laboratory Investigation 44[1], 40A. 1981.
Ref Type: Abstract
Abstract: From the 70th Annual Meeting of the International Academy of Pathology, U.S.A. -- Canadian Division, Chicago, Ill,USA, March 2-6, 1981. Abstract. "Leiomyomas of the female genital tract occurred in four Indian rhinoceroses (Rhinoceros unicoris) and three Indian elephants (Elephas maximus indicus) exhibited at the National Zoological Park, Washington, D.C., Regent's Park Zoo, London, England, San Diego Zoo, San Diego, California, and Los Angeles Zoo, Los Angeles, California.    The majority of the animals were aged without recent reproductive activity.  Tumors were mainly in the vaginal tract in the rhinoceros and the uterus of the elephants.  The tumors in two of the three rhinoceroses were discovered clinically via rectal palpation.  A postmortem examination of the rhinoceros at the National Zoo showed endometrial cysts and a large (25 cm) follicular cyst of one ovary.  Ovarian cysts were also found in one of the elephants.  The tumors consisted of circumscribed collections of interlacing, well differentiated, smooth muscle-like cells with varying amounts of connective tissue. The uterine tumors were all intramural, whereas the vaginal tumors in the rhinoceros were often pedunculated.  Although intrauterine leiomyomas (fibroids) are extremely common in women, they are very rare in domestic animals.  The role of hyperestrinism in leiomyoma induction remains controversial in humans, and is presently unknown in animals.  The prevalence of cystic ovaries and reproductive difficulties may indicate a hormonal relationship with leiomyomas in zoo animals as well."

Nambiar, M.O.R., 1981. Leptaden (vet) as a galactagogue in an elephant with deficient lactation. Indian Veterinary Journal 58, 667-668.

Poole, J.H., Moss, C.J., 1981. Musth in the African elephant, Loxodonta africana. Nature 292, 830-831.
Abstract: The phenomenon of musth in male Asian elephants, Elephas maximus, has long been recognized.  Musth, which has been likened to rutting behaviour in ungulates, refers to a set of physical and behavioural characteristics displayed periodically by adult male elephants.  The most obvious manifestations are a sharp rise in aggressive behaviour, copious secretions from and enlargement of the temporal glands, and a continuous discharge of urine.  It has been speculated that a similar phenomenon occurs in males of the African genus, Loxodonta africana, but most workers have concluded that it does not exist.  Here we show that musth does occur in the African elephant and that its manifestations are similar to those in the Asian elephant.

Ramsay, E.C., Lasley, B.L., Stabenfeldt, G.H., 1981. Monitoring the estrous cycle of the Asian elephant (Elephas maximus), using urinary estrogens. American Journal of Veterinary Research 42, 256-260.
Abstract: The estrous cycle of the Asian elephant (Elephas maximus) was monitored by analysis of urinary estrogens.  Daily morning urine samples were analyzed for estrone (E₁), estradiol (E₂), and total immunoreactive estrogen (E_T).  The E_T values were shown to correlate poorly with E₁ and E₂ and failed to reveal any patterns of reproductive cycling. Daily E₁ and E₂ values, indexed by creatinine concentrations, demonstrated cyclic profiles in those samples of sufficient concentrations.  The technique offered a simple, noninvasive method for determining ovarian function in the elephant.

Ruedi, D., Kuepfer, U. Semen collection in the African elephant (Loxodonta africana): a step towards artificial insemination. Proc.Am.Assoc.Zoo Vet.  142-143. 1981.
Ref Type: Conference Proceeding

Schmidt, M.J., Hess, D.L., Schmidt, A.M., Henneous, R.L., Groves, D.A., Haight, J.D. The estrous cycle of the Asian elephant. Proc.Am.Assoc.Zoo Vet.  91-95. 1981.
Ref Type: Conference Proceeding

Ali, S.M., 1980. Some aspects of behaviour and breeding biology of the Indian wild elephant. Tigerpaper 7, 9.

Holt, W.V., Jones, R.C., Skinner, J.D., 1980. Studies of the deferent ducts from the testis of the African elephant, Loxodonta africana.  II. Histochemistry of the epididymis. Journal of Anatomy 130, 367-379.
Abstract: The three main segments of the elephant epididymis were examined for the occurrence, in the spermatozoa and lining epithelium, of carbohydrates, neutral lipids and phospholipids, ATPase, alkaline phosphatase, succinic dehydrogenase, glucose-6-phosphate dehydrogenase, diaphorases, hydroxysteroid dehydrogenases, acid phosphatase and non-specific esterase. The most distinct feature of the carbohydrate content of the epididymis was a layer of acidic, alcian blue-positive glycoprotein over the luminal surface of the epithelium, particularly in the terminal segment. PAS-positive, diastase-resistant inclusions were also found throughout the epididymis. Neutral lipid occurred as droplets above and below the nucleus in the epithelium of the middle segment, and as supranuclear accumulations in the terminal segment. All the enzymes except the steroid dehydrogenases were detected in the epididymal epithelium, and all except the steroid dehydrogenases and acid phosphatase were detected in the spermatozoa. There was considerable variation in the intensity of the cytochemical reactions in the epithelium, but not in the spermatozoa, in different regions of the epididymis. In general, the enzymes involved in active transport showed strongest reactions in the initial and terminal segments, the reactions in the stereocilia being the most intense. The enzymes involved in energy metabolism showed strongest reactions in the middle and terminal segments, with the activity being fairly evenly distributed throughout the cytoplasm of the principal cells. However, the two lysosomal enzymes which were studied showed quite different distributions: the reactions for acid phosphatas were strongest in the initial and middle segments, whilst the reactions for non-specific esterase were strongest in the middle and terminal segments. It is suggested that the initial segment is involved in absorptive and anabolic activity, the middle segment in anabolic activity, and the terminal segment (where spermatozoa are stored ready for ejaculation) in considerable metabolic activity and active transport of substrates across the epithelium.

Jones, R.C., 1980. Luminal composition and maturation of spermatozoa in the genital ducts of the African elephant (Loxodonta africana). Journal of Reproduction and Fertility 60, 87-93.

Percival, R., 1980. The mating of elephants. Loris 15, 232.

Strazielle, L., 1980. Birth of an Asian elephant at the Paris Zoo. Mammalia 44, 592-594.

 1979. "Motty" -- Birth of an African/Asian elephant at Chester Zoo. Elephant 1, 36-40.

Chappel, S.C., Schmidt, M.J., 1979. Cyclic release of luteinizing hormone and the effects of luteinizing hormone-releasing hormone injection in Asiatic elephants. American Journal of Veterinary Research 40, 451-453.
Abstract: Cyclic changes in serum concentrations of luteinizing hormone (LH) were observed throughout the estrous cycle of Asiatic elephants (Elephas maximus).  The increase in serum LH was correlated with a slight increase in serum estradiol concentration and the onset of behavioral heat (willingness to mate).  In a second series of studies, injection of luteinizing hormone-releasing hormone after 3 days of estrone administration induced an increase in serum LH.  These studies indicate that the Asiatic elephant exhibits a cyclic LH release that can be experimentally induced by estrone and luteinizing hormone-releasing hormone administration.

Clark, H.W., Laughlin, D.C., Bailey, J.S., Brown, T.M., 1979. Isolation of mycoplasma from the genital tracts of elephants. Elephant 1(3), 9-10.

