Elephant
Bibliographic
Database

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Elephant Bibliographic Database
www.elephantcare.org

References Updated October 2007

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

     2.    Helke K.L., Mankowski J.L. and Manabe Y.C. 2007. Animal models of cavitation in pulmonary tuberculosis.Tuberculosis (Edinb) 86: 337-348.
Abstract: Transmission of tuberculosis occurs with the highest frequency from patients with extensive, cavitary, pulmonary disease and positive sputum smear microscopy. In animal models of tuberculosis, the development of caseous necrosis is an important prerequisite for the formation of cavities although the immunological triggers for liquefaction are unknown. We review the relative merits and the information gleaned from the available animal models of pulmonary cavitation. Understanding the host-pathogen interaction important to the formation of cavities may lead to new strategies to prevent cavitation and thereby, block transmission.

     3.    Lacasse C., Terio K., Kinsel M.J. et al. 2007. Two cases of atypical mycobacteriosis caused by Mycobacterium szulgai associated with mortality in captive African elephants (Loxodonta africana).Journal of Zoo and Wildlife Medicine 38: 101-107.
Abstract: Mycobacterium szulgai was associated with mortality in two captive African elephants (Loxodonta africana) housed at Lincoln Park Zoo. The first elephant presented with severe, acute lameness of the left rear limb. Despite extensive treatments, the animal collapsed and died 13 mo after initial presentation. Necropsy revealed osteomyelitis with loss of the femoral head and acetabulum and pulmonary granulomas with intralesional M. szulgai. The second elephant collapsed during transport to another institution with no premonitory clinical signs. This animal was euthanized because of prolonged recumbency. Granulomatous pneumonia with intralesional M. szulgai was found at necropsy. Two novel immunoassays performed on banked serum samples detected antibody responses to mycobacterial antigens in both infected elephants. It was not possible to determine when the infection was established or how the elephants were infected. When reviewing the epidemiology of this organism in humans, however, transmission between elephants seemed unlikely because human-to-human transmission of this organism has never been reported and a third elephant in the herd was not affected. In addition to Mycobacterium bovis and Mycobacterium tuberculosis, atypical mycobacterial organisms need to be considered potentially pathogenic in elephants.

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

     5.    Sreekumar E., Janki M.B.V., Arathy D.S. et al. 2007. Molecular characterization and expression of Interferon-gamma of Asian elephant (Elephas maximus).Vet Immunol Immunopathol 118: 75-83.
Abstract: Tuberculosis (TB) caused by Mycobacterial organisms has emerged as one of the major diseases in captive elephants. In vitro Interferon-gamma (IFN-gamma) assay is being used as an ancillary test for early detection of TB in domestic and captive wild animals. In the present study, basic sequence information and immunological cross-reactivity of this major cytokine of Asian elephants were explored. At predicted amino acid level, IFN-gamma of Asian elephant showed maximum identity to that of horse (73%). Other IFN-gamma amino acid sequences that showed high level identity were that of giant panda (72%), dog (71%), nine-banded armadillo (69%), cattle (63%) and human (62%). IFN-gamma promoter sequences of Asian elephant, human, cattle and mouse showed high level conservation of the putative transcription factor binding sites, TATA box and transcriptional start site. The functionally important human IFN-gamma promoter elements, such as AP-2IRE-BE, YY1-gammaIFN-BED, ATFCS and AP-1gammaINF binding sites, were absolutely conserved in the corresponding elephant sequence. There was only a single nucleotide variation in the other two important elements, NFAT-gammaINF and IFN-gammaPE, indicating the highly conserved regulation of IFN-gamma expression across different species. Phylogenetic analysis based on IFN-gamma protein sequences revealed a closer relation of Asian elephants and nine-banded armadillo. This shows a closer evolution of these members of Afrotheria and Xenarthra, respectively; and supports the previous reports based on mitochondrial DNA studies. In Western blot analysis, IFN-gamma of Asian elephant expressed in Escherichia coli was detected using an anti-bovine IFN-gamma monoclonal antibody, indicating immunological cross-reactivity.

