Elephant
Bibliographic
Database

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Elephant Bibliographic Database
www.elephantcare.org

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

Garner, M.M., Helmick, K., Ochsenreiter, J., Richman, L.K., Latimer, E., Wise, A.G., Maes, R.K., Kiupel, M., Nordhausen, R.W., Zong, J.C., Hayward, G.S., 2009. Clinico-pathologic features of fatal disease attributed to new variants of endotheliotropic herpesviruses in two Asian elephants (Elephas maximus)
119. Vet. Pathol. 46, 97-104.
Abstract: The first herpesviruses described in association with serious elephant disease were referred to as endotheliotropic herpesviruses (EEHV) because of their ability to infect capillary endothelial cells and cause potentially fatal disease. Two related viruses, EEHV1 and EEHV2, have been described based on genetic composition. This report describes the similarities and differences in clinicopathologic features of 2 cases of fatal endotheliotropic herpesvirus infections in Asian elephants caused by a previously unrecognized virus within the betaherpesvirus subfamily. EEHV3 is markedly divergent from the 2 previously studied fatal probosciviruses, based on polymerase chain reaction sequence analysis of 2 segments of the viral genome. In addition to ascites, widespread visceral edema, petechiae, and capillary damage previously reported, important findings with EEHV3 infection were the presence of grossly visible renal medullary hemorrhage, a tropism for larger veins and arteries in various tissues, relatively high density of renal herpetic inclusions, and involvement of the retinal vessels. These findings indicate a less selective organ tropism, and this may confer a higher degree of virulence for EEHV3

Miller, M.A., Olea-Popelka, F., 2009. Serum antibody titers following routine rabies vaccination in African elephants. Journal of the American Veterinary Medical Association 235, 978-981.
Abstract: Objective-To evaluate serum antibody titers in captive African elephants (Loxodonta africana) following routine vaccination with a commercially available, inactivated rabies vaccine. Design-Seroepidemiologic study. Animals-14 captive African elephants from a single herd. Procedures-Elephants were vaccinated as part of a routine preventive health program. Initially, elephants were vaccinated annually (2 mL, IM), and blood was collected every 4 or 6 months for measurement of rabies virus-neutralizing antibody titer by means of the rapid fluorescent focus inhibition test. Individual elephants were later switched to an intermittent vaccination schedule to allow duration of the antibody response to be determined. Results-All elephants had detectable antibody responses following rabies vaccination, although there was great variability among individual animals in regard to antibody titers, and antibody titers could be detected as long as 24 months after vaccine administration. Young animals were found to develop an antibody titer following administration of a single dose of the rabies vaccine. Age and time since vaccination had significant effects on measured antibody titers. Conclusions and Clinical Relevance-Results indicated that African elephants developed detectable antibody titers in response to inoculation with a standard large animal dose of a commercially available, inactivated rabies vaccine. The persistence of detectable antibody titers in some animals suggested that vaccination could be performed less frequently than once a year if antibody titers were routinely monitored

Parker, D.M., Bernard, R.T.F., 2009. Levels of aloe mortality with and without elephants in the Thicket Biome of South Africa. African Journal of Ecology 47, 246-251.
Abstract: Studies concerning the influence of African elephants (Loxodonta africana) on vegetation have produced contradictory results; some show minimal or no effect while others report significant elephant-induced effects. Elephants are generalist megaherbivores but will selectively feed from preferred plant species. We investigated the mortality of aloe plants (highly preferred food items for elephants) at five sites with elephants (treatment) and five paired sites without elephants (control) in the Eastern Cape Province of South Africa. A significantly higher proportion of aloes were dead at treatment sites and significantly more aloes that had lost their crown (headless) were found at treatment sites compared with controls. We conclude that although the proportions of dead aloes at treatment sites were significantly higher, it remains unclear whether there is a need to be concerned with the potential small-scale extinction of aloes from parts of the Eastern Cape Province. The observed mortality may merely be an artefact of the loss of large herbivores through disease (e.g. rinderpest) and hunting in the past.

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

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

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.

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

Zong, J.C., Latimer, E., Heaggans, S.Y., Richman, L.K., Hayward, G.S. Pathogenesis and molecular epidemiology of fatal elephant endotheliotropic disease associated with the expanding Proboscivirus genus of the betaherpesvirinae. Proceedings 2007 IEF Symposium.  23-35. 2008.
Ref Type: Conference Proceeding

Sinclair, A.R., Mduma, S.A., Hopcraft, J.G., Fryxell, J.M., Hilborn, R., Thirgood, S., 2007. Long-term ecosystem dynamics in the Serengeti: lessons for conservation. Conserv. Biol. 21, 580-590.
Abstract: Data from long-term ecological studies further understanding of ecosystem dynamics and can guide evidence-based management. In a quasi-natural experiment we examined long-term monitoring data on different components of the Serengeti-Mara Ecosystem to trace the effects of disturbances and thus to elucidate cause-and-effect connections between them. The long-term data illustrated the role of food limitation in population regulation in mammals, particularly in migratory wildebeest and nonmigratory buffalo. Predation limited populations of smaller resident ungulates and small carnivores. Abiotic events, such as droughts and floods, created disturbances that affected survivorship of ungulates and birds. Such disturbances showed feedbacks between biotic and abiotic realms. Interactions between elephants and their food allowed savanna and grassland communities to co-occur. With increased woodland vegetation, predators' capture of prey increased. Anthropogenic disturbances had direct (hunting) and indirect (transfer of disease to wildlife) effects. Slow and rapid changes and multiple ecosystem states became apparent only over several decades and involved events at different spatial scales. Conservation efforts should accommodate both infrequent and unpredictable events and long-term trends. Management should plan on the time scale of those events and should not aim to maintain the status quo. Systems can be self-regulating through food availability and predator-prey interactions; thus, culling may not be required. Ecosystems can occur in multiple states; thus, there may be no a priori need to maintain one natural state. Finally, conservation efforts outside protected areas must distinguish between natural change and direct human-induced change. Protected areas can act as ecological baselines in which human-induced change is kept to a minimum

Wellehan, J.F., Johnson, A.J., Childress, A.L., Harr, K.E., Isaza, R., 2007. Six novel gammaherpesviruses of Afrotheria provide insight into the early divergence of the Gammaherpesvirinae. Vet Microbiol 2007 Aug 19; [Epub ahead of print].
Abstract: The Afrotheria represent an early branching of placental mammals. Only two herpesviruses from Afrotheria have been previously identified, and the genus Proboscivirus in the subfamily Betaherpesvirinae has been proposed for them. Six novel gammaherpesviruses were identified in four species in the superorder Afrotheria by detection and analysis of their DNA polymerase genes. Elephantid herpesvirus 3 (ElHV3) and Elephantid herpesvirus 4 (ElHV4) were identified from conjunctival swabs from Asian elephants (Elephas maximus). ElHV3 was also found
in a vaginal swab from one elephant with vaginitis. Elephantid herpesvirus 5 (ElHV5) was identified from vaginal swabs of two Asian elephants with vaginal plaques. Elephantid herpesvirus 6 was discovered in a conjunctival swab from an African elephant (Loxodonta africana). Procavid herpesvirus 1 (PrHV1) was found in spleen and conjunctival swabs of rock hyrax (Procavia capensis). Trichechid herpesvirus 1 (TrHV1) was identified from skin and buffy coats of Florida manatees (Trichechus manatus latirostris). ElHV3 and ElHV4 form a distinct cluster, and ElHV5, ElHV6, TrHV1, and PrHV1 form a second cluster. These viruses may have codiverged with their host species. Phylogenetic analysis of these novel herpesviruses suggests that two separate groups of gammaherpesviruses may have codiverged with the Afrotheria.

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

Agnew, D.W. Brain removal in charismatic mega-vertebrates:  A not-so-charismatic chore.  2006 Proceedings American Association of Zoo Veterinarians.  2006.
Ref Type: Conference Proceeding
Abstract: Examination of the central nervous system, both grossly and histologically, is an important component of a complete necropsy.  Cerebral nematodiasis, West Nile Virus infection, rabies, distemper, and organophosphate toxicity are just a few of the possible diseases with serious herd and public health significance which may only be diagnosed by careful analysis of the brain and/or spinal cord.  Removal of the brain is strongly suggested for a complete necropsy, and though it may appear a daunting task, a few guidelines and power tools will allow efficient removal of the brain and a complete necropsy.
It is usually preferred that the brain be removed whole by removal of the skull cap.  This technique has been well documented in necropsy texts and is commonly taught in veterinary schools.  Briefly, after skinning the skull, a saw or ax may be used to cut on either side from the foramen magnum and the occipital condyles cranially and dorsally in a circular pattern (Fig. 1). This technique is useful to examine the brain in situ and remove it whole, but unfortunately requires skinning of the head, can be time-consuming, and is almost impossible to complete in rhinoceros and elephants. There are many alternative approaches to brain removal, but the author has found the following methods using commonly available tools are quick, leave a relatively intact skull, and the brain itself is removed in two parts.  Certainly, the techniques presented here can be adapted to the individual preferences of the prosector and to other similar species.  If nothing else, a discussion of brain removal techniques will reinforce the importance of collecting a complete set of tissues during a post-mortem examination.

Ehlers, B., Dural, G., Marschall, M., Schregel, V., Goltz, M., Hentschke, J., 2006. Endotheliotropic elephant herpesvirus, the first betaherpesvirus with a thymidine kinase gene
411. Journal of General Virology 87, 2781-2789.
Abstract: Endotheliotropic elephant herpesvirus (elephantid herpesvirus 1; ElHV-1) is apathogenic for African elephants (Loxodonta africana), but causes fatal haemorrhagic disease in Asian elephants (Elephas maximus). This is thought to occur through transmission from African elephants in places where both species are housed, such as zoological gardens. The virus has caused considerable losses in North American and European zoological gardens and thus severely impedes breeding of the endangered Asian elephant. Previously, the ultrastructural and genetic characterization of ElHV-1 from a male Asian elephant that died from the disease at the Berlin zoological gardens in 1998 have been reported. Here, a partial characterization of the ElHV-1 genome is presented. A 60 kbp locus, spanning 34 open reading frames, was analysed. Most of the detected genes were found to be conserved among the herpesviruses and showed an overall arrangement most similar to that of betaherpesviruses, in particular Human herpesvirus 6 and Human herpesvirus 7. Most importantly, in addition to a protein kinase gene that is homologous to the human cytomegalovirus UL97 gene, a thymidine kinase (TK) gene was found, which is generally missing in betaherpesvirus genomes. Thus, ElHV-1 is the only known betaherpesvirus to encode a TK gene. This peculiarity might contribute to the fulminant pathogenicity of ElHV-1, but also provide a crucial enzymic activity for developing an efficient antiviral therapy with currently available nucleoside analogues

Hildebrandt, T.B., Hermes, R., Ratanakorn, P., Rietschel, W., Fickel, J., RetNat, D., Frey, R., Reid, C., Goeritz, F. Ultrasonographic assessment and ultrasound-guided biopsy of the retropharyngeal lymph nodes in elephants.  2006 Proceedings American Association of Zoo Veterinarians.  117-118. 2006.
Ref Type: Conference Proceeding
Abstract: So far there are no valid diagnostic tools available for identifying latent carriers of endotheliotropic elephant herpes virus (EEHV).  For this reason, the lateral retropharyngeal lymph node complex (LARELYNOC) of elephants, identified during postmortem studies as target organ for EEHV and suitable for transcutaneous biopsy, was grossly described. Transcutaneous ultrasound (3.5 MHz) was applied behind the ear region to identify the LARELYNOC containing up to four single lymph nodes on each side. The lymph node tissue is situated 20-50 mm below the skin surface. An ultrasonographic assessment of the LARELYNOC and two biopsies were performed on 39 healthy Asian elephants (Elephas maximus). Samples were tested for EEHV via PCR. Whole blood samples were also collected and tested for active EEHV infection. Lymph nodes were ultrasonographically classified as active (calculated mean volume=17.4 ± 6.9 cm3, P>0.001), inactive (calculated mean volume=3.1 ± 0.6 cm3, P<0.001), or chronic active (calculated mean volume=10.6 ± 1.0 cm3, P<0.05). Histology confirmed not only the presence of lymph tissue but also the ultrasonographically diagnosed reactivity status of the lymph node biospies.  Although all samples including whole blood were found to be negative for the EEHV DNA particles, the successful development of this procedure in elephants could prove beneficial for the screening of not only latent EEHV infections but might also be a less dangerous alternative method for the diagnosis of zoonotic infections such as tuberculosis.

Isaza, R., Davis, R.D., Moore, S.M., Briggs, D.J., 2006. Results of vaccination of Asian elephants (Elephas maximus) with monovalent inactivated rabies vaccine. American Journal of Veterinary Research 67, 1934-1936.
Abstract: OBJECTIVE: To evaluate the humoral immune response of Asian elephants to a primary IM vaccination with either 1 or 2 doses of a commercially available inactivated rabies virus vaccine and evaluate the anamnestic response to a 1-dose booster vaccination. ANIMALS: 16 captive Asian elephants. PROCEDURES: Elephants with no known prior rabies vaccinations were assigned into 2 treatment groups of 8 elephants; 1 group received 1 dose of vaccine, and the other group received 2 doses of vaccine 9 days apart. All elephants received one or two 4-mL IM injections of a monovalent inactivated rabies virus vaccine. Blood was collected prior to vaccination (day 0) and on days 9, 35, 112, and 344. All elephants received 1 booster dose of vaccine on day 344, and a final blood sample was taken 40 days later (day 384). Serum was tested for rabies virus-neutralizing antibodies by use of the rapid fluorescent focus inhibition test. RESULTS: All elephants were seronegative prior to vaccination. There were significant differences in the rabies geometric mean titers between the 2 elephant groups at days 35, 112, and 202. Both groups had a strong anamnestic response 40 days after the booster given at day 344. CONCLUSIONS AND CLINICAL RELEVANCE: Results confirmed the ability of Asian elephants to develop a humoral immune response after vaccination with a commercially available monovalent inactivated rabies virus vaccine and the feasibility of instituting a rabies virus vaccination program for elephants that are in frequent contact with humans. A 2-dose series of rabies virus vaccine should provide an adequate antibody response in elephants, and annual boosters should maintain the antibody response in this species

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.

