Elephant
Bibliographic
Database

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Elephant Bibliographic Database
www.elephantcare.org

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

Basken, N.E., Green, M.A., 2009. Cu(II) bis(thiosemicarbazone) radiopharmaceutical binding to serum albumin: further definition of species dependence and associated substituent effects
46. Nucl. Med. Biol. 36, 495-504.
Abstract: INTRODUCTION: The pyruvaldehyde bis(N(4)-methylthiosemicarbazonato)copper(II) (Cu-PTSM) and diacetyl bis(N(4)-methylthiosemicarbazonato)copper(II) (Cu-ATSM) radiopharmaceuticals exhibit strong, species-dependent binding to the IIA site of human serum albumin (HSA), while the related ethylglyoxal bis(thiosemicarbazonato)copper(II) (Cu-ETS) radiopharmaceutical appears to exhibit only nonspecific binding to HSA and animal serum albumins. METHODS: To further probe the structural basis for the species dependence of this albumin binding interaction, we examined protein binding of these three radiopharmaceuticals in solutions of albumin and/or serum from a broader array of mammalian species (rat, sheep, donkey, rabbit, cow, pig, dog, baboon, mouse, cat and elephant). We also evaluated the albumin binding of several copper(II) bis(thiosemicarbazone) chelates offering more diverse substitution of the ligand backbone. RESULTS: Cu-PTSM and Cu-ATSM exhibit a strong interaction with HSA that is not apparent with the albumins of other species, while the binding of Cu-ETS to albumin is much less species dependent. The strong interaction of Cu-PTSM with HSA does not appear to simply correlate with variation, relative to the animal albumins, of a single amino acid lining HSA's IIA site. Those agents that selectively interact with HSA share the common feature of only methyl or hydrogen substitution at the carbon atoms of the diimine fragment of the ligand backbone. CONCLUSIONS: The interspecies variations in albumin binding of Cu-PTSM and Cu-ATSM are not simply explained by unique amino acid substitutions in the IIA binding pocket of the serum albumins. However, the specific affinity for this region of HSA is disrupted when substituents bulkier than a methyl group appear on the imine carbons of the copper bis(thiosemicarbazone) chelate

Bertelsen, M.F., Kjelgaard-Hansen, M., Grondahl, C., Heegaard, P.M., Jacobsen, S., 2009. Identification of acute phase proteins and assays applicable in nondomesticated mammals
72. J. Zoo. Wildl. Med. 40, 199-203.
Abstract: The serum concentration of acute phase proteins (APPs) increases dramatically in response to inflammation and tissue injury. APPs are clinically useful in a range of domesticated mammals; however, knowledge is limited in nondomesticated mammals. The detective ability of two assays for each of three potential APPs--serum amyloid A (SAA), C-reactive protein (CRP), and haptoglobin (Hp)--was evaluated in eight species. For SAA, a turbidimetric immunoassay (TIA) demonstrated significant detective abilities in the Asian elephant (Elaphas maximus), impala (Aepyceros melampus), musk ox (Ovibos moschatus), and chimpanzee (Pan troglodytes), as did an SAA enzyme-linked immunosorbent assay (ELISA) in the impala. For CRP, both TIA and ELISA had significant detective abilities in the chimpanzee. For Hp, a colorimetric assay demonstrated significant detective abilities in impala, musk ox, sitatunga (Tragelaphus spekeii), and chimpanzee, as did the Hp ELISA in the impala, musk ox, and sitatunga. In conclusion, these results suggest that assays for detection of relevant APPs in several nondomesticated animals are available

Knauf, S., Blad-Stahl, J., Lawrenz, A., Schuerer, U., Wehrend, A., 2009. Plasma preparation and storage for African elephants (Loxodonta africana)
74. J. Zoo. Wildl. Med. 40, 71-75.
Abstract: The use of plasma as a life-saving tool for neonatal African elephants (Loxodonta africana) that failed passive transfer of immunoglobulins is proposed. The methodology of blood sampling, plasma extraction, and plasma storage is described. Values for cellular component sedimentation and biochemical parameters of extracted plasma that was collected from 2 female elephants is presented. The proposal for a central plasma bank for elephants in European zoos is suggested

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

Carter, A.M., Miglino, M.A., Ambrosio, C.E., Santos, T.C., Rosas, F.C., Neto, J.A., Lazzarini, S.M., Carvalho, A.F., da Silva, V.M., 2008. Placentation in the Amazonian manatee (Trichechus inunguis). Reprod. Fertil. Dev. 20, 537-545.
Abstract: Evidence from several sources supports a close phylogenetic relationship between elephants and sirenians. To explore whether this was reflected in similar placentation, we examined eight delivered placentae from the Amazonian manatee using light microscopy and immunohistochemistry. In addition, the fetal placental circulation was described by scanning electron microscopy of vessel casts. The manatee placenta was zonary and endotheliochorial, like that of the elephant. The interhaemal barrier comprised maternal endothelium, cytotrophoblasts and fetal endothelium. We found columnar trophoblast beneath the chorionic plate and lining lacunae in this region, but there was no trace in the term placenta of haemophagous activity. The gross anatomy of the cord and fetal membranes was consistent with previous descriptions and included a four-chambered allantoic sac, as also found in the elephant and other afrotherians. Connective tissue septae descended from the chorionic plate and carried blood vessels to the labyrinth, where they gave rise to a dense capillary network. This appeared to drain into shorter vessels near the chorionic plate. The maternal vasculature could not be examined in the same detail, but maternal capillaries ran rather straight and roughly parallel to the fetal ones. Overall, there is a close resemblance in placentation between the manatee and the elephant

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.

Mikota, S.K., 2008. Tuberculosis in elephants. In: Fowler, M.E., Miller, R.E. (Eds.), Zoo and Wild Animal Medicine, Current Therapy 6th edition. Saunders/Elsevier, St. Louis, pp. 355-364.

Pacheco, J.M., Traulsen, A., Antal, T., Dingli, D., 2008. Cyclic neutropenia in mammals. Am. J. Hematol. 83, 920-921.
Abstract: Cyclic neutropenia (CN) has been well documented in humans and the gray collie. A recent model of the architecture and dynamics of hematopoiesis has been used to provide insights into the mechanism of cycling of this disorder. It provides a link between the cycling period and the cells where the mutated ELA2 is expressed. Assuming that the biologic defect in CN is the same in dogs, and the observation that the structure of hematopoiesis is invariant across mammals, we use allometric scaling techniques to correctly predict the period of cycling in the gray collie and extend it to other mammals from mice to elephants. This work provides additional support for the relevance of animal models to understand disease but cautions that disease dynamics in model animals are different and this has to be taken into consideration when planning experiments

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.

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

Aroch, I., King, R., Baneth, G., 2007. Hematology and serum biochemistry values of trapped, healthy, free-ranging rock hyraxes (Procavia capensis) and their association with age, sex, and gestational status. Vet. Clin. Pathol. 36, 40-48.
Abstract: BACKGROUND: The rock hyrax (Procavia capensis) is an herbivore prevalent from South Africa to Turkey, and a most common zoo animal. Although many studies of hyrax diseases and physiology are available, clinicopathologic data are limited. OBJECTIVES: The purpose of this study was to establish comprehensive hematologic and biochemical reference intervals for trapped, apparently healthy, free-ranging rock hyraxes using modern laboratory methods and to assess differences related to sex, gestation, and age. METHODS: Blood samples were obtained from 27 healthy, free-ranging hyraxes under anesthesia. Gender, body weight, and gestational status were recorded. Hematologic (n = 25) and serum biochemical (n = 22) analyses were performed using standard automated methodology. Data for male vs female, adult vs juvenile, and pregnant vs nonpregnant female hyraxes were compared using the Mann-Whitney U-test. Associations between variables were assessed using Pearson's or Spearman rank correlation tests. RESULTS: Significant age- and sex-related, but not gestation-related differences were observed in several variables. Serum alkaline phosphatase activity and phosphorus concentration were significantly higher in juveniles compared with adults. A unique type of monocyte comprised 1-3% of leukocytes in 4 hyraxes. Markedly high serum creatine kinase (CK) activity was observed in most hyraxes. CONCLUSIONS: The large number of animals and the availability of sex, age, and gestational data in this study will be useful to zoo and wildlife veterinarians working with rock hyraxes. High serum concentrations of betahydroxybutyric acid in the rock hyrax, compared with dogs, cats, and ruminants, may be related to its unique digestive system. High CK activity may have been the result of a capture myopathy-like syndrome. The unique monocytes in hyraxes resemble those of elephants and are a novel finding in this species

Clauss, M., Steinmetz, H., Eulenberger, U., Ossent, P., Zingg, R., Hummel, J., Hatt, J.M., 2007. Observations on the length of the intestinal tract of African Loxodonta africana (Blumenbach 1797) and Asian elephants Elephas maximus (Linne 1735). European Journal of Wildlife Research 53, 68-72.
Abstract: The digestive tract of elephants is surprisingly short compared to other herbivorous mammals. However, measurements relating the length of the intestine to the body mass of the respective individual are rare. In this study, we report such data for an African elephant and an Asian elephant. Our data support the hypothesis that Asian elephants have a longer intestinal tract than their African counterparts. These findings are in accord with the observation of longer retention times and higher digestion coefficients in Asian as compared to African elephants. This difference between the species could be the reflection of slightly different ecological niches, with Asian elephants adapted to a natural diet with a higher proportion of grass.

Oni, O., Sujit, K., Kasemsuwan, S., Sakpuaram, T., Pfeiffer, D.U., 2007. Seroprevalence of leptospirosis in domesticated Asian elephants (Elephas maximus) in north and west Thailand in 2004. Veterinary Record 160, 368-371.
Abstract: Serum samples from Asian elephants (Elephas maximus) in the Kanchanaburi, Chiang Mai and Lampang provinces of Thailand were tested using the microscopic agglutination test against 22 serovars of Leptospira interrogans. A titre of more than 1:100 was used as evidence of infection. In northern Thailand, the seroprevalence was 58 per cent and the prevalent serovars were Leptospira interrogans serovar Sejroe, Leptospira interrogans serovar Tarassovi, Leptospira interrogans serovar Ranarum and Leptospira interrogans serovar Shermani. In western Thailand, the seroprevalence was 57 per cent and the prevalent serovars were L Tarassovi, L Sejroe, L Ranarum, Leptospira interrogans serovar Bataviae and L Shermani. These results were similar to studies in domestic livestock and stray dogs in the Bangkok district. Among the elephants from Kanchanaburi there were significant associations between seropositivity and between the camp and between the prevalent serovars and the camp

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

Ball, R., Dumonceaux, G., Olsen, J., Burton, M.S., 2006. Comparison of trunk wash results matched to Multiantigen Print Immunoassay (MAPIA) in a group of captive Asian elephants (Elephas maximus). Proceedings International Elephant Conservation and Research Symposium 242-243.

Ball, R.L., Dumonceaux, G., Olsen, J.H., Burton, M.S., Lyashchenko, K. Comparison of trunk wash results matched to multiantigen print immunoassay (MAPIA) in a group of captive Asian elephants (Elephas maximus). 2006 Proceedings American Association of Zoo Veterinarians.  303-304. 2006.
Ref Type: Conference Proceeding
Abstract: Introduction: Between 1994 and June 2005, there were 34 confirmed cases of tuberculosis in elephants in the U.S. population. Thirty-one Asian (Elephas maximus) and three African (Loxodonta africana) elephants were affected. Mycobacterium tuberculosis was the etiologic agent in 33 cases and M. bovis in one case. Cases of tuberculosis caused by an unusual nontuberculous mycobacteria, M. szulgai have recently occurred as well.  Currently, TB in elephants remains a diagnostic dilemma. The sensitivity of trunk wash culture, the currently recommended test for diagnosis, is unknown. False negatives have been documented (trunk wash negative elephants that were subsequently found to be culture positive at necropsy).  Other non-culture techniques for TB diagnosis include ELISA, and PCR. A novel technology, MultiAntigen Print ImmunoAssay (MAPIA) and lateral-flow technology (Rapid Test)  has been evaluated and used to diagnose tuberculosis in captive elephants with encouraging results.  One concern with this serologic testing is the possibility of Mycobacterium other than tuberculosis (MOTT) cross-reacting with the antigen used in the Rapid Test or the MAPIA and leading to a false positive.  With numerous MOTT routinely cultured from trunk washes, this is a valid concern. Methods and Materials: A retrospective analysis was done at Busch Gardens Tampa Bay and Chembio, Inc. that matched trunk wash results to serum samples.  All serum was collected within 7 days of the trunk wash and analyzed with the Rapid Test and MAPIA. Four Asian elephants with a total of 18 samples met this criteria and had serum submitted for testing. Results and Discussion: Table 1 lists the results and the organisms cultured. While the sampling is limited in this pilot project, it appears that MOTT does not evoke a response when assayed with the Rapid Test or MAPIA. The recent cases of M. szulgai do demonstrate the potential usefulness for this test when a disease develops from MOTT.  The usefulness of this new technology, taken in conjunction with other clinical data including trunk washes when indicated, is a valuable tool in the healthcare of captive elephants.

LITERATURE CITED
1 Lacasse, C., K.C. Gamble, K. Terio, L.L. Farina, D.A. Travis, and M.Miller. 2005. Mycobacterium szulgai osteroarthritis and pneumonia in an African elephant (Loxdonta africana). Proc. Am. Assoc. Zoo Vet. Ann. Meet. Pp. 170-172.
2 Larsen, R.S., M.D. Salman, S.K. Mikota, R. Isaza, R.J. Montali, and J. Triantis.  2000.  Evaluation of a multiple-antigen enzyme-linked immunosorbent assay for detection of Mycobacterium tuberculosis infection in captive elephants.  J. Zoo Wildl. Med. 31:291-302.
3 Lyashchenko, K., et al.  2000. A multiantigen print immunoassay for the serological diagnosis of infectious diseases.  J. Immunol. Methods  242:91-100
4 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. Ann. Meet.  Pp. 64-65

Dangolla, A., Ekanayake, D.K., Rajapakse, R.P., Dubey, J.P., Silva, I.D., 2006. Seroprevalence of Toxoplasma gondii antibodies in captive elephants (Elephaus maximus maximus) in Sri Lanka
516. Veterinary Parasitology 137, 172-174.
Abstract: Serum samples collected during August 2003-June 2004 from 45 privately owned captive and 8 elephants from the Pinnawala Elephant Orphanage were tested for the presence of antibodies against Toxoplasma gondii using the direct modified agglutination test (MAT). Antibodies were found in sera of 14 of 45 (32%) privately owned elephants with titers of 1:25 in three, 1:50 in three, 1:100 in three, 1:200 in three, and 1:400 in three elephants. The elephants from Pinnawala Elephant Orphanage were seronegative. This is the first report of T. gondii seroprevalence in elephants in Sri Lanka

Lyashchenko, K.P., Greenwald, R., Esfandiari, J., Olsen, J.H., Ball, R., Dumonceaux, G., Dunker, F., Buckley, C., Richard, M., Murray, S., Payeur, J.B., Andersen, P., Pollock, J.M., Mikota, S., Miller, M., Sofranko, D., Waters, W.R., 2006. Tuberculosis in elephants: antibody responses to defined antigens of Mycobacterium tuberculosis, potential for early diagnosis, and monitoring of treatment
438. Clin. Vaccine Immunol. 13, 722-732.
Abstract: Tuberculosis (TB) in elephants is a re-emerging zoonotic disease caused primarily by Mycobacterium tuberculosis. Current diagnosis relies on trunk wash culture, the only officially recognized test, which has serious limitations. Innovative and efficient diagnostic methods are urgently needed. Rapid identification of infected animals is a crucial prerequisite for more effective control of TB, as early diagnosis allows timely initiation of chemotherapy. Serology has diagnostic potential, although key antigens have not been identified and optimal immunoassay formats are not established. To characterize the humoral responses in elephant TB, we tested 143 serum samples collected from 15 elephants over time. These included 48 samples from five culture-confirmed TB cases, of which four were in Asian elephants infected with M. tuberculosis and one was in an African elephant with Mycobacterium bovis. Multiantigen print immunoassay (MAPIA) employing a panel of 12 defined antigens was used to identify serologic correlates of active disease. ESAT-6 was the immunodominant antigen recognized in elephant TB. Serum immunoglobulin G antibodies to ESAT-6 and other proteins were detected up to 3.5 years prior to culture of M. tuberculosis from trunk washes. Antibody levels to certain antigens gradually decreased in response to antitubercular therapy, suggesting the possibility of treatment monitoring. In addition to MAPIA, serum samples were evaluated with a recently developed rapid test (RT) based on lateral flow technology (ElephantTB STAT-PAK). Similarly to MAPIA, infected elephants were identified using the RT up to 4 years prior to positive culture. These findings demonstrate the potential for TB surveillance and treatment monitoring using the RT and MAPIA, respectively

Mikota, S.K., Dumonceaux, G., Miller, M., Gairhe, K., Giri, K., Cheeran, J.V., Abraham, D., Lyashchenko, K., Larsen, S., Payeur, J., Waters, R., Kaufman, G., \, 2006. Tuberculosis in elephants: An update on diagnosis and treatment; implications for control in range countries. Proceedings International Elephant Conservation and Research Symposium 109-118.

Moller, T., Roken, B.O., Lewerin, S.S., Lyashchenko, K., 2006. The elephant Rapid Test (RT) the future diagnostic test for TB (M. tuberculosis) in elephants? Call for a validation study in Europe. Proceedings International Elephant Conservation and Research Symposium 119-124.

Oni, O., Wajjwalku, W., Boodde, O., Chumsing, W., 2006. Canine distemper virus antibodies in the Asian elephant (Elaphas maximus)
407. Veterinary Record 159, 420-421.

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

Shoshani, J., Kupsky, W.J., Marchant, G.H., 2006. Elephant brain. Part I: gross morphology, functions, comparative anatomy, and evolution
446. Brain Res. Bull. 70, 124-157.
Abstract: We report morphological data on brains of four African, Loxodonta africana, and three Asian elephants, Elephas maximus, and compare findings to literature. Brains exhibit a gyral pattern more complex and with more numerous gyri than in primates, humans included, and in carnivores, but less complex than in cetaceans. Cerebral frontal, parietal, temporal, limbic, and insular lobes are well developed, whereas the occipital lobe is relatively small. The insula is not as opercularized as in man. The temporal lobe is disproportionately large and expands laterally. Humans and elephants have three parallel temporal gyri: superior, middle, and inferior. Hippocampal sizes in elephants and humans are comparable, but proportionally smaller in elephant. A possible carotid rete was observed at the base of the brain. Brain size appears to be related to body size, ecology, sociality, and longevity. Elephant adult brain averages 4783 g, the largest among living and extinct terrestrial mammals; elephant neonate brain averages 50% of its adult brain weight (25% in humans). Cerebellar weight averages 18.6% of brain (1.8 times larger than in humans). During evolution, encephalization quotient has increased by 10-fold (0.2 for extinct Moeritherium, approximately 2.0 for extant elephants). We present 20 figures of the elephant brain, 16 of which contain new material. Similarities between human and elephant brains could be due to convergent evolution; both display mosaic characters and are highly derived mammals. Humans and elephants use and make tools and show a range of complex learning skills and behaviors. In elephants, the large amount of cerebral cortex, especially in the temporal lobe, and the well-developed olfactory system, structures associated with complex learning and behavioral functions in humans, may provide the substrate for such complex skills and behavior

Takahashi, H., Yamashita, M., Shigehara, N., 2006. Cranial photographs of mammals on the web: The Mammalian Crania Photographic Archive (MCPA2) and a comparison of bone image databases. Anthropological Science 114, 217-222.
Abstract: The Mammalian Crania Photographic Archive (MCPA2) is a website (http://1kai.dokkyomed.ac.jp/mammal/en/mammal.html) that includes a collection of 10,950 photographs of mammalian crania, which have been taken with a high-resolution digital camera. In the present report, we outline the characteristics of MCPA2 and how it was created, and make brief comparisons with several similar websites currently accessible via the internet. The archived MCPA2 materials include 1825 cranial specimens, ranging from insectivores to elephants, which have been macerated in Japan during the past 35 years and prepared for osteological study. Of the 16 orders represented in the database, primates comprise the major group with 704 specimens. Each cranium was placed with the orbitomeatal (Frankfort) or palatine plane horizontal, and was photographed in six perpendicular views from a long distance using a telephoto or telemacro lens. These long-distance shots decrease perspective distortion that lead to measurement errors when studying cranial profiles and landmark positions, and enable detailed observation and measurement of specific bony characteristics on a computer screen. From our website, images can be searched using (1) the taxonomic table, (2) Japanese name, (3) English name, and (4) scientific name. In the page of search results, in addition to the images, four caliper measurements and additional text (taxonomy, sex, and age) are available for every specimen.