     6.    Une Y. and Mori T. 2007. Tuberculosis as a zoonosis from a veterinary perspective.Comp Immunol Microbiol Infect Dis Aug 13; [Epub ahead of print].
Abstract: Tuberculosis is an important disease among many zoonoses, because both Mycobacterium tuberculosis and Mycobacterium bovis, which are the major causes of tuberculosis, are highly pathogenic, infect many animal species and thus are likely to be the source of infection in humans. In particular, monkeys are highly susceptible to these bacteria and are important spreaders. Recently, two outbreaks of M. tuberculosis occurred in four different kinds of monkeys and humans were also infected with the disease in Japan. In zoos, tuberculosis was reported not only in monkeys, but also in several different kinds of animals, including elephants. Pets such as dogs and cats are believed to be generally less susceptible to M. tuberculosis, but in this article we introduce a case of infection from man to dog by close contact. Japan is one of the few countries that have been able to control M. bovis infection. In other countries, however, cases of bovine tuberculosis and human M. bovis infection have been reported, and thus further attention is still required in the future.

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

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

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

     9.    Bojesen A.M., Olsen K.E. and Bertelsen M.F. 2006. Fatal enterocolitis in Asian elephants (Elephas maximus) caused by Clostridium difficile.Vet Microbiol Epub ahead of print.
Abstract: Two cases of fatal enteritis caused by Clostridium difficile in captive Asian elephants are reported from an outbreak affecting five females in the same zoo. Post mortem examination including histopathology demonstrated fibrinonecrotic enterocolitis. C. difficile was isolated by selective cultivation from two dead and a third severely affected elephant. Four isolates were obtained and found positive for toxin A and B by PCR. All isolates were positive in a toxigenic culture assay and toxin was demonstrated in the intestinal content from one of the fatal cases and in a surviving but severely affected elephant. PCR ribotyping demonstrated that the C. difficile isolates shared an identical profile, which was different from an epidemiologically unrelated strain, indicating that the outbreak was caused by the same C. difficile clone. It is speculated that the feeding of large quantities of broccoli, a rich source of sulforaphane, which has been shown to inhibit the growth of many intestinal microorganisms may have triggered a subsequent overgrowth by C. difficile. This is the first report of C. difficile as the main cause of fatal enterocolitis in elephants. The findings emphasize the need to regard this organism as potentially dangerous for elephants and caution is recommended concerning antibiotic treatment and feeding with diets containing antimicrobials, which may trigger an expansion of a C. difficile population in the gut.

   10.    Dumonceaux G. and Mikota S. 2006. Tuberculosis treatment protocols and complications for elephants.Proceedings International Elephant Conservation and Research Symposium: 84-85.

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

   12.    Michel A.L., Bengis R.G., Keet D.F. et al. 2006. Wildlife tuberculosis in South African conservation areas:Implications and challenges.Veterinary Microbiology 112: 91-100.
Abstract: Tuberculosis, caused by Mycobacterium bovis, was first diagnosed in African buffalo in South Africa's Kruger National Park in 1990. Over the past 15 years the disease has spread northwards leaving only the most northern buffalo herds unaffected. Evidence suggests that 10 other small and large mammalian species, including large predators, are spillover hosts. Wildlife tuberculosis has also been diagnosed in several adjacent private game reserves and in the Hluhluwe-iMfolozi Park, the third largest game reserve in South Africa. The tuberculosis epidemic has a number of implications, for which the full effect of some might only be seen in the longterm. Potential negative long-term effects on the population dynamics of certain social animal species and the direct threat for the survival of endangered species pose particular problems for wildlife conservationists. On the other hand, the risk of spillover infection to neighboring communal cattle raises concerns about human health at the wildlife-livestock-human interface, not only along the western boundary of Kruger National Park, but also with regards to the joint development of the Greater Limpopo Transfrontier Conservation Area with Zimbabwe and Mozambique. From an economic point of view, wildlife tuberculosis has resulted in national and international trade restrictions for affected species. The lack of diagnostic tools for most species and the absence of an effective vaccine make it currently impossible to contain and control this disease within an infected free-ranging ecosystem. Veterinary researchers and policy-makers have recognized the need to intensify research on this disease and the need to develop tools for control, initially targeting buffalo and lion.