Reid, C.E., Hildebrandt, T.B., Marx, N., Hunt, M., Thy, N., Reynes, J.M., Schaftenaar, W., Fickel, J., 2006. Endotheliotropic elephant herpes virus (EEHV) infection. The first PCR-confirmed fatal case in Asia
436. Vet. Q. 28, 61-64.
Abstract: Since 1995, 4 suspected cases of Endotheliotropic Elephant Herpes Virus (EEHV) infection, i.e. based on clinical presentation, have occurred in Asia without resulting in epidemic outbreaks as expected. In order to confirm the presence of EEHV on the continent of Asia, viral DNA particles from liver samples of a wild-caught 3-year-old elephant found dead at a Cambodian elephant sanctuary and clinically diagnosed with EEHV, were PCR processed using known EEHV strain primers. The presence of EEHV viral nucleic acids was confirmed and the nucleic acids had a 99% sequence similarity to the U.S.A strain (gene bank locus: AF117265) and 97% sequence similarity to the European strain (gene bank locus: AF354746) assigning this case to the EEHV-1 cluster. More than the confirmation of EEHV on the continent of Asia, is the phylogenic relationship to the USA and European strains with no corresponding contact or transport of USA or European elephants to Asia. Thus, this brings many of the traditional theories into question. Although almost forgotten, this disease is still ramped in captive elephant populations worldwide and continues to devastate particularly the neonatal and weaning-age population. Special attention and continued research are needed specifically in the area of basic virology and epidemiology

Sharam, G., Sinclair, A.R.E., Turkington, R., 2006. Establishment of broad-leaved thickets in Serengeti, Tanzania: The influence of fire, browsers, grass competition, and elephants. Biotropica 38, 599-605.
Abstract: The role of Euclea divinorum in the establishment of broad-leaved thickets was investigated in Serengeti National Park, Tanzania. Thickets are declining due to frequent fires, but have not reestablished when fires have been removed. Seedlings of E. divinorum, a fire-resistant tree, were found in grassland adjacent to thickets and as thicket canopy trees and may function to facilitate thicket establishment. Seedlings of thicket species were abundant under E. divinorum canopy trees but not in the grassland, indicating that E. divinorum can facilitate forest establishment. We examined E.divinorum establishment in grassland by measuring survival and growth of seedlings with respect to fire, browsers, elephants, and competition with grass. Seedling survival was reduced by fire (50%), browsers (70%), and competition with grass (50%), but not by elephants. Seedling growth rate was negative unless both fire and browsers, or grass was removed. Establishment of thickets via E. divinorum is not occurring under the current conditions in Serengeti of frequent fires, abundant browsers, and dense grass in riparian areas. Conditions that allowed establishment may have occurred in 1890-1920s during a rinderpest epizootic, and measurements of thicket canopy trees suggest they established at that time.

Wellehan, J.F.X., Johnson, A.J., Isaza, R. Identification of two novel herpesviruses associated with ocular inflammation in Asian elephants (Elephas maximus).
2006 Proceedings American Association of Zoo Veterinarians.  173. 2006.
Ref Type: Conference Proceeding
Abstract: Disease caused by a herpesvirus (EEHV) is a serious concern in Asian elephant (Elephas maximus) calves.  Herpesviruses are known for latency and life-long infections, with periodic shedding from mild inflammatory lesions in adapted adult hosts, and ocular disease has been seen with other herpesviruses in other species.  Ocular inflammation is not uncommonly seen in Asian elephants. Degenerate PCR primers targeting a conserved region of herpesvirus DNA-dependent DNA polymerase were used to amplify products from eye swabs of eight Asian elephants with epiphora, blepharitis, and conjunctivitis.  Nucleotide sequencing of the PCR products showed two novel herpesviruses distinct from EEHV.  Comparative sequence analysis shows that these viruses are probable members of the subfamily Gammaherpesvirinae. The sequence phylogeny of these viruses has implications for both viral and host evolution.  Further understanding and characterization of these viruses is needed to understand their role in elephant health.

Zuba, J.R., Oosterhuis, J.E., Pessier, A.P. The toenail "abscess" in elephants: treatment options including cryotherapy and pathologic similarities with equine proliferative pododermatitis (canker).  2006 Proceedings American Association of Zoo Veterinarians.  187-190. 2006.
Ref Type: Conference Proceeding
Abstract: Foot problems potentially represent the single most important clinical disease of captive elephants.  Predisposing factors include obesity, lack of exercise, nail or sole overgrowth, improper foot care, poor hygiene, inappropriate enclosure surfaces, poor conformation, malnutrition and secondary skeletal disorders such as degenerative joint disease.  Furthermore, factors such as elephant management philosophy, disposition of elephants, facilities and competency of staff in caring for elephant feet will contribute significantly to the foot health of captive animals.  It is important to note that these conditions are rarely reported in free-ranging elephants. The elephant toenail abscess is characterized grossly by proliferative outgrowth of "crab meat-like" tissue that may acutely rupture through the surface of the nail wall and/or adjacent cuticle or sole. True abscess formation with localized collections of suppurative material is not a consistent clinical feature.  In most cases, the inciting cause of these lesions are typically not found and are likely due to one or more of the predisposing factors listed above.  Once established, these frustrating lesions require extensive, intensive and prolonged medical attention.  If not cared for properly, these wounds may progress to phalangeal osteomyelitis and the need for surgical intervention.  Sole abscesses are equally frustrating and difficult to manage with proposed etiologies similar to toenail lesions. There are no reports in the literature describing the pathology of the classic proliferative abscess tissue of the elephant nail abscess.  Although variously interpreted as fibrous or granulation tissue, the authors are unaware of previous histologic descriptions of this tissue.  Biopsy samples of toenail abscess tissue from two Asian elephants (Elephas maximus) at the San Diego Wild Animal Park (SDWAP) consisted of stratified squamous epithelium arranged in columns resembling horn tubules.  The predominant histologic finding was marked, near diffuse, hydropic degeneration of keratinocytes.  There were multifocal areas of suppurative inflammation with admixed bacterial colonies.  Inflammatory foci comprised only a small portion of the lesion and were interpreted as the external surfaces of the biopsy with likely secondary bacterial colonization. Because descriptions of the normal histology of the elephant toenail could not be located, a grossly normal toenail from a different Asian elephant was obtained to compare histologic features with those of the toenail abscesses.  Sections demonstrated formation of the toenail in a manner similar to that of the hoof of the horse and cattle with tubular, intertubular and laminar horn.  Primary and secondary epidermal laminae were identified.  Proliferative lesions of horn-producing epithelium associated with ballooning degeneration and inadequate keratinization of keratinocytes, have been described in horses as equine "canker" and coronary band dystrophy.  Equine canker is most commonly observed in the hind feet of draft horses and begins in the frog sometimes with extension to the sole and hoof wall.  Grossly, lesions are characterized by soft white papillary to "cauliflower-like" tissue associated with a foul odor. Similar to what is noted in elephant foot problems, predisposing factors for the development of equine canker include poor hygiene or wet environmental conditions. There is a lack of gross and histologic description of the normal nail and sole tissue of the elephant and further investigations are warranted.  A review of the anatomy and histology of the normal equine hoof may provide a basic understanding of the elephant nail until more specific and detailed elephant information is available.  From our investigation, the authors offer that a more accurate description of the elephant toenail abscess would be proliferative pododermatitis, the term synonymous with equine canker.  A more colloquial term such as "elephant canker" may be appropriate, as well. Canker in the horse is an uncommon but difficult to treat disease of the hoof.  Historically, treatment options for elephant toenail abscesses include corrective trimming, superficial debridement and application of topical disinfectants or antibiotics. Others have constructed innovative sandals to treat and protect the affected sole or nail with success. The use of regional intravenous perfusion of the affected limb with antibiotics has also been successful. Since the elephant nail abscess now appears to be histologically and clinically comparable to equine canker, this novel characterization of an old disease may offer unique insight for treatment.  In the least, it has provided our practice with a new list of treatment options and experienced equine clinicians for consultation who have been managing patients with a similar disease for many years. One of the Asian elephants at the SDWAP has had chronic toenail abscesses for over 2 yr. Radiographs of the affected digits, as reported by others to assess degree of involvement, have fortunately been negative for evidence of osteomyelitis.  Several bacterial and fungal cultures of deep tissue biopsies and swabs of affected lesions have resulted in a mixture of organisms with no consistent single etiologic agent.  Biopsies were found negative for presence of viral DNA (elephant papillomavirus and herpesvirus) by PCR.  Typical elephant foot care at the SDWAP includes trimming and debriding with hoof knives, foot soaks and topical antibiotics.  Although difficult, attempts are made in keeping the affected foot clean and dry.  Following recommendations for the treatment of equine canker, we recently implemented the routine use of cryotherapy in all elephants with proliferative pododermatitis with improved success in the control and recession of exuberant nail lesions. The proliferative tissue of the nail is first cleaned then disinfected, debrided, trimmed with hoof knives and allowed to dry. Modified brass branding tools with contact surfaces of variable size (2-5 cm diameter) and shape (round or ovoid) are placed into liquid nitrogen (-196 C) for several minutes and then placed directly on the cankerous tissue for 30-60 sec.  This process is then repeated 4-5 min later, following a complete thaw of tissue.  Within 24 hr, the cryoburned tissue becomes macerated and necrotic and is readily removed with gentle scrubbing.  Cryotherapy offers the advantage of destroying tissue to a deeper level than trimming alone and provides hemostasis, as well.  Because of decreased sensation at the cryotherapy treatment site, a memorable painful event is avoided and the elephant patient is more routinely accepting of this technique. With the use of hoof knives, we typically remove 2-3 mm of proliferative tissue before the patient refuses further treatment, presumably due to discomfort.  With cryotherapy, we are able to remove an additional 3-5 mm of tissue by cell freezing and necrosis.  The result is quicker resolution of cankerous lesions without the need for aggressive, and potentially painful, interventions. In conclusion, it appears that elephant nail abscesses can best be described as proliferative pododermatitis, or canker, as is seen in other species.  Further gross and microscopic descriptions of normal and pathologic nail or sole lesions are necessary.  Routine cryotherapy has shown promise in the treatment of these chronic, frustrating and potentially devastating lesions of our captive elephants.

Hildebrandt, T.B., Hermes, R., Ratanakorn, P., Rietschel, W., Fickel, J., Frey, R., Wibbelt, G., Reid, C., Goritz, F., 2005. Ultrasonographic assessment and ultrasound-guided biopsy of the retropharyngeal lymph nodes in Asian elephants (Elephas maximus)
552. Veterinary Record 157, 544-548.
Abstract: Endotheliotropic herpesvirus causes a fatal disease in young Asian elephants, but there are no methods for identifying latent carriers of the virus. During the postmortem study of one female African elephant and three male and two female Asian elephants, a lymph node located bilaterally caudoventral to the parotid gland, approximately 1.5 to 5 cm below the skin, was identified as suitable for transcutaneous ultrasound-guided biopsy. An ultrasonographic assessment and two biopsies were performed on 39 Asian elephants, and these lymph nodes were classified ultrasonographically as active, inactive or chronically active. The calculated mean (se) volume of 10 active lymph nodes was 17.4 (6.9) cm(3), and that of three chronically active lymph nodes was 10.6 (1.0) cm(3), whereas the mean volume of 17 inactive lymph nodes was 3.1 (0.6) cm(3). The presence of lymph node tissue in samples obtained by ultrasound-guided biopsy from three animals that were maintained under conditions that allowed for additional sampling was confirmed histologically. The dna extracted from the lymphoid tissue and the whole blood of all the elephants was negative for endotheliotropic herpesvirus by PCR.

Wise, D.J., Carter, G.R., Flores, E.F. Laboratory Diagnosis of Viral Infections. Carter, G. R., Wise, D. J., and Flores, E. F. A Concise Review of Veterinary Virology. Ithaca: International Veterinary Information Service (www. ivis. org), 2005; Document No. A3407. 0305.  2005.
Ref Type: Electronic Citation

Greenwood, A.D., Englbrecht, C.C., MacPhee, R.D., 2004. Characterization of an endogenous retrovirus class in elephants and their relatives
667. BMC. Evol. Biol. 4, 38.
Abstract: BACKGROUND: Endogenous retrovirus-like elements (ERV-Ls, primed with tRNA leucine) are a diverse group of reiterated sequences related to foamy viruses and widely distributed among mammals. As shown in previous investigations, in many primates and rodents this class of elements has remained transpositionally active, as reflected by increased copy number and high sequence diversity within and among taxa. RESULTS: Here we examine whether proviral-like sequences may be suitable molecular probes for investigating the phylogeny of groups known to have high element diversity. As a test we characterized ERV-Ls occurring in a sample of extant members of superorder Uranotheria (Asian and African elephants, manatees, and hyraxes). The ERV-L complement in this group is even more diverse than previously suspected, and there is sequence evidence for active expansion, particularly in elephantids. Many of the elements characterized have protein coding potential suggestive of activity. CONCLUSIONS: In general, the evidence supports the hypothesis that the complement had a single origin within basal Uranotheria

Musser, J.M.B., 2004. A practitioner's primer on foot-and-mouth disease. Journal of the American Veterinary Medical Association 224, 1261.
Abstract: Foot-and-mouth disease (FMD) is caused by an RNA virus of the genus Aphthovirus; 7 immunologically distinct serotypes of the virus have been identified. Susceptible species are mainly domestic and wild even-toed ungulates, such as cattle, sheep, goats, pigs, bison, and deer. All body fluids of infected animals can contain the virus and are considered infective. The primary mode of transmission is animal-to-animal transmission through inhalation or ingestion of aerosois containing the virus. The virus can also be spread mechanically by contaminated organic debris and fomites and can survive for 48 hours on human oral and nasal mucosa and be spread to uninfected animals in the manner. There is a rapid progression of clinical signs after an animal becomes infected, and the virus spreads rapidly throughout a herd. Clinical signs include excessive salivation; fever; vesicles and erosions of the oral and nasal mucosa, coronary band, interdigital area, and teats; lameness; sloughing of claws; reluctance to move; anorexia; mastitis; decreased milk production; and abortion or weak newborns. In mature animals, FMD has high morbidity and low mortality rates. Infected animals can become inapparent carriers of the virus.

Chakraborty, A., 2003. Diseases of elephants (Elephas maximus) in India-A Review. Indian Wildlife Year Book 2, 74-82.

Fickel, J., Liekfeldt, D., Richman, L.K., Streich, W.J., Hildebrandt, T.B., Pitra, C., 2003. Comparison of glycoprotein B (gB) variants of the elephant endotheliotropic herpesvirus (EEHV) isolated from Asian elephants (Elephas maximus). Vet Microbiol 91, 11-21.
Abstract: The recently described elephant endotheliotropic herpesviruses (EEHV) have been associated with the deaths of numerous captive elephants. A proposed tool for the detection of EEHV infection in elephants is the PCR-based screening for EEHV-DNA in whole blood samples. Unfortunately, this detection method has only been successful in post-mortem analyses or in animals already displaying clinical signs of EEHV disease, thus rendering this method unsuitable for identification of carrier elephants. Here, we focus on glycoprotein B (gB) for serologic assay development, since gB is an envelope protein known to induce a neutralising antibody response in other herpesvirus infections. We sequenced the entire gB gene from five Asian elephants with EEHV, representing four different gB variants. Computer-aided methods were used to predict functionally important regions within EEHVgB. An extra-cytoplasmic region of 153 amino acids was predicted to be under positive selection and may potentially contain antigenic determinants that will be useful for future serologic assay development.