Kirkwood, T.B., Holliday, R., 1979. The evolution of aging and longevity. Proc R Soc Lond B Biol Sci 205, 531-546.
Abstract: Aging is not adaptive since it reduces reproductive potential, and the argument that it evolved to provide offspring with living space is hard to sustain for most species. An alternative theory is based on the recognition that the force of natural selection declines with age, since in most environments individuals die from predation, disease or starvation. Aging could therefore be the combined result of late-expressed deleterious genes which are beyond the reach of effective negative selection. However, this argument is circular, since the concept of 'late expression' itself implies the prior existence of adult age-related physiological processes. Organisms that do not age are essentially in a steady state in which chronologically young and old individuals are physiologically the same. In this situation the synthesis of macromolecules must be sufficiently accurate to prevent error feedback and the development of lethal 'error catastrophes'. This involves the expenditure of energy, which is required for both kinetic proof-reading and other accuracy promoting devices. It may be selectively advantageous for higher organisms to adopt an energy saving strategy of reduced accuracy in somatic cells to accelerate development and reproduction, but the consequence will be eventual deterioration and death. This 'disposable soma' theory of the evolution of aging also proposes that a high level of accuracy is maintained in immortal germ line cells, or alternatively, that any defective germ cells are eliminated. The evolution of an increase in longevity in mammals may be due to a concomitant reduction in the rates of growth and reproduction and an increase in the accuracy of synthesis of macromolecules. The theory can be tested by measuring accuracy in germ line and somatic cells and also by comparing somatic cells from mammals with different longevities.

Nair, P.G. Reprodcutive behavior of elephants. State Level Workshop on Elephants.  36-49. 1979. India, College of Veterinary and Animal Sicences, Kerala Agricultural University.
Ref Type: Conference Proceeding

Ralls, K., Brugger, K., Ballou, J., 1979. Inbreeding and juvenile mortality in small populations of ungulates. Science 206, 1101-1103.

Adams, J., Garcia III, A., Foote, C.S., 1978. Some chemical constituents of the secretion from the temporal gland of the African elephant (Loxodonta africana). Journal of Chemical Ecology 4, 17-25.
Abstract: The temporal glands of African elephants were examined microscopically and histologically, and were found to be modified apocrine sweat glands.  The secretion from thse glands was analyzed by gas chromotography and mass spectrometry, and some of the major volatile components have been identified as phenol and m- and p-cresol.

Clark, H.W., Bailey, J.S., Laughlin, D.C., Brown, T.M., 1978. Isolation of mycoplasma from the genital tracts of elephants. Zentralblatt fur Bakteriologie,Parasitenkunde,Infektionskrankheiten und Hygiene 1. Abt. Originale 241, 262.

Eriksen, E., 1978. The birth of an Asiatic elephant Elephas maximus in the Copenhagen Zoo. Zoologische Garten 45, 421-432.

Gruenberg, K., Jarofke, D., 1978. Surgical removal of excessive callous growth from the vulva of an Indian elephant (Elephas maximus). Erkrankungen der Zootiere 14, 301-304.

Jones, R.C., 1978. Studies on handling spermatozoa from the African elephant, Loxodonta africana. In: Watson, P.F. (Ed.), Artificial breeding of non-domestic animals. Symposium of the Zoological Society of London No. 43. Academic Press, NY, pp. 261-269.
Abstract: The motility of elephant spermatozoa was reduced by dilution into Krebs-Henseleit-Ringer (KHR) at 10 and 5 degrees C rather than 30 degrees C.  However, elephant spermatozoa were not as susceptible to cold shock as spermatozoa from many scrotal mammals.  Spermatozoa freshly collected form the epididymis were immotile or only a small portion showed weak motility.  A 50-fold dilution of freshly collected samples with epididymal plasma slightly increased the proportion of motile cells, but a 50-fold dilution with phosphate-buffered saline or KHR induced motility in a high proportion of cells.  However, only a small proportion of cells were motile in epididymal semen stored for one hour before dilution.  Dilution rates higher than 50-fold with KHR reduced the survival of spermatozoa during incubation at 37 degrees C.  Spermatozoa survived best in diluents with a high sodium to potassium ratio, but varying the ratio did not seem to affect the induction of motility by diluting semen.  In a factorial experiment the effects on the survival of spermatozoa at 37 degrees C of diluent pH (values of 5.5, 7.0 and 8.5) and osmotic pressure (150, 225, 330, 375 and 450 mosmol/kg) were tested and it was found that these factors did not have independent effects.  In general spermatozoa survived best in media at a pH of 8.5.  However, at a pH of 5.5 the best survival occurred in diluents with the lowest osmotic pressure, at pH 7.0 the optimal osmotic pressure was about 250 mosmol/kg and at pH 8.5 the optimal osmotic pressure was about 275 mosmol/kg.

Mollel, C.L., 1978. Cervico-vaginal prolapse in an African elephant. East African Wildlife Journal 16, 59.

Douglas-Hamilton, O., 1977. Twins are light relief for Manyara's elephant - perhaps! Africana 6, 10-11.

Effron, M., Griner, L., Benirschke, K., 1977. Nature and rate of neoplasia found in captive wild mammals, birds, and reptiles at necropsy. Journal of the National Cancer Institute 59, 185-198.
Abstract: The nature and rate of neoplasia found at necropsy of captive wild animals of the Zoological Society of San Diego collection were studied.  Neoplasia was present at necropsy in 2.75% of 3,127 mammals, 1.89% of 5,957 birds, and 2.19% of 1,233 reptiles.  Neoplasms were not detected during 198 necropsies of amphibians.  Gross and histologic examinations were performed on the 92 mammalian, 111 avian, and 28 reptilian neoplasms.  The lesions were diagnosed.  The findings included a high frequency of lymphosarcomas in birds and reptiles, multiple endocrine tumors in 2 European mouflons (Ovis musimon), and proliferative lessions of the billiary duct and pancreatic ductal systems in several species. Note: Only elephant tumor is Asian female--Papillomatous tumor of vulva.

Eisenberg, J.F., Kleiman, D.G., 1977. The usefulness of behavior studies in developing captive breeding programmes for mammals. International Zoo Yearbook 17, 81-89.

Hanks, J., 1977. Comparative aspects of reproduction in the male hyrax and elephant. In: Calaby, J.H., Tyndale-Biscoe, C.H. (Eds.), Reproduction and evolution. Australian Academy of Science, pp. 155-164.

Longo, L.D., Hill, E.P., 1977. Carbon monoxide uptake and elimination in fetal and maternal sheep. Am. J Physiology 232, 324-330.

Robinson, P.T., Meier, J.E., 1977. Surgical removal of a tumor from an Asian elephant. Veterinary Medicine Small Animal Clinician 72, 1638-1640.

von Elke Scheurmann, G., 1977. "Musth" in the Asiatic elephant. Giessener Beitr. Entwicklingforsch 1, 87-92.