Wemmer, C., Krishamurthy, V., Shrestha, S., Hayek, L.A., Thant, M., 2006. Assessment of Body Condition in Asian Elephants (Elephas maximus). Zoo Biology 25, 187-200.
Abstract: A method of assessing body condition of Asian elephants (Elephas maximus) is presented. The method uses visual assessment to assign numerical scores to six different regions of the body, which are totaled to give a numerical index ranging from 0-11. The relationship between the index and morphometric variables is compared for a sample of 119 juvenile and young adult elephants from southern India, Nepal, and Myanmar. Mean ages of males and females were similar. Mean index of body condition (with standard error [SE]) was 7.370.2 points. No significant correlation was found between index of body condition and age over both sexes (r50.01, n550). Results were equivalent when sexes were treated separately (females: r50.03, n524; males: r50.01, n526). Sexes did not differ in height of the shoulder or body condition in our sample, but there was significant sexual dimorphism in breadth of the zygomatic arch and three measures of subcutaneous fat: girth of neck, thickness of cervical fold, and thickness of anal flap. These three measures were also significantly correlated with each other. Our assessment method should prove a practical tool for ecologic studies, but the relationship of the index topercentage of body fat should be determined using heavy water dilution
methodology.

Wiedner, E., Alleman, R., Isaza, R. Urinalysis in Asian elephants (Elephas maximus).
2006 Proceedings American Association of Zoo Veterinarians.  265. 2006.
Ref Type: Conference Proceeding
Abstract: Elephants have been reported with renal disease, and arthritic elephants are often maintained on long-term administration of non-steroidal anti-inflammatory drugs which have been associated with renal injury in other species.  Therefore, establishment of the reference values of normal urine would be a useful tool to elephant health care. Routine urinalysis was performed on urine collected from healthy female adult Asian elephants (Elephas maximus, n=30). Elephants were included in the study if they had normal serum urea nitrogen and creatinine serum concentrations, no history of urinary tract disease, and had not received any medication within the past 3 mo.  Urinalysis included gross description of urine color and clarity, measurement of pH and specific gravity, biochemical analysis and sediment evaluation.   Presence of protein was assessed by the sulfosalicylic acid tubidimetric test.  All urine samples were submitted for aerobic bacterial culture. Initial findings included identification of calcium carbonate crystals in the urine of most of the animals in this study; some (n=5) contained small numbers of struvite crystals (magnesium ammonium phosphates). Most samples were alkaline and clarity ranged from clear to flocculent. Specific gravity tended to be fairly low, and isosthenuria was frequent. Trace bilirubin was detected in some samples.  No ketonuria or proteinuria was found.  Glucosuria, which has been reported as a transient and normal finding in captive elephants, was not detected in any samples.

Agnew, D.W., Hagey, L., Shoshani, J., 2005. The elephants of Zoba Gash Barka, Eritrea: part 4. Cholelithiasis in a wild African elephant (Loxodonta africana). J. Zoo. Wildl. Med. 36, 677-683.
Abstract: A 4.0-kg cholelith was found within the abdominal cavity of a dead wild African elephant (Loxodonta africana) in Eritrea. Analysis of this cholelith by histochemistry, electron microscopy, electrospray mass spectroscopy, and energy-dispersive x-ray spectroscopy revealed it was composed of bile alcohols but no calcium, bilirubin, or cholesterol. Bacteria were also found in the cholelith. Similar, but smaller, bile stones have been identified previously in other wild African elephants and an excavated mammoth (Mammuthus columbi). Choleliths have been reported only once in a captive Asian elephant (Elephas maximus). Elephants, along with hyraxes (Procavia capensis) and manatees (Trichechus manatus), are unique among mammals in producing only bile alcohols and no bile acids, which may predispose them to cholelithiasis, particularly in association with bacterial infection. Dietary factors may also play an important role in cholelith formation.

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

Hove, T., Mukaratirwa, S., 2005. Seroprevalence of Toxoplasma gondii in farm-reared ostriches and wild game species from Zimbabwe. Acta Trop 94, 49-53.
Abstract: One hundred and seventy one serum samples from 10 game species from Zimbabwe were tested for IgG antibodies to Toxoplasma gondii infection using the modified agglutination test (MAT). Significantly higher seroprevalences were found in the felidae (Panthera leo) (92% of 26), bovidae (Tragelaphus species) (55.9% of 34)and farm-reared struthionidae (Struthio camelus) (48% of 50) compared to the other groups tested. Among the bovidae, the nyala (Tragelaphus angasii) had the highest seroprevalence of 90% (9/10). Anti-Toxoplasma antibody prevalences in browsers [greater kudu (Tragelaphus strepsiceros) (20% of 10), giraffe (Giraffa camelopardalis) (10% of 10) and elephant (Loxodonta africana) (10% of 20)] were generally in the lower range. No antibodies were detected in the wild African suidae [warthog (Phacochoerus africanus) and bushpig (Potamochoerus larvatus)]. Attempts to isolate T. gondii from the heart muscles of seropositve ostriches by subinoculation in BALB/c mice were unsuccessful.

Larsen, R.S., Kay, M., Triantis, J., Salman, M.D. Update on serological detection of Mycobacterium tuberculosis infection in Asian elephants. 2005 Proceedings AAZV, AAWV, AZA Nutrition Advisory Group.  62-63. 2005.
Ref Type: Conference Proceeding
Abstract: Tuberculosis has become an important disease in captive elephants, particularly Asian elephants (Elephas maximus). Diagnosing tuberculosis in elephants has been problematic as many tests have inadequate sensitivity or specificity.2-4 A multiple-antigen enzyme-linked immunosorbent assay (ELISA) was previously investigated for detecting infection in Asian elephants and African elephants (Loxodonta africana); this test had excellent sensitivity and specificity, but needed further evaluation.1 Modifications to the multiple-antigen ELISA panel have since been made. Valuable antigens were retained, other antigens were removed, and new ones were added.  This modified ELISA was re-evaluated, using serum from 68 Asian elephants. Sixteen had M. tuberculosis -positive trunk cultures, while 52 were either culture negative at necropsy or had a history of negative trunk cultures and no contact with infected elephants. Seven elephants were evaluated over time. The test was 100% (95% CI; 95-100%) specific and 94% (95% CI; 79-100%) sensitive using two of the six antigens (M. bovis strain AN5 culture filtrate and M. tuberculosis early secretory antigenic target 6). "Effectively-treated" elephants had decreasing seroreactivity, but those that were culture-positive post-treatment were more consistently seroreactive.  Although "effectivelytreated" elephants had declining seroreactivity, they still usually had higher values than animals that had never been infected. Serology continues to show great promise in detecting tuberculosis in elephants, often detecting infection months-to-years sooner than trunk wash culture.  Advances in techniques may soon make serology even more practical.  While serology should not replace trunk-wash culture, it is a useful adjunct for early detection of infection in elephants and for monitoring treatment.
ACKNOLWEDGMENTS We thank the many veterinarians, owners, caretakers, and managers of elephant-owning institutions that participated in this investigation, as well as Drs. Michele Miller and Susan Mikota for helping to coordinate sample collection. We also thank Kimberly Deines and other laboratory personnel who processed ELISA samples.  The study was partially funded by a grant from USDA, CSREES to Colorado State University Program of Economically Important Infectious Animal Diseases.
LITERATURE CITED
1.Larsen, R.S., M.D. Salman, S.K. Mikota, R. Isaza, R.J. Montali, and J. Triantis. 2000.  Evaluation of a multiple-antigen enzyme-linked immunosorbent assay for detection of Mycobacterium tuberculosis in captive elephants.  J. Zoo Wildl. Med. 31: 291-302.
2. Mikota, S.K., L. Peddie, J. Peddie, R. Isaza, F. Dunker, G. West, W. Lindsay, R.S. Larsen, M.D. Salman, D. Chatterjee, J. Payeur, D. Whipple, C. Thoen, D.S. Davis, R.J. Montali and J. Maslow.  2001. Epidemiology and diagnosis of Mycobacterium tuberculosis in six groups of elephants.  J. Zoo Wildl. Med. 32: 1-16.
3. Mikota, S.K., R.S. Larsen, and R.J. Montali.  2000.  Tuberculosis in elephants in North America.  Zoo Biol. 19: 393-403.
4. U.S. Department of Agriculture.  2003.  Guidelines for the control of tuberculosis in elephants.  Animal and Plant Health Inspection Service; Animal Care. Washington, D.C. http://www.aphis.usda.gov/ac/TBGuidelines2003.pdf.

Loannidis, J.P.A., 2005. Why most published research findings are false. PLoS Med 2, e124.

Lyashchenko, K., Miller, M., Waters, W.R. Application of MAPIA (Multiple antigen print immunoassay) and rapid lateral flow technology for tuberculosis testing of elephants. 2005 Proceedings AAZV, AAWV, AZA Nutrition Advisory Group.  64-65. 2005.
Ref Type: Conference Proceeding
Abstract: Tuberculosis (TB) remains a serious re-emerging disease in wildlife and zoo animals. Mycobacterium tuberculosis has been isolated from 30 captive Asian elephant (Elephas maximus within 14 herds in the United States (1994-2004) and Mycobacterium bovis has been isolated from one African elephant (Loxodonta africana) (Mikota, pers. comm.).3 There are several challenges with elephant TB diagnosis. Culture of trunk wash has relatively poor sensitivity and is subject to contamination.  Skin test is not validated in elephants and there is little reliability in these results.4   Serologic tests are appealing because  samples can be stored for future analysis, archived samples can be analyzed, various assay platforms can be directly compared, and these assays are amenable to serial analysis (e.g., to monitor therapy).  There is currently a multiple antigen ELISA test available for experimental use in elephants.1

To improve tuberculosis control, new diagnostic tools should be rapid, accurate, and host species-independent. Two novel serologic methods, MultiAntigen Print ImmunoAssay (MAPIA) and lateral-flow technology (Rapid Test), have been adapted for use in white-tailed deer, European badger, cattle, and Asian and African elephants for the detection of TB-specific antibody. Serologic markers of diagnostic importance have been identified for each host tested so far. With MAPIA, a machine prints specific antigens horizontally on a nitrocellulose membrane which can be cut into strips and used in Western blot.2   Strips are incubated with test serum samples, then an anti-Ig conjugate and color developer.  Using this assay, an antibody response to multiple mycobacterial antigens has been observed in sera from M. tb-infected elephants. No antibody response was detected to any antigens in non-infected elephant sera.  Additionally, the kinetics of antibody responses by elephants undergoing antibiotic therapy indicates that the MAPIA could be used for monitoring treatment and to determine recrudescence of infection. 

Using selected antigens, a lateral-flow test was developed for rapid antibody detection that can be used in multiple species. The Rapid Test can use serum, plasma, or whole blood and provides results within 15 min.  These tests are similar to in-clinic tests for FIV/FeLV detection (snap test, IDDEX). If a band is present in the test strip, it indicates a positive reaction (antibody present).
A panel of sera from healthy and TB infected elephants showed good correlation between the MAPIA and the rapid test (Table 1).

In summary, it appears that TB-infected elephants produce a robust antibody response that can be detected in serologic assays.  Of special significance is the kinetics of the response, which may permit earlier detection of infection than current diagnostic methods.  While initial results are promising, additional studies are required to validate these two assays.  A relatively small set of serum samples from documented infected and non-infected elephants was used, and more samples are needed to further validate the tests. MAPIA has been used to optimize antigen selection in order to make the most sensitive and specific Rapid Test. This strategy may also allow for identification of "treatment-sensitive" antigens that could be used in the MAPIA format to monitor TB therapy.  While elephants will be used as an initial "proof of concept" species for test development, additional samples from other species will also be evaluated to determine applicability to other species (i.e., a host species-independent test), thus benefiting other groups such as primates, rhinos, cervids, etc.

ACKNOWLEDGMENTS
The authors thank the zoos and individuals that have provided samples and assistance with this research, including Ray Ball, Carol Buckley, Jenifer Chatfield, Genny Dumonceaux, Javan Esfandiary, Rena Greenwald, Scott Larsen, Susan Mikota, Torsten Moller, Dick Montali, Mike Richards, Heidi Riddle, Mo Salman, Scott Terrell, and many others.  This research was supported by Chembio Diagnostics, Inc.
LITERATURE CITED
1 Larsen, R.S., M.D. Salman, S.K. Mikota, R. Isaza, R.J. Montali, and J. Triantis. 2000.  Evaluation of a multiple-antigen enzyme-linked immunosorbent assay for detection of Mycobacterium tuberculosis  
  infection in captive elephants.  J. Zoo Wildl. Med. 31:291-302.
2 Lyashchenko, K., et al.  2000. A multiantigen print immunoassay for the serological diagnosis of infectious diseases.  J. Immunol. Methods  242:91-100. 
3 Mikota, S.K., and J. Maslow.  2002.  Epidemiology and treatment of tuberculosis in elephants:  2002.  Proc. Am. Assoc. Zoo Vet. Annu. Meet.  Pp. 384-387.

Moller, T., Roken, B., Petersson, L., Vitaud, C., Lyashchenko, K. Preliminary results of a new serological test for detection of TB-infection (Mycobacterium tuberculosis) in elephants (Elephas maximus and Loxodonta africanum) - Swedish Case studies. Verh.ber.Erkrg.Zootiere. 42, 173-181. 2005.
Ref Type: Conference Proceeding

Natiello, M., Lewis, P., Samuelson, D., 2005. Comparative anatomy of the ciliary body of the West Indian manatee (Trichechus manatus) and selected species. Vet. Ophthalmol. 8, 375-385.
Abstract: OBJECTIVE: To examine the anatomy of the ciliary body in the West Indian manatee (Trichechus manatus), paying close attention to its vascularization and to compare to those of its distant relative, the African elephant (Loxodonta africana), the amphibious hippopotamus (Hippopotamus amphibius) and the aquatic short-finned pilot whale (Globicephala macrorhynchus). PROCEDURE: Specimens from each species were preserved in 10% buffered formalin, and observed stereomicroscopically before being embedded in paraffin, sectioned and stained by Masson trichrome, hematoxylin and eosin, and periodic acid-Schiff for light microscopic evaluation. RESULTS: The network of blood vessels in the ciliary processes of the West Indian manatee appear to have an intricate pattern, especially with regard to venous outflow. Those of the elephant are slightly less complex, while those of the hippopotamus and whale have different vascular patterns within the ciliary body. Musculature within the ciliary body is absent within the manatee and pilot whale. CONCLUSIONS: In general, there appears to be a direct relationship between the increased development of vasculature and the loss of musculature within the ciliary bodies of the aquatic and amphibious mammals presently studied. Specifically, the ciliary body of the West Indian manatee has a comparatively unique construction, especially with regard to its vasculature.

Sanna, M.T., Manconi, B., Castagnola, M., Giardin, B., Masia, D., Messana, I., Olianas, A., Patamia, M., Petruzzelli, R., Pellegrini, M., 2005. Functional and structural characterization of the myoglobin from the polychaete Ophelia bicornis. Biochem J 389, 497-505.
Abstract: The myoglobin of the polychaete annelid Ophelia bicornis was isolated, purified to homogeneity and characterized. The primary structure, obtained from cDNA and protein sequencing, consists of 139 amino acid residues. The alignment with  other globin sequences showed that O. bicornis myoglobin misses the pre-A helix and the first six residues of the A helix. The presence of a PheB10-GlnE7 haem distal residue pair is in agreement with the measured oxygen affinity (P50=0.85 mmHg; 1 mmHg=0.133 kPa) and the only slightly higher autoxidation rate constant (0.28 h(-1)) with respect to that of the sperm whale myoglobin mutant E7  His-->Gln (0.21 h(-1)) and to elephant myoglobin (0.1 h(-1)). Oxygen-binding co-operativity was found to be absent under all the examined experimental conditions. The resistance of O. bicornis myoglobin towards autoxidation seems to confirm the important role of part of the A helix in the stability of the globin. The higher pKa of the acid-alkaline ferric transition of O. bicornis with respect to Asian elephant myoglobin, as well as the higher absorbance ratio  of its ferric form to the oxy form measured in the Soret region (gammamet/gammaoxy) with respect to that of the African elephant myoglobin, suggested a stronger interaction between the distal glutamine and the water molecule at the sixth co-ordinate position.

Suedmeyer, W.K., Oosterhuis, J., Kollias, G., Fagan, D., Hornoff, B., Dodam, J., Shafford, H. Elephant restraint device assisted anesthesia in an African elephant (Loxodonta africana). 2005 Proceedings AAZV, AAWV, AZA Nutrition Advisory Group.  189-191. 2005.
Ref Type: Conference Proceeding
Abstract: Modern elephant management programs often include the use of protected contact. This allows improved safety for the elephant staff but may limit access to medical conditions occurring in elephants.
A 27-yr-old female African elephant (Loxodonta africana) weighing an estimated 3,700 kg was anesthetized for evaluation of a chronic, progressive, fistulous tract of the left ventral mandible. The mandible was routinely cultured, flushed with diluted peroxide, chlorhexidine, betadine solution, or alternating antibiotics, based on microbial sensitivities. To properly assess the left mandible, the elephant had to be placed in right lateral recumbency, which was accomplished with the use of a commercially available rotational elephant restraint device (ERD). Because of the protected contact management program, right lateral recumbency could not be guaranteed at the time of immobilization. Malpositioning, tusk fracture and/or related injury could occur upon recumbency without the additional control afforded by the ERD. The ERD is a hydraulically operated unit that comfortably restrains an elephant, minimizing safety risks to the animal and staff. The ERD consists of one solid wall, three side panels, and hinged floor. The ends of the restraint are closed with moveable shift doors. The three side panels can be moved independently depending upon the size of the animal and are further subdivided with moveable "subpanels" to allow direct access to various areas of the animal. In addition, support straps help gently stabilize limbs when performing medical procedures. The unit is positioned within the elephant holding facility at the Kansas City Zoo. The unit was installed in 1994 during renovation of the elephant exhibit, whereupon the elephant management program was changed from free-contact to protected contact. The ERD is utilized for reproductive assessments, semen collection, transabdominal ultrasound, evaluation of integumentary wounds, ophthalmic and aural examination, and administration of injectable medications. However, no elephant had been anesthetized and rotated in the restraint. The affected animal could not be guaranteed to re-enter the ERD once rotated, but would enter and station in the ERD on a daily basis. Because of this, a conspecific was conditioned to allow rotation without the use of sedatives or tranquilizers, to prepare for the actual immobilization. Adjustments in strap placement, cushioning, critical evaluation of mechanical stability, and placement of hydraulic panels allowed staff to prepare for the actual immobilization, minimizing complications. The elephant was conditioned to enter and station in the ERD. After strapping the distal limbs, thorax and caudal abdomen for support, the elephant was immobilized with a combination of 3,000 IU of hyaluronidase (O'Brien Pharmacy, Kansas City, MO USA), 10 mg acepromazine maleate, and 7 mg etorphine hydrochloride (Wildlife Pharmaceuticals Inc., Fort Collins, CO USA) via pole syringe. Close monitoring of induction was performed and when stage III anesthetic plane was achieved, the elephant was rotated into right lateral recumbency, elevating the elephant 6 feet above the floor. No voluntary movement of the animal was noted while the restraint was in motion. Direct arterial blood pressure, indirect oscillometric blood pressure, blood gases, respiratory rate, excursion characteristics, cardiac rate and rhythm, and pulse oximetry was routinely monitored during the procedure. Anesthesia was maintained with intermittent boluses of etorphine hydrochloride. Intravenous physiologic fluids (lactated Ringers solution) were maintained via an i.v. aural catheter, and insufflation with oxygen was provided on a continual basis. Oral examination and palpation demonstrated an incomplete transverse fissure of the left mandibular molar, intact gingival, and proper dental occlusion with the upper arcade.  Digital radiographs of the left mandible were performed based on exposures obtained with a set of skeletonized jaws. Advantages of this diagnostic modality are the immediate imaging results, portability, and digital imaging and storage, and does not require a developer or fixative. Adjustments in radiographic angle and technique were made to obtain the best diagnostic image. Radiographic imaging demonstrated a sequestrum consisting of a fractured enamel plate  ²of the mandibular molar with a fistulous tract that coursed ventrally to communicate through the skin. The elephant was elevated 6 feet above the ground, which presented unique challenges. Because of the relatively small operating space, intubation was not possible, but insufflation was readily achieved and successful based on pulse oximetry trends. A commercial lift was utilized to elevate two large-animal circle anesthetic units to the level of the elephant's head. During immobilization the legs were cushioned and restraint straps removed to lessen the potential for occlusive damage to the tissues. The ERD allows an elephant to be positioned in either right or left lateral recumbency.
Upon completion of diagnostic procedures, the narcotic agent was reversed with 1,400 mg naltrexone hydrochloride (Zoopharm, Laramie, WY USA) administered 25% intravenously and 75% subcutaneously. The elephant awoke within 90 sec and was rotated to a standing position within the restraint. Thereafter, the elephant was confined in the restraint for approximately 45 min, until no untoward effects were likely to occur. The elephant was released from the restraint and resumed normal eating and drinking within 8 hr, and voluntarily entered the restraint within 2 wk following the procedure. The elephant was stable throughout the procedure; however, a predetermined objective for mean arterial blood pressures (<200 MAP) was not achieved. Hyaluronidase was utilized to promote rapid absorption of the narcotic and neuroleptic agents.³ Acetylpromazine was used to maintain peripheral perfusion by reducing the hypertensive effects of etorphine,¹ which has been documented in previous immobilizations of African elephants.^3-5 Etorphine hydrochloride, a powerful narcotic agent, has been successfully used as an immobilizing agent in both wild and captive African elephants.^3-5 Use of an ERD allowed full control of the immobilization, increasing safety for personnel, preventing injury to the elephant, and positioning the left mandible on the dorsal plane. Disadvantages are the elevated height of the elephant, relatively small operating space, and disrupted line of sight communication. A second procedure will be performed in the near future to address the fracture and subsequent sequestrum diagnosed during the first immobilization. The elephant is currently being conditioned to allow restraint in a holding stall that will allow greater access to the oral cavity and surgical manipulation of the affected mandible.
ACKNOWLEDGMENTS
We thank the staff of the Kansas City Zoological Park for their care, concern, and expertise in helping make this procedure a success.
LITERATURE CITED
1 Booth, N.H. Psychotropic agents. In: Booth, N.H., and R.E. McDonald (eds.). Veterinary Pharmacology and Therapeutics.  W.B. Saunders, Co., Philadelphia, PA. P. 329.
2 Fagan, V.D.A., J.E. Oosterhuis, and A. Roocraft. 2001. Captivity disorders in elephants: impacted molars and broken tusks. Der Zoologische Garten 71:281-303.
3 Honeymoon, V.L., G.R. Pettifer, and D.H. Dyson. 1992. Arterial blood pressure and blood gas values in normal standing and laterally recumbent African (Loxodonta africana) and Asian (Elephas maximus)    elephants. J. Zoo Wildl. Med. 23:205-210.
4. Kock, R.A., P. Morkel, and M.D. Kock. 1993. Current immobilization procedures used in elephants. In: Fowler,
M.E. (ed.).  Zoo and Wild Animal Medicine: Current Therapy 3. W.B. Saunders Co., Philadelphia, PA.  Pp. 436-441.
5 Raath, J.P. 1999. Relocation of African elephants. In: Fowler, M.E., and R.E. Miller (eds.). Zoo and Wild Animal Medicine: Current Therapy 4. W.B. Saunders, Co., Philadelphia, PA.  Pp. 525-533.