Isaza, R., Hunter, R.P., Richman, L.K., Montali, R.J., Schmitt, D.L., Koch, D.E., Lindsay, W.A. Famciclovir pharmacokinetics in young Asian elephants (Elephas maximus). Proc. American Assoc. of Zoo Veterinarians.  82-83. 2003.
Ref Type: Conference Proceeding
Abstract: Asian elephants (Elephas maximus) are susceptible to a unique infection caused by elephant endotheliotropic herpesvirus (EEHV).3,4 Worldwide, between the years 1983 and 2000, there have been 26 confirmed deaths from this virus in Asian elephants.2 Although most cases have been fatal, treatment with famciclovir (Famvir, SmithKline Beecham Pharmaceuticals, Philadelphia,PA 19101 USA) has been associated with survival in three cases of six cases of EEHV infection proven by PCR.2,5,6 Dose selections for surviving elephants (5.5 - 8.0 mg/kg, p.o. every 8 hr) were made without the benefit of elephant pharmacokinetics and were a direct extrapolation from recommended human dosages (7 mg/kg, p.o. every 8 hr).5,6  In this study, famciclovir was administered both orally and rectally in healthy young Asian elephants. The doses tested in this study were 5 mg/kg orally, 5 mg/kg rectally, and 15 mg/kg rectally. Blood samples were analyzed for famciclovir and penciclovir using a validated LC/MS assay. Famciclovir was absorbed well by both routes and underwent rapid biotransformation to the active compound penciclovir. None of the plasma samples had detectable famciclovir. Pharmacokinetic parameters for penciclovir were determined using non-compartmental analysis. After a single oral dose of 5 mg/kg the Cmax was 1.3 ìg/mL with a Tmax at 1.1 h. After a rectal dose of 5 mg/kg the Cmax  was 1.2 ìg/mL with a Tmax at 0.34 hr. After a rectal dose of 15 mg the t½ was 2.6 h, with a Cmax of 3.6 ìg/mL at Tmax 0.66 h. These results were similar to those reported in humans where an oral dose of 500 mg (7 mg/kg) had a t½ of about 2 h with a Cmax of 3.3 ìg/mL. A dose range of 8 -15 mg/kg given orally or rectally every 8 hours should produce penciclovir concentrations in Asian elephants that are considered therapeutic in humans.
LITERATURE CITED
1. Hardman, J.G., L.E. Limbird and A.G. Gilman. 2001. In: Goodman and Gilman's The pharmacological basis of therapeutics, Tenth edition. McGraw-Hill Co. New York, New York. USA. Pp. 1322-1324.
2. Montali, R.J., Richman, L.K., Mikots,S.K., Schmitt,D.L., Larsen, R.S., Hildebrandt,T.B., Isaza,R., Lindsay,W.A. Management Aspects of Herpesvirus Infections and Tuberculosis in Elephants. International Elephant and  Rhino Research Symposium, Vienna, June 7-11, 2001.
3. Richman, L.K., R.J. Montali, R.L. Garber, M.A. Kennedy, J. Lehnhardt, T. Hildebrandt, D. Schmitt, D. Hardy, D.J. Alcendor, G.S. Hayward. 1999a. Novel endotheliotropic herpesvirus fatal for Asian and African elephants. Science. 283:1171-1176.
4. Richman, L.K., R.J. Montali, R.C. Cambre, D. Schmitt, D. Hardy, T. Hildebrandt, F.M. Hamzeh, A. Shahkolahi, G.S. Hayward. 2000.  Clinical and pathological findings of a newly recognized disease of elephants caused by endotheliotrophic herpesvirus.  J. Wild. Dis. 36:1-12.
5. Schaftenaar, W., J.M.C.H. Mensink, A.M. de Boer, T.B. Hildebrandt, and J. Fickel. 2001. Successful treatment of a subadult Asian elephant bull (Elephas maximus) infected with the endotheliotropic elephant herpes virus. Verhber. Erkrg. Zootiere. P. 40.
6. Schmitt, D., D.A. Hardy, R.J. Montali, LK Richman, Lindsay WA, R. Isaza, G. West. 2000. Use of famciclovir for the treatment of endotheliotropic herpesvirus infections in Asian elephants (Elephas maximus). J. Zoo Wild. Med. 31:518-522.

Mahanta, P.N., 2003. Health monitoring and common diseases in free ranging elephants. In: Das, D. (Ed.), Healthcare, Breeding and Management of Asian Elephants. Project Elephant. Govt. of India, New Delhi, pp. 130-136.

Rahman, T., 2003. Infectious and non-infectious disease of elephants. In: Das, D. (Ed.), Healthcare, Breeding and Management of Asian Elephants. Project Elephant. Govt. of India, New Delhi, pp. 108-118.

Rehman, A., 2003. Disease control program of elephants. In: Das, D. (Ed.), Healthcare, Breeding and Management of Asian Elephants. Project Elephant. Govt. of India, New Delhi, pp. 152-156.

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

Chandrasekharan, K., 2002. Specific diseases of Asian elephants. Journal of Indian Veterinary Association Kerala 7, 31-34.
Abstract: The earliest writing describing the diseases of elephants in ancient literature said to be the works on "Gajasastra" (Elephantology) written in Sanskrit by authors like Gautama, Narada, Mrigacharma, Rajaputra and Vyasa. "Hasthyayurveda" a legendary book in Sanskrit written by a safe Palakapya deals with some diseases, treatment, desirable and undesirable points of selection, management practices and some mythological aspects on the origin of elephants. The earliest book in English dealing with diseases of elephants seems to be that of W. Gilchrist "A practical treatise on the treatment of diseases of elephants" published in 1848. Later Slym (1873), Sanderson (1878), Steel (1885), Evans (1910), Herpburn (1913), Milroy (1922), Ptaff (1940), Ferrier (1947), Utoke Gale (1974), Chandrasekharan (1979) and Panicker (1985) have documented their findings on the incidence, etiology and control of diseases of Asian elephants.

Essbauer, S., Meyer, H., Kaaden, O.R., Pfeffer, M., 2002. Recent cases in the German poxvirus consulting laboratory. Revue de Medecine Veterinaire 153, 635-642.
Abstract: Designated as poxvirus consulting laboratory by the Robert-Koch-Institute (Berlin), we provide scientific advice regarding any aspects of poxviruses affecting different animals e.g. cats, elephants, swines, birds and men. Human smallpox was eradicated in the 1980s, and consequently with diminishing vaccination a generation susceptible for other zoonotic poxviruses grows up. Although the epidemiology of orthopoxvirus infections in Germany remains unclear, in the last few years we observed a drastic increase of zoonotic poxvirus infections with three case reports on human 'cowpox' infections presented here. In the two cases, we could only retrospectively trace the source of the orthopoxvirus to cats based on seroconversion. In one case, a young cat transmitted the virus to three humans; all developed clinical pox lesions. Underlying the zoonotic potential of cowpoxviruses (CPXV), these viruses exhibit a broad host range. In the year 2000, two elephants (Elephas maximus) of a German travelling circus revealed a fatal orthopoxvirus infection. The animals exhibited many poxviral lesions and died. Thus, we provide the modified vaccinia virus Ankara (MVA) for vaccination of exotic or expensive animals. Classical virological and serological methods as well as molecular-biological techniques including PCR, sequencing and restriction fragment patterns of the newly isolated poxviruses show a very close relationship of the investigated CPXV isolates irrespective of their host species. These findings and our long-term data give evidence of an increase in orthopoxviruses infections in animals and men; thus, highlighting the importance of further investigations on virus transmission and orthopoxvirus reservoirs.

McCallum, H., Dobson, A., 2002. Disease, habitat fragmentation and conservation. Proc. R. Soc. Lond. B 269, 2041-2049.
Abstract: Habitat loss and the resultant fragmentation of remaining habitat is the primary cause of loss of biological diversity. How do these processes affect the dynamics of parasites and pathogens? Hess has provided some important insights into this problem using metapopulation models for pathogens that exhibit 'S-I' dynamics; for example, pathogens such as rabies in which the host population may be divided into susceptible and infected individuals. A major assumption of Hess's models is that infected patches become extinct, rather than recovering and becoming resistant to future infections. In this paper, we build upon this framework in two different ways:  first, we examine the consequences of including patches that are resistant to infection; second, we examine the consequences of including a second species of host that can act as a reservoir for the pathogen. Both of these effects are likely to be important from a conservation perspective. The results of both sets of analysis indicate that the benefits of corridors and other connections that allow species to disperse through the landscape far outweigh the possible risks of increased pathogen transmission. Even in the commonest case, where harmful pathogens are maintained by a common reservoir host, increased landscape connectance still allows greater coexistence and persistence of a threatened or endangered host.

Schaftenaar, W., 2002. Use of vaccination against foot and mouth disease in zoo animals, endangered species and exceptionally valuable animals. Rev. sci. tech. Off. int. Epiz. 21, 613-623.
Abstract: A historical review of foot and mouth disease (FMD) in non-domestic species is given and the use of FMD vaccines to protect those species is described. Several non-domestic species are susceptible to FMD. Legislation in many countries, based on the definition of FMD-free status as determined by the Office International des Epizooties (OIE: World organisation for animal health), forms an important barrier against the use of vaccines. National authorities may even feel obliged to slaughter animals of threatened species protected by international
agreements during an outbreak of FMD to preserve their FMD-free status. The importance of international breeding programmes for endangered species is forcing the international community to reconsider the role that vaccination against FMD should play in animal health prevention programmes of captive populations. Much research is still required in regard to vaccine types and diagnostic procedures. Species-specific differences in susceptibility to FMD make this a challenging research topic for zoological institutions.  Use of vaccination against foot and mouth disease in zoo animals, endangered species and exceptionally valuable animals

Burkhardt, S., Goltz, M., Bergmann, V., Ochs, A., Weiler, H., Hentschke, J., 2001. Genetic and ultrastructural characterization of a European isolate of the fatal endotheliotropic elephant herpesvirus. Journal of General Virology 82, 475-482.
Abstract: A male Asian elephant (Elephas maximus) died at the Berlin zoological gardens in August 1998 of systemic infection with the novel endotheliotropic elephant herpesvirus (EIHV-1). This virus causes a fatal haemorrhagic disease in Asian elephants, the so-called endothelial inclusion body disease, as reported from North American zoological gardens. In the present work, EIHV-1 was visualized ultrastructurally in affected organ material. Furthermore, a gene block comprising the complete glycoprotein B (gB) and DNA polymerase (DPOL) genes as well as two partial genes was amplified by PCR-based genome walking and sequenced. The gene content and arrangement were similar to those of members of the Betaherpesvirinae. However, phylogenetic analysis with gB and DPOL consistently revealed a very distant relationship to the betaherpesviruses. Therefore, EIHV-1 may be a member of a new genus or even a new herpesvirus subfamily. The sequence information generated was used to set up a nested-PCR assay for diagnosis of suspected cases of endothelial inclusion body disease. Furthermore, it will aid in the development of antibody-based detection methods and of vaccination strategies against this fatal herpesvirus infection in the endangered Asian elephant.

Ehlers, B., Burkhardt, S., Goltz, M., Bergmann, V., Ochs, A., Weiler, H., Hentschke, J., 2001. Genetic and ultrastructural characterization of a European isolate of the fatal endotheliotropic elephant herpesvirus. J Gen Virol 82 (Pt 3), 475-482.
Abstract: A male Asian elephant (Elephas maximus) died at the Berlin zoological gardens in August 1998 of systemic infection with the novel endotheliotropic elephant herpesvirus (ElHV-1). This virus causes a fatal haemorrhagic disease in Asian elephants, the so-called endothelial inclusion body disease, as reported from North American zoological gardens. In the present work, ElHV-1 was visualized ultrastructurally in affected organ material. Furthermore, a gene block comprising the complete glycoprotein B (gB) and DNA polymerase (DPOL) genes as well as two partial genes was amplified by PCR-based genome walking and sequenced. The gene content and arrangement were similar to those of members of the Betaherpesvirinae. However, phylogenetic analysis with gB and DPOL consistently revealed a very distant relationship to the betaherpesviruses.Therefore, ElHV-1 may be a member of a new genus or even a new herpesvirus subfamily. The sequence information generated was used to set up a nested-PCR assay for diagnosis of suspected cases of endothelial inclusion body disease. Furthermore, it will aid in the development of antibody-based detection methods and of vaccination strategies against this fatal herpesvirus infection in the endangered Asian elephant.

Fickel, J., Richman, L.K., Montali, R., Schaftenaar, W., Goritz, F., Hildebrandt, T.B., Pitra, C., 2001. A variant of the endotheliotropic herpesvirus in Asian elephants (Elephas maximus) in European zoos. Veterinary Microbiology 82, 103-109.
Abstract: Newly discovered, lethal elephant endotheliotropic herpesviruses (EEHV) have been identified in both Asian (Elephas maximus) and African (Loxodonta africana) elephants. Carried by otherwise healthy African elephants, they can be fatal, mainly for young Asian elephants. Since zoos often harbour both elephant species, we conducted a survey on the presence of EEHV in elephants (Asian elephants, n=57; African elephants, n=17) from 12 zoos and 3 circuses in Europe (Germany, Switzerland and the Netherlands), and 1 zoo in Israel [date not given]. Six of the 57 Asian elephants were positive for EEHV. Five elephants died of the infection, while one survived. EEHV was not detected in any of the 17 African elephants. All EEHV that affected the Asian elephants belonged to the EEHV1 group. We described the detection and the partial sequencing of an endotheliotropic herpesvirus variant (named EEHV1b) in Asian elephants, being either an EEHV endogenous to Asian elephants or indicating different sources (African elephants) of infection.

Greenwood, A.D., Lee, F., Capelli, C., DeSalle, R., Tikhonov, A., Marx, P.A., MacPhee, R.D., 2001. Evolution of endogenous retrovirus-like elements of the woolly mammoth (Mammuthus primigenius) and its relatives. Mol Biol Evol 18, 840-870.
Abstract: Endogenous retrovirus-like elements characterizable by a leucine tRNA primer (ERV-Ls) are reiterated genomic sequences known to be widespread in mammals, including humans. They may have arisen from an ancestral foamy virus-like element by successful germ line infection followed by copy number expansion. However, among mammals, only primates and rodents have thus far exhibited high copy number amplification and sequence diversification. Conventionally, empirical studies of proviral amplification and diversification have been limited to extant species, but taxa having good Quaternary fossil records could potentially be investigated using the techniques of "ancient" DNA research. To examine evolutionary parameters of ERV-Ls across both time and taxa, we characterized this proviral class in the extinct woolly mammoth (Mammuthus primigenius) and living elephants, as well as extant members of the larger clade to which they belong (Uranotheria, a group containing proboscideans, sirenians, hyraxes, and their extinct relatives). Ungulates and carnivores previously analyzed demonstrated low copy numbers of ERV-L sequences, and thus it was expected that uranotheres should as well. Here, we show that all uranothere taxa exhibit unexpectedly numerous and diverse ERV-L sequence complements, indicating active expansion within this group of lineages. Selection is the most parsimonious explanation for observed differences in ERV-L distribution and frequency, with relative success being reflected in the persistence of certain elements over a variety of sampled time depths (as can be observed by comparing sequences from fossil and extant elephantid samples).