Buss, I.O., Estes, J.A., Rasmussen, L.E., Smuts, G.L., 1976. The role of stress and individual recognition in the function of the African elephant's temporal gland. Mammalia 40, 437-451.
Abstract: Biochemical measurements were made from a sample of temporal gland secretion from each of five wild African elephant bulls (23 to 38 years of age) collected in Kruger National Park, South Africa between November 1974 and April 1975.  Total protein content was high (26-57 mg/ml), acid phosphatase ranged between 1.9 and 6.3 mM/h/mgm protein, and lactic dehydrogenase levels were undetectable.  Total lipid content in the secretion averaged 80 mg% and ranged from 75 to 87 mg%.  Triglycerides were just detectable, varying from 2 to 8 mg%, and phospholipids ranged from 9 to 11 mg% (ave. 10 mg%).  Cholesterol content was surprisingly high, measuring 12, 19, 26, 36, and 70 mg% for five samples of secretion.    Field observations indicated that stress triggers liberation of temporal gland secretion.  Among 116 elephants collected in Uganda, secretory activity of their temporal glands was more frequent during dry (probably more stressful) than during wet seasons.  Among 62 elephants driven by helicopter to roadways for collection in Kruger National Park, 23 driven relatively far and fast were in prominent musth; most of those driven slower and shorter distances showed no evidence of musth.  The matriarchal leader of an elephant family near Lake Albert, Uganda developed very prominent temporal gland activity after an hour and 45 minutes of vigorously defending three of her family members.    Chemical individuality of cholesterol levels in temporal glands of five adult bulls suggests a pheromone-producing function which serves for individual recognition by the African elephant.  Direct observations of wild elephants also suggest that the temporal gland functions as a scent gland helping to recognize other members of the group or to find them.

Darin-Bennett, A., Morris, S., Jones, R.C., White, I.G., 1976. The glycerylphosphorylcholine and phospholipid pattern of the genital duct and spermatazoa of the African elephant, Loxodonta africana. Journal of Reproduction and Fertility 46, 506-507.

Longo, L.D., 1976. Carbon monoxide effects on oxygenation of the fetus in utero. Science 19, 523-525.

Nevill, G.F., Crompton, W.G., Hennessy, M.A., Watson, P.F., 1976. Instrumentation for artificial insemination in the African elephant  Loxodonta africana. International Zoo Yearbook 16, 166-171.

Pillay, K.R., 1976. A note on the diagnosis of pregnancy in elephants. Indian Veterinary Journal 53, 19-21.

Rowlands, I.W., 1976. Artificial insemination of mammals in captivity. International Zoo Yearbook 16, 230-233.

Gombe, S., Heap, R.B., Sale, J.B., 1975. Endocrinology of pregnancy in the hyrax; plasma progesterone concentration and erythrocyte metabolism. Proc. Physiol. Soc. September, 13P-14P.

Heap, R.B., Gombe, S., Sale, J.B., 1975. Pregnancy in the hyrax and erthrocyte metabolism of progesterone. Nature 257, 809-811.

Jones, R.C., Bailey, D.W., Skinner, J.D., 1975. Studies on the collection and storage of semen from the African elephant, Loxodonta africana. Koedoe 18, 147-164.

Leuthold, W., Leuthold, B.M., 1975. Parturition and related behavior in the African elephant. Zeitschrift fur Tierpsychologie 39, 75-84.
Abstract: The behaviour of a female African elephant before and after parturition was observed and photographed in the Isiolo Game Reserve, Kenya.  Birth was very rapid (not actually seen). Post-partum activity was characterized by intensive care-giving behaviour in which a few other animals cooperated with the mother.  Observations are compared with the few other accounts on the subject.

Plotka, E.D., Seal, U.S., Schobert, E.E., Schmoller, G.C., 1975. Serum progesterone and estrogens in elephants. Endocrinology 97, 485-487.
Abstract: Serum progesterone and estrogens were measured by radioimmunoassay in the serum of immature, mature, and pregnant African and Asian elephants.  Progesterone was elevated from 26 to 215 pg/ml in nonpregnant animals and up to 480pg/ml in late pregnancy animals.  No relationship to reproductive state was evident for the low levels of estrogens, which ranged from 9 to 37 pg/ml.

Smith, N.S., Buss, I.O., 1975. Formation, function and persistence of the corpora lutea of the African elephant (Loxodonta africana). Journal of Mammalogy 56, 30-43.
Abstract: Between 1956 and 1967, 146 female African elephants were collected in Uganda and 23 sets of ovaries from pregnant animals were collected in Zambia.  These were examined for number and diameter of corpora lutea, corpora rubra, and Graafian follicles.  Histological sections were made of corpora lutea, corpora rubra, and ovarian stroma.  In each corpus luteum 15 microscopic fields were examined.  In each field 100 points were examined and cells at each point were classified as one of four classes.  Presence of vascularity and cytoplasmic vacuolation was noted.  Follicular development is repressed during gestation, especially in the 5 to 16 months of gestation.  Lactation has little apparent influence on follicular development.  Ovaries of all pregnant elephant contained more than one corpus luteum.  In the Zambian collection the mane number of corpora lutea with stigmata of oculation (2.4 was significantly different, P < .01) than the mean number without stigmata (3.5).  Mean diameter of the corpora lutea with stigmata was significantly (P < .01) larger than those without stigmata.  Corpora lutea resulting from ovulation are formed in the first few months of pregnancy, whereas corpora lutea formed by luteinization of nonovulated follicles can be formed at all stages.  The elephants of this sample had approximately 2.2 estrous cycles before becoming pregnant.  The mean number of corpora lutea from 41 pregnant elephants was 6.0.  There was no difference in the number or dieter of corpora lutea from the first or second halves of pregnancy.  The mean number of corpora rubra in pregnant animals was 10.8 and the mean maximum diameter  was 7.1 millimeters.  In the ovaries of some animals in the late stages of pregnancy or shortly after parturition, there were no corpora rubra.  The number of corpora rubra in the last half of pregnancy was not significantly greater than those in the first half.  The average time of persistence of corpora rubra was about 77 months.  Cell types within the corpora lutea were heterogeneously distributed within individual corpora lutea.  Within a single elephant there may be two or more groups of similar corpora lutea.  Typical lutein cells were less abundant in the late stages of pregnancy.  Cell types were not correlated with the presence of absence of stigmata or ovulation.  Cytoplasmic vacuolation was more abundant in the second to thirteenth month of gestation.  The corpora lutea of elephants are probably secreting hormones most actively during 2 to 14 months of gestation.

Watson, P.F., D'Souza, F., 1975. Detection of oestrus in the African elephant (Loxodonta africana). Theriogenology 4, 203-209.
Abstract: Swabs of mucus and cells from the reproductive tract of a 15 year old female African elephant in captivity were examined. Daily samples were obtained over a 1-year period by means of a probe designed to penetrate the urogenital sinus to a depth of 90 cm.  Dried smears of mucous material showed ferning patterns at intervals of approximately 16 days.  Dried spots of supernatant from washing of the swabs also showed intense ferning at 16 day intervals, but with greater regularity. Smears were stained and examined for the presence of squamous cells over a 4-month period.  Results indicate a regular occurrence of certification at approximately 15-day intervals. These observations indicate that the oestrous cycle of this elephant has a duration of approximately 16 days.  This is the first detailed study of the oestrous cycle in the African elephant, knowledge of which is essential for artificial breeding.

Gale, U.T., 1974. Burmese timber elephant. Trade Corporation, Rangoon, Burma.

Jones, R.C., Rowlands, I.W., Skinner, J.D., 1974. Spermatozoa in the genital ducts of the African elephant, Loxodonta africana. Journal of Reproduction and Fertility 41, 189-192.

Jones, R.C., Skinner, J.D., Rowlands, I.W., 1974. The role of the urogenital ducts of the African elephant, Loxodonta africana. Journal of Reproduction and Fertility 36, 441-442.