Waters, W.R., Palmer, M.V., Bannantine, J.P., Greenwald, R., Esfandiari, J., Andersen, P., McNair, J., Pollock, J.M., Lyashchenko, K.P., 2005. Antibody responses in reindeer (Rangifer tarandus) infected with Mycobacterium bovis. Clinical and Diagnostic Laboratory Immunology 12, 727-735.
Abstract: Despite having a very low incidence of disease, reindeer (Rangifer tarandus) are subject to tuberculosis (TB) testing requirements for interstate shipment and herd accreditation in the United States. Improved TB tests are desperately needed, as many reindeer are falsely classified as reactors by current testing procedures. Sera collected sequentially from 11 (experimentally) Mycobacterium bovis-infected reindeer and 4 noninfected reindeer were evaluated by enzyme-linked immunosorbent assay (ELISA), immunoblotting, and multiantigen print immunoassay (MAPIA) for antibody specific to M. bovis antigens. Specific antibody was detected as early as 4 weeks after challenge with M. bovis. By MAPIA, sera were tested with 12 native and recombinant antigens, which were used to coat nitrocellulose. All M. bovis-infected reindeer developed responses to MPB83 and a fusion protein, Acr1/MPB83, and 9/11 had responses to MPB70. Other antigens less commonly recognized included MPB59, ESAT-6, and CFP10. Administration of purified protein derivatives for skin testing boosted serum antibody responses, as detected by each of the assays. Of the noninfected reindeer, 2/4 had responses that were detectable immediately following skin testing, which correlated with pathological findings (i.e., presence of granulomatous lesions yet the absence of acid-fast bacteria). The levels of specific antibody produced by infected reindeer appeared to be associated with disease progression but not with cell-mediated immunity. These findings indicate that M. bovis infection of reindeer elicits an antibody response to multiple antigens that can be boosted by skin testing. Serological tests using carefully selected specific antigens have potential for early detection of infections in reindeer.

Windberger, U., Plasenzotti, R., Voracek, T., 2005. The fluidity of blood in African elephants (Loxodonta africana)
543. Clin. Hemorheol. Microcirc. 33, 321-326.
Abstract: The large cellular volume of erythrocytes and the increased plasma concentration of proteins in elephants are factors which potentially affect blood rheology adversely. To verify blood rheology, routine hemorheologic variables were analyzed in four African elephants (Loxodonta africana), housed in the zoo of Vienna. Whole blood viscosity at three different shear rates (WBV at low shear rate: WBV 0.7 s(-1) and WBV 2.4 s(-1); WBV at high shear rate: WBV 94 s(-1) done by LS30, Contraves) and erythrocyte aggregation (aggregation indices AI by LS30; aggregation indices M0, M1 by Myrenne aggregometer) were high (WBV 94 s(-1): 5.368 (5.246/5.648); WBV 2.4 s(-1): 16.291 (15.605/17.629); WBV 0.7 s(-1): 28.28 (25.537/32.173) mPa s; AI 2.4 s(-1): 0.25 (0.23/0.30); AI 0.7 s(-1): 0.24 (0.23/0.28); M0: 7.8 (6.4/8.4); M1: 30.2 (25/31)). Plasma viscosity (PV) was increased as well (1.865 (1.857/1.912) mPa s) compared to other mammalian species. These parameters would indicate a decrease in blood fluidity in elephants. However, erythrocyte rigidity (LORCA, Mechatronics) was decreased, which in contrast, has a promotive effect on peripheral perfusion. Blood rheology of the elephants was determined by a high whole blood and plasma viscosity as the result of pronounced erythrocyte aggregation and high plasma protein concentration. Thus, in the terminal vessels the resistance to flow will be increased. The large erythrocytes, which might impede blood flow further due to geometrical reasons, however, had a pronounced flexibility. We conclude that the effect of the increased inner resistance to peripheral blood flow was counteracted by the decreased rigidity of the erythrocytes to enable an adequate blood flow in African elephants

Principles of validation of diagnostic assays for infectious diseases.  2004.
Ref Type: Electronic Citation

Dangolla, A., Ekanayake, D.K., Rajapakse, R., Dubey, J.P., Silva, I.D. Presence of Toxoplasma gondii antibodies in captive elephants (Elephaus maximus maximus)) in Sri Lanka. Proceedings of the Peradeniya University Research  Sessions, Sri Lanka, Vol 9 November 10, 2004.  209-214. 2004.
Ref Type: Conference Proceeding
Abstract: Forty-five privately owned captive, apparently healthy elephants were blood sampled in order to detect the presence of antibodies against Toxoplasma gondii during August 2003- June 2004. The elephants were from 5 of the 11 districts in which, captive elephants live at present. Blood samples were transported in ice to the laboratory, serum separated immediately and frozen until analysed. A direct modified agglutination test was performed on serum. Sera of 14 of 45 (32%) elephants were found to be positive for the test. Highest titre reported was 1:400. A significantly high number of females (11/14) were positive for T.gondii antibodies compared with the male elephants (P = 0.02). The differences in titres among the age groups of elephants were not significant (P > 0.05). It appears that Toxoplasmosis is common among the Sri Lankan captive Elephants. Considering the nature of the infection, the sero-positive animals could become clinically diseased when immune-compromised, such as observed in the case of tuberculosis. Elephants, may have contacted this disease through faecal contamination from wild, semi wild or domestic animals. The fate of the parasites within elephant's body and investigating the possibility of elephants infecting humans or other animals may be of importance. This is the first report on presence of antibodies to T.gondii infection in elephants in Sri Lanka

Lazar, J., Rasmussen, L.E., Greenwood, D.R., Bang, I.S., Prestwich, G.D., 2004. Elephant albumin: a multipurpose pheromone shuttle
691. Chem. Biol. 11, 1093-1100.
Abstract: (Z)-7-dodecenyl acetate (Z7-12:Ac) is present in the urine of female Asian elephants (Elephas maximus) approaching ovulation and functions as a female-to-male sex pheromone. Here we show that a significant fraction of the pheromone in the urine is bound to a protein, elephant serum albumin (ESA), and provide evidence for key physiological functions of urinary ESA. Our biochemical and behavioral experiments suggest a three-fold role of ESA in pheromone signaling: (1) transporting Z7-12:Ac from serum into urine; (2) extending the presence of the pheromone in the environment without hampering detection; and (3) targeting pheromone delivery to chemosensory organs through localized release of the ligand induced by a pH change. The exploitation of albumin in pheromone transport clearly distinguishes the elephant from other mammals studied, and complements the uniqueness of elephant anatomy, physiology, and behavior

Loomis, M.R., Loomis, J.M. Equipment for use in monitoring anesthetized animals in remote geographic locations. 2004 PROCEEDINGS AAZV, AAWV, WDA JOINT CONFERENCE.  499-501. 2004.
Ref Type: Conference Proceeding
Abstract: Monitoring anesthetized animals in remote geographic locations with no electrical power source can be accomplished with the use of commercially available equipment or with modifications of available equipment. The use of portable solar panels to recharge batteries can supply adequate power to operate most equipment. Equipment for monitoring oxygenation, ventilation, cardiac rhythm and rate, blood pressure and core temperature have been successfully used in areas without an electrical grid or electrical generators.

Criteria for Choice of Equipment for Field Use
Size, weight, power requirements, durability and the ability to operate in harsh environmental conditions should be considered when choosing monitoring equipment for field use. Of concern are the power requirement and the source of the power, particularly in areas where there is no power grid or generator available.

Power Source
There are a number of types of rechargeable batteries on the market.1 Nickel metal halide batteries (NiMH) were chosen for use in monitoring equipment in this study (MAHA Powerx 2100mAh, Thomas Distributing, 128 East Wood, Paris, IL 61944). NiMH batteries have several features that make them attractive for remote use.1 They can be recharged 500 to 1000 times, have no memory, have a fairly steady discharge curve and have the least negative environmental impact when disposed of than other available batteries.1 One disadvantage of NiMH batteries is that they have a self discharge rate of 2-3% per day when not in use. AA NiMH batteries produce 1.2 volts.
Battery energy output is measured in milliamp hours (mAh)2. A battery rated at 1700 mAh will produce 1700 mA for 1 hr. Different manufacturers produce batteries with different power outputs. AA NiMH batteries are rated at up to 2400 mAh. The higher the mAh, the greater the output of the battery.

Batteries are charged using fast, smart chargers attached to portable solar panels (iPowerUS fast smart charger, iPower corporation, CA, USA). A fast charger delivers the amount of current necessary to recharge the battery in 1 hr or less. In general, a slower charge rate will extend the overall life of the battery.3 To overcome the deleterious effects of rapidly charging a battery, a smart charger has a current-limiter built into it that reduce the current as the battery is charged, thereby preventing most of the deterioration.3 The fast smart charger is attached to a portable solar panel (Sun Catcher Expedition solar charger, PowerQwest, Inc.3400 Corporate Way, Suite C Duluth, GA 30096 USA) via a 12 volt "cigarette lighter" type plug. The panel produces 25 watts of power, which is more than enough power to charge 8 AA NiMH batteries at a time. Equipment that uses AA or AAA batteries is preferred so that a large number of different sized rechargeable batteries are not required in the field.

Monitoring Equipment
Oxygenation is measured with a pulse oximeter or by arterial blood gas determination using a portable clinical analyzer. Several brands of pulse oximeters have been successfully used and recharged in the field. An Invacare model 3402NV (Sims BCI, Inc., Waukesha, WI 53186) is relatively small, light weight and operates on 6 AA batteries. This oximeter is durable and operates well on rechargeable AA NiMH batteries.

An I-Stat portable clinical analyzer (Heska Corp. 1613 Prospect Parkway, Fort Collins, CO 80525 USA) has been successfully used in the field using rechargeable 9-volt NiMH batteries. A challenge of using the I-Stat in the field is the analyzer's normal operating temperature of 16-30°C (61-86°F). The I-Stat has been kept in the proper operating temperature range by placing it in a 12-volt thermoelectric cooler (Coleman, Spirit Lake, IA 51360, USA). The thermoelectric cooler runs directly off of the solar panel.

Ventilation is measured using capnography or arterial blood gas determination. The criteria for choice of a capnograph include a waveform display, mainstream and sidestream capabilities and powered by rechargeable AA batteries. The Novametrix Tidal Wave model 615 (Novametrix Medical Systems, INC., Wallingford, and CT USA 06492) meets these criteria. The Tidal Wave comes standard with a rechargeable computer-type battery, but can be ordered with a battery tray, which holds 7 AA batteries. This instrument is durable and operates well on rechargeable NiMH batteries. The sidestream capability allows a large gauge needle to be placed in the lumen of a large endotracheal tube for sampling.

Cardiac rate and rhythm are monitored by use of an electrocardiograph (ECG). A compact ECG unit (Heska Vet/ECG 2000, Heska Corp., 1613 Prospect Parkway, Fort Collins, CO 80525 USA) that operates on 3 AAA rechargeable NiMH batteries is durable and dependable in the field. Blood pressure is measured by a direct arterial line or by indirect methods. Of the indirect methods, automated oscillometry has been successfully used in the field. No automated oscillometric blood pressure machine that runs on replaceable batteries could be found. A compact, durable instrument, Oscillomate 9300 (CAS Medical Systems, Inc., 44 East Industrial Blvd., Branford, CT 06405), was modified for field use. A transformer was manufactured which is inserted between the internal battery of the blood pressure monitor and the solar panel. This allows the internal battery of the blood pressure monitor to be recharged directly from the solar panel.

All monitoring equipment, battery chargers and rechargeable NiMH batteries are transported into the field in a backpack that is designed for photographic equipment (Lowepro Supertrecker AW II, Lowepro USA, P.O. Box 6189, Santa Rosa, CA 95406). All of the above equipment has been dependably used to monitor immobilized elephants in a variety of remote habitats in Cameroon, including dry, hot habitat,2 hot humid habitat.

LITERATURE CITED
1. New technology batteries guide: available battery types. http://www.nlectc.org/txtfiles/batteryguide/batype. htm, March, 2004.
2. New technology batteries guide: performance, economics and tradeoffs.http://www.nlectc.org/txtfiles/batteryguide/ba-type.htm, March, 2004.
3. New technology batteries guide: battery chargers and adapters.http://www.nlectc.org/txtfiles/batteryguide/ba-char.htm, March, 2004.
4. Horne, W.A., M.N. Tchamba, and M.R. Loomis. 2001. A simple method of providing intermittent positivepressureventilation to etorphine-immobilized elephants (Loxodonta africana) in the field. J. Zoo Wildl.Med. 32: 519-522.

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

Clauss, M., Wang, Y., Ghebremeskel, K., Lendl, C.E., Streich, W.J., ., 2003. Plasma and erythrocyte fatty acids in captive Asian (Elephas maximus) and African (Loxodonta africana) elephants. Vet Rec 153, 54-58.
Abstract: The fatty acid components of the plasma triglycerides and the phospholipid fractions of the red blood cells of a captive group of two African (Loxodonta africana) and four Asian (Elephas maximus) elephants were investigated. All the animals received the same diet of hay, fruits and vegetables, and concentrates. A comparison with data from free-ranging African elephants or Asian work-camp elephants showed that the captive elephants had lower proportions of polyunsaturated fatty acids (PUFAs), and for several lipid fractions a higher n-6:n-3 ratio, than their counterparts in the wild or under the more natural, in terms of diet, work-camp conditions. The difference in PUFA content was smaller in the African than in the Asian elephants. The captive Asian elephants tended to have lower levels of n-3 and total unsaturated fatty acids in their red blood cells than the captive African elephants.  Institute of Animal Physiology, Physiological Chemistry and Animal Nutrition, Veterinary Faculty, Ludwig-Maximilians University of Munich, Veterinarstrasse 13, D-80539 Munich, Germany.

Dutta, D., 2003. Physiology of elephant. In: Das, D. (Ed.), Healthcare, Breeding and Management of Asian Elephants. Project Elephant. Govt. of India, New Delhi, pp. 17-22.

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

Isaza, R., Behnke, B.J., Bailey, J.K., McDonough, P., Gonzalez, N.C., Poole, D.C., 2003. Arterial blood gas control in the upright versus recumbent Asian elephant. Respir Physiolo Neurobiol 134, 169-176.
Abstract: In the elephant, there is concern that lateral recumbency (LR) impairs respiratory muscle and lung function resulting in clinically significant arterial hypoxemia. Using healthy adult female Asian elephants (Elephas maximus, n=6), the hypothesis was tested that, given the O₂ binding characteristics of elephant blood, substantial reductions in arterial O₂ pressure PaO₂   in LR could be tolerated without lowering arterial O₂ content appreciably. Fifteen minutes of LR decreased PaO₂ from 103+/-2 (upright, U) to 77+/-4 mmHg (P<0.05) and hemoglobin O₂ saturation (U, 97.8+/-0.1, LR, 95.3+/-0.5%, P<0.05). However, due to a recumbency-induced hemoconcentration, arterial O₂  content was unchanged (U, 18.2+/-2.4, LR, 18.3+/-2.1 ml O₂  per 100 ml). In addition, there was a mild hyperventilation in LR that reduced arterial CO₂ pressure (PCO₂₎ from 39.4+/-0.3 to 37.1+/-1.0 mmHg (P<0.05). These data indicate that the Asian elephant can endure at least short periods of LR without lowering arterial O₂  content.

Mahato, G., 2003. Signs of health and sicknes, preventive healthcare, deworming and quarantine of elephants. In: Das, D. (Ed.), Healthcare, Breeding and Management of Asian Elephants. Project Elephant. Govt. of India, New Delhi, pp. 123-129.

Rezaian, M., Yamashiro, S., Thanawongnuwech, R.ed., Ingkaninun, P. Comparison of elephant and bovine platelet ultrastructure. Proceedings 11th International Symposium of the World Association of Veterinary Laboratory Diagnosticians and OIE Seminar on Biotechnology.  19. 2003.  Thai Veterinary Medical Association; Bangkok; Thailand. 11-9-2003.
Ref Type: Conference Proceeding

Physiological reference values for elephants.  2002. Apple Valley, MN.
Ref Type: Electronic Citation

 2002. Large Animal Internal Medicine. Mosby, St.Louis.

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

du Plessis, L., Stevens, K., 2002. Blood platelets of the African elephant. J Comp Pathol 127, 208-210.
Abstract: Platelet counts of 22 elephants varied between 229 x 10(9) and 622 x 10(9)/L.Light microscopy showed the platelets to be small, while electron microscopy revealed well preserved but activated platelets with some morphological differences from other mammalian platelets. Dense bodies, alpha-granules, glycogen and mitochondria were present in most sections. Microtubules and the surface connecting canalicular system appeared to be absent, but numerous filamentous structures and Golgi bodies were observed.

Pitts, N.I., Mitchell, G., Raath, C., 2002. Succinylcholine overdose in the African elephant (Loxodonta africana) and impala (Aepyceros melampus): pharmacokinetics, pharmacodynamics and physiological responses. South African Journal of Science 98, 581-588.
Abstract: We investigated the mechanism of the delayed effect of succinylcholine (SuCh) in elephants, by correlating the plasma concentration of SuCh with alterations in respiratory and cardiovascular function and with changes in plasma markers of metabolism. These changes were compared with those in impalas, following a lethal SuCh dose in each species. Total entry of SuCh into the circulation (cumulative dose) and total exposure of neuromuscular receptors to unhydrolysed SuCh (area under curve of plasma, SuCh vs. time), were determined. Absorption of intramuscular SuCh was slower, and the cumulative dose lower in elephant than impala, but exposure to intact SuCh was similar in both. SuCh produced apnoea, a fall in PaO2 and pH, and rises in the PaCO2 and plasma catecholamine and cortisol concentrations, and variable cardiovascular responses. These changes took longer to develop in elephant than impala, but in both species death was associated with metabolic consequences of severe hypoxia. We conclude that the delayed effect of SuCh in elephant does not arise from differences in SuCh pharmacodynamics between the species but can be attributed to different pharmacokinetics, the lower mass-specific metabolic rate of the elephant, and its greater tolerance of severe metabolic changes before death results.

Tresamol, P.V., 2002. Elephant surra. Journal of Indian Veterinary Association Kerala 7, 60.