Hildebrandt, T.B., Fickel, J., Goritz, F., Rietschel, W., Liekfeldt, D., Montali, R.J., Richman, L.K., Ratanakorn, P. Survey on presence of the endotheliotropic elephant herpesvirus (EEHV) in Thai camp 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.  183-184. 2001.  American Association of Zoo Veterinarians. 9-18-2001.
Ref Type: Conference Proceeding

Hinke, A., Wipplinger, J. A Severe Case of Pox Disease in Two Asian Elephants (Elephas maximus) of a Small Travelling Circus Overwintering Near Erfurt. A Research Update on Elephants and Rhinos; Proceedings of the International Elephant and Rhino Research Symposium, Vienna, June 7-11, 2001.  53-56. 2001. Vienna, Austria, Schuling Verlag. 2001.
Ref Type: Conference Proceeding
Abstract: A 32 and a 35 year old Asian elephant (Elephas maximus) belonging to a small travelling circus showed symptoms of a severe pox disease in September 2000 during overwintering near Erfurt. From mucous conjunctival excretions and typical lesions of the mucosa of the mouth a poxvirus strain was isolated which showed the biological characteristics of cowpox virus (Orthopoxvirus bovis). Because of the fact that the elephants were treated about 5 weeks the wrong way from another veterinarian who had no experience with pox disease in elephants medical treatment started to became a bit difficult. After weeks of intensive medical care the condition improved, however due to massive cycles of further virus development it deteriorated, and the animals had to be euthanised after about 5 weeks of treatment.

McLelland, D., Kirkpatrick, J.F., Rose, K., Dixon, R. Studies on encephalomycarditis virus (EMCV) in a zoologic context. AAZV,AAWV,ARAV,NAZWV Joint Conf.  337. 2001.
Ref Type: Conference Proceeding

Montali, R.J., Richman, L.K., Mikota, S.K., Schmitt, D.L., Larsen, R.S., Hildebrandt, T.B., Isaza, R., Lindsay, W.A. Management Aspects of Herpesvirus Infections and Tuberculosis in Elephants. A Research Update on Elephants and Rhinos; Proceedings of the International Elephant and Rhino Research Symposium, Vienna, June 7-11, 2001.  87-95. 2001. Vienna, Austria, Schuling Verlag. 2001.
Ref Type: Conference Proceeding
Abstract: Elephant endotheliotropic herpesvirus (EEHV) infections and tuberculosis have emerged as causes of illness and mortality in captive elephants. Twenty-six confirmed EEHV cases are documented. Since 1995, 7 have occurred in North America, 10 in Europe and 2 in Asia. A PCR test was used to detect the virus in symptomatic animals; a serological test to identify carrier elephants is under development. The African elephant is a potential source of the EEHV that is lethal for Asian elephants. Fatal infections have also occurred in Asian elephants without African elephant contacts. Three of 6 elephants recovered after treatment with antiviral famciclovir; however, more research is needed to improve the usefulness of this drug. Asian elephants that are less than 10-years old and have been moved to another facility and/or have had contact with African elephants are at increased risk for contracting EEHV. Animals traveling between facilities with a history of EEHV cases may be at greater risk. All young elephants should be monitored daily for anorexia, lethargy, body swellings and blue discoloration (bruising) of the tongue, and be trained for blood sampling and potential oral and rectal treatment with famciclovir.
Since 1996, Mycobacterium tuberculosis has affected about 3% of Asian elephants in North America. Most were from 5 U.S. States with some contacts between private herds. Mandatory annual testing for tuberculosis by trunk wash cultures was established in 1998, and 22 culture-positive M. tuberculosis elephants were identified between 1996-2001. Fifteen were treated with anti-tuberculosis drugs and 7 that died or were euthanized were proven to have tuberculosis at necropsy. Antemortem sera was available from 4/7 4 (75%) were strongly ELISA positive. Tuberculosis is uncommon in African elephants but was recently associated with M. bovis in the U.S. and M. tuberculosis in Germany. Conversely, M. bovis tuberculosis, apparently unrecognized in Asian elephants, recently occurred in Germany. Management issues of elephant tuberculosis will be discussed relative to its complex epidemiology and clinical-pathological correlations.

Ratanakorn, P. Elephant Health Problems and Management in Cambodia, Lao and Thailand. A Research Update on Elephants and Rhinos; Proceedings of the International Elephant and Rhino Research Symposium, Vienna, June 7-11, 2001.  111-114. 2001. Vienna, Austria, Schuling Verlag. 2001.
Ref Type: Conference Proceeding

Richman, L.K., Montali, R.J., 2001. Elephant herpesvirus infections. In: Williams, E.S., Barker, I.K. (Eds.), Infectious Diseases of Wild Mammals, 3rd edition. Iowa State University Press, Ames, Iowa, USA, pp. 170-178.

Ryan, S.J., Thompson, S.D., 2001. Disease risk and inter-institutional transfer of specimens in cooperative breeding programs: Herpes and the elephant species survival plans. Zoo Biology 20, 89-101.
Abstract: Managers of cooperative breeding programs and re-introduction projects are increasingly concerned with the risk of disease transmission when specimens are transferred among facilities or between facilities and the natural environment. We used data maintained in North American studbooks to estimate the potential risks of disease transmission by direct and indirect contact of specimens in the American Zoo and Aquarium Association's Elephant Species Survival Plan. Histological evidence for a novel herpesvirus disease transmitted between and within elephant species housed in North American facilities prompted an examination of the scope of possible transmission routes within the captive population. We found that, compared with other species managed through Species Survival Plans, elephants experience relatively few transfers between zoos. Nevertheless, the number of direct contacts with other elephants born during the study period of 1983-1996 (excluding stillbirths) was much higher than we had anticipated (&mgr; = 25 +/- 27; N = 59) and the number of potential indirect contacts was surprisingly large (&mgr; = 143 +/- 92; N = 59). Although these high rates of potential contacts complicate exact identification of infection pathways for herpesvirus, we were able to propose potential routes of transmission for the histologically identified cases. Furthermore, the extraction of data from studbooks allowed us to readily identify other specimens that did not succumb to the disease despite similar exposure. Moreover, we were able to identify other possible cases to recommend for histological examination. Herein we reveal the possibilities of multiple disease transmission pathways and demonstrate how complex the patterns of transmission can be, confounded by the unknown latency of this novel herpesvirus. This emphasizes the need for zoo veterinarians and cooperative breeding programs to consider the full potential for disease transmission associated with each and every inter-zoo transfer of specimens.

Schaftenaar, W., Mensink, J.M.C.H., Deboer, A.M., Hildebrandt, T.B., Fickel, J. Successful treatment of a subadult Asian elephant bull (Elephas maximus) infected with elephant herpes virus. Proc. of the  International Symposium for diseases of Zoo and Wildlife Animals (Rotterdam).  141-146. 2001.
Ref Type: Conference Proceeding

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

Calle, P.P., Ludwig, G.V., Smith, J.F., Raphael, B.L., Clippinger, T.L., Rush, E.M., McNamara, T., Manduca, R., Linn, M., Cook, R.A., et, al. Clinical aspects of West Nile virus infection in a zoological collection. Proc. AAZV and IAAAM Joint Conf.  2000.
Ref Type: Conference Proceeding

Fickel, J., Richman, L.K., Reinsch, A., Montali, R., Schaftenaar, W., Goritz, F., Hildebrandt, T.B. Survey on the occurrence of the endotheliotropic elephant herpesvirus (EEHV) in Asian (Elephas maximus) and African (Loxodonta africana) elephants in Europen zoos. European Association of Zoo and Wildlife Veterinarians Third Scientific Meeting, Paris, France, May 31-June 4, 2000.  2000.
Ref Type: Conference Proceeding

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.

Richman, L.K., Montali, R.J., Hayward, G.S., 2000. Review of a newly recognized disease of elephants caused by endotheliotropic herpesviruses. Zoo Biology 19, 383-392.
Abstract: There are two newly recognized herpesviruses that cause a fatal disease syndrome in elephants. They are known as the elephant endotheliotropic herpesviruses, of which one is fatal for Asian elephants (Elephas maximus) and the other for African elephants (Loxodonta africana) The disease syndrome affects predominantly young elephants and has been described in North America, Europe, and Israel. The predominant clinical signs for both species include lethargy, oedematous swellings of the head, neck, and thoracic limbs, oral ulceration, cyanosis of the tongue, and death of most elephants in 1-7 days. Three affected young Asian elephants recovered after a 3-4-week course of therapy with the anti-herpesvirus drug famciclovir. Additional reported herpesvirus-associated lesions in otherwise healthy elephants include localized skin papillomas in African elephants, proliferative vulval lymphoid patches in African elephants, and pulmonary nodules in African elephants. Recent findings suggest that these localized herpesvirus-associated lesions in healthy African elephants may be one source of the herpesvirus that causes disseminated disease and death in the Asian species and the African species. These findings have implications for management practices in facilities keeping both African and Asian elephants and in protecting natural elephant habitats from virulent forms of the virus.

Richman, L.K., Montali, R.J., Cambre, R.C., Schmitt, D., Hardy, D., Bengis, R.G., Hamzeh, F.M., Shahkolahi, A., Hayward, G.S., 2000. Clinical and pathological findings of a newly recognized disease of elephants caused by endotheliotropic herpesviruses. Journal of Wildlife Diseases 36, 1-12.
Abstract: The unique clinical and pathological findings in nine Asian elephants (Elephas maximus) and two African elephants (Loxodonta africana) from North American Zoos with a highly fatal disease caused by novel endotheliotropic herpesviruses are described. Consensus primer polymerase chain reaction combined with sequencing yielded molecular evidence that confirmed the presence of 2 novel but related herpesviruses associated with the disease, one in Asian elephants and the other in African elephants. Disease onset was acute, with lethargy, edema of the head and thoracic limbs, oral ulceration and cyanosis of the tongue followed by death of most animals in 1 to 7 days. Pertinent laboratory findings in 2 of 3 clinically evaluated animals included lymphocytopenia and thrombocytopenia. Two affected young Asian elephants recovered after a 3- to 4-week course of therapy with famciclovir. PM examination in the fatal cases revealed pericardial effusion and extensive petechial hemorrhages in the heart and throughout the peritoneal cavity, hepatomegaly, cyanosis of the tongue, intestinal hemorrhage and ulceration. Histologically, there were extensive microhemorrhages and edema throughout the myocardium and mild, subacute myocarditis. Similar hemorrhagic lesions with inflammation were evident in the tongue, liver and large intestine. Lesions in these target organs were accompanied by amphophilic to basophilic intranuclear viral inclusion bodies in capillary endothelial cells. Transmission electron microscopy of the endothelial inclusion bodies revealed 80 to 92 nm diameter viral capsids consistent with herpesvirus morphology. The short course of the herpesvirus infections, with sudden deaths in all but the 2 surviving elephants, was ascribed to acute cardiac failure attributed to herpesvirus-induced capillary injury with extensive myocardial hemorrhage and edema.

Schmitt, D.L., Hardy, D.A., Montali, R.J., Richman, L.K., Lindsay, W.A., Isaza, R., West, G., 2000. Use of famciclovir for the treatment of endotheliotrophic herpesvirus infections in Asian elephants (Elephas maximus). Journal of Zoo and Wildlife Medicine 31, 518-522.
Abstract: Two juvenile Asian elephants (E. maximus) presented with an acute onset of facial oedema and lethargy. Examination of the oral cavity of each animal revealed cyanosis of the tip and distal margins of the tongue suggestive of endothelial inclusion body disease (EIBD) of elephants. Whole-blood samples were obtained, and polymerase chain reaction tests confirmed the presence of elephant herpesvirus. The animals were administered famciclovir (Flamvir; 500 mg/70 kg body weight, with a loading dose of 1000 mg/70 kg body weight) a potent human anti-herpesvirus drug, in the course of their disease, and recovery followed a treatment regime of 3-4 wk. These are the first known cases of elephants surviving EIBD.

Backues, K.A., Hil, M., Palmenberg, C., Miller, C., Soike, K.F., Aguilar, R., 1999. Genetically engineered Mengo virus vaccination of multiple captive wildlife species. Journal of Wildlife Diiseases 35, 384-387.

Barman, N.N., Sarma, D.K., Das, S., Patgiri, G.P., 1999. Foot-and-mouth disease in wild and semi-domesticated animals of the north-eastern states of India. Indian Journal of Animal Sciences 69, 781-783.
Abstract: The outbreaks (n=23) of foot and mouth disease (FMD) in the northeastern states of India for 14 years (1974 to 1997) were reported. The outbreaks were recorded in 7 different species of wild and semi-domesticated animals. The highest number of outbreaks was recorded in mithun, followed by yak and elephant. Contact with migratory cattle possibly played an important role in the spread of the disease. The morbidity rates in mithun and yak were 22.90 and 24.51%, respectively. About 6.5% of the affected mithun died during the outbreaks. Three FMD cases were recorded in the elephant, and baby elephants were affected with severe erosive lesions in the foot pad and trunk. Sources of infection in elephants were contaminated water and feed. In sambar deer, morbidity and mortality were 35.57 and 10.81%, respectively. In spotted and barking deer, the morbidity was 18.75%. Sources of infection were suspected to be the feed and attendants from nearby villages where FMD outbreaks in cattle was observed. In wild buffaloes, 3 out of 67 were affected and the source of infection was similar to deer. Of the 23 clinical samples typed for the presence of FMD virus, 11 were positive for FMD virus type O, 2 for type A, and each for A22 and Asia.