Ogilvie, P.W., Moody, A., Seitz, S., 1974. Vital statistics of a self-perpetuating elephant herd. AAZPA Regional Conference Proceedings 157-162.
Abstract: Successful reproduction of elephants has continued in the Portland Zoo since April of 1962.  The collection currently contains two adult males, six adult females, and one infant female.  One of the males and three of the females were born in the Portland collection.  Several births have been observed, photographed, and described together with other behaviors and dynamics of the collection.

Perry, J.S., 1974. Implantation, foetal membranes and early placentation of the African elephant, Loxodonta africana. Philos. Trans. R. Soc. Lond. [Biol] 269, 109-135.

Sacher, G.A., Staffeldt, E.F., 1974. Relation of gestation time to brain weight for placental mammals: Implications for the theory of vertebrate growth. The American Naturalist 108, 593-615.

Fawcett, D.W., Neaves, W.B., Flores, M.N., 1973. Comparative observations on intertubular lymphatics and the organization of the interstitial tissue of the mammalian testis. Biology of Reproduction 9, 500-532.

Fowler, M.E., Hart, R., 1973. Castration of an Asian elephant, using etorphine anesthesia. Journal of the American Veterinary Medical Association 163, 539-543.
Abstract: A 9-year-old Asian elephant was castrated, using etorphine HCl for anesthesia.  The intraabdominal surgery was completed in 2 stages.  Respiratory and heart rates were normal throughout each surgical procedure.  Normal PaCO₂ and PaO₂ were maintained without the need of intermittent positive pressure ventilation.

Fowler, M.E., 1973. Castration of an elephant. Journal of Zoo and Wildlife Medicine 4, 25-27.

Hanks, J., 1973. Growth and development of the ovary of the African elephant, Loxodonata africana. Puku 7, 126-131.
Abstract: Aspects of growth and development of the ovary of the African elephant are described.  There was a pronounced hypertrophy of foetal ovarian interstitial tissue in the second half of gestation.  The left ovary was larger than the right in the majority of prepubertal and foetal elephants.  There was a gradual increase in the mean number of macroscopically visible follicles from the age of six years up to the mean age of first ovulation at 14 years.

Jones, R.C., 1973. Collection, motility and storage of spermatozoa from the African elephant, Loxodonta africana. Nature 243, 38-39.

Ogle, T.F., Braach, H.H., Buss, I.O., 1973. Fine structure and progesterone concentration in the corpus luteum of the African elephant. Anatomical Record 175, 707-724.
Abstract: Corpora lutea from one recently post-partum and six pregnant African elephants (Loxodonta africana) were fixed for electron microscopy.  Progesterone concentration was determined for five of these corpora lutea utilizing the competitive protein binding technique.  Lutein cell size and progesterone concentration increased from one and two months of pregnancy to a maximum at three months (P< 0.01).  After eight months of pregnancy, lutein cell size decreased to the one-month level. Maximum lutein progesterone was 4.1 ng/mg, much lower than that reported for other mammals.  Lutein cell fine structure exhibited characteristics typical of lutein cells from other species but with some variation.  Unusual features include peripheral distribution of organelles, large stores of lipid and lipofuscin throughout pregnancy, and mitochondria sparsely populated with lamellar cristae.  The data indicate that the corpus luteum of the African elephant has significant but limited steroidogenic capabilities.

Smith, N.S., Buss, I.O., 1973. Reproductive ecology of the female African elephant. Journal of Wildlife Management 37, 524-534.
Abstract: During 1957 through 1964, 146 female African elephants (Loxodonta africana) were collected in Uganda; data were also obtained from 23 pregnant elephants collected in Zambia in 1967.  The reproductive status of 142 adult and subadult female elephants in Uganda was 17.6% pregnant, 79.6% not pregnant, 2.1% estrus, and 0.7% status unknown; 45.1% lactating and 5.6% lactation status unknown.  Female elephants reach sexual maturity between 7 and 15 years in age.  The reproductive rate of elephants in Uganda is decreasing.  The duration of lactation was about 4.8 years.  The postpartum conception interval has increased from 24.1 months in 1947-1950 (Perry 1953) to 77.0 months in 1964.  Elephants in Uganda may breed during all months of the year; however breeding peaks do occur because there are significantly more young born in the rainy seasons than the dry seasons.  About 2 months after the onset of the rains, the physiological state of the female is apparently enhanced, and increased reproductive activity ensues.  The increased activity continues about 2 months beyond cessation of the rains.  In Zambia, breeding increases as the rainy season progresses and as the dry season progresses, breeding decreases.

Fowler, M.E., 1972. Castration of an elephant. Proceedings American Association of Zoo Veterinarians 25-27.

Hanks, J., 1972. Reproduction of elephant, Loxodonta africana, in the Luangwa Valley, Zambia. Journal of Reproduction and Fertility 30, 13-26.
Abstract: Aspects of reproduction in the African elephant in the Luangwa Valley in Zambia were studied in relation to the population dynamics of the species.  The fetal and secondary sex ratio up to 16 years of age did not depart significantly from equality.  Males left family units soon after 16 years of age and joined bachelor herds.  From 1964 to 1968, 88% of conceptions were in the rains, but in 1969 there was a shift in the breeding season peak to dry months of the year.  There was no evidence of seasonal breeding in the male elephant.  Females reached maturity at 14 years, and males at 15 years, when the combined weights of the testes reached 650 to 700 g, and the mean seminiferous tubule diameter reached 90 to 120 micrometers.  The mean calving interval was 3.5 to 4.0 years.  In the population, 6% of the elephant were less than 1 year old. Apparent cycles of recruitment were considered to be artefact caused by slight inaccuracies of the aging technique used. Corpora albicantia accumulated at the approximate mean rate of 0.6/year, and the significance of this was examined in relation to comparative studies of population fertility.  Reproductive senescence was a consequence of a combination of uterine defects and a reduction of oocyte number.

Hanks, J., Short, R.V., 1972. The formation and function of the corpus luteum in the African elephant, Loxodonta africana . Journal of Reproduction and Fertility 29, 79-89.
Abstract: The uterus and ovaries of 617 elephants shot in Zambia were examined.  Corpora lutea seem to be necessary for the development of endometrial glands, and before conception can occur, a certain critical mass of luteal tissue has to be achieved by accumulating crops of CL from successive cycles. The elephant can be either monovular or polyovular, and ovulation is spontaneous.  New ovulations do not occur during pregnancy, and the presence of an embryo prolongs the life of the CL.  There is great variability in luteal size, small CL being commonest in non-pregnant animals and large ones in pregnant animals.  The CL do not enlarge during gestation, and some of the smaller ones may regress.  The number of CL in pregnant elephants varied with the age of the cow, the younger elephants having a significantly higher number.  Larger CL (>20mm in diameter) predominated in older animals.  Very little progesterone appears to be secreted by the corpora lutea, and the hormone could not be detected in the peripheral blood during gestation.  If progesterone is necessary for pregnancy, the elephant must be extremely sensitive to it, and may be forced to accumulate a large mass of relatively inactive CL before sufficient hormone is available to enable the animal to become pregnant.

Jainudeen, M.R., Katongole, C.B., Short, R.V., 1972. Plasma testosterone levels in relation to musth and sexual activity in the male Asiatic elephant, Elephas maximus. Journal of Reproduction and Fertility 29, 99-103.
Abstract: Testosterone was measured in the peripheral blood plasma of eleven male Asiatic elephants, using a competitive protein-binding assay.  When the animals showed no signs of musth, the testosterone levels were low (<0.2 to 1.4 ng/ml); as they began to come into musth and the temporal glands started to enlarge, the testosterone levels rose (4.3 to 13.7 ng/ml), and when the animals were in full musth, with discharging temporal glands and an aggressive temperment, the levels were extremely high (29.6 to 65.4 ng/ml).  Musth may therefore be comparable to the rutting behavior of some seasonally breeding mammals, although, in the elephant, there is some indication that it may be induced by sexual activity.