Tuntasuvan, D., Theeraphan, A., Phoengpong, N., Jitnupong, W., Lungka, G., 2002. Comparison of serum chemistry values and serum mineral values between captive and free-ranging elephants in Thailand. 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. 213-218.
Abstract: For copies write to: Forest Resources Officer, FAO Regional Office for Asia and the Pacific, Maliwan Mansion, Phra Atit Road, Bangkok 10200, Thailand; Email: masakazukashio@fao.org

Harr, K., Isaza, R., Harvey, J. Clinicopathological findings in Mycobacterium tuberculosis culture-positive elephants (Elephas maximus) in comparison to clinically normal 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.  209-211. 2001.  American Association of Zoo Veterinarians.
Ref Type: Conference Proceeding

Tuntasuvan, D., Mohkaew, K., Dubey, J.P., 2001. Seroprevalence of Toxoplasma gondii in elephants (Elephas maximus indicus) in Thailand. Journal of Parasitology 87, 229-230.
Abstract: Serum samples from 156 captive elephants (Elephas maximus indicus) collected between 1994 and 1999 in Thailand were examined for antibodies to T. gondii using the modified agglutination test (MAT) and the latex agglutination test (LAT). Antibodies to T. gondii were found in 45.5% of 156 elephants by MAT (_1:50) and 25.6% of 156 elephants by LAT (_1:64). This is the first report of T. gondii infection in E. maximus indicus from Asia.

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.

Aird, B., 2000. Clinical and hematological manifestations of anemia. In: Feldman, B., Zinkl, J.G., Jain, N.C. (Eds.), Schalm's Veterinary Hematology. Lippinicott, Williams & Wilkins, Baltimore,  Maryland, USA, pp. 140-142.

Aird, B., 2000. Acute blood loss anemia. In: Feldman, B., Zinkl, J.G., Jain, N.C. (Eds.), Schalm's Veterinary Hematology. Lippinicott, Williams & Wilkins, Baltimore,  Maryland, USA, pp. 151-153.

Enoe, C., Georgiadis, M.P., Johnson, W.O., 2000. Estimation of sensitivity and specificty of diagnostic tests and disease prevalence when the true disease state is unknown. Preventive Veterinary Medicine 45, 61-81.

Freeman, K., 2000. Bone marrow evaluation. In: Feldman, B., Zinkl, J.G., Jain, N.C. (Eds.), Schalm's Veterinary Hematology. Lippinicott, Williams & Wilkins, Baltimore,  Maryland, USA, pp. 29-32.

Greiner, M., Gardner, I., 2000. Epidemiologic issues in the validation of veterinary diagnostic tests. Preventive Veterinary Medicine 45, 3-22.

Hohenhaus, A.E., (Lungka, G., 2000. Transfusion reactions. In: Feldman, B., Zinkl, J.G., Jain, N.C. (Eds.), Schalm's Veterinary Hematology. Lippinicott, Williams & Wilkins, Baltimore,  Maryland, USA, pp. 864-868.

Horne, W.A., Tchamba, M.N., Loomis, M.R. A simple method of providing intermittent positive-pressure ventilation to etorphine-immobilized elephants (Loxodonta africana) in the field. Proceedings AAZV and IAAAM Joint Conference.  27-30. 2000.
Ref Type: Conference Proceeding

Kramer, J.W., 2000. Normal hematology of the horse. In: Feldman, B., Zinkl, J.G., Jain, N.C. (Eds.), Schalm's Veterinary Hematology. Lippinicott, Williams & Wilkins, Baltimore,  Maryland, USA, pp. 1069-1074.

Larsen, R.S., Salman, M.D., Mikota, S.K., Isaza, R., Montali, R.J., Triantis, J., 2000. Evaluation of a multiple-antigen enzyme-linked immunosorbent assay for detection of Mycobacterium tuberculosis infection in captive elephants. Journal of Zoo and Wildlife Medicine 31, 291-302.
Abstract: Mycobacterium tuberculosis has become an important agent of disease in the captive elephant population of the United States, although current detection methods appear to be inadequate for effective disease management. This investigation sought to validate a multiple-antigen enzyme-linked immunosorbent assay (ELISA) for screening of M. tuberculosis infection in captive elephants and to document the elephant's serologic response over time using a cross-sectional observational study design. Serum samples were collected from 51 Asian elephants (Elephas maximus) and 26 African elephants (Loxodonta africana) from 16 zoos and circuses throughout the United States from February 1996 to March 1999. Infection status of each animal was determined by mycobacterial culture of trunk washes. Reactivity of each serum sample against six antigens was determined, and the linear combination of antigens that accurately predicted the infection status of the greatest number of animals was determined by discriminant analysis. The resulting classification functions were used to calculate the percentage of animals that were correctly classified (i.e., specificity and sensitivity). Of the 77 elephants sampled, 47 fit the criteria for inclusion in discriminant analysis. Of these, seven Asian elephants were considered infected; 25 Asian elephants and 15 African elephants were considered noninfected. The remaining elephants had been exposed to one or more infected animals. The specificity and sensitivity of the multiple-antigen ELISA were both 100% (91.9-100% and 54.4-100%, respectively) with 95% confidence intervals. M. bovis culture filtrate showed the highest individual antigen specificity (95%; 83.0-100%) and sensitivity (100%; 54.4-100%). Serum samples from 34 elephants were analyzed over time by the response to the culture filtrate antigen; four of these elephants were culture positive and had been used to calculate the discriminant function. Limitations such as sample size, compromised ability to ascertain each animal's true infection status, and absence of known-infected African elephants suggest that much additional research needs to be conducted regarding the use of this ELISA. However, the results indicate that this multiple-antigen ELISA would be a valuable screening test for detecting M. tuberculosis infection in elephant herds.

Lin, Y.N., Wong, W.K., 2000. Sero-prevalence of Japanese Encephalitis virus in various species of animals in Singapore - a preliminary study. Singapore Journal of Primary Industries 28, 57-61.
Abstract: A study was conducted to determine exposure of local animals to Japanese encephalitis virus (JEV) 9 years after the removal of its main amplifying host animals in Singapore. 295 serum samples from local (40 dogs, 10 cats, 10 wild boars, 40 chickens, 10 dairy cattle, 10 dairy goats, 55 crows, 44 domestic pigs) and imported (10 ducks, 28 chickens, 5 babirusas, 11 pot-bellied pigs, 5 bearded pigs, 1 Ankole cattle, 1 Asian elephant, 2 orangutans, 3 long-tailed macaques, 1 springbok, 2 reticulated pythons, 1 mouflon, 1 red-eared terrapin, 1 Cape hunting dog, 1 siamang, 1 leopard cat, 1 puma and 5 bats) were screened for JEV antibodies. Imported exotic animals like babirusas, pot-bellied pigs, bearded' pigs, Ankole cattle, Asian elephant and orangutans were seropositive for JEV. High titres and exposure rates were observed in 72.7% of pot-bellied pigs and 100% of the babirusas and bearded pigs with titres of 430.5, 256 and 256, respectively. Antibodies were not detected in the other species of wildlife tested. Among local animals, 100% of goats had moderate antibody levels (GMT=34) while 70% of local cattle had low (GMT=9,8) titres. Ten pet dogs were seronegative while military and stray dogs had 20 and 40% exposure rates, respectively. Local ducks and chickens had low exposure rates (10%, GMT=32 and 0%, respectively) while imported ducks and chickens from Malaysia had high rates (80%, GMT_90.5 and 35.7%, GMT=48.5%, respectively). The cats and crows were seronegative for JEV antibodies. 89% of domestic sows (GMT=217) and 20% of gilts (GMT=16) were seropositive while weaners were seronegative. Wild boars from Pulau Tekong also had high exposure rates (100%, GMT=238.9). It is concluded that a low level of JEV is currently maintained in certain animal populations in Singapore.

Lyashchenko, K., Singh, M., Colangeli, R., Gennaro, M.L., 2000. A multi-antigen print immunoassay for the development of serological diagnosis of infectious disease. Journal of Immunological Methods 242, 91-100.

Mikota, S.K., Kahn, M.J., 2000. Hematology of the elephant. In: Feldman, B., Zinkl, J.G., Jain, N.C. (Eds.), Schalm's Veterinary Hematology. Lippinicott, Williams & Wilkins, Baltimore,  Maryland, USA, pp. 1191-1195.

Morrisey, J.K., 2000. Blood transfusions in exotic species. In: Feldman, B., Zinkl, J.G., Jain, N.C. (Eds.), Schalm's Veterinary Hematology. Lippinicott, Williams & Wilkins, Baltimore,  Maryland, USA, pp. 855-860.

Sellon, D.C., 2000. Blood transfusions in large animals. In: Feldman, B., Zinkl, J.G., Jain, N.C. (Eds.), Schalm's Veterinary Hematology. Lippinicott, Williams & Wilkins, Baltimore,  Maryland, USA, pp. 849-854.

Subhachalat, P., Panichkriangkrai, W., Mahasawangkul, S., Angkawanich, T., 2000. Serum cholinesterase levels in elephants. Thai Journal of Veterinary Medicine 30, 63-68.
Abstract: Serum cholinesterase (ChE) levels were determined in adult elephants in the summer, rainy and winter seasons by a colorimetric method using acetylthiocholine as the substrate. ChE levels were found to be lower in adult males than in adult females at all 3 seasons (P < 0.05). ChE level was highest in the rainy season. It became lower in the summer and winter seasons. The difference was found in both sexes. This study is the first report on the measurement of ChE levels in elephants.

Tveden, H., Weiss, D.J., 2000. Classification and laboratory evaluation of anemia. In: Feldman, B., Zinkl, J.G., Jain, N.C. (Eds.), Schalm's Veterinary Hematology. Lippinicott, Williams & Wilkins, Baltimore,  Maryland, USA, pp. 143-15.

Valli, V.E., Jacobs, R.M., 2000. Structure and function of the hemopoietic system. In: Feldman, B., Zinkl, J.G., Jain, N.C. (Eds.), Schalm's Veterinary Hematology. Lippinicott, Williams & Wilkins, Baltimore,  Maryland, USA, pp. 225-239.

Wardrop, K.J., 2000. Clinical blood typing and crossmatching. In: Feldman, B., Zinkl, J.G., Jain, N.C. (Eds.), Schalm's Veterinary Hematology. Lippinicott, Williams & Wilkins, Baltimore,  Maryland, USA, pp. 795-798.

Weiss, D.J., 2000. Clinical hematology. In: Feldman, B., Zinkl, J.G., Jain, N.C. (Eds.), Schalm's Veterinary Hematology. Lippinicott, Williams & Wilkins, Baltimore,  Maryland, USA, pp. 57-60.

Welles, E.G., 2000. Clinical interpretation of equine leukograms. In: Feldman, B., Zinkl, J.G., Jain, N.C. (Eds.), Schalm's Veterinary Hematology. Lippinicott, Williams & Wilkins, Baltimore,  Maryland, USA, pp. 405-410.

Bhat, M.N., Manickam, R., Ramkrishna, J., 1999. Screening of captive wild animals for tuberculosis. Indian Veterinary Journal 76, 959-961.
Abstract: The passive haemagglutination (PHA) test was used to test 109 captive elephants (Elephas maximus), and spotted deer (Cervus axis), blackbuck (Antilope cervicapra) and common langurs (Semnopithecus entellus?) (4 of each) for tuberculosis; 51 of the elephants and the 4 langurs were also assessed by the tuberculin test. PHA titres of 1:16 or 1:32 were found in 4 elephants, 1 deer and 2 langurs, but all were apparently healthy except 1 langur that had clinical signs indicative of tuberculosis. There were 4 positive reactors in the tuberculin tests, all elephants, but these animals did not have significant PHA titres. It is concluded that the procedures and reagents used for the diagnosis of tuberculosis in domestic animals are not reliable for testing wild animals.

Hietala, S.K., Gardner, I.A., 1999. Validity of using diagnostic tests that are approved for use in domestic animals for non-domestic species. In: Fowler, M.E., Miller, R.E. (Eds.), Zoo and Wild Animal Medicine, 4^th ed. W.B. Saunders, Philadelphia, pp. 55-57.

Hove, T., Dubey, J.P., 1999. Prevalence of Toxoplasma gondii antibodies in sera of domestic pigs and some wild game species from Zimbabwe. Journal of Parasitology 85, 372-373.
Abstract: Serum samples from 97 domestic pigs (Sus scrofa) slaughtered in Harare, Zimbabwe, in 1995, and from 19 elands (Taurotragus oryx [Tragelaphus oryx]), 67 sable antelopes (Hippotragus niger), 3 warthogs (Phacochoerus aethiopicus), 3 bushpigs (Koiropotamus [Potamochoerus] koiropotamus), 2 white rhinos (Ceratotherium simus [C. simum]), 18 African buffaloes (Syncerus caffer), 69 wildebeest (Connochaetes taurinus), and 19 African elephants (Loxodonta africana) in Zimbabwe, were tested for Toxoplasma gondii IgG antibodies by the modified agglutination test (MAT) with whole formalized tachyzoites and mercaptoethanol. Sera were diluted at 1:25, 1:50, and 1:500 for MAT testing; sera with antibodies in a 1:25 dilution were considered to have T. gondii infection. Antibodies were found in 9 domestic pigs (9.3%), 7 elands (36.8%), 8 sable antelopes (11.9%), no warthogs, no bush-pigs, one white rhino (50%), one buffalo (5.6%), 10 wildebeest (14.5%), and 2 elephants (10.5%).

Jayathangaraj, M.G., John, M.C., Ramesh, S., Ahamed, N.M., 1999. Haematological and biochemical findings in captive Asiatic elephants (Elephas maximus). International Journal of Animal Sciences 14, 245-247.
Abstract: The haematological and biochemical parameters were estimated using blood samples obtained from apparently healthy four elephants maintained in the temples and the values were analysed.

Morris, D.D., Barton, M.H., 1999. Techniques for evaluating the hemostatic system. In: Colahan, P.T., Merritt, A.M., Moore, J.N., Mayhew, I.G. (Eds.), Equine Medicine and Surgery. Mosby, St. Louis MO USA, pp. 1986-1987.

Sarmah, B.C., Kalita, D.J., Pathak, S.C., 1999. Mineral status of elephant. Indian Veterinary Journal 76, 661-662.
Abstract: The serum concentrations of Ca, Mg, Fe, Zn, Cu, Mo and Mn were measured by atomic absorption spectrometry in blood samples collected from 20 elephants in 2 reserves (Kaziranga and Manas) in Assam; the results are tabulated. In general, the levels in animals on the 2 reserves were similar, except that the Fe and Cu concentrations were significantly higher in the animals at Kaziranga; differences in the mineral contents of the soil and the plants were suggested as a possible explanation for the variations.

Savage, C.J., 1999. Diseases of the Liver. In: Colahan, P.T., Merritt, A.M., Moore, J.N., Mayhew, I.G. (Eds.), Equine Medicine and Surgery. Mosby, St. Louis MO USA, pp. 816-833.

Senthilkumar, A., Jayathangaraj, M.G., John, M.C., Srinivasan, S.R., 1999. Serum creatinine level in captive Asiatic elephants (Elephas maximus). Indian Journal of Animal Health 38, 181.

Sood, R., 1999. Medical Laboratory Technology: Methods and Interpretation. Jaypee Brothers, New Delhi, India.

Tell, L., Kabbur, M.B., Smith, W.L., Gage, L., Cullor, J.S., 1999. Oxygen radical production by Asian elephant (Elephas maximus) heterophils and Holstein cattle (Bos taurus) neutrophils. Journal of Zoo and Wildlife Medicine 30, 402-407.
Abstract: Oxidative metabolic burst activity by Asian elephant (n=15) heterophils and Holstein cattle (n=16) neutrophils was indirectly evaluated by measuring the oxidation of non-fluorescent intracellular 2',7'-dichlorofluorescin (DCFH) to fluorescent 2', 7'-dichlorofluorescein (DCF) with flow cytometry. The results were recorded as mean channel fluorescence intensity. Phorbol myristate acetate at 50 ng/ml was used to activate the cells. Elephant heterophils and bovine neutrophils exhibited similar abilities to oxidize intracellular DCFH to DCF. A wide range of mean channel fluorescence intensity (1803-7393) was observed among individual elephants, and the range of intensities was similar to that observed in bovine neutrophils, supporting the concept of functional heterogeneity of heterophils among elephants.

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

Bhat, M.N., Manickam, R., Aruni, W., 1998. Detection of bluetongue antibody and antigen in Indian elephants, spotted deer and blackbucks. Indian Journal of Animal Sciences 68, 135.

Bhat, M.N., Manickam, R., Nedunchelliyan, S., Jayakumar, V., 1998. Detection of leptospirial antibodies in the sera of elephants. Indian Veterinary Journal 75, 201-203.
Abstract: Leptospirosis is an infectious disease of man and animals, caused by antigenically distinct members of the genus  Leptospira.  Upadhya et al (1979) detected antibody against Leptospira valbuzzi and L. pyrogenes in the sera of elephants.  Arora (1994) detected antibodies against L. pomona in a sambar and black buck.  In the present study, a serological survey was undertaken to detect antibodies against seven serovars of Leptospira sp. in elephants (Elephas maximus), spotted deer (Axis axis), and blackbucks (Antelope cervicapra). Serum samples were collected from 109 elephants in 15 elephant camps in Madras, Karnataka, and the Andaman and Nicobar Islands, and from 4 spotted deer and 4 blackbuck in a zoo in Madras. In 23 (21%) of the elephants, antibodies, at titres of 1:100 to 1:200, were found by microscopic agglutination tests, to 6 serovars of Leptospira interrogans: pomona, icterohaemorrhagiae, grippotyphosa, hebdomadis, hardjo, and canicola. No leptospiral antibodies were found in the serum of the other animals. Clinical signs were present only in the elephants with L. grippotyphosa; the other serovars are reported for the first time in Indian elephants.

Cheryk, L.A., Gentry, P.A., Bast, T., Yamashiro, S., 1998. Alterations in blood platelet morphology during aggregate formation in the Asian elephant (Elephas maximus).  Journal of Zoo and Wildlife Medicine 29, 177-182.
Abstract: The ultrastructure of Asian elephant (Elephas maximus) platelets before and after activation with the agonist platelet activating factor (PAF) was studied.  The unactivated platelet has a distinct ultrastructural appearance:  the cytoplasm contains large randomly distributed granules but lacks the internal cristae that typify the open canalicular system in many types of mammalian platelets.  Following PAF stimulation, large platelet aggregates form, but many platelets remain discrete, with little evidence of pseudopod formation or fusion of membranes.  Two types of platelets are visible within the aggregates:  those that are morphologically intact and those with gaplike features on the outer membrane and that have become degranulated, appearing as empty swollen sacs. The lack of platelet membrane fusion within the aggregates may permit the reversal of aggregation that is a characteristic response of elephant platelets to PAF.

Jacobson, R.H., 1998. Validation of serological assays for diagnosis of infectious diseases. Rev. sci. tech. Off. int. Epiz. 17, 469-486.

Kelly, P.J., Carter, S.D., Azwai, S.M., Cadman, H.F., 1998. Isolation and characterization of immunoglobulin g and IgG subclasses of the African elephant (Loxodonta africana). Comparative Immunology, Microbiology and Infectious Diseases 21, 65-73.
Abstract: Immunoglobulins were precipitated from sera pooled from 5 African elephants (killed for population control) with ammonium sulfate and separated by gel filtration and fast protein liquid chromatography (ion exchange). Analysis of the fractions by SDS-PAGE showed IgG of 150 kDa with up to 5 subclasses, each having heavy chains of 57 kDa and light chains of 27 kDa. Three monoclonal antibodies against human IgG and polyclonal antibodies against canine, bovine, cameline, equine, phocine and feline IgG showed strong cross-reactivity with the African elephant IgG subclasses. No serum molecules corresponding to IgM or IgA could be detected, even when ammonium sulfate precipitation was used at 50% saturation.

Wright, P.F., 1998. International standrads for test methods and reference sera for diagnostic tests for antibody detection. Rev. sci. tech. Off. int. Epiz. 17, 527-533.

 1997. Laboratory Procedures for Veterinary Technicians. Mosby, St. Louis MO.

Barnard, B.J.H., 1997. Antibodies against some viruses of domestic animals in southern African wild animals. Onderstepoort Journal of Veterinary Research 64, 95-110.
Abstract: Twenty-four species of South African wild animals were tested for the presence of antibodies to the viruses of 16 common diseases of domestic animals around 1993-5. Positive results were obtained for African horse sickness, equine encephalomyelitis virus, equine herpesvirus-1, bovine herpesvirus-1, Allerton disease (Herpes mammillitis; bovine herpesvirus 2), lumpy skin disease, parainfluenza, encephalomyocarditis, bluetongue, Wesselsbron disease, bovine ephemeral fever, and Akabane disease complex. No antibodies could be demonstrated against the viruses of equine influenza, equine infectious anaemia, equine viral arteritis or Rift Valley fever. The negative results support observations that the latter diseases, with the exception of equine viral arteritis, are absent in South Africa. The number of animal species found positive for a specific virus, ranged from 0-16. No antibodies were found in crocodiles or warthogs, whereas antibodies against Wesselsbron and bovine herpesvirus-1 were present in 16 species. Antibodies against viruses of horses were found almost exclusively in zebras and, although elephants reacted to African horse sickness, no neutralizing antibodies against it could be demonstrated in their sera. Zebras were also found to be positive for Wesselsbron and Akabane, which are usually regarded as viruses of ruminants. Antibodies against most viruses were encountered in all vegetation zones in South Africa, but most viruses were more prevalent in the high-rainfall zone in KwaZulu-Natal.