Burkhardt, S., Hentschke, J., Weiler, H., Ehlers, B., Ochs, A., Walter, J., Wittstatt, U., 1999. Elephant herpes virus - a problem for breeding and housing of elephants. Berliner und Munchener Tierarztliche Wochenschrift. 112, 174-179.
Abstract: Herpesvirus infections which take a fatal turn on African elephants as well as on Asian elephants seem to occur increasingly not only in the USA but also in European stocks. The endotheliotropic herpesvirus causes a rapidly progressing and severe disease which makes any therapeutical effort unsuccessful and finally results in death of the animal, especially in young Asian elephants. As all attempts to culture the virus failed up to now, molecular biological procedures have to be used more often for diagnostic purpose together with the common methods of pathology, virology, and electronmicroscopical evaluation. This is a report on the case of 'KIBA', an eleven year old male elephant at the Zoological Garden Berlin, infected with the endotheliotropic elephants herpesvirus. 'KIBA' was born at the Zoo in Houston, Texas, and raised within his herd. Upon arriving in Berlin in November 1997 he adapted to the new premises and climate and new social circumstances without any problems. In June 1998 he already serviced three females of his new herd several times. In August 1998 he died after passing a peracute progression of the disease after residenting in Berlin for only 9 months. The dissection of the animal revealed some evidence on an agent damaging the endothelium. Major signs indicating this agent were bleedings in several serous membranes, mucosa and on the right atrium, as well as other parts of the myocardium. Furthermore there have been ulcerations at various localizations of the whole digestive tract. Slightly basophilic intranuclear inclusion bodies have been found histologically in endothelial cells of different organ samples. An examination of altered organ-material by electronmicroscopy made some herpesvirus-like particles visible. A virological investigation first revealed evidence of giant cell formations with solitary basophilic intranuclear inclusion bodies in different cell cultures, however, without any distinct cytopathogenic effect. Supported by molecular biological procedures the infection of 'KIBA' could be verified as the elephants herpesvirus. By means of PCR and subsequent sequence analysis a DNA-sequence typical for the elephants herpesvirus could be obtained which showed an identity of 97% with the terminase sequence of the elephant herpesvirus described by American authors. The deduced amino acid-sequences were 100% identical. To the terminase of the human cytomegalovirus, the elephant sequence had an identity of 53% (similarity: 74%). Based on the cooperation of ILAT, Institute of Veterinary-Pathology/Free University Berlin, Robert-Koch-Institut Berlin, and Zoological Garden Berlin, the cause of 'KIBA's' death could be discovered immediately. Possible implications of this case especially on breeding and keeping elephants are discussed briefly.

Ferber, D., 1999. Virus suspect identified in elephant deaths. Science 283, 1093-1094.

Kuntze, A., 1999. Poxvirus infections in elephants. In: Fowler, M.E., Miller, R.E. (Eds.), Zoo and Wild Animal Medicine: Current Therapy 4. W.B. Saunders, Philadelphia, PA, USA, pp. 547-550.

Meyer, H., Schay, C., Mahnel, H., Pfeffer, M., 1999. Characterization of orthopoxviruses isolated from man and animals in Germany. Archives of Virology 144, 491-501.
Abstract: Fourteen orthopoxvirus strains isolated from humans, cats, a dog, a cow, and an elephant in Germany between 1985 and 1991 were characterized. All were classified as cowpox virus based on haemorrhagic lesions induced on the chorioallantoic membrane of chicken eggs and reactivity of a 160 kDa protein with anti-A-type inclusion protein hyperimmune serum in a Western blot. More detailed comparison of the isolates by restriction endonuclease mapping using HindIII and XhoI demonstrated a close relationship between all isolates and confirmed them as cowpox viruses. Some minor differences between the isolates were detected which proved to be of epidemiological value. One group consisting of 5 closely related isolates contained a unique 4.0 kb HindIII fragment. In a Southern blot this fragment failed to hybridize with other cowpox virus isolates including the reference strain.

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

Richman, L.K., Montali, R.J., Garber, R.L., Kennedy, M.A., Lehnhardt, J., Hildebrandt, T., Schmitt, D., Hardy, D., Alcendor, D.J., Hayward, G.S., 1999. Novel endotheliotropic herpesviruses fatal for Asian and African elephants. Science 283, 1171-1176.
Abstract: A highly fatal haemorrhagic disease was identified in 10 young Asian (Elephas maximus) and African (Loxodonta africana) elephants at zoos in the USA between 1983 and 1997. In the affected animals there was ultrastructural evidence for herpesvirus-like particles in endothelial cells of the heart, liver, and tongue. Consensus primer polymerase chain reaction combined with sequencing yielded molecular evidence that confirmed the presence of 2 novel but related herpesviruses associated with the disease, one in Asian elephants and another in African elephants. Otherwise healthy African elephants with external herpetic lesions yielded herpesvirus sequences identical to that found in Asian elephants with endothelial disease. It is suggested that the Asian elephant deaths were caused by cross-species infection with a herpesvirus that is naturally latent in, but normally not lethal to, African elephants. A reciprocal relationship may exist for the African elephant disease.

Richman, L.K., Montali, R.J., Cambre, R.C., Schmitt, D., Hardy, D. Clinical and pathologic aspects of a fatal herpesvirus disease in Asian (Elephas maximus) and African (Loxodonta africana) elephants. Proceedings of the American Association of Zoo Veterinarians.  263-266. 1999. 10-9-1999.
Ref Type: Conference Proceeding

Richman, L.K., Montali, R.J., Hildebrandt, T., Fickel, J., Schmitt, D., Hayward, G.S. Status of a new, fatal herpesvirus disease in elephants in North America and Europe.  Verhandlangsbericht des 39 International Symposium uben Erkrankungen der Zoo und Wildtiere, Wien. 39:17-21. Verh.ber Erkrg. Zootiere 39.  17-21. 1999.
Ref Type: Conference Proceeding

Wimalaratne, O., Kodikara, D.S., 1999. First reported case of elephant rabies in Sri Lanka. Veterinary Record 144, 98.
Abstract: An 84-year-old female domesticated elephant presented with a 4-day history of lethargy. Appetite and water intake was normal but the following day she was unsteady, aggressive and restless. There were secretions from both temporal glands. On the sixth day she was completely anorectic, had developed paralysis of the trunk and was unable to stand, falling each time she tried to stand up, and she was noticed to be blind. She died on the ninth day after the first symptoms were observed. PM examination showed the brain to be more vascular than normal and a brain smear was positive for rabies antigen. A serum sample went to the WHO Collaborating Center for Rabies in Bangkok, Thailand, which determined a rabies virus neutralizing antibody titre of 0.68 IU/ml by the rapid fluorescent focus inhibition test. Antigenic typing and genetic sequencing showed the virus to be similar, but not identical, to the common Sri Lankan dog rabies variant, although there was no history of an animal bite to the elephant.

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.

Hunter, P., Swanepoel, S.P., Esterhuysen, J.J., Raath, J.P., Bengis, R.G., Van der Lugt, J.J., 1998. The efficacy of an experimental oil-adjuvanted encephalomyocarditis vaccine in elephants, mice and pigs. Vaccine 16, 55-61.
Abstract: An oil-adjuvanted inactivated encephalomyocarditis (EMC) vaccine was developed to protect a wild population of elephants against a natural outbreak of disease. The experimental vaccine was initially tested for efficacy by challenging mice and pigs. Mice showed protection against challenge and pigs developed high antibody levels. Since both vaccinated and control pigs failed to develop clinical disease, apparently due to the low virulence of the strain in this species, protection in pigs could not be evaluated. Three wild elephants and 12 captive elephant calves given the vaccine developed high antibody titres. All of the captive elephants were protected from a challenge 2 months after vaccination, whereas 6 controls (not vaccinated but challenged) developed fatal or sub-clinical myocarditis. This is apparently the first report of an inactivated EMC vaccine inducing high antibody titres in domestic and wild animals. Due to the potency of this vaccine and the acceptability of the oil adjuvant used, it has potential for use in animals in zoological collections as well as in the pig industry.

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

Schmitt, D.L., Hardy, D.A., 1998. Use of famciclovir for the treatment of herpesvirus in an Asian elephant. Journal of the Elephant Managers' Association 9, 103-104.

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.

Backues, K.A., Aguilar, R.A., Hill, M., Palemberg, A.C. A new modified live virus vaccine for encephalomyocarditis (EMC) virus protection, preliminary trials at the Audubon Zoo. Proc. Amer.Assoc. of Zoo Vet.  166-167. 1997.
Ref Type: Conference Proceeding

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., 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.

Bhat, M.N., Manickam, R., 1997. Foot and mouth disease virus infection associated (VIA) antibodies in wild herbivores. Indian Veterinary Journal 74, 827-820.

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

Cambre, R.C., Buick, W.W., 1996. Special challenges of maintaining wild animals in captivity in North America. Rev Sci Tech 15, 251-266.
Abstract: The maintenance of wild animals in captivity in North America is regulated by a number of different laws and government agencies in each country. Member institutions of zoo and aquarium associations in Canada, the United States of America and Mexico experience an extra tier of regulation in the form of industry standards, which are sometimes stricter than those imposed by government. Climate, natural disasters and harmful pest species all contribute to the challenge of keeping animals in certain locales. Vigilance against zoonotic disease transmission is maintained through industry and government-mandated sanitation standards, which are fortified by reporting regulations of local, regional and Federal health agencies. Current controversies in the keeping of particular taxa in North America include the threat to non-human primate breeding programmes precipitated by strict new import regulations, the fear of herpesvirus B infection, and commercial airline transport bans. Successive human fatalities among elephant handlers have prompted the industry and governments to re-examine the manner in which these potentially dangerous creatures are maintained in captivity.

Emerson, C.L., Wagner, J.L., 1996. Antibody responses to two encephalomyocarditis virus vaccines in rhesus macaques (Macaca mulatta). J Med Primatol 25, 42-45.
Abstract: Two groups of rhesus macaques (Macaca mulatta) housed in rodent-controlled outdoor corrals were inoculated with two different encephalomyocarditis virus (EMCV) vaccines. One group (n = 45) received a vaccine made from an inactivated field isolate of virus cultured during an outbreak at a zoo in Florida. This vaccine produced fourfold increases in the titers of 28 animals (62%); the increases persisted for at least 18 months (last test) after a single injection of the vaccine. The other group (n = 51) received a vaccine made from an inactivated porcine field strain of the virus. This vaccine did not produce titers in any of the vaccinees.

Osorio, J.E., Hubbard, G.B., Soike, K.F., Girards, S., van der Werf, S., Moulin, J., Palmenberg, A.C., 1996. Protection of non-murine mammals against encephalomyocarditis virus using a genetically engineered Mengo virus. Vaccine 14, 155-161.

Richman, L.K., Montali, R.J., Cambre, R.C., Lehnhardt, J., Kennedy, M., Kania, S., Potgieter, L. Endothelial inclusion body disease:  a newly recognized fatal herpes-like infection in Asian elephants. Proceedings American Association of Zoo Veterinarians.  483-486. 1996.
Ref Type: Conference Proceeding

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.

Chandrasekharan, K., Radhakrishnan, K., Cheeran, J.V., Nair, K.N.M., Prabhakaran, T., 1995. Review of the Incidence, Etiology and Control of Common Diseases of Asian Elephants with Special Reference to Kerala. In: Daniel, J.C. (Ed.), A Week with Elephants; Proceedings of the International Seminar on Asian Elephants. Bombay Natural History Society; Oxford University Press, Bombay, India, pp. 439-449.
Abstract: Incidence, etiology, symptoms and control of specific and non-specific diseases of captive and wild elephants have been reviewed. Asian elephants have been observed to be susceptible to various parasitic diseases such as helminthiasis, trypanosomiasis and ectoparasitic infestations, bacterial diseases such as tetanus, tuberculosis, haemorrhagic septicemia, salmonellosis and anthrax, viral diseases such as foot and mouth disease, pox and rabies and non-specific diseases like impaction of colon, foot rot and corneal opacity. A detailed study extending over two decades on captive and wild elephants in Kerala, revealed high incidence of helminthiasis (285), ectoparasitic infestation (235), impaction of colon (169) and foot rot (125). Diseases such as trypanosomiasis (21), tetanus (8), tuberculosis (5) pox (2) and anthrax (1) were also encountered. The line of treatment against the diseases mentioned, have been discussed in detail.

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.

Raath, J.P., Bengis, R.G. The evaluation of a vaccine against encephalomyocarditis infection in elephants (Loxodonta africana) under controlled conditions. Proceedings, American Association of Zoo Veterinarians, Wildlife Disease Association, American Association of Wildlife Veterinarians. Joint Conference, East Lansing, Michigan, August 12-17, 1995.  304-308. 1995.
Ref Type: Conference Proceeding
Abstract: Encephalomyocarditis killed 64 elephants in the Kruger National Park between October 1993 and November 1994. An inactivated vaccine was inoculated into 14 elephants aged 6-8 years, 6 of which were challenged later with virulent virus. 3 of 4 infected, unvaccinated elephants developed the clinical disease and 2 died. Vaccinated elephants developed antibodies at 1-4 weeks after vaccination, and the 6 challenged animals remained healthy.

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.

Meiswinkel, R., Braack, L.E.O., 1994. African horsesickness epidemiology:  five species of Culicoides (Diptera: Ceratopogonidae) collected live behind the ears and at the dung of the African elephant in the Kruger National Park, South Africa. Oderstepoort Journal of Veterinary Research 61, 155-170.
Abstract: During the culling of elephants (Loxodonta africana) at 5 sites in the Kruger National Park, South Africa, 682 Culicoides of 5 species of the subgenus Avaritia were found either living behind the ears of elephants or attracted to the freshly disemboweled intestinal dung of elephants. The species were Culicoides tororoensis, C. kanagai, C. loxodontis, and 2 undescribed species, Culicoides sp. £50 and Culicoides sp. £54 pale form (p.f.). Of 511 female midges found behind ears, 39.9% were nulliparous, 57.3% empty parous, 2.5% freshly bloodfed and 0.2% gravid. The age composition of this subpopulation indicates that the Culicoides were behind the ears to suck blood and, furthermore, would do so in broad daylight. The age composition of 171 Culicoides of 3 species attracted to dung was entirely different: 1.8% nulliparous, 14.6% empty parous, and 83.0% gravid, indicating that the great majority of midges captured at dung were about to oviposit or had just oviposited. Immediately after culling, light traps were operated at 2 of the sites. Of 4023 Culicoides of 21 species captured, 93% were of the same 5 species found on the ears and at the dung of elephants. Using these and other unpublished data pertaining to the rearing of these 5 Avaritia species from elephant dung over the past 7 years, the life cycle of these Culicoides is broadly sketched, the first for any Afrotropical species of the genus. The implications that the close association between elephant and midge has for the dispersal and geographic distribution of the latter, and how it may influence the involvement of midges in the transmission of diseases such as African horse sickness, are also discussed. Owing to difficulties in identifying species of the subgenus Avaritia in the Afrotropical Region, the taxonomy of each of the 5 above-mentioned species is briefly appraised. Of the remaining 16 species (7%) captured in light traps, 15 (6%) belong to that sector of the genus Culicoides whose immature stages develop in groundwater habitats and include C. imicola, which comprised only 2% of the light-trap collections. The large disparity in the adult abundance patterns of the "dung" and "groundwater" species in the middle of dry bushveld is probably the result of differences in host and larval habitat preferences, and is briefly discussed. Finally, the few reports extant on the wild-host preferences of Afrotropical Culicoides are reviewed.