Jainudeen, M.R., Feranado, S.T., Sentheshanmuganathan, S., 1972. Certain biochemical constituents of seminal plasma of elephant (Elephas maximus). American Journal of Veterinary Research 33, 649-651.

Jainudeen, M.R., McKay, G., Eisenberg, J.F., 1972. Observations on musth in the domesticated Asiatic elephant (Elephas maximus). Mammalia 36, 247-261.

Lockhart, M., 1972. Birth of an elephant. Loris 12, 259-262.

Maberry, M.B., 1972. Diagnosis of pregnancy in Asiatic elephant. Journal of Zoo and Wildlife Medicine 3, 31-33.

Peters, J.M., Maier, R., Hawthorne, B.E., Storvik, C.A., 1972. Composition and nutrient content of elephant (Elephas maximus) milk. Journal of Mammalogy 53, 717-724.
Abstract: Gross composition and nutrient content were determined for milk samples from four cows of a captive family of Indian elephants during the course of eight calvings at the Portland Zoo in Oregon.  The milk had a lower concentration of fat than that frequently reported for this species and showed a unique fatty acid composition with respect to the large amounts of capric acid (10:0) present.  Data for amino acid analyses are given as well as values for ascorbic acid, thiamin, riboflavin, and pyridoxine.  Vitamin A and carotene were shown to be almost non-existent.

von Elke Scheurmann, G., Jainudeen, M.R., 1972. "Musth" beim asiatischen elefanten, Elephas maximus. Zoologische Garten 42, 131-142.

Davidar, P., 1971. The Teppakadu twins. Journal of the Bombay Natural History Society 68, 819-820.

Eisenberg, J.F., McKay, G.M., Jainudeen, M.R., 1971. Reproductive behavior of the Asiatic elephant (Elephas maximus maximus L.). Behavior 38, 193-225.

Fujimoto, E., Koto, N., Imori, T., Nakama, S., 1971. Gonadotropin in the urine of a pregnant Indian elephant -- a case report. Zoologica 55, 73.
Abstract: In 1963, at Takarazuka Zoo, Japan, a young female Indian elephant became pregnant, and in May, 1965, she gave birth to a very large stillborn calf (weighing 133.3 kg, male). The time of conception was problematical, but it was assumed as April or May of 1963, hence the gestation period may have been 24 or 25 months, a little longer than average.  Pregnancy diagnosis was attempted during the early and middle gestation period.  For exploration, an urinary gonadotropin was checked by the Friedman and Aschheim-Zondek tests on the whole urine samples collected twice in August 1963.  Results showed apparently positive results in both tests.  However, the samples collected in May and September, 1964, showed negative in three tests, including a male frog (Rana) reaction which was subjected to the concentrated urine samples.  So, probably a gondotropic substance many have been excreted in urine of this elephant at some time of the early pregnancy, and this may be more like FSH than LH in its activity.

Hanks, J. The reproductive physiology of the African elephant, Loxodonta africana.  1971.  University of Cambridge.
Ref Type: Thesis/Dissertation

Jainudeen, M.R., Eisenberg, J.F., Jayasinghe, J.B., 1971. Semen of the Ceylon elephant, Elephas maximus. Journal of Reproduction and Fertility 24, 213-217.
Abstract: Summary.  The procedure employed for the collection of semen from a captive male elephant is described.  Sperm-rich and sperm-free ejaculates were obtained.  Seven sperm-rich ejaculates were evaluated.    Seminal characteristics investigated in the elephant compared favourably with those of other domestic animals of normal fertility.  The mean concentration of spermatozoa was 1200 x 10(6)/ml in the sperm-rich ejaculates.  Individual spermatozoa measured 58.5 micrometers (average) in length.

Jainudeen, M.R., Bongso, T.A., Perera, B.M.O.A., 1971. Immobilisation of aggressive working elephants (Elephas maximus). Veterinary Record 89, 686-688.
Abstract: The capture of aggressive working elephants, Elephas maximus, by the drug immobilisation technique is described. Doses of 5 mg to 8 mg etorphine hydrochloride alone, satisfactorily immobilised four adult elephants.  Cyprenorphine hydrochloride reversed the immobilising effects almost immediately and completely.  Recovery was uncomplicated.  The value of this method of capture is discussed in relation to aggressive working elephant.

Jainudeen, M.R., Eisenberg, J.F., Tilakertne, N., 1971. Oestrous cycle of the Asiatic elephant, Elephas maximus, in captivity. Journal of Reproduction and Fertility 27, 321-328.
Abstract: During the course of a programme to breed the Asiatic elephant, Elephas maximus, in captivity, the oestrous cycles of eleven adult females were studied.  Two methods were used for detecting oestrus: (1) daily testing with a male elephant, and (2) urogenital smear cytology.  Overt signs of oestrus were not observed but "standing" oestrus was observed in ten animals. The duration of oestrus ranged from 2 to 8 days with a mode of 4 days.  Oestrous cycles in six animals ranged from 18 to 27 days with a mean of 22 days.  Urogenital smear cytology failed to indicate accurately the onset of behavioural oestrus but increases in the number of cornified cells may occur before, during and slightly after behavioural oestrus.  Considerable mating activity occurred during oestrus.  A description of mating behaviour is presented.  The findings are discussed in relation to breeding elephants in captivity and to the phenomenon of temporal gland activity.

Estes, J.A. Observations on the temporal gland of the African elephant.  1970. Washington, USA, Washington State University.
Ref Type: Thesis/Dissertation

Jainudeen, M.R., 1970. The use of etorphine hydrochloride for restraint of a domesticated elephant (Elephas maximus). Journal of the American Veterinary Medical Association 157, 624-626.
Abstract: A domestic male Asian elephant (Elephas maximus) in "musth" (aggressive state) was successfully immobilized with 8 mg. of etorphine hydrochloride (M.99).  The clinical signs of immobilization were comparable to those reported in the African elephant (Loxodonta africana).  Cyprenorphine hydrochloride (M.285) reversed the immobilizing effects almost immediately and completely.  Recovery was uncomplicated.

Kurt, F., 1970. A comparison of reproduction in tame and wild elephants. IUCN Publications,New Series (Morges) 17, 148-154.

McCullagh, K.G., Widdowson, E.M., 1970. The milk of the African elephant. British Journal of Nutrition 24, 109-117.
Abstract: 1. Analyses have been made of milk collected from thirty wild African elephants immediately after they were shot. 2.  The milk contained an average of 5.1% protein, 9.3% fat and 3.6% lactose.  The concentration of lactose decreased and the concentration of protein and fat increased with advancing lactation.  Inorganic constituents were present in approximately the same proportions as in bovine milk.  3.  The contribution of capric acid to the total fatty acids, previously shown to be extremely high, increased with advancing lactation.  4.  The significance of these findings to the preparation of milk for rearing young elephants by hand is discussed.

Elapata, S.A.I., 1969. The sexual behavior of wild elephants in Ceylon. Loris 11, 246-247.

Krishne Gowda, C.D., 1969. A brief note on breeding Indian elephants, Elephas maximus, at the Mysore Zoo. International Zoo Yearbook 9, 99.