Bhat, M.N., Manickam, R., 1997. Detection of rinderpest antibodies in elephants, spotted deer, and blackbucks. International Journal of Animal Sciences 12, 201-203.

Bhat, M.N., Manickam, R., Kumanan, K., 1997. Serological evidence of bovine herpesviruses 1 and 2 in Asian elephants. Journal of Wildlife Diseases 33, 919-920.
Abstract: Antibodies were detected against bovine herpesviruses 1 (BHV 1) and 2 (BHV 2) in Asian elephants (Elephas maximus) using the passive haemagglutination (PHA) test. The study was conducted during May to December 1994 using sera collected from zoos and national parks in India. Four (4%) of 109 elephant sera had PHA titres ranging from 1:8 to 1:32 against BHV 1. 25 (23%) of the 109 elephant sera had PHA titres ranging from 1:8 to 1:64 against BHV 2. It is concluded that Asian elephants appear to be better reservoirs for herpesviruses which are serologically related to BHV 2.

Kania, S.A., Richman, L.K., Kennedy, M., Montali, R.J., Potgleter, L.N.D., 1997. The isolation, detection, and cross-reactivity of Asian elephant IgG for the development of serological diagnostic tests. Journal of Veterinary Allergy and Clinical Immunology 5, 125-128.
Abstract: Asian elephant (Elephas maximus) IgG was isolated and purified using a recombinant protein A and proteins G affinity matrix and DEAE cibacron blue chromatography. Rabbits were inoculated with elephant IgG to produce anti-Asian elephant IgG. Using an ELISA, it was determined that the anti-Asian elephant sera has strong reactivity with Asian elephant IgG and African elephant (Loxodonta africana) IgG, moderate reactivity with manatee (Trichechus manatus latirostris) IgG and weaker reactivity with IgG from hyrax (Procavia capensis) and black rhinoceros (Diceros bicornis). Commercially available antisera produced against cattle, deer, and rabbit IgG react weakly with Asian elephant IgG.

 1996. Principles of validation of diagnostic assays for infectious diseases. Manual of Standards for Diagnostic Tests and Vaccines. Office International des Epizooties (O.I.E.), Paris, pp. 8-15.

Briggs, M., Phillips, L.G., Kimball, V.P., Valli, V.E. Bone marrow collection in the Asian elephant (Elephas maximus). Proceedings American Association of Zoo Veterinarians.  431-433. 1996.
Ref Type: Conference Proceeding
Abstract: The collection of bone marrow is a common practice for the diagnosis of a variety of disease in mammals and birds.  These include disease which can cause any type of blood dyscrasias, anemias, thrombocytopenias, and/or leukopenias.  Examples of a few of these diseases include feline leukemia, feline lymphosarcoma, lymphoma, bovine leukosis, aplastic anemias of many etiologies, equine infectious anemia, and various types of leukemia.  Dependent upon the species involved, marrow is routinely collected from a variety of locations, such as the crest of the tibia in psittacines, or the ribs, iliac crest, or sternum in equids, to the dorsal spinous processes or sternum in the cow. Although in these species there is seldom a  problem with collection, obtaining marrow from some of the megavertebrates such as the elephant or rhinoceros proves to be problematic due to both the restraint of the animals and their size.  This paper describes a simple and effective way to obtain quality bone marrow samples from an Asian elephant (Elephas maximus) and thus likely applicable to other megavertebrate species.

Gardner, I.A., Hietala, S., Boyce, W.M., 1996. Validity of using serological tests for diagnosis of diseases in wild animals. Rev. Sci Tech. Off. Int. Epiz. 15, 323-335.

Gentry, P.A., Ross, M.L., Yamada, M., 1996. Blood coagulation profile of the Asian elephant (Elephas maximus). Zoo Biology 15, 413-423.
Abstract: The coagulation profile of seven Asian elephants was assessed using human reference plasma as a standard.  The plasma values for the majority of the coagulation proteins evaluated, including Factors VII, IX, X, and XI, and antithrombin, were similar to that of human plasma.  The average Factor VIII:C value was 1.95 units/ml, approximately twice that of the human value.  Human recombinant tissue factor was effective as an activator of the tissue factor-factor VII pathway as measured by the prothrombin time assay.  The elephant plama effectively corrected the clotting defect of human Factor XI-deficient plasma but failed to do so with bovine Factor Xi-deficient plasma.  However, elephant plama Factor XII was not readily activated by the commercial activated partial thromboplastin time (APTT) reagent formulated with a soluble activator, and consequently the activity of this protein could not be precisely determined.  The average (+/-SD) APTT result of 65.6+/-9.2 sec was twice as long as that of the human reference plasma. Despite the presence of relatively high levels of fibrinogen, 4.61+/-0.49 gm/l, no fibrinolytic activity was detected in any of the elephant plasma samples using a standard fibrin plate assay system.

Porter, C.A., Goodman, M., Stanhope, M.J., 1996. Evidence on mammalian phylogeny from sequences of exon 28 of the von Willebrand factor gene. Mol Phylogenet Evol 5, 89-101.
Abstract: Phylogenetic relationships among 27 extant mammalian species (representing 15 placental orders) were studied using sequences of exon 28 of the gene encoding von Willebrand Factor (vWF), a glycoprotein which functions in blood clotting. Analysis of sequences coding for vWF revealed evidence for several subordinal and superordinal groupings, but the earliest branching sequence of placental mammals was left largely unresolved. Strong support was found for a monophyletic clade consisting of elephants, sea cows, hyraxes, aardvarks, and elephant shrews. This systematic placement of the elephant shrews agrees strongly with two other molecular data sets (interphotoreceptor retinoid binding protein and alpha-lens crystallins) and is consistent with analysis of fossil elephant shrews recently discovered in north Africa. Evidence from vWF sequences agrees with a number of previous molecular and morphological studies in providing strong support for the monophyly of both bats and rodents. The orders Primates, Proboscidea, Carnivora, Perissodactyla, and Artiodactyla were represented by more than one species which joined in each case to form a monophyletic order.

Sarma, K.K., Kalita, D., Dutta, B., Barua, S.K., 1996. Determination of mean arterial pressure (MAP) in Asian elephant (Elephas maximus). Indian Veterinary Journal 73, 777-778.

Barnard, B.J.H., Bengis, R.G., Keet, D.F., Dekker, E.H., 1995. Epidemiology of African horsesickness: antibodies in free-living elephants (Loxodonta africana) and their response to experimental infection. Onderstepoort Journal of Veterinary Research 62, 271-275.
Abstract: Serum samples were obtained from blood collected from elephants during a culling operation in Kruger National Park, South Africa, in 1993. Sera from 63/80 (79%) elephants reacted positively in an ELISA for African horse sickness virus (AHSV). The titres of almost 65% of the positive samples were less than 10 000. In comparison, 34/34 zebra samples reacted positively and their ELISA titres were significantly higher, with more than 84% having a titre of 10 000 or higher. 26% of 14 sera from elephants tested for the 9 types of AHSV, reacted positively with virus-neutralizing titres of 20 or higher. Experimental infection of 6 elephant calves resulted in conflicting results. No detectable viremia nor virus could be demonstrated in the organs of the calves and none of them mounted significant levels of neutralizing antibodies against the virus. On the other hand, all calves showed a slight rise in ELISA titres. This rise, however, was modest when compared with the rise in experimentally infected zebra. The presence of low levels of group- and type-specific antibodies in the serum of some free-living elephants was judged to be the result of natural hyper-immunization due to frequent exposure to infected biting insects. It is concluded that, despite the presence of low levels of antibodies, elephants should be regarded as poorly susceptible and unlikely to be a source of AHSV.

Gentry, P.A., Cheryk, L.A., Yamashiro, S., 1995. Absence of pseudopod formation in activated elephant platelets. Thrombosis and Haemostasis 73, 1071.

Grobler, D.G., Raath, J.P., Braack, L.E.O., Keet, D.F., Gerdes, G.H., Barnard, B.J.H., Krick, N.P.J., Jardine, J., Swanepoet, R., 1995. An outbreak of encephalomyocarditis-virus infection in free ranging African elephants in the Kruger National Park. Onderstepoort Journal of Veterinary Research 62, 97-108.
Abstract: An increase in unexplained elephant mortality was seen in the Kruger National Park (KNP) from December 1993 to November 1994, concurrent with a wide-spread increase in the KNP rodent population.  The majority of animals were found dead.  Examination of carcasses ruled out common causes of death, including poaching, anthrax, intraspecific fighting, and intoxication.  Sixty-four animals died from unexplained causes during the perceived outbreak, 83% of which were adult bulls.  Eight carcasses were in sufficiently good condition for tissues to be collected for diagnostic testing.  Cardiac failure appeared to be the most likely cause of death in seven of the animals, with gross findings of pulmonary edema, hepatic congestion, ascities, and hydrothorax.  Myocarditis and necrosis of myocytes were the most striking findings on histopathological examination.  Heart tissue from three animals was submitted for virus isolation; all three yielded encephalomyocarditis (EMC) virus.  Serologic testing for EMC virus antibody was performed on the KNP between 1984 and 1994.  Results demonstrated that the virus has  been present in the KNP from 1987 on.  EMC virus antibody was not detected in preserved rodent tissues until 1993, prior to the rodent population explosion and the outbreak of disease in elephants.  It is unclear whether rodents play a role in transmitting the virus to other animals or if they reflect a general circulation of the virus in multiple species in a given environment.  One lion cub which was found dead with bacterial pneumonia had a serum neutralizing antibody titer to EMC virus of 128.  It is hypothesized that this animal may have been predisposed to pneumonia through the formation of lung edema as a result of EMC virus infection.  Three lions that were seen feeding on the carcass of an elephant with lesions compatible with EMC virus infection were monitored for seroconversion, which did not occur.  EMC virus disappears rapidly from most tissues after death and probably was not present in the tissues consumed by the lions.  The predilection for male elephants could not be explained, although increased mortality among males has also been demonstrated with EMC virus in mice.

Ratnasooriya, W.D., Amarasinghe, A.B.C., Kodikara, D.S., 1995. Total serum cholesterol levels of Sri Lankan elephants (Elephas maximus maximus). Ceylon Journal of Science, Biological Sciences 24, 11-15.
Abstract: Total serum cholesterol level was determined in blood samples from 43 captive elephants using an enzymatic technique. The mean total serum cholesterol level was 45.23 ± 1.37 mg/dl (means ± SEM). The serum cholesterol level of male elephants (48.42 ± 1.90 mg/dl) was significantly higher than that of female elephants (43.34 ± 1.81 mg/dl). There was no significant difference in the serum cholesterol level amongst the 3 morphological 'types' of males (Atha: 46.42 ± 4.22 mg/dl; Aliya: 51.00 ± 2.77 mg/dl; and Pussa: 45.23 ± 2.98 mg/dl) and the two morphological 'types' of females (Athinna: 47.79 ± 3.95 mg/dl and Alidena: 41.98 ± 1.98 mg/dl). The serum cholesterol level of non working elephants (45.70 ± 2.6 mg/dl) was not significantly different from that of working elephants (45.14 ± 1.51 mg/dl). However, there was a significant difference in the serum cholesterol level of the Aliya (51.00 ± 22.77 mg/dl) and Alidena (41.98 ± 1.98 mg/dl).

Schumacher, J., Heard, D.J., Caligiuri, R., Norton, T., Jacobson, E.R., 1995. Comparative effects of etorphine and carfentanil on cardiopulmonary parameters in juvenile African elephants (Loxodonta africana).  Journal of Zoo and Wildlife Medicine 26, 503-507.
Abstract: Fourteen African elephants (Loxodonta africana) were immobilized with either etorphine hydrochloride (3.2 ± 0.5 µg/kg i.m.) or carfentanil citrate (2.4 µg/kg i.m.). Induction time with etorphine was significantly longer (30 ± 21 min) than with carfentanil (8 ± 2 min).  Immediately following immobilization all elephants were placed in lateral recumbency and respiratory rate, heart rate, and rectal body temperature were monitored every 5 min throughout the immobilization period.  Arterial blood samples, collected from an auricular artery, were taken 10 min after immobilization and every 15 min thereafter for up to 1 hr.  At the first sampling, mean values for arterial blood gas variables for etorphine immobilized elephants were pHa, 7.29 ± 0.03; PaCO₂, 53.4 ± 5.2 mmHg; PaO₂, 71.8 ± 13.8 mmHg; standard base excess (SBE), -1.6 ± 2.9 mEq/L; and HCO3, 25.7 ± 2.7 mEq/L. After 1 hr of immobilization, mean arterial blood gas values were pHa, 7.32 ± 0.06; PaCO₂ , 57.2 ± 9.6 mm Hg; and PaO₂ , 53.8 ± 10.5 mm Hg; SBE, 2.7 ± 1.4 mEq/L; and HCO_3-, 30.6 ± 1.6 mEq/L. For carfentanil immobilized elephants, blood gas values at the first time of collection were pHa, 7.28 ± 0.04; PaCO₂, 52.1 ± 2.8 mmHg; PaO₂, 78.3 ± 14.7 mmHg; SBE, -2.3 ± 24 mEq/L; and HCO_3-, 24.3 ± 2.1 mEq/L.  Sixty minutes after the first sampling, blood gas values of one elephant were pHa, 7.38; PaCO₂, 48.7 mmHg; PaO₂, 52 mmHg; SBE, 3.4 mEq/L, and HCO_3-, 28.8 mEq/L.  Over time there was a progressive decline in arterial PO2 in all elephants.  It is concluded that elephants immobilized with either etorphine HCl or carfentanil developed hypoxemia (PaO₂ < 60 mmHg) after 30 min of immobilization.  It is recommended that the administration of one of these opioid drugs be accompanied by supplemental oxygen, or followed by an inhalant anesthetic in 100% oxygen for prolonged procedures.  Diprenorphine or nalmefene reversal was rapid and uneventful in both the etorphine and carfentanil group.  No cases of renarcotization were noted. Additional excerpt: All elephants in the etorphine group (n=8) received diprenorphine at a mean dosage of 8.3 ± 1.1 µg/kg IV. Two elephants in the carfentanil group (n=6) were administered diprenorphine at a dosage of 8.9 µg/kg IV and IM.  Three elephants in this group received nalmefene hydrochloride.  One of the three elephants was given nalmefene 166.7 µg/kg both IV and SC. Two of the three elephants were given nalmefene IV and IM. The dosage was 88.9 µg/kg IV and IM in one elephant and 53.3 µg/kg IV and IM in the other. One elephant in the carfentanil group was administered nalmefene (88.9 µg/kg IV) followed by diprenorphine (8.9 µg/kg IM).

 1994. Veterinary Laboratory Medicine. Iowa State University Press, Ames, IA.

Brown, J.L., Schoeneman, H.M., Raath, J.P., de Vos, V., Bush, M., 1994. Serum concentrations of cationic minerals in free-ranging elephants (Loxodonta africana) in the Kruger National Park. Journal of Zoo and Wildlife Medicine 25, 495-499.
Abstract: Blood samples were collected from 22 free-ranging adult African elephants (Loxodonta africana) in the Kruger National Park, Republic of South Africa, and analyzed for concentrations of serum sodium, potassium, calcium, magnesium, copper, zinc, and iron. Fourteen of the animals (males) were anesthetized at the time of sample collection; the remaining samples were obtained from male (n = 4) and female (n = 4) elephants during an annual population control program.  There were no gender differences in mineral concentrations among culled elephants.  Serum calcium, magnesium, copper, and sodium concentrations also were similar between anesthetized and culled animals.  In contrast, serum potassium and zinc concentrations were higher and serum iron concentration was lower in culled elephants than in anesthetized males.

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.

Komoin-Oka, C., Truc, P., Bengaly, Z., Formenty, P., Duvallet, G., Lauginie, F., Raath, J.P., N'-Depo, A.E., Leforban, Y., 1994. A study of the prevalence of trypanosome infections in different species of wild animals in Comoe National Park Cote d'Ivoire: preliminary results of a comparison of three diagnostic methods. Revue d'Elevage et de Medecine Veterinaire des Pays Tropicaux 47, 189-194.
Abstract: Microscopic examination of thin smears, an ELISA, and kit for in vitro isolation of trypanosomes (KIVI) were compared for diagnosis of trypanosomiasis in 3 elephants, 53 African buffaloes, 12 roan antelope (Hippotragus equinus), 9 hartebeest (Alcelaphus buselaphus), 19 waterbuck (Kobus ellipisiprymnus), 61 Kob (Kobus kob), and 6 wart hogs (Phacochoerus aethiopicus). In the 82 animals on which all 3 tests were carried out, 20% were positive by the thin smear method, 50% by the ELISA and 80% by the KIVI test. The trypanosomes were probably T. brucei, T congolense and T. vivax but these were not identified.

Kelly, P.J., Tagwira, M., Matthewman, L., Mason, P.R., Wright, E.P., 1993. Reactions of sera from laboratory, domestic and wild animals in Africa with protein A and A recombinant chimeric protein AG. Comparative Immunology, Microbiology and Infectious Diseases 16, 299-305.
Abstract: An ELISA was developed to determine the reactivity of peroxidase labelled Protein A and a recombinant Protein A + Protein G construct, to sera from a variety of laboratory, domestic and wild animals from Africa. There was variability in the binding capacity of sera from individuals of the same species, but 4 groups could be recognized. Sera from birds and crocodiles were at most weakly reactive with either Protein A or the chimeric construct. Sera from some domestic animals such as horse, goat and cat, and sera from some wild ungulates including buffalo, wildebeest, waterbuck and impala were reactive with Protein A, but reacted to a much greater degree with the chimeric construct. Sera from larger wild animals such as elephant, rhinoceros and giraffe were strongly reactive with the chimeric protein and moderately reactive with Protein A. Sera from primates and dog, pig, guinea pig and rabbit reacted strongly with both proteins. It was concluded that as chimeric proteins that combine the IgG binding capacities of Protein A and Protein G can be used to detect immunoglobulin from a wide variety of African wild animal species, they may be of great value in seroepidemiological investigations of these animal populations.

Kock, M.D., Martin, R.B., Kock, N., 1993. Chemical immobilization of free-ranging African elephants (Loxodonta africana) in Zimbabwe, using etorphine (M99) mixed with hyaluronidase, and evaluation of biological data collected soon after immobilization. Journal of Zoo and Wildlife Medicine 24, 1-10.
Abstract: Sixteen adult female free-ranging elephants were immobilized in July 1990, using a mean (±SE) dose per animal of 11.6 ± 0.3 mg of etorphine (M99) mixed with a standard dose of hyaluronidase (4500 IU), at the Sengwa Wildlife Research Area, Zimbabwe, to attach telemetry and infrasound detection collars. The 16 elephants were reimmobilized in December 1990, using higher doses of etorphine (standardized at 15 mg total dose) with hyaluronidase (4500 IU), to remove the collars. The higher doses of etorphine produced more rapid inductions. Biological data were collected on both occasions. Significant differences in selected measures indicative of stress, including lactic dehydrogenase and aspartate transaminase, were seen between immobilizations. Comparisons were made of selected health measures between samples collected in the early winter and late winter/early spring season. Significant differences were seen with total protein, albumin, urea nitrogen, creatinine, calcium, magnesium, inorganic phosphorus, chloride, and alanine transaminase.

Silva, I.D., Kuruwita, V.Y., 1993. Hematology, plasma, and serum biochemistry values in domesticated elephants (Elephas maximus ceylonicus) in Sri Lanka. Journal of Zoo and Wildlife Medicine 24, 440-444.
Abstract: This study was conducted to further strengthen the knowledge of hematology and blood chemistry of Asian elephants (Elephas maximus ceylonicus).  Blood samples collected from 108 healthy, domesticated elephants, without chemical immobilization and with a male:female ratio of 1.16, were analyzed for cellular and biochemical measures using standard hematologic techniques.  The mean values obtained for these measures were comparable to those reported for domesticated Asian elephants in other countries.  The range of erythrocyte measures were: packed cell volume, 25-45%; erythrocyte count, 1.7-5.0 x 10⁶/ul; hemoglobin concentrations, 7.4-15.4 g/dl; mean corpuscular volume, 81-160 fl; mean corpuscular hemoglobin, 24-56 pg; mean corpuscular hemoglobin concentration 23-39 g/dl; and sedimentation rate, 64-148 mm/hr.  A higher mean and a wider range were observed for the erythrocyte count when compared to other reports.  The leukocyte count varied from 4 to 21 x 10³/ul, and lymphocytes were present in largest proportions (38%).  Equal proportions (approximately 25%) of heterophils and moncytes were found in the peripheral blood, and both of these leukocytes exhibited peroxidase activity in their cytoplasmic granules, whereas lymphocytes did not.  The nuclei of the granulocytes were poorly segmented, and heterophils had reddish cytoplasmic granules.  Approximately 80% of the monocytes had segmented nuclei.  The platelet count ranged from 0.8 to 4 x 10⁵/ul, a range that was narrower when compared with other reports.  Total plasma protein concentration ranged from 6 to 13 g/dl, with a fibrinogen concentration of 0.04-2.8 g/dl.  There were no significant differences between values for males and females. Likewise, there were no differences in blood values between these domesticated elephants and the free-ranging, chemically immobilized elephants that were studied separately.