Meiswinkel, R. Six species easily confused with Culicoides imicola in Africa: notes on their life histories. Foot and mouth disease, African horse sickness and contagious bovine pleuropneumonia: OIE Scientific Conference, Gaborone, Botswana, 20-23 April, 1994: summaries and conclusions.  45-46. 1994. Paris; France, Office International des Epizooties.
Ref Type: Conference Proceeding
Abstract: Studies have shown that there are at least 6 species related to, and easily confused with, C. imicola. These together form the imicola group within the subgenus Avaritia. Five of these species have only very recently been described or remain undescribed. Each of these species has a unique life history. Distinct larval habitats and differences in adult prevalence, abundance, host preference and geographic distribution have an important bearing on their potential to transmit AHS virus. Three of the new imicola group species live exclusively in the dung of the elephant, rhino, zebra, buffalo and wildebeest. In certain places and seasons these Avaritia species are far commoner than C. imicola. The risk from translocation of unvaccinated wild herbivores (potential virus reservoirs) into the farming land is discussed, with particular reference to the establishment of some of these species in dung of cattle.

Mikota, S.K., Sargent, E.L., Ranglack, G.S., 1994. Medical Management of the Elephant. Indira Publishing House, West Bloomfield MI.

Sharma, D.K., Islam, S., Hazarika, A.K., 1994. Foot and mouth disease in a baby elephant (Elephas maximus). Indian Journal of Veterinary Pathology 18, 55-56.

Bengis, R., 1993. Care of the African elephant Loxodonta africana in captivity. The capture and care manual : capture, care, accommodation and transportation of wild African animals. Pretoria : Wildlife Decision Support Services : South African Veterinary  Foundation, Pretoria, pp. 506-511.

Berry, H.H., 1993. Surveillance and control of anthrax and rabies in wild herbivores and carnivores in Namibia. Revue Scientifique et Technique Office International des Epizooties 12, 137-146.
Abstract: Anthrax has been studied intensively in Etosha National Park, Namibia since 1966; in addition, since 1975, mortality due to rabies and all other causes has been recorded, totaling 6190 deaths. Standard diagnostic procedures demonstrated that at least 811 deaths (13%) were due to anthrax and 115 deaths (2%) were caused by rabies. Of the total number of deaths due to anthrax, 97% occurred in zebra (Equus burchelli), elephant (Loxodonta africana), wildebeest (Connochaetes taurinus) and springbok (Antidorcas marsupialis) while 96% of rabies deaths occurred in kudu (Tragelaphus strepsiceros), jackal (Canis mesomelas), bat-eared fox (Otocyon megalotis) and lion (Panthera leo). Anthrax deaths were highest in the rainy season for zebra, wildebeest and springbok, while elephant mortality peaked during dry seasons. No statistical relationship existed between seasonal rainfall and overall incidence of either anthrax or rabies. Control of anthrax is limited to prophylactic inoculation when rare or endangered species are threatened. Incineration of anthrax carcasses and chemical disinfection of drinking water are not feasible at Etosha. Rabies control consists of the destruction of rabid animals and incineration of their carcasses when possible.

Ebedes, H., 1993. The use of long-acting tranquilizers in captive wild animals. The capture and care manual : capture, care, accommodation and transportation of wild African animals. Pretoria : Wildlife Decision Support Services : South African Veterinary  Foundation, Pretoria.

Prins, H.H.T., Van-der-Jeugd, H.P., 1993. Herbivore population crashes and woodland structure in East Africa. Journal of Ecology Oxford 81, 305-314.
Abstract: From 1985 to 1991, bush encroachment was serious in Lake Manyara National Park, northern Tanzania. Shrub cover increased by about 20%. The increase was independent of initial (1985) shrub cover. Since 1987 there has been a steep decline in the number of African elephant (Loxodonta africana) in the Park due to poaching. Elephant density decreased from about 6 per km2 to about 1 per km2. However, shrub establishment, as determined from counting tree-rings, preceded poaching. Shrub establishment in two areas of the Park coincided with anthrax epidemics that drastically reduced the impala [Aepyceros melampus] population. In the northern section of the Park this occurred in 1984, in the southern section in 1977. The diameter increment of Acacia tortilis was 5.24 mm/yr, irrespective of the size of the trees. Size measurements indicated that an even-aged stand of A. tortilis established in 1961, which coincided with another anthrax outbreak among impala. Size measurements of old A. tortilis trees indicated that another even-aged stand established at the end of the 1880s. The size of trees of this stand was not significantly different from a stand in Tarangire National Park, nor from a stand near Ndutu (on the boundary between Serengeti National Park and Ngorongoro Conservation Area), also in northern Tanzania. All three stands are likely to have originated from bush establishment caused by the rinderpest pandemic at the end of the 1880s. It is suggested that seedling establishment of A. tortilis is a rare event under the prevailing conditions of high browsing pressures by ungulates such as impala. Punctuated disturbances by epidemics among these ungulates create narrow windows for seedling establishment, which may explain the occurrence of even-aged stands.

Arora, B.M., 1992. An overview of infectious diseases and neoplasms of the elephants (Elephas maximus) in India. In: Silas, E.G., Nair, M.K., Nirmalan, G. (Eds.), The Asian Elephant: Ecology, Biology, Diseases, Conservation and Management (Proceedings of the National Symposium on the Asian Elephant held at the Kerala Agricultural University, Trichur, India, January 1989). Kerala Agricultural University, Trichur, India, pp. 159-161.

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.

Chakraborty, T., Majumdar, B.K., 1992. Foot and mouth disease in captive 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.

Chandrasekharan, K., 1992. Prevalence of infectious diseases in elephants in Kerala and their treatment. 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. 148-155.

Jacoby, F. Contribution to the epidemiology of cowpox virus in the Federal Republic of Germany. Untersuchungen zur Epidemiologie des Kuhpockenvirus in der Bundesrepublik Deutschland.  1-140. 1992. Giessen, Germany, Fachbereich Veterinarmedizin, Justus-Liebig-Universitat.
Ref Type: Thesis/Dissertation
Abstract: The indirect immunofluorescence test for antibodies to cowpox orthopoxvirus was positive in 218 of 303 wild rodents (Microtus agrestis, M. arvalis, Apodemus flavicollis, Clethrionomys glareolus and Rattus norvegicus). Attempts to isolate the virus failed. 202 of 277 cats from 58 of 67 locations in Germany, also 61 of 106 cattle and 13 of 38 zoo or circus elephants were also positive.

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.

Pilaski, J., Kulka, D., Neuschulz, N., Ippen, R.ed., Schroder, H.D. Outbreak of pox among African elephants in Thuringer Zoo at Erfurt, Germany. Erkrankungen der Zootiere. Verhandlungsbericht des 34. Internationalen Symposiums uber die Erkrankungen der Zoo- und Wildtiere, Santander-Spain 1992.  111-118. 1992.
Ref Type: Conference Proceeding

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).

Brahmasa, A., 1991. Cryosurgery of cutaneous papilloma in two Asian elephants: a case report. Thai Journal of Veterinary Medicine 21, 151-159.

Maity, P.K., Nandi, B., Chatterjee, U., Sarkar, C.R., Mazumdar, R., Bhattacharya, A.K., 1991. Foot and mouth disease transmitted to elephants (Elephas maximus) from infected cattle. Indian J Comp Micro Immunol Infect Dis 12, 68-78.

Chakraborty, T., Majumdar, B.K., 1990. Foot and mouth disease in an elephant at Calcutta zoological garden: a case report. Indian Veterinary Medical Journal 14, 213-214.

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.

Ossent, P., Guscetti, F., Metzler, A.E., Lang, E.M., Rubel, A., Hauser, B., 1990. Acute and fatal herpesvirus infection in a young Asian elephant (Elephas maximus). Vet. Pathol. 27, 131-133.
Abstract: Infections with herpesvirus may cause papillomatous lesions in the Asian and African elephant.  In both species, the virus has been reported to localize only in the skin.  Disseminated nodules of epithelial cells were found in the lungs of a high percentage of wild African elephants.  In these cases, the proliferated cells contained intranuclear inclusion bodies in which herpesvirus particles were observed by electron microscopy.  The virus in those cases caused no illness.  This report documents the necropsy findings of a juvenile Asian elephant dying peracutely from massive generalized hemorrhage due to lesisons in the endothelial cells of the capillaries.  The cell nuclei frequently contained inclusion bodies in which herpesvirus particles were demonstrated.  This has not been described in elephants before.

Pade, K., Ruedi, D., Pilaski, J., Heldstab, A., Muller, M. Lethal outbreak of pox among five Asian elephants of a German travelling circus. Erkrankungen der Zootiere. Verhandlungsbericht des 32. Internationalen Symposiums uber die Erkrankungen der Zoo und Wildtiere vom 23. Mai bis 27. Mai 1990 in Eskilstuna. Erkrankungen der Zootiere. Verhandlungsbericht des 32. Internationalen Symposiums uber die Erkrankungen der Zoo- und Wildtiere vom 23. Mai bis 27. Mai 1990 in Eskilstuna , 147-155. 1990. Berlin, German Democratic Republic, Akademie Verlag.
Ref Type: Conference Proceeding

Pilaski, J., 1990. Pox in European zoos. Foreign Animal Disease Report 18, 6-7.

Sironi, G., Caniatti, M., Caniatti, M., 1990. Immunohistochemical detection of papillomavirus structural antigens in animal hyperplastic and neoplastic epithelial lesions. Journal of Veterinary Medicine Series A 37 , 760-770.
Abstract: One hundred and seventy two hyperplastic and neoplastic epithelial lesions from 8 different mammalian and 1 avian species were tested with an immunohistochemical technique to detect papillomavirus structural antigens. Selected lesions were diagnosed histologically as papilloma, fibropapilloma, equine sarcoid, squamous cell carcinoma, basalioma, epulis, keratoacanthoma, trichoepithelioma, pilomatrixoma, epidermal inclusion cyst, and hyperkeratotic or acanthotic epidermal lesions. Positive nuclear staining was detected in 14 out of 23 papillomas, 8 out of 32 fibropapillomas and in 1 out of 3 hyperplastic epidermal lesions. Positive samples were found in 5 of 8 mammalian species. Selected samples were also examined by transmission electron microscopy. In 4 samples papillomavirus was seen. In two other samples, negative with immunoperoxidase technique, papovavirus-like particles were observed.

Hegel, G.V., Hanichen, T., Mahnel, H., Wiesner, H., 1989. Warts (papilloma/sarcoid) in elephant. Erkrankungen der Zootiere 31, 201-205.
Abstract: Warts ( Papilloma, Sarcoid) in  Elephants  ( Hegel,G.)1989; translated from German by Gerda Martin. Papilloma virus - from the group Papova virus - is considered  an etiological agents of wart- like skin changes in cattle, sheep, mountain goat, and rabbit. (ROSENBERGER,1970; ROLLE and MAYR, 1984). Equine sarcoid (PALMER. 1985) found in horses is most likely caused by bovine papilloma virus. The alternate name is based on clinical and morphological differences in the actual papilloma. In the initial stage, the sarcoid is similar to that of the papilloma; however in later stages, tumorous decay on the surface of the epidermis, and proliferation of the mesenchymal part of the tumor in the subcutis dominate (DIET and WIESNER, 1982). Wart- like changes in the skin of elephants as described by PILASKI et al (1987, 1988), proved to be caused by Herpes virus.  Such skin changes in elephants are not rare and require treatment since size and volume of the excrescences  may cause functional disturbances in the patient. Even if the animal's  general   well being  is not impaired, the importance of esthetics and hygiene should not be disregarded in a place where there are spectators and visitors (zoo, circus). The following paper reports findings of wart- like skin changes in elephants. Observations and Therapy In the elephants  kept in the Hellabrunn  Zoo, no case of papilloma or similar skin tumors had occurred since 1972. First case: In 5-28 - 1987, a ca. 18 month old female L.a. named " Sabi" arrived In Hellabrunn. This animal had a wart- like thickening of 1 cm at the dorsal end of the trunk. After 8 weeks, more of those such skin changes appeared on trunk and lower lip without  impairment in general well being. Treatment consisted of  one daily, subcutaneous injection of 1 amp. Chelidonium D7 (DHU Chelidonium majus L.), and application of fresh ??Schoellkraut  juice dabbed onto the warts but was unsuccessful.  After a change of treatment was made: 10 drops of Thuja D4 (DHU Thuja occidentalis L) and 20 drops Acidum nitricum D12 (DHU Acidum nitricum), orally, once a day, at separate times of the day, there remained, after 2 weeks, a wart on the lower lip the size of a cherry pit, and the before mentioned  wart on the dorsal end of  the trunk had now grown to the size of a cherry. Even the strength of Thuja LM 6 (DHU Thuja occidentalis L.)  20 drops, oral, the growth of the wart on the dorsal end of the trunk, now with a diameter of 5 cm, could not be stopped: Exstirpation had to be performed. Frequent  sucking had promoted strong ulceration.  A secondary infection  had set in,  the surface showed granular  tissue exuding blood and pus.

On 10 -  6 -  1987 the growth was exstirpated and tissue was sent for virolog. and histolog. examination. In addition, tissue was removed from a fresh small wart for vaccine. During the operation the animal was immobilized (anesthetic: 0.3 ml Immobilon* (large animal Immobilon Rc* - Vet. Ltd.),  10mg Xylacin, 150 IE Hyaluronidase i. m.). There were no complications during recovery. Two  weeks post op., the first vaccination was given, followed by a second vacc. four weeks there after, of 5.0 ml, subcut.., of an auto vaccine developed by the Institute for Medical Microbiology, Dept. of Infectious and Epidemic Medicine. In February 1988, there occurred another bout with wart- like growth on the ventral part of the trunk, lower jaw, shoulders and feet, some with a diameter of 15 mm.  From the sedated young animal tissue was taken from several newly grown warts for the manufacture of auto vaccine (sedation: "Hellabrunner Mischung" / 150 IE Hyaluronidase). After 10 days, the first vaccination was given, and by the time of the second vacc." Sabi" was free of externally visible skin changes.