Krumrey, W.A., Buss, I.O., 1969. Observations on the adrenal gland of the African elephant. Journal of Mammalogy 50, 90-101.
Abstract: Forty-nine female and 32 male African elephants (Loxodonta africana) were collected from July 19558 to May 1959 in Bunyoro District, Uganda.  Reproductive status, body weight, and measurements were recorded and ages estimated. Gross morphology and histology of the adrenal gland are described: histology of the elephant adrenal corresponds with generalized descriptions for those of other eutherian mammals. The fetal cortex is well developed in an elephant 2 months of age, but gradually degenerates and is completely absorbed by the fourth year of postnatal life without giving rise to another transitory zone.  No significant increase in relative adrenal weight related to Uganda's December-to-March dry season was detected.  Relative adrenal weight of elephants is higher in females than males of comparable age, in immature males that mature males, and in immature females than nonpregnant or nonlactating females.  Pronounced increase in relative adrenal weight is related to pregnancy and lactation.

Laws, R.M., 1969. Aspects of reproduction in the African elephant, Loxodonta africana. Journal of Reproduction and Fertility Suppl. 6, 193-217.
Abstract: This report is based on material from some 3000 elephants, including over 1500 females, examined in East Africa. Five different unit populations are represented which are at different stages of a generalized population cycle.  They range in density from about 2.5 elephants per square mile to over 10 per square mile, and adjustments to the reproductive cycles are correlated with population density.  The mean age of attainment of sexual maturity increases through the ranked density series from 11 to 20 years in a density-dependent way.  Evidence on mean calving intervals, obtained from the proportion of pregnant to non-pregnant females, and from age-specific placental scar counts, is presented.  These also appear to be density-dependent.  From the analysis of a sample 222 embryos and foetuses, seasonal patterns of reproduction are identified within populations.  The seasonal peak in conceptions has been retarded by 5 months in a high-density population, possibly due to nutritional deficiencies.  Long-term cycles of recruitment to the populations are also demonstrated.  They are of large amplitude, have a cycle length of 6 to 8 years, and are correlated with rainfall preceding conception.  The ovarian cycle has been investigated and data on corpora lutea numbers, luteal tissue weights, corpora albicantia numbers and weights, and follicle diameters are presented.  It is concluded that there is no replacement or augmentation of corpora lutea by ovulation or luteinization of unovulated follicles in mid-pregnancy, and that the elephant is probably seasonally poly-oestrous and poly-ovular, also producing large numbers of accessory corpora lutea by luteinization of unovulated follicles in early pregnancy.  Growth in height and weight, sex-ratio, incidence of twins and duration of suckling are also briefly discussed.

Molamure, A.H.E., 1969. Elephants -- Marginal notes on musth and mating. Loris 11, 345-346.

Short, R.V., 1969. Notes on the teeth and ovaries of an African elephant of known age. Journal of Zoology (Lond) 158, 421-425.
Abstract: A captive female African elephant, known to be 27 years old, died as a result of trauma.  Her growth rate was similar to that of other captive African elephants, and slightly greater than that of wild animals.  The 5th molar was in full wear, and the 6th was just coming into wear.  There was extensive dental caries of the labial, lingual and occlusal surfaces of the 5th molars, presumably due to the unnatural diet.  The ovaries contained a large number of cystic follicles, and at least 50 regressing corpora lutea.  These abnormalities are probably related to the fact that the elephant had never been mated.

Smith, J.G., Hanks, J., Short, R.V., 1969. Biochemical observations on the corpora lutea of the African elephant, Loxodonta africana. Journal of Reproduction and Fertility 20, 111-117.

Smith, N.S. The persistance and functional life of the corpus luteum in the African elephant.  1969. Pullman, Washington, USA, Washington State University.
Ref Type: Thesis/Dissertation

Buss, I.O., Johnson, O.W., 1967. Relationships of Leydig cell characteristics and intratesticular testosterone levels to sexual activity in the African elephant. Anatomical Record 157, 191-196.
Abstract: Histological characteristics of testis tissues from 25 African elephants (Loxodonta africana) collected in Uganda, showed no consistent relationships among the following variables: Leydig cell size, cytoplasmic characteristics, and abundance; testicular testosterone content; and age.  From these findings, plus field observations of sexual behavior, emerges the hypothesis that individual cyclicity in Leydig cell function was inherent in the elephant population studied.  Testosterone content of testes from 32 elephants (including the 25 studied histologically) suggested that lone bulls were not of a senile nature since they contained relatively large quantities of testosterone and were relatively young (from about 12 to 25 years of age).  Also, lone bulls were observed searching out estrous females.  Among bulls collected from family units and herds, testosterone levels and behavior differed conspicuously. Behavior appeared to be directly related to testosterone content in several instances.  Non-aggressive behavior among members of bull herds, plus the high proportion of such individuals with low testosterone content, suggest that some of these animals were in a depressed phase of sexual activity whereas others were undergoing pubertal development.

Dittrich, L., 1967. Contribution about the propagation and raising of the Indian elephant Elephas maximus in captivity with a review of elephant births in European zoos and circuses. Zoologische Garten 34, 56-92.

Johnson, O.W., Buss, I.O., 1967. The testis of the African elephant (Loxodonta africana). I. Histological features. Journal of Reproduction and Fertility 13, 11-21.

Johnson, O.W., Buss, I.O., 1967. The testis of the African elephant (Loxodonta africana). II. Development, puberty and weight. Journal of Reproduction and Fertility 13, 23-30.

Lang, E.M., 1967. The birth of an African elephant, Loxodonta africana, at Basle Zoo. International Zoo Yearbook 7, 154-157.

Laws, R.M., 1967. Occurrence of placental scars in the uterus of the African elephant (Loxodonta africana). Journal of Reproduction and Fertility 14, 445-449.
Abstract: Placental scars were recorded in uteri of forty-four non-pregnant mature elephants.  Some 159 scars were observed averaging 3-6/female (range 1 to 9).  They were almost equally distributed between the right and left cornua, and the distribution of implantation sites is illustrated.  The relation between estimating ages of the elephants and the number of placental scars they show indicates a mean calving interval of about 4 years but there is evidence that the calving interval has increased in recent years.

Riegel, K., Bantels, H., Buss, I.O., Wright, P.G., Kleihauer, E., Luck, C.P., Parer, J.T., Metcalfe, J., 1967. Comparative studies of respiratory functions of mammalian blood. IV. Fetal and adult African elephant blood. Respiratory Physiology 2, 182.

Seth-Smith, A.M.D., Parker, I.S.C., 1967. A record of twin foetuses in the African elephant. East African Wildlife Journal 5, 167.

Short, R.V., Mann, T., Hay, M.F., 1967. Male reproductive organs of the African elephant, Loxodonta africana. Journal of Reproduction and Fertility 13, 517-536.

 1966. Family planning among elephants. New Scientist 519, 215.

Buss, I.O., Smith, N.S., 1966. Observations on reproduction and breeding behavior of the African elephant. Journal of Wildlife Management 30, 375-388.
Abstract: Observations on the breeding behavior of the African elephant (Loxodonta africana) in Uganda show that some wild females begin to breed at approximately 7 years and all females breed by approximately 11 years of age.  A declining population and lower annual increment of elephants south of the Victoria Nile River in Murchison Falls National Park are associated with an average postpartum reconception interval longer than the 24.1-month period reported in 1953 by Perry.  The change in length of this interval is possibly a function of self-regulation in population numbers.  At least in some parts of its range, the wild African elephant breeds throughout the year with a distinct seasonal acceleration.  Corpora lutea present during early pregnancy are not replaced during the subsequent stage of pregnancy but are maintained throughout gestation and for about 2 months after parturition.  Old or degenerate corpora lutea persist for at least 4.5 years postpartum.  Apparently in some animals corpora lutea develop before conception, whereas in others, conception apparently occurs when corpora are established.  At least two Graafian follicles may ovulate and develop into corpora lutea within a short time.  Cows have multiple mates, there is no prolonged male-female relationship, and frequently there is no fighting by bulls over females.