Silva, I.D., Kuruwita, V.Y., 1993. Hematology, plasma, and serum biochemistry values in free-ranging elephants in Sri Lanka. Journal of Zoo and Wildlife Medicine 24, 434-439.
Abstract: This study was conducted to broaden the knowledge of the hematology of healthy free-ranging Asian elephants (Elephas maximus celonyicus). Blood cell measurements and plasma and serum biochemistries were determined using standard hematologic techniques.  Blood samples were collected from 37 elephants following chemical immobilization.  When compared to other domestic species such as bovids, equids, canids and felids, the elephants' packed cell volumes (31-44%), hemoglobin concentrations (9.8-15.8g/dl), and mean corpuscular hemoglobin concentrations (28-40g/dl) were similar.  Erythrocyte counts (2.3-5.4 x 10⁶/ul) were lower, and mean corpuscular volumes (81-158 fl), mean corpuscular hemoglobins (30-50 pg), and leukocyte counts (8-26 x 10³/ul) were higher.  Lymphocytes comprised the largest proportion of (44%) of leukocytes.  The nuclei of granulocytes were poorly segmented, and the neutrophils (heterophils) had reddish cytoplasmic granules with demonstrable peroxidase activity.  Heterophils and moncytes were present in similar proportions.  There were two types of monocytes, typical monocytes with unsegmented nuclei and monocytes with segmented nuclei.  The cytoplasmic granules of both types had peroxidase activity, but unlike heterophils, these granules were not visible with Romanowsky-type stains.  Most of the biochemical measures were comparable to values for bovine, equine, canine and feline species, with slight differences in the ranges. Plasma protein concentrations ranged from 6 to 11 g/dl, and 10% of the elephants had concentrations above 10 g/dl.  Wider ranges were observed in elephants for concentrations of plasma fibrinogen (0.13-2.70g/dl), creatinine (1.0-3.8 mg/dl), and total and conjugated bilirubin (0.23-0.94 mg/dl and 0.07-0.76 mg/dl, respectively), although the icterus index was 2.  Values for plasma urea nitrogen (3-18 mg/dl), serum glutamic:pyruvate transaminase activity (4-40 U/L), serum calcium (5.5-11.0 mg/dl), and serum calcium:phosphorus ratio (1.09-3.68) were lower in the elephant than in bovine, equine, canine and feline species.

Binepal, V.S., Wariru, B.N., Davies, F.G., Soi, R., Olubayo, R., 1992. An attempt to define the host range for African horse sickness virus (Orbivirus, Reoviridae) in east Africa, by a serological survey in some Equidae, Camelidae, Loxodontidae and Carnivore. Vet. Microbiol. 31, 19-23.
Abstract: A survey was carried out in horse, zebra, elephant, camel, sheep and goat and wild carnivore sera for virus-serum neutralizing antibody to the nine type strains of African horse sickness virus. Antibody was found amongst the horse, zebra and elephant sera to all nine different strains. No antibody was detected in any sera from camels, sheep and goats. None was found in sera from hyaena and jackals in this series but had been detected earlier.

Dathe, H.H., Kuckelkorn, B., Minnemann, D., 1992. Salivary cortisol assessment for stress detection in the Asian elephant (Elephas maximus): A pilot study. Zoo Biology 11, 285-289.
Abstract: Effects of introducing an unfamiliar female into an Asian elephant herd at Tierpark Berlin were monitored by means of salivary cortisol assessment.  Saliva samples were obtained from a second female for comparative purposes.  The period of familiarization was characterized by an enhanced cortisol level in both animals, with a maximum on the second day after joining.  Cortisol returned to normal on the following day. Manipulations of the keepers caused a transitory increase on two other days.   Possibilities for the use of this noninvasive method of stress monitoring in various management situations are indicated.

George, S., Nirmalan, G., 1992. Kinetic parameters of lactate dehydrogenase in the Indian elephant. 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. 54-56.

Honeyman, V.L., Pettifer, G.R., Dyson, D.H., 1992. Arterial blood pressure and blood gas values in normal standing and laterally recumbent African (Loxodonta africana) and Asian (Elephas maximus) elephants. Journal of Zoo and Wildlife Medicine 23, 205-210.
Abstract: Normal cardiopulmonary data in seven African (Loxodonta africana) and eight Asian (Elephas maximus) elephants were documented in conscious animals standing and in left lateral (LL) recumbency.  In the standing position, arterial blood pressures, heart and respiratory rates, and blood gas values did not differ (P > 0.05) over time or between species. Systolic, diastolic, and blood pressure means (+/- SEM) were 178.6 (+/- 2.94), 118.7 (+/- 3.10), and 144.6 (+/- 2.90) mm Hg, respectively, in unsedated standing elephants.  Arterial blood pressures increased (P </= 0.05) with time in LL recumbency and were highest (179.83 +/- 9.32) by the last reading at 16.5 +/- 0.41 min.  Arterial PO2 values decreased (P </= 0.05) from 96.2 (+/- 1.55) mm Hg while standing to 83.8 (+/- 3.37) mm Hg by 13.6 (+/- 6.8) min in LL recumbency.  Lateral recumbency increased (P < 0.05) arterial pH, adjusted base excess, and HCO3- content; however, these changes were not considered clinically significant.  Clinically healthy unsedated laterally recumbent elephants may be at risk of developing clinically significant hypoxemia and hypertension in the absence of alteration in more readily measured cardiopulmonary parameters.

Kalanidhi, A.P., Nagaish, K., Palanissamy, R., Srinivasan, V.A., 1992. Screening of Indian elephants, cattle and sheep for antibodies to foot and mouth disease virus-infection associated antigen. Indian Veterinary Journal 69, 390-393.
Abstract: All of the 24 serum samples taken from Indian elephants in a wildlife sanctuary in Theppakadu were negative in the double immunodiffusion test for antibodies to virus infection associated antigen (VIA) and showed insignificant titres in serum neutralization tests. Antibodies to VIA were detected in the serum of vaccinated as well as in foot and mouth disease-infected cattle and sheep sampled from the Nilgiris District of Tamil Nadu; 13 of 21 cattle and 5 of 6 sheep were positive.

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.

Schroder, H.D., Fischer, M., Ippen, R.ed. Contribution to the occurrence of infection of zoo mammals with influenzavirus type A. Erkrankungen der Zootiere. Verhandlungsbericht des 34. Internationalen Symposiums uber die Erkrankungen der Zoo- und Wildtiere, Santander-Spain 1992.  119-125. 1992.
Ref Type: Conference Proceeding
Abstract: Serum samples from 65 of 203 mammals possessed antibodies to 7 type A influenza viruses in HI titres between 1:8 and 1:1024, comprising 5 of 19 carnivores, 4 of 10 elephants, 10 of 26 Perissodactyla and 46 of 137 Artiodactyla [details tabulated]. Titres were highest against A/Philippines/2/82 (H3N2).

Sreekumar, K.P., Nirmalan, G., 1992. Normal values for certain serum enzymes of clinical value in Indian elephants. Veterinary Research Communications 16, 411-414.
Abstract: Blood samples were collected in the morning before feeding from clinically healthy baby elephants (5), adult males (10) and adult non-pregnant non-lactating females (13) from the ear vein. Aspartate amino transferase levels were 10.2 ± 3.8 u/litre, 15.7 ± 2.7 and 18.5 ± 2.4 in baby elephants, males and females respectively. Alanine aminotransferase levels were 5.6 ± 0.9, 4.8 ± 0.5 and 5.0 ± 0.5, lactic dehydrogenase levels were 366.7 ± 90.0, 468.8 ± 55.1 and 398.8 ± 78.0, and creatine kinase levels were 30.3 ± 12.0, 51.2 ± 7.4 and 43.8 ± 12. No influence of age or sex could be detected for any of the parameters studied.

Sreekumar, K.P., Nirmalan, G., 1992. Fatty acid composition of plasma lipids in the Indian elephant (Elephas maximus). 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. 51-53.

Wemmer, C., Krishnamurthy, V., 1992. Methods for taking standard measurements of live domestic 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. 34-37.

Yathiraj, S., Choudhuri, P.C., Rao, D.S.T., Reddy, P.K., 1992. Clinico-haematological observations on Indian elephant (Elephas maximus indicus). Indian Veterinary Journal 69, 995-997.
Abstract: In 3 apparently healthy elephants (a male aged 40, and 2 females aged 20 and 60) the mean values for heart rate, respiratory rate and body temperature, respectively, were 34.66±1.08/min, 7.88±0.09/min and 35.25±0.07°C in the mornings, and 36.22±1.07/min, 8.33±0.15/min and 35.75±0.06°C in the afternoons. Haemoglobin values averaged 11.65±0.49 g%, and PCV 33.25±0.46%. Various erythrocyte and leukocyte counts and indices are presented.

Hattingh, J., Pitts, N.I., De-Vos, N.I., Moyes, D.G., Ganhao, M.F., 1991. The response of animals to suxamethonium (succinyldicholine) and succinylmonocholine. Journal of the South African Veterinary Association 62 , 126-129.
Abstract: The time which elapses before cessation of breathing, and blood pressure and blood gas changes after the i.m. administration of suxamethonium, or a mixture of suxamethonium and hexamethonium, was compared in immobilized African elephants (Loxodonta africana) and buffaloes (Synceros caffer). In addition, the respiratory responses of elephants and other animals to i.v. administration of suxamethonium and succinylmonocholine are reported, as are the effects of darting animals with succinylmonocholine. Respiration was affected in a similar fashion in all species investigated. However, the characteristic gradual decrease in respiratory rate seen in elephants during culling, using suxamethonium, resembles the effects observed when succinylmonocholine is administered. It is suggested that elephants are killed by this first breakdown product of suxamethonium during culling and/or that unique acetylcholine receptors may be involved.

Loypetjra, P., 1991. Electrocardiography of the wildlife animals. Thai Journal of Veterinary Medicine 21, 175-186.
Abstract: The electrocardiogram of wildlife animals was recorded using hexaxial lead system.  The animals were seventeen crocodiles, one gibbon, three lorises, three tigers, four elephants and one binturong.  All of them were conscious during the measurement.      The Lead II of electrocardiogram was used in evaluating heart rate, rhythm and measuring the amplitude, time interval and segment.  Standard limb leads were employed to calculate mean electrical axis of the ventricles.      Electrocardiographic features of P, QRS and T waves in all species were normal without slurring or notching.  The values of heart rate per minute of crocodile, gibbon, loris, tiger, elephant, and binturong were 43-65, 166, 125-214, 90-154, 35-49 and 150, respectively.  The duration of P wave in crocodile was between 0.051-0.179 sec, QRS wave was 0.108-0.158 sec, P-R interval was between 0.24-0.42 sec, and Q-T interval was between 0.282-0.454 sec.  P wave duration in gibbon, loris, tiger, elephant and binturong were nearly the same which were 0.04 to 0.08 sec.  The mean electrical axes were between 66 deg-72 deg, 57 deg, 110 deg, 70 deg-85 deg, 40 deg-90 deg and 120 deg in crocodile, gibbon, loris, tiger, elephant and binturong, respectively.      The large variation of ECG values within species was thought to be the unrestrained and exciting effects.  However, this was considered to be normal and could be found in each species.

Fithian, C.L., Briggs, M.B. Leukocyte counts in African elephants. AAZPA Reg.Conf.Proc.  189-193. 1990.
Ref Type: Conference Proceeding

George, S., Nirmalan, G., 1990. Kinetic parameters of lactate dehydrogenase in the Indian elephant. Indian Journal of Animal Science 6 , 25-28.

Haagsma, J., Eger, A. ELISA for diagnosis of tuberculosis and chemotherapy in zoo and wildlife animals. Proceedings of American Association of Zoo Veterinarians , 107-110. 1990.
Ref Type: Conference Proceeding
Abstract: The aim of this study was to improve the diagnosis of bovine tuberculosis in zoo and wildlife animals, in particular by using an Enzyme-Linked Immunosorbent Assay (ELISA). In addition, suspected cases of tuberculosis (TB) with a positive skin test and /or ELISA were treated with antituberculosis drugs. The diagnosis of TB in animals is based primarily on the intradermal tuberculin test, corresponding with cellular immune response. Although this test has practical disadvantages in zoo animals, the application is still of high value. For this purpose tuberculins with a well controlled high potency and specificity should be used. In order to diagnose hypergic or anergic animals it is recommended to use PPD tuberculin with double strength (2 mg tuberculoprotein per ml) or to double the dose (0.2 ml instead of 0.1 ml), so that about 10,000 I.U. are applied. A strict interpretation scheme can increase the efficacy of the test, in particular in the comparative test. In order to improve the diagnosis, we have studied for some years the use of the ELISA which corresponds with humoral immunity.

Hattingh, J., Pitts, N.I., Ganhao, M.F., Moyes, D.G., de Vos, V., 1990. Blood constituent responses of animals culled with succinyldicholine and hexamethonium. Journal of the South African Veterinary Medical Association 61, 117-118.
Abstract: Blood constituent responses of elephants and buffaloes culled in the Kruger National Park, using a mixture of succinyldicholine and hexamethonium, were compared to those of animals culled with succinyldicholine only. The results show a decreased physiological response in the animals culled with the mixture, characterized by lower total catecholamine, cortisol and glucose concentrations. Neither a delay of up to 30 min in obtaining blood samples from culled animals, nor a delay of up to 30 min in processing samples obtained immediately after cessation of respiration, gave any significant difference in the blood constituents which were measured.

Hill, F.W.G., Smith, D.A., 1990. Clinical chemistry values for free-ranging elephants (Loxodonta africana) in Hwange National Park, Zimbabwe. Zimbabwe Veterinary Journal 21,  33-42.
Abstract: Data are given for urea, cholesterol, creatinine, calcium, inorganic phosphorus, magnesium, albumin and total proteins in blood samples collected from 92 wild elephants, of which 42 were males and 50 were females. Elephants were grouped by age, from 0-4 years, 5-10, 11-20 and >21 years. Ca, Mg, creatinine, proteins and albumin were not affected by age or sex. P, cholesterol and urea values were highest in the youngest group of elephants.

Mehrotra, M.L., Shukla, D.C., 1990. Seroprevalence, diagnosis and differential diagnosis of bluetongue virus disease in India. Indian Journal of Virology 6, 98-103.
Abstract: A serological survey, using the agar gel precipitation test, for bluetongue virus disease (BT) in sheep (mainly) and other domestic and wild animals was carried out in 11 states of India. Antibodies against the virus were detected in 8 states among sheep, buffalo and baby elephant. About 25% of the sera were positive to the test. The disease was diagnosed by isolation and identification of the virus from clinical cases. The BHK21 cell line was used for the isolation of virus. Viruses indistinguishable from BT virus were isolated from 4 states including Madhya Pradesh, Maharashtra, Kashmir, Uttar Pradesh. The disease was differentiated from rinderpest and foot-and-mouth disease.

Metzler, A.E., Ossent, P., Guscetti, F., Rubel, A., Lang, E.M., 1990. Serological evidence of herpesvirus infection in captive Asian elephants (Elephas maximus). Journal of Wildlife Diseases 26, 41-49.
Abstract: In mid 1988 a 3-yr-old Asian elephant (Elephas maximus) from a circus in Switzerland died following generalized manifestation of a herpesvirus infection. In an effort to determine prevalence of infection with the herpesvirus, and due to lack of a corresponding virus isolate, it was decided to evaluate contact animals and elephants from a second herd for antibody to bovine herpesvirus 1 (BHV1) and bovine herpesvirus 2 (BHV2). Of 15 sera tested four displayed low neutralizing antibody titers to BHV2. None of the sera neutralized BHV1. However, as evidenced by protein A-mediated immunoprecipitation of metabolically radio- labeled virus-infected and mock-infected cell antigens, followed by separation of precipitation products in SDS-polyacrylamide gels, the 15 sera precipitated multiple antigens from both viruses. Similar results were obtained when using BHV4 antigens. The extent of reaction was most distinct with respect to BHV2 antigens, less prominent with BHV1 antigens, and least with BHV4 antigens. The respective protein patterns, although less marked, matched well with those obtained with bovine reference sera. Additional evaluation of sera from six elephants from two zoos in the Federal Republic of Germany gave essentially identical results. It was concluded that at least one herpesvirus, immunologically related to BHV2, may be widely distributed among captive Asian elephants, and that this virus apparently does not cause overt disease in the majority of animals

Mushi, E.Z., Hill, F.W.G., Dawe, P., Riess, R., 1990. Antibodies to bluetongue and African horse sickness viruses in the sera of elephants in Zimbabwe. Bulletin of Animal Health and Production in Africa 38, 475.
Abstract: Of 92 elephants [Loxodonta africana] culled in the Hwange and Gonarezhou National Parks in 1985, 13 had low CF titres (1:4-1:16) to bluetongue virus and 74 had CF titres of 1:4-1:18 to African horse sickness virus.

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.

Ratnasooriya, W.D., Fernando, S.B.U., Manatunga, A.M.V.R., Caldera, H.S., Liyanage, G.K., Premakumara, G.A.S., 1990. Haematological values for adult Asian elephants (Elephas maximus maximus) at the Pinnawala Elephant Orphanage, Sri Lanka. Med. Sci. Res. 19, 899-902.

Sreekumar, K.P., Nirmalan, G., 1990. The fatty acid composition of plasma lipids in the Indian elephant (Elephas maximus indicus). Veterinary Research Communications 14, 427-431.
Abstract: Blood samples were taken from 9 adult Indian elephants of both sexes and ranging from 21-56 years of age. None of the females were pregnant or lactating. A table is given of the percentage distribution of the individual fatty acids in the plasma lipids of the animals. The relative distribution varied from animal to animal and some of the fatty acids were only present in traces or were absent in some animals. The percentage of unsaturated fatty acids (35-69%) was significantly greater than that of the saturated fatty acids. (25-39%). The sex of the animal was not found to have any effect. The proportion of mono-unsaturated fatty acids ranged from 11.76 to 23.17% while that of the poly-unsaturated fatty acids ranged from 22.03% to 52.72%.

Sreekumar, K.P., Nirmalan, G., 1990. Certain physical characteristics of the blood of Indian elephants (Elephas maximus indicus). Indian Journal of Animal Science 60, 1061-1064.

Stephanos, J.J., Addison, A.W., 1990. Spectroscopic and kinetic aspects of Elephas maximus hemoglobin. Eur. J Biochem. 189, 185-191.
Abstract: In comparison with myoglobin and human and Glycera dibranchiata hemoglobins, the heme distal side amino acid exchanges within the heme environment of elephant tetrameric hemoglobin (Hbe) only slightly affect the electronic and ESR spectra of Hbe(III) and Hbe(II) derivatives, several of which were prepared and characterized by optical and ESR spectroscopy. Addition of 2,3- bisphosphoglycerate or inositol hexakisphosphate to Hbe(II)NO causes tension in the Fe-N(proximal His) bond, although the behaviour differs in detail from that of HbA(II)NO. There are two equilibrium states of Hbe having significantly different kinetics for the Hbe(III)----Hbe(II) reaction of Hbe(III)NO. This autoreduction occurs in the form of two parallel processes, which collapse into one intermediate rate in the presence of Gri(2, 3)P2. The temperature dependences of the rates enable deduction of delta H0 and delta S0 for the linked equilibrium, and yield linear Eyring plots for Hbe(III)NO, from which activation parameters were estimated on the basis of a previously described mechanism

 1989. Normal physiological data. International Species Information System, Apple Valley, MN.

Gentry, P.A., Niemuller, C., Ross, M.L., Liptrap, R.M., 1989. Platelet aggregation in the Asian elephant is not dependent on thromboxane B₂ production. Comparative Biochemistry and Physiology [A] 94, 47-51.
Abstract: 1. The platelet aggregation response to several known platelet agonists was evaluated in four Asian elephants. The platelets were highly responsive to stimulation with platelet-activating factor (PAF) and collagen, less responsive to adenosine diphosphate (ADP) and non-responsive to arachidonic acid, serotonin and epinephrine. 2. Arachidonic acid (1 x 10(-4) M), while inducing no aggregation, caused the release of 1248 +/- 1147 pg/ul (mean +/- SD) of thromboxane B2 (TXB2), the stable metabolite of thromboxane A2 from stimulated platelet. The addition of 1 x 10(-4) M ADP to platelets caused suboptimal aggregation and the release of only 25 +/- 10 pg TXB2/microliters. 3. The calcium channel blocker, verapamil, produced a dose-dependent inhibition of PAF-induced but not collagen-induced aggregation. The cyclooxygenase inhibitor, acetylsalicylic acid, produced no inhibition of either collagen- or PAF-induced aggregation

Pattnaik, B., Venkataramanan, R., 1989. Detection of virus-infection-associated (VIA) antibody in serum of animals susceptible to foot-and-mouth-disease virus. Indian Journal of Animal Sciences 59, 356-357.
Abstract: Foot and mouth disease virus (FMDV) infection in tissue culture and in animals lead to the production of a small heat-labile virus infection associated (VIA) antigen. Sera from two elephants tested, were positive for VIA antibodies in the double-immunodiffusion test. Of 180 cattle serum samples from animals without known vaccination history, 64 were positive for VIA antibody. Of the VIA negative 59 cattle all were negative for FMDV neutralizing antibody. Virus carriers can be detected within regularly vaccinated herds by monitoring the VIA antibody.