On 6. 6. 1988, "Sabi" fell ill again. Over night she was covered with 48 warts,  with  diameters from 2mm - 15 mm on trunk and head, and 10 more on the chest.The attempt to "ice"  the warts with liquid nitrogen was not successful. Instead, coagulation of ca. 20 of the larger warts was used. The monopolar coagulation electrode of the Erbotom F 2 (Erbe Elektromedizin) coagulates reaching deeply  into the healthy zone of the surrounding tissue. As before, tissue for the manufacture of the auto vaccine was taken, as well as  0.5 ml of blood from the ear vein for the manufacture of a "own- blood"  nosode. (Large animal, premedication: 20 mg Xylazin i.m., 20 minutes later : 0.5 ml Immobilon R (large animal Immobilon R c - Vet Ltd.) and 150 IE Hyaluronidase i.m. The following day, "Sabi" was given the "own- blood" nosode at a strength of C5 (20 drops daily).In addition, she was vacc. once again. Since "Sabi" was free of warts at the time of the second vaccination -   given 4 weeks after the first - the "own- blood" treatment was discontinued. Shortly there after, however, several new warts cropped up (diameter ca. 1 cm), so that the "own- blood"  treatments were continued. Since that time "Sabi" has had no recurrences.Second case : The Indian elephant cow (E. maximus) , named "Dirndl" , age ca. 22 years, had been kept in the box next to "Sabi" since "Sabi's" arrival. They kept trunk contact. On 5-2-1988, "Dirndl" showed on the distal trunk a substantially increased raised area ca. 2 x 2 cm oozing blood. It seemed to be an injury from a metal rope used in off limiting. The wound was disinfected and treated twice a day with chloromycetin spray with Gentian violetR (Parke Davis).  After  one week  the growth had increased substantially  and on the surface,  it had a cauliflower-like ulcerated  appearance.Upon light touch or movement of the dorsal  trunk, blood appeard spontaneously.   Four days later, the growth was exstirpated, while the animal was standing. (Sedation: 2.2 ml Hellabrunn mixture / 150 Hyaluronidase i. m.) . The attempt to close the skin of the trunk over the wound failed because the tension in that area was too great.  The surface of the wound was cauterized and treated with ChloromycetinSpray with Gentian violet R (Parke Davis). Tissue for pathological and histological examination was sent out.  One week after the operation,  the area of the wound was  highly swollem and the wound was infected. Treatment: Several times a day, an  ablution with a 0.1 % Rivanol solutionnR (Asid - 2 Aethoxy-6.9-diamin  acridinlactat)  and application of Sulfonamid-Codliver oil salve (WDT = Sulfadimidin- Sodium- cod liver oil).  In addition, analogous to "Sabi" , once daily 20 drops of "own-blood" nosode,  potency C 5  given orally. Three weeks post. op., there could be clearly distinguished a limited relapse, an area of  6 x 9 cm rising  ca. 2 cm  above the healthy skin of the trunk. The surface looked like the first growth.  It was extirpated under general anesthetic  (Premed.:80 mg Xylazin i. m., 20 min. later: 1.8 ml ImmobilionR and 150 IE Hyaluronidase). In addition, the whole wound was coagulated  by monopolar coagulation electrode as above. Daily for 4 weeks, the wound  was brushed with a 1:5 wood tar -alcohol - solution.There were no complications during recovery. After 5 weeks , all that could be seen was a ca. 1.5 cm long small scar on the skin of the trunk.

Histomorphological Findings:  Fixation with formalin, embedding in paraffin; stain: Hemalaun-Eosin, connective tissue stain  in the manner of Masson. The histomorphological  findings based on the tissue samples of "Sabi" and "Dirndl"  are the  same, and agree  with the findings of 3 other skin tumor tissue taken from elephants of different origin (tab. 1). The tumors  consist mainly of fibroplastic cells  with more or less  abundant collagen fibers and blood vessels. The boundaries from the adjacent corium and lower skin is largely indistinct.  In all larger neoplasties , the covering epidermis has been preserved  at margins only due to superficial ulceration.  Here the P. acuta aseptica diffusa borders  are irregular and strongly profiled, the epithelium is acanthoid and hyperkeratotic. The nuclei of tumor cells  are considerably anisomorphic, some have gigantic nuclei. Mitosis is frequent. Due to the ulcerated epidermis , there is deep infiltration with infectious cells. Virological findings: From the extirpated tissue taken from the African Elephant "Sabi" ca. 3 g was homogenated, in addition, the cells were "opened" by defrosting and ultrasound, and the "cleared" tissue suspension was analyzed  for free virus particles after concentration and negative-contrasting with  electron microscopy .  At the same time, small tissue samples of 2 mm  from deeper epidermis layers  were fixed as usual for the ultrahistological exam , embedded in epoxy  resin, and ultra thin slices were scanned by the electron microscope. No papilloma virus was found  in the concentrated, cell free tissue extract or  the ultrathin slices of tissue samples .No virus particle of  any kind was found.

Discussion
To  show papilloma by culturing cannot be done since  no species of this genus can  be propagated  in cell cultures  with the exception of its original host. The failed  attempt to prove their presence with the electronmicroscope does not exclude a papilloma virus etiology in tumors. When virus particles are viewed in higher concentrations, the electron microscopic  proof is successful. Using ultrahistologyical methods the particles in cell nuclei can only be found when the few cells  of specific skin cells  are in the virus propagation stage. In the case of virally induced papilloma however, a true virus propagation is not necessary. In the last few years, it was found that equine sarcoid  can be caused  by bovine papilloma virus. But it was only the genome of the virus which could be isolated by means of gene technology (ALTMANN, 1980; HAUSEN, 1980); the virus itself could not.  The oncogenetic potency of the virus in heterologic hosts , without true virus production, has been established.  A broader spectrum of hosts  for , at least , the papilloma virus in cattle seems to be the case.  And a bovine papilloma induced skin fibromatose in  (a) horse has been reported (LANCASTER, 1979). This virus can also appear in wild 'cud chewers, perhaps even carnivores. It is in part also related to the human papilloma virus. The possibility of transfer to humans (LANCASTER 1982) as well as other mammals such as elephants  has not been proved but is probable. In comparing the histological findings of the 5 skin growths with those of the viral fibropapilloma in cattle and horse (called equine sarcoid here), the relative immaturity of the tumorous tissue is evident. It compares to the so- called sarcoid in horses. The sarcoid-like structure and the indistinct  separation from healthy tissue  speaks for a virus etiology and  morphologically a relapse can be expected. This occurred in both of the clinically described cases.

A differential diagnosis excludes a Herpes virus infection, as described by PILASKI et al. (1987, 1988) in elephants on the basis of different histological findings. Inclusions could not be found in any of the cases. The warts on the elephants were clinically similar to the well known sarcoids in horses (DIETZ and WIESNER, 1982). The two sick animals were in "trunk contact" occupying adjacent boxes. Almost one year after the arrival of  "Sabi" who had warts, "Dirndl" fell sick. That points to the infectious nature of warts. The relapse after the first operation on "Dirndl" suggests that the extirpation of the growths was not complete. This may be related to the fact that the animal was standing  and only sedated. In contrast , the extirpation of the "relapse" was carried out on a fully immobilized animal and with the use of the Erbotom F 2  for coagulation including the adjacent tissue.  We know of various 'wart therapies' in human medicine with differing success. The various treatments employed in the one and one half years of "Sabi's" illness can be labled neither successful, nor unsuccessful. The use of auto vaccine which is analogous  to a "stable specific " vaccine in the treatment of papilloma in cattle, could  perhaps have triggered the recurrence of warts at the  conclusion of the vaccination treatments. That would favor the etiology of a virus 'picture.' The influence of the 'burn' or extirpation of a single or more growths which returned, in the surrounding growths cannot be determined. It remains inconclusive if the use of the "own- blood" nosode C 5 aided the successful therapy , since the necessity  to fight a recurrence had not yet occurred.

Kuntze, A., 1989. Dermatopathies in elephants and their treatment. Kleintierpraxis 34, 405-415.

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.

von Hegel, G., Hanichen, T., Mahnel, H., Wiesner, H. Warts (papilloma/sarcoid) in two elephants. Erkrankungen der Zootiere. Verhandlungsbericht des 31. Internationalen Symposiums uber die Erkrankungen der Zoo- und Wildtiere, Dortmund 1989.  201-205. 1989. Berlin, German Democratic Republic, Akademie-Verlag.
Ref Type: Conference Proceeding

Wells, S.K., Gutter, A.E., Soike, K.F., Baskin, G.B., 1989. Encephalomyocarditis virus: Epizootic in a zoological collection. Journal of Zoo and Wildlife Medicine 20, 291-296.
Abstract: Encephalomyocarditis virus (EMCV) was isolated from eight nonhuman primates, one Thomson's gazelle (Gazella thomsoni), and one dromedary camel (Camelus dromedarius) that died peracutely between January 1985 and October 1987 at Audubon Park Zoo, New Orleans, Louisiana.  Gross pathology consisted of excessive pericardial fluid, epicardial hemorrhages, and pale foci within the myocardium.  Microscopic changes included myofiber necrosis, edema, and mononuclear cell infiltration within the myocardium.      Anti-EMCV antibody was found in a variety of species including a capybara (Hydrochoerus hydrochaeris), which subsequently died of a necrotizing myocarditis but from which virus was not isolated.  Although one hospital staff member had a high anti-EMCV antibody titer, all primate keepers were seronegative.      Encephalomyocarditis virus was recovered from 38 wild rodents, one opposum (Didelphis virginiana), and one rabbit (Sylvilagus sp.) collected on the zoo grounds.  Fifty-five percent of the positive samples were found in areas where confirmed deaths had occurred or antibody-positive animals were housed.  A killed vaccine was developed and administered to six domestic cats, 12 primates, and one camel.  Antibody response to vaccination was variable.

Gaskin, J.M. Encephalomyocarditis: A potentially fatal virus infection of elephants. Proc.Ann.Elephant Workshop 9.  133-136. 1988.
Ref Type: Conference Proceeding

Pilaski, J., Rosen-Wolff, A., 1988. Poxvirus infection in zoo-kept mammals. In: Darai, G. (Ed.), Virus Diseases in Laboratory and Captive Animals pp. 83-100.

Pilaski, J., Hentscheke, J., Sinn, D., Francke, R., Rosenbruch, M., Olberding, P., Molle, G., 1988. Two virus diseases of different aetiology in Asian elephant (Elephas maximus) in samll traveling circus. Erkrankungen der Zootiere 30, 263-269.

Rahman, H., Dutta, P.K., Dewan, J.N., 1988. Foot and mouth disease in elephant (Elephas maximus). Zentralbl. Veterinarmed. [B]. 35, 70-71.
Abstract: A natural case of foot and mouth disease in an Indian elephant (Elephas maximus) is recorded.  The virus isolated was typed as Asian 1.  The source of infection possibly transmitted indirectly was traced to an outbreak of FMD-Asia 1 in cattle and buffalos of the district.

Gaskin, J.M., Andresen, T.L., Olsen, J.H., Schobert, E.E., Buesse, D., Lynch, J.D., Walsh, M., Citino, S., Murphy, D., 1987. Encephalomyocarditis in zoo animals: Recent experiences with the disease and vaccination. Proceedings of the 1st International Conference on Zoological and Avian Medicine 491.
Abstract: Encephalomyocarditis (EMC), a specific viral infection caused by a group of antigenically related viruses in the family Picornaviridae, a genus of Cardiovirus, continues to be a source of sporadic mortality loss in zoo animals in Florida.  Deaths in a young Nyala antelope, 2 chimpanzees, 3 llamas, a two-toed sloth, 3 ringtail lemurs, a ruffed lemur, and an orangutan have recently been confirmed by virus recovery.  Experimental vaccine trials were initiated in pygmy goats, Barbados sheep, and white mice using B-propiolactone inactivated virus preparations.  Various adjuvants, including aluminum hydroxide, mineral oil, and dimethyl dioctadecyl ammonium bromide (DDAB) were used to enhance the immune responses to inactivated virus.  The vaccine preparations produced varying levels of hemagglutinations-inhibition (HI) antibodies in the immunized animals.  Experimental challenge of unvaccinated weaned pigs, pygmy goats, and Barbados sheep demonstrated that, although they seroconverted, they did not become ill when exposed to the virulent EMC virus strains used in this study. Laboratory mice, however, proved to be very susceptible when exposed to these same strains, and either died acutely or developed posterior paresis and paralysis subsequent to challenge.  All experimental vaccine preparations protected mice against challenge.  In vaccinated goats and sheep, the oil-emulsion-adjuvanted and DDAB-adjuvanted vaccines produced the highest and most persistent HI antibody titers.  Sera obtained from African elephants were screened for HI antibodies to EMC virus.  Ninety-three African elephant sera from the Kruger National Park in the Republic of South Africa had titers of less than 10 hemagglutination-inhibition units (HIU) while 4 of 76 imported juvenile African elephants had titers from 10-40 HIU and the rest had no titer.  EMC virus infections are apparently acquired in Florida from reservoir hosts and HI titers of 40 HIU or higher indicate subclinical infection with the virus.  Experimental vaccines may help prevent EMC in susceptible species; HI responses to vaccination in various exotic species are being evaluated.

Pilaski, J., Rosenbruch, M., Gelderblom, H., Olberding, P., Hagenbeck, C., 1987. Herpes virus infectionin an Asian elephant (Elephas maximus). Erkrankungen der Zootiere 29, 179-184.

Rosen, A., Pilaski, J., Darai, G., 1987. Genomic characterization of a poxvirus isolated from a child. Med Microbiol Immunol (Berl) 176, 181-188.
Abstract: A poxvirus was isolated from a six-year-old girl. The comparative analyses of the genome of this isolate (H-CP-LSax) which were carried out using the restriction endonucleases BamHI, HindIII, KpnI, MluI, NcoI, SacI, and SmaI revealed that this isolate is a member of the genus orthopoxvirus. Since the girl had never been vaccinated against smallpox, and had close contact to domestic animals, including cats, rabbits and guinea pigs, the genome of H-CP-LSax virus was genetically analysed in comparison with other known orthopoxviruses. The analysis demonstrates clearly that the HindIII cleavage pattern of H-CP-LSax DNA is different from the HindIII DNA cleavage patterns of vaccinia virus, cowpox virus, rabbit poxvirus, cat poxvirus, ectromelia virus, and okapi poxvirus. Surprisingly, it was found that the HindIII and SmaI cleavage patterns of the DNA of one out of six elephant poxviruses which were analysed under the same conditions were virtually identical to the HindIII and SmaI cleavage patterns of H-CP-LSax DNA. Although SmaI and HindIII digestion of both virus genomes gave the same fragment patterns, the viral DNAs can be distinguished from each other by the restriction endonucleases SacI, BamHI, and KpnI, which also show high similarities in the fragmentation patterns of both viruses. The results obtained in this study indicate three possibilities concerning the origin of H-CP-LSax virus. Firstly that the H-CP-LSax virus originated from an unknown animal species. Secondly, that this virus is a variant of elephant poxvirus in which the HindIII and SmaI sites are extremely conserved, and finally that H-CP-LSax can be a recombinant virus of unknown origin.

Seaman, J.T., Finnie, E.P., 1987. Acute myocarditis in a captive African elephant (Loxodonta africana). Journal of Wildlife Diseases 23, 170-171.