Dittrich, L., 1966. Breeding Indian elephants, Elephas maximus, at the Hanover Zoo. International Zoo Yearbook 6, 193-196.

Roth, H.H., Austen, B., 1966. Twin calves in elephants. Saeugeteirkundliche Mitteilungen 14, 342-345.

Short, R.V., 1966. Oestrous behaviour, ovulation and the formation of the corpus luteum in the African elephant (Loxodonta africana). East African Wildlife Journal 4, 56-68.

Kleihauer, E., Buss, I.O., Luck, C.P., Wright, P.G., 1965. Haemoglobins of adult and foetal African elephants. Nature 207, 424-425.

Short, R.V., Buss, I.O., 1965. Biochemical and histological observation on the corpora lutea of the African elephant (Loxodonta africana). Journal of Reproduction and Fertility 9, 61-67.
Abstract: Progesterone determinations were carried out on eleven corpora lutea obtained from six African elephants in various stages of the oestrous cycle and pregnancy.  In no case was it possible to detect any progesterone whatsoever, even when using an extremely sensitive gas chromatographic technique, coupled with an internal isotope standard of ⁴C₁₄-progesterone to correct for extraction losses.  The histological appearance of many of the corpora lutea in these elephants suggested that they might be degenerate structures. It is therefore postulated that the functional life of the elephant's corpus luteum may be relatively short, even though it persists structurally for long periods of time.  However, we cannot entirely exclude the possibility that the elephant's corpus luteum never secretes progesterone at all.

Werksman, R., 1965. The birth of an elephant. African Wild Life 19, 335.

Amoroso, E.C., Perry, J.S., 1964. The foetal membranes and placenta of the African elephant. Philos. Trans. R. Soc. Lond. [Biol] 248, 1-34.

Cooper, R.A., Connell, R.S., Wellings, S.R., 1964. Placenta of the Indian elephant, Elephas indicus. Science 146, 410-412.
Abstract: The placenta of the Indian elephant is incompletely annular and zonary microscopically and occupies the equator of an ovoid chorioallantoic sac.  The amnion is fused with the chorion over the zone.  Microscopically, the placenta is labyrinthine and endotheliochorial with a rudimentary marginal hematoma. Both macroscopically and microscopically it resembles the placentas of the carnivores, particularly the raccoon, the cat and the dog.

Johnson, O.W. Histological and quantitative characteristics of the testes, observations on the teeth and pituitary gland, and the possibility of reproductive cyclicity in the African elephant (Loxodonta africana).  1-124. 1964. Pullman, Washington, USA, Washington State University.
Ref Type: Thesis/Dissertation

Levin, A.H., 1964. Elephants mate in Orpen Dam. African Wild Life 18, 202-203.

Norris, C.E., 1964. Mating of elephant. Loris December , 88-89.

Perry, J.S., 1964. The structure and development of the reproductive organs of the female African elephant. Philos. Trans. R. Soc. Lond. [Biol] 248, 35-52.

Wright, A., 1964. Mating habits of the elephant. African Wild Life 18, 301-302.

McGaughey, C.A., 1963. Musth. Ceylon Veterinary Journal 11, 105-107.

Nel, I.Z.G., 1963. ...and the elephants make love. African Wild Life 17, 157-158.

Venkatasubba, R.S.R., Prasad, M.R.N., 1963. The nuclear sex in the Indian elephant, Elephas maximus L. Naturwissenechaften 50, 313.

Anghi, C.G., 1962. Breeding Indian elephants, Elephas maximus, at the Budapest Zoo. International Zoo Yearbook 4, 83-86.

Maberry, M.B., 1962. Breeding Indian elephants at Portland Zoo. International Zoo Yearbook 4, 80-83.

West, L.J., Pierce, C.M., 1962. Lysergic acid diethylamide: Its effects on a male Asiatic elephant. Science 138, 1100-1103.
Abstract: Summary:Researchers gave LSD to a zoo elephant in order to "induce a behavioral abberation that might resemble the phenomenon of going on musth."  Elephant cause of death was asphixiation secondary to laryngeal spasm.

Anghi, C.G., 1961. Quinba, the fifth elephant calf. International Zoo News 8, 78-79.

Evans, G.H., 1961. Elephants and Their Diseases: A Treatise on Elephants. Government Printing, Rangoon, Burma.

Simon, K.J., 1959. Preliminary studies on composition of milk of Indian elephants. Indian Veterinary Journal 36, 500-503.

Poppleton, F., 1957. The birth of an elephant. Oryx 4, 180-181.

Poppleton, F., 1957. An elephant birth. African Wild Life 11, 106-108.

Pillai, P.B.K., 1956. Elephant foetus aborted by recently captured ceylon elephant. Ceylon Veterinary Journal 4, 14-16.

Friant, M., 1954. [Form of the brain of elephant (Loxodonta africana Blum.) during prenatal life.]
2306. C. R. Hebd. Seances Acad. Sci. 238, 1534-1535.

Hill, W.C.O., 1953. The reproduction of the African elephant. In: Ward, R. (Ed.), The Elephant in East Central Africa. Rowland Ward Ltd., London and Nairobi, pp. 61-67.

Osman-Hill, W.C., 1953. The reproduction of the African elephant. In: Ward, R. (Ed.), The elephant in East Central Africa -- a monograph. Rowland Ward, Ltd., London, pp. 61-67.

Perry, J.S., 1953. The reproduction of the African Elephant, Loxodonta africana. Philos. Trans. R. Soc. Lond. [Biol] 237, 93-149.

Friant, M., 1951. [The form of the brain, during ontogeny, in elephants (Elephantidae).]. C. R. Hebd. Seances Acad. Sci. 232, 2137-2138.

Ayer, A.A., Mariappa, D., 1950. External characters of three fetuses of the Indian elephant. Proceedings of the Indian Academy of Science XXXI(B), 193-209.

Hindle, E.M., 1950. Birth of an elephant in the Rome Zoo. Zoo Life 5, 7-9.

Ferrier, A.J., 1947. The care and management of elephants in Burma. Steel Brothers, London.

Burne, E.C., 1943. A record of gestation periods and growth of trained Indian elephant calves in the Southern Shan States, Burma. Proc. Zool. Soc. London Ser. A 113, 27-43.

Flower, S.S., 1943. Notes on age at sexual maturity, gestation period and growth of the Indian elephant, Elephas maximus. Proc. Zool. Soc. London Ser. A 113, 21-26.

Hill, W.C.O., 1938. The external and radiological anatomy of a foetal Asiatic elephant. Ceylon Journal of Science 21, 31-43.

Neuville, H., 1937. Recherches comparatives sur l'organe femelle des elephants avec remarques sur les formations dites hymenales. Annales des Scinces Naturelles,Zoologies et Biologie Animale 20 10, 245-295.

Schulte, T.L., 1937. The genito-urinary system of the  Elephas indicus male. American Journal of Anatomy 61 , 131-157.