Sreekumar, K.P., Nirmalan, G., 1989. Serum proteins in Indian elephants. Kerala Journal of Veterinary Science 20, 88-93.

Sreekumar, K.P., Nirmalan, G., 1989. Mineral status in the blood of Indian elephants. Indian Journal of Animal Sciences 59, 1253-1258.
Abstract: Sodium, potassium, calcium, magnesium, iron, copper and zinc have been estimated in the sera of Indian elephants (babies, tuskers and adult females) and the iron:copper ratio calculated. But for the higher level of calcium in baby elephants compared to that in adult females, no influence of age or sex could be detected in the distribution pattern of major elements. Baby elephants had a lower level of copper than adult females. The iron:copper ratio was higher in baby elephants than in tuskers.

Wissdorf, H., Poulsen-Naatrup, C., 1989. Presentation of the Vena caudalis centralis, a further important anatomical structure for venipuncture in Asian elephant situated dorsally in the tail. Anatomia Histologia Embryologia. 13, 279-280.

Gromadzka-Ostrowska, J., Jakubow, K., Zalewska, B., Krzywicki, Z., 1988. Haematological and blood biochemical studies in female domesticated Indian elephants (Elaphas maximus L.). Comparative Biochemistry and Physiology [A] 89, 313-315.
Abstract: 1. Some haematological and biochemical blood parameters in female Indian elephants were investigated. 2. Haematological data were as follows: Ht = 39.2 +/- 2.36%, Hb = 10.1 +/- 0.54 g%, RBC = 2.66 +/- 0.32 x 10(6)/mm3, WBC = 5.43 +/- 0.48 x 10(3)/mm3. Lymphocytes, determined on blood smears were mainly leucocytes from (67.0 +/- 1.59%). Data for MCV, MCH and MCHC are also given. 3. Blood plasma was separated into 5 main fractions, total plasma protein concentration was 6.98 +/- 0.53 g%, A/G ratio was 0.69 +/- 0.1. 4. Plasma minerals concentration was as follows: Na, 3044 +/- 194 micrograms/ml; K, 529 +/- 38.5 micrograms/ml; Mg, 33.0 +/- 3.43 micrograms/ml; Ca, 181.0 +/- 17.8 micrograms/ml; InP, 44.6 +/- 6.1 micrograms/ml. Ca: P ratio was 3.25 +/- 0.34. 5. Some seasonal differences in investigated parameters were observed. Ht values, WBC and neutrophils number as well as Ca, and Mg concentrations were higher during winter, whereas RBC and Na and InP concentrations were lower in winter.

Mihm, F.G., Machado, C., Snyder, R., 1988. Pulse oximetry and end-tidal CO₂ monitoring of an adult Asian elephant. Journal of Zoo and Wildlife Medicine 19, 106-109.
Abstract: The adequacy of ventilation during etorphine anesthesia of a 20-yr-old Asian elephant (Elephas maximus) was monitored with a pulse oximeter to measure arterial hemoglobin oxygen saturation (SaO₂) and a CO₂ analyzer to measure end-tidal CO₂ concentrations (PetCO₂).  Immediately after the first anesthetic induction, SaO₂ values of 45% were noted while the animal was breathing room air at a rate of 6/min.  The SaO₂ readings increased to 93% 15 min after administration of 5 liters/min of oxygen via the trunk.  Seven arterial blood gas samples obtained during two anesthetics, and once while unanesthetized, provided PaO₂ and PaCO₂ values which compared favorably with SaO₂ and PetCO₂.  In the anesthetized animal, PaO₂ ranged between 31 and 70 mmHg while SaO₂ values were 70-95%.  At the same time, measurements of PaCO₂ ranged from 42 to 57 mmHg while values of PetCO₂ ranged from 35 to 57 mmHg.  Pulse oximetry and end-tidal CO₂ monitoring are easy to apply and should increase the safety of anesthesia for these animals.

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.

Morris, P.J., Held, J.P., Jensen, J.M. Clinical pathologic features of chronic renal failure in an African elephant (Loxodonta africana). Proc.1st.Intl.Conf.Zool.Avian Med.  468-472. 1987.
Ref Type: Conference Proceeding

Sharma, V.S., Traylor, T.G., Gardiner, R., Mizukami, H., 1987. Reaction of nitric oxide with heme proteins and model compounds of hemoglobin. Biochemistry 26, 3837-3843.
Abstract: Rates for the reaction of nitric oxide with several ferric heme proteins and model compounds have been measured. The NO combination rates are markedly affected by the presence or absence of distal histidine. Elephant myoglobin in which the E7 distal histidine has been replaced by glutamine reacts with NO 500-1000 times faster than do the native hemoglobins or myoglobins. By contrast, there is no difference in the CO combination rate constants of sperm whale and elephant myoglobins. Studies on ferric model compounds for the R and T states of hemoglobin indicate that their NO combination rate constants are similar to those observed for the combination of CO with the corresponding ferro derivatives. The last observation suggests that the presence of an axial water molecule at the ligand binding site of ferric hemoglobin A prevents it from exhibiting significant cooperativity in its reactions with NO.

Sreekumar, K.P., Nirmalan, G., 1987. Studies on the haemoglobin and albumin polymorphism in Indian elephants (Elephas maximus indicus). Kerala Journal of Veterinary Science 18, 69-72.

Brown, I.R.R., 1986. More on the relation between magnesium and calcium in serum and plasma. Clinical Chemistry 32, 221.

Kleinschmidt, T., Czelusniak, J., Goodman, M., Braunitzer, G., 1986. Paenungulata: a comparison of the hemoglobin sequences from elephant, hyrax, and manatee. Molecular Biology and Evolution 3, 427-435.
Abstract: Inspection of the amino acid differences among hemoglobin sequences of a wide range of mammalian species suggested that at alpha 19, alpha 110, alpha 111, beta 23, beta 44, and beta 56, synapomorphies group manatee (Trichechus inungius, Sirenia), Indian and African elephant (Elephas maximus and Loxodonta africana, Proboscidea), and rock hyrax (Procavia habessinica, Hyracoidea) into a monophyletic clade. Results obtained by parsimony analysis provide evidence for this grouping--and thus support for the genealogical validity of Simpson's superorder Paenungulata, which contains as the extant orders Proboscidea, Sirenia, and Hyracoidea. All of the 39 most, or nearly most, parsimonious of 10,395 trees constructed from a tandemly combined alpha- and beta-hemoglobin sequence for 103 vertebrate species (of which 79 were mammals from 16 extant orders), depicted Paenungulata as one of the most anciently separated branches of Eutheria. It was found on examining thousands of alternative trees that to not group Proboscidea, Hyracoidea, and Sirenia in a monophyletic clade required at least four additional substitutions.

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.

Yu, N.T., Thompson, H.M., Mizukami, H., Gersonde, K., 1986. The cobalt-nitrosyl stretching vibration as a sensitive resonance Raman probe for distal histidine-nitrosyl interaction in monomeric hemoglobins. Eur. J Biochem. 159, 129-132.
Abstract: The Co-NO stretching vibration has been assigned in the resonance Raman spectra of various cobalt-substituted monomeric hemoglobins by employing isotope-labeling of nitrosyl (14N16O, 15N16O, 14N18O). Monomeric hemoglobins with a distal histidine (sperm whale myoglobin and leghemoglobin) exhibit this vibration at 573-575 cm-1, whereas hemoglobins without distal histidine (elephant myoglobin and insect hemoglobin from Chironomus thummi thummi, CTT III) show this vibration in the range of 553-558 cm- 1. The Fe-NO stretching vibration which occurs in the range of 554-556 cm-1 does not reflect the distal histidine-ligand interaction. Therefore, the Co-NO moiety which is isoelectronic with the Fe-O2 moiety is a good monitor for distal effects on the exogenous ligand of hemoglobins, especially due to the fact that in hemoglobins with distal histidine the Fe-O2 stretching vibration (567-572 cm-1) is similar to the Co-NO stretching vibration

Allen, J.L., Jacobson, E.R., Harvey, J.W., Boyce, W., 1985. Hematologic and serum chemical values for young African elephants (Loxodonta africana) with variations for sex and age. Journal of Zoo and Wildlife Medicine 16, 98-101.

Kerr, E.A., Yu, N.T., Bartnicki, D.E., Mizukami, H., 1985. Resonance raman studies of CO and O₂ binding to elephant myoglobin (distal His(E7)----Gln). Journal of Biological Chemistry 260, 8360-8365.
Abstract: Carbon monoxide and dioxygen were employed as resonance Raman- visible ligands for probing the nature of the heme-binding site in elephant myoglobin, which has glutamine in the distal position (E7) instead of the usual histidine. The distal histidine (E7) residue has been thought to be responsible for weakening carbon monoxide binding to hemoproteins. It is of interest to see how the His(E7)----Gln replacement affects such parameters as nu(Fe-N epsilon), nu(Fe-CO), delta(Fe-C-O), nu(C-O), delta(Fe-O-O), and nu(O-O) vibrational frequencies and relative intensities. Elephant myoglobin has a CO affinity approximately 6 times higher than that for human/sperm whale myoglobin (Mb). If this enhanced affinity were solely due to the removal of some of the steric hindrance that normally tilts the CO off the heme axis, one would expect the nu(Fe-CO) frequency to decrease and the nu(C-O) frequency to increase relative to the corresponding values in sperm whale Mb. However, the opposite was found. In addition, strong enhancement of the Fe-C-O bending mode was observed. These results suggest that the Fe-C-O linkage remains distorted. In elephant Mb, new interactions resulting from the conformational change accompanying ligand binding may be responsible for the increased CO binding. Similar spectra were obtained for elephant and sperm whale oxymyoglobin. This suggests that the interactions of bound O2 are not markedly affected by the glutamine replacement

Braunitzer, G., Stangl, A., Schrank, B., Krombach, C., Weisner, H., 1984. Phosphate-haemoglobin interaction. The primary structure of the haemoglobin of the African elephant (Loxodonta africana, Proboscidea): asparagine in position 2 of the beta-chain. Hoppe-Seyler's Z. Physiol. Chem. 365, 743-749.
Abstract: The primary structure of the haemoglobin of the African Elephant (Loxodonta africana) is reported.  The sequence was determined by means of a sequenator.  The haemoglobin differs in 26 amino acids in the alpha-chains in and 27 in the beta-chains from that of adult human hemoglobin.  The haemoglobin of the African Elephant, like that of the Indian Elephant and the llama, has only 5 binding sites for polyphosphate.  This finding explains the low p(O₂)₅₀ value in whole blood as a result of the lower 2,3-bisphosphoglycerate-haemoglobin interaction.  This is discussed in relation to aspects of respiratory physiology; some points are also of interest with regard to the Second Punic War and Hannibal's crossing of the Alps.

Chatterjee, A., 1984. Association of a Stephanofilaria indistinguishable form S. assamensis with lesions on the feet of Indian elephant (Elephas maximus). Indian Journal of Animal Health 23, 29-35.

Hattingh, J., 1984. Effects of etorphine and succinyldicholine on blood composition in elephant and buffalo. South African Journal of Zoology 19, 286-290.

Hattingh, J., Wright, P.G., de Vos, V., McNairn, I.S., Ganhao, M.F., Silove, M., Wolverson, G., Cornelius, S.T., 1984. Blood composition in culled elephants and buffaloes. Journal of the South African Veterinary Medical Association 55, 157-164.
Abstract: Blood composition of succinyldicholine culled elephants and buffaloes was compared with that of undisturbed animals shot in the brain. The results show statistically significant differences in a number of variables including plasma ACTH and cortisol concentrations. The observed changes are attributed to stress induced by a combination of herding and darting with succinyldicholine and asphyxia. Extrapolation from blood oxygen tensions suggests that this stress may be perceived for an undetermined period which is probably longer in elephants than buffaloes.

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.

Jacobson, E.R., Heard, D.J., Brock, K.A. Effects of oxygen supplementation on blood gas values in African elephants. Proc. Amer. Assoc. Zoo Vet.  60. 1984.
Ref Type: Conference Proceeding

Kleinschmidt, T., Braunitzer, G., 1983. The primary structure of hemoglobins of the rock hyrax (Procavia habessinica, Hyracooidea): insertion of glutamine in the alpha chains. Hoppe. Seylers. Z. Physiol. Chem. 364, 1303-1313.
Abstract: The chromatography of the hemoglobin of the rock hyrax (Procavia habessinica) gives two components (73% HbI and 27% HbII). The amino-acid analysis and the sequences of the globin chains elucidated with the phenylthiohydantoin method, did not show any differences between the alpha I and alpha II or beta I and beta II chains, respectively. The different chromatographical behaviour cannot be explained. After chain separation by chromatography on CM-52 cellulose, all four primary structures were elucidated automatically in a sequenator on the chains and the tryptic peptides. In 20% of the beta I chains the N-terminal valine was blocked by acetyl. The alignment was performed by homology with the chains of human adult hemoglobin. The alpha chain of the rock hyrax has 142 amino-acid residues, i.e. one residue more than normal mammalian alpha chains, caused by an insertion of glutamine in the GH region supposed between positions 115 and 116. A comparison of human and hyrax hemoglobins shows an exchange of 21 amino-acid residues in the alpha chains and of 24 in the beta chains. Some substitutions in alpha 1 beta 1 contacts and in the surrounding of the heme are not supposed to effect the function of the hemoglobin. The phylogenetic relationship between the rock hyrax and the Indian elephant (Elephas maximus) on the one hand and with some Perissodactyla on the other, is discussed. Up to now the exchanges of alpha 110(G17)Ala leads to Ser and beta 56(D7)Gly leads to His have only been found in hyrax and elephant. This indicates a certain relationship between Hyracoidea and Proboscidea.

Pichaicharnarong, A., Loypetjra, P., Chaiyabutr, N., Komonmena, V., Usanakornkul, S., Boonnamsiri, V., Devakul, C., 1983. The thyroid activity of Asiatic elephants in Thailand. Acta Veterinaria (Beograd) 33, 73-80.
Abstract: Values of serum T3 and T4 were determined by radioimmunoassay in 58 normal Asiatic elephants (Elephas maximus, Linnaeus 1758) with ages ranging from 1 to 80 years. The mean concentrations of T4 and T3 were 113.6 ± 27.0 umol/l and 1.8 ± 0.7 umol/l respectively.  The RT3U values (resin triiodothyronine125 uptake) were assessed in 65 elephants.  The mean was 30.1 ± 3.8%.  Both serum T4 and T3 decreased with age.  The RT3U of 1-10 year old elephants was higher than that of older elephants, but there was not statistical difference between any age range.  Details of the effects of sex and age of the Asiatic elephant on the values of T3 (RIA), T4 (RIA) and RT3U are discussed.

Braunitzer, G., Jelkmann, W., Stangl, A., Schrank, B., Krombach, C., 1982. Hemaglobins, XLVIII: the primary structure of hemoglobin of the Indian elephant (Elephas maximus, Proboscidae): beta 2 = Asn. Hoppe. Seylers. Z. Physiol. Chem. 363, 683-691.
Abstract: The primary structure of the hemoglobin of the Indian Elephant (Elephas maximus) is given. The sequence was determined automatically in a sequenator. By homologous comparison with adult human HbA, the alpha-chains differ by 24 exchanges and the beta-chains by 27 exchanges. Furthermore, we report p(O2)50 values with regard to altered contact sites with 2,3- bisphosphoglycerate in Indian elephant hemoglobin. Our findings explain the low p(O2)50 and the reduced interaction with 2,3- bisphosphoglycerate. Elephant hemoglobin has, like that of the Llama, only five phosphate binding sites. In addition, we have made an attempt to relate these results to aspects of respiratory physiology. Some implications of these biochemical and physiological results, concerning the Second Punic War and Hannibal's Alp transition, are given.

Sulochanna, S., Pillai, R.M., Abdulla, P.K., 1981. Erythrocyte rosette forming cells in the peripheral blood of Indian elephants. Ind. J. Comp. Micro, Imm, & Inf. Dis. 2, 42-44.

Brown, I.R.F., White, P.T., 1980. Elephant blood haematology and chemistry. Comparative Biochemistry and Physiology [B] 65, 1-12.

Hattingh, J., de Vos, V., Bomzon, L., Marcus, E., Jooste, C., Chertkow, S., 1980. Comparative physiology of colloid osmotic pressure. Comparative Biochemistry and Physiology [A] 67, 203-206.
Abstract: 1. Colloid osmotic pressure (C.O.P.) and other related parameters were measured in the serum of 20 species of wild animals.  2.  No significant statistical correlations could be found on an inter- or intraspecies basis between C.O.P. and albumin concentrations, total serum protein concentration and the A/G ratio and no theoretical or empirical formulae accurately predicted this value.  3.  The results are discussed in relation to which conponents in serum influence C.O.P. and it is stressed that this value can only be accurately determined by measurement.

White, P.T., 1980. Blood parasites in free-living African elephants. Zoologische Garten 50, 45-48.

Brown, I.R.F., White, P.T., 1979. Serum electrolytes, lipids, and cortisol in the African elephant, Loxodonta africana. Comparative Biochemistry and Physiology [A] 62, 899-901.
Abstract: 1. Serum electrolytes were measured in the African elephant from the game parks of Uganda.  The overall mean for sodium was 136.5 ± 5.6 mmol/l (n = 132) and for potassium 6.24 ± 0.86 mmol/l (n = 105).  There was, however, some evidence for a seasonal variation in these values.  2. Serum lipid levels were considerably lower in the elephant than in man.  The overall mean cholesterol was 1.58 ± 0.50 mmol/l (n = 84) and triglyceride 0.59 ± 0.29 mmol/l (n = 61).  Some geographical variation was observed in the triglyceride results.  3.  Serum cortisol levels were variable and ranged from 66 to 825 mmol/l in 33 elephants.

Gordon, D.H., Isaacson, M., Isaacson, M., 1979. Plague antibody in large African mammals. Infect Immun 26, 767-769.
Abstract: Plague hemagglutinating antibodies to a titer of 1:1,024 were demonstrated in 6.6% of buffalo and 0.3% of elephant sera tested 1 year after a plague epidemic in the same area.

Kuntze, A., Hunsdorff, P., 1979. Further haematological and biochemical findings (Ca, inorganic P, Mg, Na, K, Fe, glucose, enzymes) with reference to iron deficiency anaemia in young elephants (Elephas maximus)]. Erkrankungen der Zootiere 13, 147-150.
Abstract: Summary:  Regular haematological checks were applied to young elephants with anaemia to study the therapeutic effects of oral and parenteral iron medication.  Additional serum electrolyte values are reported, with some of them discussed.  They are relating to calcium, phosphorus, magnesium, sodium, potassium, glucose, SGOT, SGPT, and alkaline phosphatase

Nirmalan, G. Physiological peculiarties of elephants. State Level Workshop on Elephants.  21-24. 1979. India, College of Veterinary and Animal Sicences, Kerala Agricultural University.
Ref Type: Conference Proceeding

Upadhya, A.S., Krishnappa, G., Ahmed, S.N., Keshavamurthy, B.S., 1979. Serological evidence of leptospiral antibodies in elephants. Current Science 48, 733.

Woodford, M.H., 1979. Blood characteristics of the African elephant (Loxodonta africana cyclotis). Journal of Wildlife Diseases 15, 111-113.
Abstract: Mean corpuscular volume, hematocrit, hemoglobin concentration and red and white cell counts from five domesticated African elephants were measured.  The results obtained are tabulated and compared with those from shot and chemically immobilized African elephants.