Jacobson, E.R., Sundberg, J.P., Gaskin, J.M., Kollias, G.V., O'Banion, M.K., 1986. Cutaneous papillomas associated with a herpesvirus-like infection in a herd of captive African elephants. Journal of the American Veterinary Medical Association 189, 1075-1078.
Abstract: Proliferative cutaneous lesions developed in a herd of captive African elephants (33 from an animal importer in Texas, and 63 young elephants collected in Zimbabwe). Group-1 elephants were purchased 8 months before the arrival of the group-2 elephants. On arrival, 7 group-1 elephants had raised nodular fibrous growths, located predominantly on their trunks. Lesions were not observed in the group-2 elephants until approximately 3 months after they were acquired. Lesions on group-2 elephants began as small focal proliferative growths that regressed or that progressed into large nodular fibrous growths that were similar in appearance to those seen in the group-1 elephants. Lesions at various stages of development were biopsied and examined. Histologically, early lesions were inverted papillomas, with hyperplastic and hypertrophic epithelial cells containing amphoteric intranuclear inclusions in the lesion center. Older, large, nodular fibrous growths were ulcerated and were composed predominantly of a thickened dermis containing fibroblasts, collagen, and a mixed inflammatory cell infiltrate; inclusions were not observed in adjacent epidermal cells. Using a peroxidase- antiperoxidase technique, we did not detect group-specific papillomavirus antigens. Southern blot hybridization analysis of DNA from lesion specimens did not indicate papillomavirus- specific genomes. Electron-microscopically, inclusions consisted of aggregates of virus particles. The particles had electron- dense and electron-lucent cores and were 95 to 103 nm in diameter. Virions developed envelopes from nuclear membranes. Mature particles were seen within the cytoplasm and filled the intercellular spaces. On the basis of size, location, conformation, and envelopment, the particles most closely resembled those of herpesviruses.

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

Pilaski, J., Rosen, A., Darai, G., 1986. Comparative analysis of the genomes of orthopoxviruses isolated from elephant, rhinoceros, and okapi by restriction enzymes. Brief report. Archives of Virology 88, 135-142.
Abstract: Orthopoxviruses from different zoo-kept mammalian species including Elephas maximus (8 isolates), Ceratotherium simum (1 isolate), and Okapia johnstoni (2 isolates) were characterized by restriction enzyme analysis of the viral genome. The four enzymes BamHI, MluI, NcoI, and SalI were found to be optimal for strain differentiation.

Gopal, T., Rao, B.U., 1984. Rabies in an Indian wild elephant calf. Indian Veterinary Journal 61, 82-83.

Pilaski, J., Foster, J.W., Matern, B., Kloppel, G., Schaller, K. Epidemiology of diseases similar to cowpox in zoo animals. Proc.Int.Symp.Dis.Zoo Anim. 26, 349-354. 1984.
Ref Type: Conference Proceeding

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

Wallach, J.D., Boever, W.J., 1983. Perissodactyla (equids, tapirs, rhinos), Proboscidae (elephants), and Hippopotamidae (hippopotamus). In: Wallach, J.D., Boever, W.J. (Eds.), Diseases of exotic animals. W.B. Saunders Company, Philadelphia, pp. 761-829.

Baxby, D., 1982. The surface antigens of orthopoxviruses detected by cross-neutralization tests on cross-absorbed antisera. J Gen Virol 58 (Pt 2), 251-262.
Abstract: Cross-neutralization tests were done on accepted species and recently isolated members of the genus Orthopoxvirus using antisera which had been separately absorbed with the various viruses. The results provided evidence for the involvement of four neutralizing antigens, and their distribution among 13 virus strains was determined. Monkeypox (Congo-8-Lombe), camelpox (Gorgan), ectromelia  (Mill Hill), 'Lenny' and elephant poxviruses had distinctive antigenic formulae. Lister and Wyeth vaccines were indistinguishable but different from Copenhagen and EM63 vaccines which were themselves distinct. Cowpox (Brighton), buffalopox (BP4), MK 10, and Moscow poxviruses were indistinguishable. Examples were found where viruses shared surface antigens but were not all neutralized by antibody to them. This reduced the practical value of the technique for virus identification. Evidence was also obtained for the existence in some viruses of a fifth antigen, antibody to which could block neutralization by antibody to one particular antigen.

Hedger, R.S., 1981. Foot-and-mouth disease. In: Davis, J.W., Karstad, L.H., Trainer, D.O. (Eds.), Infectious diseases of wild mammals. The Iowa State University Press, Ames, Iowa.

Karstad, L., 1981. Miscellaneous viral infections. In: Davis, J.W., Karstad, L.H., Trainer, D.O. (Eds.), Infectious diseases of wild mammals. The University of Iowa Press, Ames, Iowa.

Murname, T.G., 1981. Encephalomyocarditis. In: Steele, J.H. (Ed.), CRC Handbook Series in Zoonoses, Section B: Viral Zoonoses. The Iowa State University Press, Ames, Iowa, pp. 137-147.

Plowright, W., 1981. Herpesvirus of wild ungulates, including malignant catarrhal fever virus. In: Davis, J.W., Karstad, L.H., Trainer, D.O. (Eds.), Infectious diseases of wild mammals. Iowa State University Press, Ames, Iowa.

Scott, G.R., 1981. Rinderpest. In: Davis, J.W., Karstad, L.H., Trainer, D.O. (Eds.), Infectious diseases of wild mammals. The Iowa State University Press, Ames, Iowa.

Sundberg, J.P., Russell, W.C., Lancaster, W., 1981. Papillomatosis in Indian elephants. Journal of the American Veterinary Medical Association 179, 1247-1249.

Gaskin, J.M., Jorge, M.A., Simpson, C.F., Lewis, A.L., Olson, J.H., Schobert, E.E., Wollenman, E.P., Marlowe, C., Curtis, M.M., 1980. The tragedy of encephalomyocarditis virus infection in zoological parks of Florida. Proceedings American Association of Zoo Veterinarians 1-7.

Malecki, G., Zuchowska, E., 1980. Pox-like disease in  Elephas maximus. Medycyna Weterynaryjna 36, 667-669.

Baxby, D., Shackleton, W.B., Wheeler, J., Turner, A., 1979. Comparison of cowpox-like viruses isolated from European zoos. Archives of Virology 61, 337-340.
Abstract: Poxvirus isolated from captive carnivores in Russia (Moscow virus) and elephants in Germany (elephant virus) were very closely-related to cowpox virus.  Immunological analysis with absorbed sera separated elephant virus but not compox and Moscow virus, whereas polypeptide analysis separated compox but not elephant and Moscow virus.  A combination of biological test separated all three.  The epidemiological implications are briefly reviewed.

Chandrasekharan, K. Common diseases of elephants. State Level Workshop on Elephants.  51-61. 1979. India, College of Veterinary and Animal Sicences, Kerala Agricultural University.
Ref Type: Conference Proceeding

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.

Gehring, H., Mayer, H., 1978. Beitrag zur diagnostik und bekampfung der pockeninfection bein elefanten. Prakt. Tieraerztl. 2, 106.

Tesh, R.B., Wallace, G.D., 1978. Observations on the natural history of encephalomyocarditis virus. American Journal of Tropical Medicine and Hygiene 27, 133-143.

Baxby, D., Ghaboosi, B., 1977. Laboratory characteristics of pox-viruses isolated from captive elephants in Germany. Journal of General Virology 37, 407-414.
Abstract: Poxviruses isolated from captive elephants in Germany have been characterized.  Although related to vaccinia and even more closely to cowpox virus, the separate identity of elephantpox virus was established by both biological and serological methods.  Elephantpox virus produces A-type inclusions in infected cells, as did cowpox, but had a lower ceiling temperature, and was more heat resistant and affected rabbits differently.  Cross neutralization tests on absorbed sera indicted that elephantpox, cowpox and vaccinia shared one surface antigen, that elephantpox and vaccinia shared an antigen absent from cowpox, and that vaccinia virus had a surface antigen absent from elephantpox and cowpox viruses.

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

Marennikova, S.S., Maltseva, N.N., Korneeva, V.I., Garanina, N., 1977. Outbreak of pox disease among carnivora (felidae) and edentata. J Infect Dis 135, 358-366.
Abstract: An outbreak of pox disease in Carnivora of the family Felidae occurred in the Moscow Zoo. Two forms of the disease were found: (1) fatal, fulminant pulmonary without skin lesions and (2) dermal with rash. The severity of the dermal form varied from subclinical to lethal. The pulmonary form was characterized by pneumonia and exudative pleuritis, and large concentrations of virus were observed in the lungs and exudate. In addition to Carnivora of the family Felidae, two giant anteaters had a severe form of the disease (dermal with hemorrhages) and died. The agent of the outbreak appeared to be very closely related to cowpox virus; however, pocks developed at a lower temperature than do those that result from infection with cowpox virus. Strains isolated from sick animals were identical to the virus previously isolated from an outbreak of pox among elephants and okapi. The most probable sources of infection were rats that were fed to some of the animals. During the outbreak, a female attendant at the zoo became infected.

Simpson, C.F., Lewis, A.L., Gaskin, J.M., 1977. Encephalomyocarditis virus infection of captive elephants. Journal of the American Veterinary Medical Association 171, 902-905.
Abstract: Four Asian elephants at each of 2 widely separated zoologic gardens in Florida died following a fulminating illness.  Tissue suspensions obtained from an elephant from each of the zoologic gardens were inoculated into newborn mice, 3- to 4-week-old mice, buffalo green monkey and baby hamster kidney cell cultures.  Encephalitis and myocarditis developed in the mice.  The cell cultures were destroyed within 24 to 72 hours, and intracytoplasmic viral inclusions were observed in infected cells by electron microscopy.  The viral agent was neutralized by known antiserum to encephalomyocarditis virus.

Hedger, R.S., Brooksby, J.B., 1976. FMD in an Indian elephant (letter). Veterinary Record 99, 93.

Pyakural, S., Singh, U., Singh, N.B., 1976. An outbreak of foot-and-mouth disease in Indian elephants (Elephas maximus). Veterinary Record 99, 28-29.
Abstract: Type O virus has been isolated from a natural outbreak of foot-and-mouth in Indian elephant.  The history, symptoms, and epizootology of this disease in these animals have also been described.

Hoff, G.L., Griner, L.A., Trainer, D.O., 1973. Blue-tongue virus in exotic ruminants. Journal of the American Veterinary Medical Association 163, 565-567.

Howell, P.G., Young, E., Hedger, R.S., 1973. Foot-and-mouth disease in the African elephant (Loxodonta africana). Onderstepoort Journal of Veterinary Research 40, 41-52.
Abstract: A strain of SAT 2 foot-and-mouth disease virus which was experimentally inoculated into the epidermis of the tongues of captive African elephants produced vesicular lesions at the site of inoculation.  After a short period of viraemia, secondary lesions developed in the mouth and on the feet giving rise to extensive tissue damage and the separation of the soles.  In spite of close contact there was no spread of the disease to other elephants and by conventional sampling techniques no carrier virus could be demonstrated.  The neutralizing antibody response was of a low order and this finding together with the observations made during the course of the experimental disease are discussed in relation to the possible role of the elephant in the epizootiology of foot-and-mouth disease in Africa.

Mayer, J., 1973. Vaccinia in humans caused by generalized infection of a circus elephant. Zentralblatt fur Bakteriologie,Parasitenkunde,Infektionskrankheiten und Hygiene 1. Abt. Originale 224, 448-452.

Gehring, H., Mahnel, H., Mayer, H., 1972. Elephant pox. Zentralbl. Veterinarmed. [B]. 19, 258-261.

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.

Erasmus, B.J., McCully, R.M., Pienaar, J.G., Young, E., Pieterse, L.M., Els, H.J. The isolation of a herpes virus from the African elephant [Loxodonta africana (Blumenbach, 1797)]. Journal of General Virology . 1971.
Ref Type: Unpublished Work
Abstract: Same case as McCully, 1969, 1971;  Some references cite this article as  J. Gen. Virol. "in press" 1971, however the article was apparently never actually published.

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.

McCully, R.M., Basson, P.A., Pienaar, J.G., Erasmus, B.J., Young, E., 1971. Herpes nodules in the lung of the African elephant [Loxodonta africana (Blumenbach, 1797)]. Onderstepoort Journal of Veterinary Research 38, 225-236.
Abstract: Lymphoid nodules associated with Cowdry Type A intranuclear inclusions in epithelial and syncytial cells were found in the lungs of 74% of 50 African elephants in the Kruger National Park.  Subsequent studies proved these were caused by a herpes virus (Erasmus,McCully,Pienaar,Young, Pieterse & Els, 1971).  The disease appears to be subclinical or latent.  This virus, in common with other herpes viruses, might be more pathogenic in some other host.  The pathogenesis of the lymphoid nodules and the various stages of their formation are given and the detailed characteristics are illustrated.

Piragino, S., 1970. An outbreak of foot and mouth disease in circus elephant. Zooprofilani 25, 17-22.

Gainer, J.H., 1969. Encephalomyocarditis virus infections in Florida, 1960-1966. Journal of the American Veterinary Medical Association 151, 421-425.

McCully, R.M., Basson, P.A., Pienaar, J.G., Erasmus, B.J., Young, E., Pieterse, L.M., 1969. Herpes nodules in elephants. Journal of the South African Veterinary Medical Association 40, 422.
Abstract: Full text:  The lungs of the majority of 50 elephants examined recently in the Kruger National Park had several lymphoid nodules varying in size from 3 to 30 mm (Fig. 1 arrow).  The alveolar lining cells within these nodules were metaplastic and contained Type A intranuclear inclusions (Fig. 2).  Electronmicroscopy revealed the presence of a virus in the epithelial cells.  It was subsequently successfully isolated in tissue culture and characterized as a herpes virus.

Gainer, J.H., Sandifur, J.R., Bigler, W.J., 1968. High mortality in a Florida swine  herd infected with encephalomyocarditis virus.  An accompaning epizootiologic survey. The Cornell Veterinarian 58, 31-47.

McGaughey, C.A., 1962. Diseases of elephants. Part 4. Ceylon Veterinary Journal 10, 3-9.

Gainer, J.H., Murchison, T.E., 1961. Encephalomyocarditis virus infection of swine. Vet. Med. 56, 173-175.

Gupta, V., 1945. Rabies in an elephant. Indian Veterinary Journal May.

D'Costa, J., 1936. Rinderpest: Its symptoms. Indian Journal of Veterinary Science and Animal Husbandry 13, 7.

Ramiah, B., 1935. An outbreak of foot and mouth disease in elephants. Indian Veterinary Journal 12, 28.

Beckett, J., 1932. Death of an elephant from rabies. Journal of the Bombay Natural History Society 36, 242-243.

Ramiah, B., 1932. Paralytic rabies in an elephant. Indian Veterinary Journal 9, 142.

Lapin, A., 1901. Maul-und-Klauenseuche bereinem elephanten. Prezg. Wet. 250, 1.

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