Driak, F., 1935. Studien der zahnanlagen an einem foetus von Elephas indicus. Morph. J. 75, 1-14.
Abstract: .

Foot, A.E., 1935. Age of puberty in the Indian elephant. Journal of the Bombay Natural History Society 38, 392.

Morris, R.C., 1935. Death of an elephant (Elephas maximus Linn.) while calving. Journal of the Bombay Natural History Society 37, 722.

Tutein-Nolthenius, A.C., 1935. Birth of an elephant calf. Journal of the Bombay Natural History Society 34, 183-184.

Robinson, G.C., 1934. Time of sexual maturity of the elephant (Elephas maximus L.). Journal of the Bombay Natural History Society 37, 950.

Friant, M., 1933. La regression de la levre superieure au cours de l'otogenie individual chez l'elephant. C. R. Acad. Sci. Paris 878-879.

Seth-Smith, D., 1932. Remarks on the age at which the Indian elephant attains sexual maturity. Proc. Zool. Soc. London Ser. A 102, 816.
Abstract: The entire article is as follows:  Mr. D. Seth-Smith, F.Z.S., made the following remarks upon the age at which the Indian Elephant attains sexual maturity: -- "Lt.-Col. G.H. Evans, quoting Sanderson, gives the usual age at which the female elephant produces her first calf at sixteen years, but he quotes an instance, on the authority of W.A. Bell, of a cow dropping a calf when she was only nine years and one month old.  This cow subsequently died. "Herr Heck, director of the Zoological Garden at Munich, informs me that a cow elephant under his charge, known to be only eight years old, has produced a calf.  This was apparently premature, and for the first twelve days of its life, the young animal was unable to suck, and the cow had to be milked by hand and the calf fed with its mother's milk from a bottle.  At twelve days old it began to suck.  The father of this calf is now ten years old and mating took place when the bull was eight and the cow six years old.  Another cow in the Munich gardens which is nine years old is expected to produce a calf in a few months' time."

Gowers, W.F. Exhibitions and notices.  On the gestation period of the African elephant (Loxodonta africana). Procedings of the Zoological Society of London , 77-78. 1931.
Ref Type: Conference Proceeding

Eales, N., 1929. The anatomy of a foetal African elephant, Elephas africanas (Loxodonta africana). Part III.  The contents of the thorax and abdomen, and the skeleton. Transactions of the Royal Society of Edinburgh 56, part I, 203-246.

Eales, N., 1928. The anatomy of a foetal African elephant, Elephas africanas (Loxodonta africana) Part II.  The body muscles. Transactions of the Royal Society of Edinburgh 55, part III, 609-642.

Bor, N.L., 1927. Musth in elephant. Journal of the Bombay Natural History Society 32, 594-596.

Eales, N., 1926. The anatomy of the head of a foetal African elephant, Elephas africanas (Loxodonta africana). Transactions of the Royal Society of Edinburgh 54, part III, 491-551.

Eales, N.B., 1925. External characteristics, skin, and temporal gland of a foetal African elephant. Procedings of the Zoological Society of London 2, 445-456.

Petit, G., 1924. Sur l'abouchement des canaux deferents et de vesicules seminales au verumontanum de l'elephant. Bull. Mus. Hist. Nat. , Paris 30, 441.

Hundley, G., 1922. The breeding of elephants in captivity. Journal of the Bombay Natural History Society 28, 537-539.

Wilson, J.C.C., 1922. The breeding of elephants in captivity. Journal of the Bombay Natural History Society 28, 1128-1129.

Adloff, P., 1919. Zur otogenie des elefantengebisses. Anatomischer Anzeiger 52, 534-540.

Bolk, L., 1917. Anatomische bemerkungen ueber einen fetus von Elephas africanas. Verh. Koninkl. Akad. Wet. Amsterdam 19, 1-40.

Bolk, L., 1917. Anatomical notes on a fetus of Elephas africanus. J. Muller, Amsterdam.

Allan, C.W., 1911. The birth of a wild elephant calf. Journal of the Bombay Natural History Society 21, 239-240.

Evans, G.H., 1910. Elephants and Their Diseases: A Treatise on Elephants. Government Printing, Rangoon, Burma.

Assheton, R., Stevens, T.G., 1905. Notes on the structure and the development of the elephant's placenta. Quarterly Journal of Microscopical Science 49, 1-37.

Slade, H., 1903. On the mode of copulation of the Indian elephant. Procedings of the Zoological Society of London 111-113.

Beddard, F.E., 1902. Birth of an elephant. Procedings of the Zoological Society of London 2, 320-322.

Shaw, W., 1900. Castration of an elephant. Veterinary Journal of London,N. S. 2, 151-152.

Paterson, A.M., 1898. The genito-urinary organs of the female Indian elephant. Journal of Anatomy and Physiology 12, 582-604.

Brauer, F.M., 1897. The oestrid of the Indian elephant. Ent. Mon. Mag. 33, 13.

Watson, M., 1883. Additional observations on the structure of the female organs of the Indian elephant (Elephas indicus). Procedings of the Zoological Society of London 517-521.

Watson, M., 1881. On the anatomy of the female organs of the Proboscidea. Transactions of the Zoological Society of London 11, 111-130.

Brown, A.E., 1880. Mr. Sclater on the birth of an elephant. Procedings of the Zoological Society of London XV, 222-223.

Chapman, H.C., 1880. The placenta and generative apparatus of the elephant. Journal of the Academy of Natural Sciences,Philadelphia 8, 413-422.

Heysham, 1880. Col. Heysham on the breeding of elephants. Procedings of the Zoological Society of London 23-24.

Miall, L.C., Greenwood, F., 1879. The anatomy of the Indian elephant. Part III alimentary canal and its appendages. Journal of Anatomy and Physiology 13, 17-50.

Mojsisovics, A., 1879. Zur kenntris der afrikanischen elefanten. Arch. Naturgesch. 45, 56.

Watson, M., 1872. Contributions to the anatomy of the Indian elephant (Elephas indicus), Part II. Urinary and generative organs. Journal of Anatomy and Physiology 7, 60-74.

Gray, J.E., 1868. Notes on the foetus of an Elephant, etc. Procedings of the Zoological Society of London 491.

Owen, 1857. Description of the foetal membranes and placenta of the elephant (Elephas indicus, Cuv.), with remarks on the value of placentary characteristics in the classification of the Mammalia. Philos. Trans. R. Soc. Lond. [Biol] 147, 347-353.

Gulliver, G., 1847. Note on the spermatozoa and on the elevator muscles of the penis of the Indian elephant. Procedings of the Zoological Society of London 105.

Mayer, J.F.C., 1847. Beitrage zur anatomie des elefanten und der urbrigen pachydermen. Nova Acta Academia Caesaeae Leopoldino-Carolinae Germanicae Nature Curiosorum 22, 1-88.

Perrault, C., 1734. Memoires pour servir a l'historie naturelle des Animaux. Academie Royal des Sciences, Paris.

Blair, P., 1710. Osteographia elephantina: or, a full and exact description of all the bones of an elephant which dy'd near Dundee, April the 27th, 1706, with their several dimensions, etc. Part II. Philos. Trans. R. Soc. Lond. [Biol] 27, 117-168.

Blair, P., 1710. Osteographia elephantina: or, a full and exact description of all the bones of an elephant which dy'd near Dundee, April the 27th, 1706, with their several dimensions, etc. Part I. Philos. Trans. R. Soc. Lond. [Biol] 27, 51-116.

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