Brown, I.R.F., White, P.T., Malpas, R.C., 1978. Proteins and other nitrogenous constituents in the blood serum of the African elephant, Loxodonta africana. Comparative Biochemistry and Physiology [A] 59, 267-270.
Abstract: 1.  Electrophoresis of blood serum of the African elephant separated 5 protein components designated albumin, alpha-, beta 1-, beta 2- and gamma-globulin.  Quantitation of these protein fractions showed that there were significant seasonal and geographic variations in the levels.  2.  Similar seasonal or geographic variations were found in the levels of urea, thyroxine, T₃ uptake and creatinine.  3.  The significance of these changes is discussed.

Caple, I.W., Jainudeen, M.R., Buick, T.D., Song, C.Y., 1978. Some clinicopathologic findings in elephants (Elephas maximus) infected with Fasciola jacksoni. Journal of Wildlife Diseases 14, 110-115.
Abstract: Severe submandidular and ventral abdominal edema was observed in an Asian elephant (Elephas maximus) in which liver flukes ( Fasciola jacksoni) were recovered from the bile ducts at post-mortem examination.  Clinico-pathologic examination of blood samples and serum from this elephant and another 8 elephants showed that most had anemia and hypoproteinemia. Fecal samples from 6 of the elephants contained from 6 to 83 eggs per gram.  Treatment of elephants with nitroxynil (10 mg/kg) by subcutaneous injection produced severe local reactions at the injection site.  Feces collected 2 and 4 months after treatment were free of trematode eggs.  Hematologic values measured 4 months after treatment showed that the hemoglobin concentration, packed cell volume, erythrocyte count and plasma protein concentration had increased to within the normal range.

Cmelik, S.H.W., Ley, H., 1978. Neutral lipids from the temporal gland of the African elephant (Loxodonta africana). Lipids 13, 195-198.

Eramus, B.J., Young, E., Pieterse, L.M., Boshoff, S.T., 1978. The susceptibility of zebra and elephants to African horsesickness virus. In: Bryans, J.T., Berger, H. (Eds.), Equine infectious diseases. Veterinary Publications, Princeton, N.J., pp. 409-413.

Kuntze, A., Hunsdorff, P., 1978. Haematological and biochemical findings (Ca, P, Mg , Fe, glucose, enzymes) in Asiatic female elephants. Erkrankungen der Zootiere 14, 309-313.
Abstract: Summary.  The hematological finding of ten clinically intact female elephants (Elephas maximus) are discussed, being compared with those of three newly imported young elephants (two years old) suffering from anemia caused by iron shortage.  Some serum electrolyte values (Ca, P, Mg, Fe, glucose, SGOT, SGPT, alkaline phosphatase) are stated.

White, P.T., Brown, I.R.F., 1978. Haematological studies on wild African elephants, Loxodonta africana. Journal of Zoology (Lond) 185, 491-503.

Brown, I.R.F., White, P.T., 1977. Serum calcium, magnesium, phosphorous and alkaline phosphatase in the African elephant, Loxodonta africana. Comparative Biochemistry and Physiology [B] 56, 159-162.
Abstract: 1. Calcium, magnesium, inorganic phosphorous and alkaline phosphatase have been measured in the serum of the African elephant.  2.  The overall mean serum calcium was found to be 11.23 mg/100 ml (range 9.4-13.1, N = 102), magnesium 4.41 mg/100 ml (range 2.6-6.2, N = 87) and phosphorous 8.62 mg/100 ml (range 3.9-13.3, N = 100).  Some seasonal and geographical variation was observed in the calcium and phosphorous results. 3.  Alkaline phosphatase levels were higher in young than in mature animals.  The values obtained were similar to those in humans.

Cmelik, S.H.W., Ley, H., 1977. A further contribution to the knowledge of the blood lipid fractions from the African elephant Loxodonta africana. Comparative Biochemistry and Physiology [B] 58, 205-209.
Abstract: 1. Plasma lipids from 5 African elephants were extracted and fractionated into cholesterol esters, free fatty acids, triglycerides, phosphatidylcholine, phosphatidylethanolamine, phophatidylinositol, sphingomyelin, and glycosphingolipids.  The fatty acids of various individual fractions were investigated by gas-chromatography.  2.  All animals, except one, had a high linoleic acid content in cholesterol esters indicating an adequate supply of linoleic acid in the diet.  3.  Phosphatidylcholine had a strong saturated character originating from the presence of unusually high quantities of stearic acid.  4.  Phosphatidylethanolamine was present in small quantities and was characterized by a low content of arachidonic acid.  5.  Sphingomyelin did not contain any long chain saturated acids.  Instead it contained 10.2-47.0% of a long chan acid which was most likely monounsaturated.  6. The presence of significant quantities of glycosphingolipids was established.

Davies, F.G., Otieno, S., 1977. Elephants and zebras as possible reservoir hosts for African horse sickness virus. Veterinary Record 100, 291-292.

Brown, I.R.F., White, P.T., 1976. Serum enzyme activities in the African elephant, Loxodonta africana. Experientia 32, 980-982.
Abstract: The serum activities of aspartate aminotransferase, alanine aminotransferase, alpha-hydroxybutyrate dehydrogenase and creatine phosphokinase have been measured in the African elephant.  In general, the values were broadly comparable with those of man except that alanine aminotransferase was much lower and creatine phosphokinase higher.  No variation due to age, sex, season or location was observed.

Skinner, R.K., Brown, I.R.F., White, P.T., 1976. Serum amylase in the African elephant, Loxodonta africana. Comparative Biochemistry and Physiology [B] 55, 363-366.
Abstract: 1.  The alpha-amylase activity in the serum of the African elephant has been measured by 2 different methods. Consistantly high values were obtained, about 10 times those of normal human serum measured by the same methods.  2. Electrophoretic separation of amylase isoenzymes revealed a singel band in gamma-globulin region and a group of 4 bands in the beta-globulin region.  3.  Both sets of findings are discussed in reation to results in other mammals.

Debbie, J.G., Clausen, B., 1975. Some haematological values of free-ranging African elephants. Journal of Wildlife Diseases 11, 79-82.
Abstract: Blood samples were collected from 23 free ranging African elephants (Loxodonta africana) in Tanzania.  Red and white blood cell counts, white blood cell differential, hemoglobin, blood copper, and blood inorganic phosphate were deterimined.  No correlation of any of these measurements with age and sex was discernible, except that the total white count was significantly higher in females than in males in all age groups.  These findings are compared with those previously reported for captive and free-ranging African elephants.

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

Riemann, G.P., Burridge, M.J., Behymer, D.E., Franti, C.E., 1975. Toxoplasma gondii antibodies in free-living African mammals. J Wildl Dis 11, 529-533.
Abstract: Twelve species of free-living African mammals from Kenya, Tanzania, Uganda and Zambia were tested for antibodies to Toxoplasma gondii using the indirect hemagglutination test. Of 157 animals sampled, 20 (13%) were seropositive. T. gondii antibodies were detected in Burchell's zebra, (Equus burchelli), hippopotamus (Hippopotamus amphibius), African elephant (Loxodonta africana), defassa waterbuck (Kobus defassa), lion (Panthera leo), and rock hyrax (Procavia capensis), The highest titers were found in elephants, two having titers of 1:4096 and one of 1:8192. These results are discussed in relation to the maintenance of T. gondii among African wildlife.

Schalm, O.W., Jain, N.C., Carroll, E.J., 1975. Normal values in blood of miscellaneous animals (Elephantidae). Veterinary Hematology. Lea and Febiger, Philadelphia, pp. 275-276.

Lewis, J.H., 1974. Comparative hematology: Studies on elephants, Elephas maximus. Comparative Biochemistry and Physiology [A] 49, 175-181.
Abstract: 1. Elephant blood clots rapidly and the clots retract only slightly.  The partial thromboplastin time is faster than human and most coagulation factors are much higher when assayed in systems designed for human plasma.  2.  Thrombin time was long but antithrombin III was essentially absent.  3. Platelet counts were in the high-normal human range and aggregation with ADP and collagen was less marked than in human platelet rich plasma.  4.  Red cells were large, discoid in shape, but fewer in number than in human blood.  5.  Leukocytes were present in greater numbers than in human blood and showed some morphological differences.  6.  Some biochemical and protein differences were also observed.

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.

Dhindsa, D.S., Sedgwick, C.J., Metcalfe, J., 1972. Comparative studies of the respiratory functions of mammalian blood.  VIII. Asian elephant (Elephas maximus) and African elephant (Loxodonta africana africana). Respiratory Physiology 14, 332-342.
Abstract: Respiratory characteristics of blood from four Asian and three African elephants were studied.  Oxygen dissociation curves of whole blood were constructed at 37 C and corrected to a plasma pH of 7.40.  The mean blood P50 values were 25.2 ± 0.5 and 23.2 ± 1.3 mm for Asian and African elephants, respectively, and these values are significantly different (p< 0.01).  The Bohr factors for both species were similar and averaged -0.351 ± 0.029 log PO2/ pH.  The Haldane effect was similar in both species (5.5 vol% C CO2 at PCO2 = 40 mmHg). The concentration of 2,3-diphosphoglycerate in elephant blood is similar to that found in normal human blood.  The blood morphology of both species was similar except that the leukocyte count was significantly higher in Asian elephants.  Starch gel electrophoresis showed that hemoglobin of Asian elephants travels at a slower rate than hemoglobin of African elephants, but both migrate faster than human A hemoglobin.

Albl, P., 1971. Studies on assessment of physical condition in African elephants. Biological Conservation 3, 134-140.
Abstract: Series of external measurements were taken from 240 carcasses of African Elephants during the dry season of 1967 in Zambia, in order to investigate fluctuations of subcutaneous fat and muscles.  In addition, the ratio of the weight of the kidneys to kidney-fat, and the contents of fat in the bone-marrow, were determined.  From these investigations are deduced and described simple criteria for assessment of the physical condition of African Elephants, which criteria allow objective classification of representative population samples. Extensive individual variations of external anatomical features complicate assessment of the condition.  Most of the investigated external physical features are more age- than nutrition-dependent.  Only the shape of the lumbar region and the kidney-fat index give a fairly reliable indication of the physical condition of the African elephant.

Basson, P.A., McCully, R.M., de Vos, V., Young, E., Kruger, S.P., 1971. Some parasitic and other natural diseases of the African elephant in the Kruger National Park. Onderstepoort Journal of Veterinary Research 38 , 239-254.

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., Jayasinghe, J.B., 1971. Hemogram of the domesticated Asiatic elephant (Elephas maximus). Journal of Zoo and Wildlife Medicine 2, 5-11.
Abstract: Baseline hematologic values were determined for 25 male and 17 female domesticated Asiatic elephants, Elephas maximus.  Age and sex-related influences on the different values are evaluated and found to be statistically not significant.  Mean values were as follows: erthrocytes 3.18 x 10⁶/cmm; hemoglobin 12.1 gm/100 ml; packed cell volume 33.7%; mean corpuscular volume 117.9 cµ; mean corpuscular hemoglobin 42.1 µµg; mean corpuscular hemaglobin concentration 36.1%; and leucocytes 14.7 x 10³/cmm.  Erythrocyte diameters ranged from 8.8 to 10.6 microns. The number of each type of leucocytes per 100 cells identified for the differential count was: lymphocytes 52.7; neutrophils 40.3; monocytes 5.3; eosinophils, 2.4; and basophils 0 to 2.  Sedimentation rate and total serum proteins were also determined.

Kalter, S.S., Heberling, R.L., Claussen, B., 1971. Antibody in wild animal (African) sera to human and simian viruses. Laboratory Animal Science 21, 829-831.

Nirmalan, G., Nair, S.G., 1971. Plasma proteins and certain non-protein nitrogenous constituents in the blood of Indian elephants (Elephas maximus). British Veterinary Journal 127, 207-213.
Abstract: An analysis of the blood of Indian elephants from plasma proteins and some of the non-protein nitrogenous constituents as influenced by age, sex and lactation revealed that elephant plasma has higher levels of total protein and globulins and a very low albumin/globulin ratio compared to that of other mammals.  The non-lactating adult females had higher levels of total protein and globulins in their plasma than baby elephants.  The blood of non-lactating adult females differed from that of lactating ones in being richer in plasma fibrinogen content and in having higher amounts of urea mitrogen and uric acid.  The non-lactating adult females also showed a higer level of blood urea nitrogen than tuskers.  The values for total non-protein nitrogen, urea nitrogen, uric acid, creatinine and free amino nitrogen in the blood of elephants were similar to those reported in the literature for other mammals.

Dellenback, R.J., Chien, S., 1970. The extinction coefficient of fibrinogen from man, dog, elephant, sheep and goat at 280 micrometers. Procedings of the Society for Experimental Biology and Medicine 134, 353-355.

Dillman, J.S., Carr, W.R., 1970. Observations on arteriosclerosis, serum cholesterol and serum electrolytes in the wild African elephant. Journal of Comparative Pathology 80, 81-87.

Gregersen, M.I., Usami, S., Bryant, C.A., Chien, S., Magazinovic, V., 1970. Rheological significance of species differences in erythrocyte deformability. Biorheology 6, 249.

Nirmalan, G., Nair, S.G., 1969. Biochemical studies on the blood of Indian elephants (Elephas maximus). Research in Veterinary Science 10, 176-180.

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.

Usami, S., Chien, S., Gregersen, M.I., 1969. Viscometric characteristics of blood of the elephant, man, dog, sheep and goat. American Journal of Physiology 217, 884-890.
Abstract: Viscosity (n) was determined on plasma, serum, and Ringer suspensions of erythrocytes from five mammalian species with different mean corpuscular volume (MCV): elephant 112 mu³, man 90 mu³, dog 74 mu³, sheep 37 mu³, and goat 18 mu³.  Samples with cell percentages (H) ranging from 5% to maximum packing were tested with a modified version of GDM viscometer at 37 C and shear rates (g) from 52 to 0.052 sec^-1.  In all species studied, plasma and serum behave as Newtonian fluids, but the n of erythrocyte suspensions increases with rising H and reduced shear rate.  The shear dependence in elephant, human and dog erythrocyte suspension is strikingly increased by the presence of serum proteins and further with the inclusion of fibrinogen.  These proteins induce the interaction between cells and plasma (e.g., aggregation).  With sheep and goat erythrocytes, which do not form significant aggregation even in the presence of proteins, the addition of serum proteins and fibrinogen does not increase the shear dependence of suspension viscosity.  In the absence of proteins, shear dependence in all species is not significantly increased until the direct interaction among cells (i.e., involving erythrocyte deformability) becomes enhanced with high H (above 65%).  The data suggest that the species difference in n is only partially due to the variations in MCV, but is more directly related to the interaction of cells with the suspending medium.

Nirmalan, G., Nair, S.G., 1968. A study of the free amino acids in the blood of Indian elephants (Elephas maximus) by paper partition chromatography. Ceylon Veterinary Journal 16, 16-18.

Moore, J.H., Sikes, S.K., 1967. The serum and adrenal lipids of the African elephant (Loxodonta africana). Comparative Biochemistry and Physiology [A] 20, 779-792.
Abstract: 1. The serum and adrenal lipids of the African elephant were fractionated by chromatography on columns of silicic acid into cholesterol esters, cholesterol, triglycerides, unesterified fatty acids and phospholipids.  The fatty acid compositions of the various lipid fractions were determined by gas-liquid chromatography.  2.  The results obtained from the African elephant were compared with the results reported in the literature for other species of mammals.  In many respects the composition of the serum lipids of the African elephant was similar to that of the rat and rabbit but was markedly different from that of the ox and man.  3.  Unlike the serum cholesterol esters and phospholipids of other animals, these two lipid fractions in the serum of these elephants contained appreciable concentrations of delta-8,11,14-eicosatrienoic acid.  4.  The total lipid content of the African elephant adrenal galnd was particularly high (63 per cent of the dry tissue).  Cholesterol esters accounted from almost half of the adrenal lipid.  Delta-8,11,14-eicosatrienoic acid was present in substantial amounts in the adrenal cholesterol esters and phospholipids.

Nirmalan, G., Nair, S.G., Simon, K.J., 1967. Hematology of the Indian elephant (Elephas maximus). Canadian Journal of Physiology and Pharmacology 45, 985-991.
Abstract: Blood samples from 11 baby elephants, 14 tuskers, and 16 females comprising 5 pregnant and 11 nonpregnant nonlactating elephants (Elephas maximus) were analyzed to assess the hematological values and the influence of age, sex and pregnancy on these. The results obtained show that, compared with other mammals, the erythrocyte count in the blood of elephants is low and the mean corpuscular volume (m.c.v.), mean corpuscular hemoglobin (m.c.h.), and erythrocyte sedimentation rate (e.s.r.) are high. The very high values for m.c.v. and m.c.h. and the low erythrocyte count seen in elephants suggest that in the evolution of blood through they are more advanced than reptiles and birds in having eliminated the nuclei from the erythrocytes, they are still in the primitive state compared with the other mammals inasmuch as their erythrocytes have not attained the efficiency in the transportation of blood gasses that results from a reduction in size to facilitate numerical increase.  A low erythrocyte count and packed cell volume, a high e.s.r., an increase in neutrophils, and a decrease in the percentage of lymphocytes and eosinophils characterize pregnancy.  In baby elephants there is an increase in the total leucocyte and lymphocyte counts and a decrease in eosinophils.

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.

Seneviratna, P., Hayasinghe, J.B., Jainudeen, M.R., 1967. Filariasis of elephants in Ceylon. Veterinary Record 81, 716-717.

Young, E., Lombard, C.O., 1967. Physiological values of the African elephant (Loxodonta africana). The Veterinarian 4, 169-172.
Abstract: The following physiological values of African elephants determined in the Kruger National Park are recorded in this article: Red cell count, hematocrit, mean corpuscular volume, mean corpuscular hemoglobin, mean corpuscular hemoglobin concentration, total hemoglobin, erythrocyte sedimentation rate, total white cell count, differential white cell count, blood urea nitrogen, body temperature, and pH of the blood plasma, saliva, urine and faeces.

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

Buettner-Janusch, J., Buettner-Janusch, V., Sale, J.B., 1964. Plasma proteins and haemoglobins of the African elephant and the hyrax. Nature 201, 510-511.

Schmitt, J., 1964. Haematological studies in elephants. Vet. Med. Rev. Leverkusen 2, 87-95.

Bartels, H., Hilpert, P., Barbey, K., Betke, K., Riegel, K., Lang, E.M., Metcalfe, J., 1963. Respiratory functions of blood of the yak, llama, camel, Dybowski deer and African elephant. American Journal of Physiology 205, 331-336.
Abstract: Blood samples from a yak, llama, camel, deer, and African elephant were analyzed for oxygen capacity, "standard bicarbonate" content, oxygen dissociation curve, and the magnitude of the Bohr and Haldane effects.  These parameters of the respiratory function of the blood have been related to the morphology of the red cells, to the weights of the animals, and to the important electrolytes in the erythrocytes and in the plasma.  The high affinity for oxgen described previously for llama blood is shared by its relative, the camel.  Both of these animals have a high concentration of hemoglobin within their erythrocytes.  Blood from the African elephant showed the greatest affinity for oxygen among the subjects studied.

Brocklesby, D.W., Campbell, H., 1963. A babesia in the African elephant. East African Wildlife Journal 1, 119.

Riggs, A., 1963. The amino acid composition of some mammalian hemoglobins: mouse, guinea pig and elephant. Journal of Biological Chemistry 238, 2983-2987.

Simon, K.J., 1961. Haematological studies on elephants. Indian Veterinary Journal 38, 241-245.

Kohira, E., 1960. The spleen of the Elephas indicus. Acta Anatomica Nipponica 35, 253-260.

Rao, S.R., Hiregaudar, L.S., 1954. Schistosomiasis in elephants in Bombay state. Indian Veterinary Journal 30, 241-242.

Weitz, B., 1953. Serological relationships of hyrax and elephant. Nature 171, 261.

Pfaff, G., 1940. Diseases of Elephants. Superintendent, Govt. Printing and Stationary, Burma, Rangoon.

Benedict, F.G., Lee, R.C., 1938. Further observations on the physiology of the elephant. Journal of Mammalogy 19, 175-194.

Benedict, F.G., 1936. The physiology of the elephant. Carnegie Institution of Washington, Washington.

Benedict, F.G., Fox, E.L., Baker, M.L., 1921. The skin temperature of pachyderms. Proceedings of the National Academy of Sciences of the United States of America 7, 154-156.

Gautrelet, J., Neuville, H., 1914. Sur le sang de mammouth. C. R. hebd. Seanc. Acad. Sci. ,Paris 158, 593-595.

Bruce, D., Hamerton, A.E., 1909. A note on the occurrence of a trypanosome in the African elephant. Proc. Roy. Soc. Lond. [B] Biol. Sci. 81, 414-416.

Schultze, C.H., 1839. Ueber das elephantenblut. Arch. Anat. Physiol. 252-254.

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