   13.    Mikota S.K., Miller M., Dumonceaux G. et al. 2006. Elephant tuberculosis diagnosis: implications for elephant management in Asian range countries. 
2006 Proceedings American Association of Zoo Veterinarians, pp. 142-143.
Abstract: Serologic tests including the ELISA, MAPIA (Multi-Antigen Print Immunoassay), and a rapid test, VetTB StatPak® (Chembio Diagnostic Systems, Inc., Medford, New York 11763 USA) have recently been developed and show great promise for the diagnosis of tuberculosis (TB) in elephants. These serologic tests detect antibodies to antigens of  Mycobacterium tuberculosis complex organisms and in some cases have detected infection years in advance of active disease and mycobacterial shedding. The diagnosis of active TB (by culture) or serologic conversion presents management challenges for captive elephants in Asian range countries.  Of the 2 billion humans world-wide infected with TB, fewer than 10% will develop active disease. This figure is unknown for elephants. The identification and management of infected elephants has ramifications for elephants and humans alike and issues such as public health and tourism may be impacted. TB is endemic among humans in Asia and where there is intermingling of elephants and humans, both species may act as reservoirs for disease transmission.  The various situations in which elephants are kept in Asia (government-owned, privately-owned, festivals, temples, zoos, etc.) make it difficult to develop a management strategy that will address all circumstances.  Other concerns are the cost of treatment for an elephant (~ $50,000 USD) and appropriate monitoring in resource-poor countries. The authors have recently undertaken the screening of 120 elephants in Nepal to further evaluate the above-mentioned (and other) diagnostic tests.  To our knowledge, this is the first organized, large-scale initiative to screen Asian elephants within a range country.  Preliminary discussions regarding the management of both culture and serologically positive government-owned and privately-owned elephants in Nepal have been initiated and may serve as a starting point for other countries as more elephants are screened within Asia.  Basic options for active (culturepositive) cases include (1) treatment, (2) segregation or (3) euthanasia.  Options for latent disease (culture-negative, serologically positive) cases include (1) treatment, (2) segregation and monitoring for active disease and (3) euthanasia.  The particular ownership/husbandry system, available resources and cultural constraints may dictate final management choices in range countries.

   14.    Mikota S.K., Dumonceaux G., Miller M. et al. 2006. Tuberculosis in elephants: An update on diagnosis and treatment; implications for control in range countries.Proceedings International Elephant Conservation and Research Symposium: 109-118.

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

   16.    Peloquin C.A., Maslow J.N., Mikota S.K. et al. 2006. Dose selection and pharmacokinetics of rifampin in elephants for the treatment of tuberculosis.J Vet Pharmacol Ther. 29: 1-6.

   17.    Riley L.W. 2006. Of mice, men, and elephants: Mycobacterium tuberculosis cell envelope lipids and pathogenesis.J Clin Invest 116: 1475-1478.
Abstract: Comment on: J Clin Invest. 2006 Jun;116(6):1660-7. Mycolic acids and structures attached to them constitute a major part of the protective envelope of Mycobacterium tuberculosis, and for this reason, their role in tuberculosis pathogenesis has been extensively studied. In this issue of the JCI, Rao et al. examine the effect of trans-cyclopropanation of oxygenated mycolic acids attached to trehalose dimycolate (TDM) on the murine immune response to infection (see the related article beginning on page 1660). Surprisingly, they found that an M. tuberculosis mutant lacking trans-cyclopropane rings was hypervirulent in mice. The recent recognition of a hypervirulence phenotype in mice associated with laboratory and clinical M. tuberculosis strains with altered cell wall components has provided new insights into how M. tuberculosis may establish persistent infection. However, to date, characterization of these bioactive products in pathogenesis has been largely reductionistic; the relationship of their effects observed in mice to the persistent infection and tuberculosis caused by M. tuberculosis observed in humans remains obscure.

   18.    Rothschild B.M. and Martin L.D. 2006. Did ice-age bovids spread tuberculosis?Naturwissenschaften 93: 565-569.
Abstract: Pathognomonic metacarpal undermining is a skeletal pathology that has been associated with Mycobacterium tuberculosis in bovids. Postcranial artiodactyl, perissodactyl, and carnivore skeletons were examined in major university and museum collections of North America and Europe for evidence of this and other pathology potentially attributable to tuberculosis. Among nonproboscidean mammals from pre-Holocene North America, bone lesions indicative of tuberculosis were restricted to immigrant bovids from Eurasia. No bone lesions compatible
with diagnosis of tuberculosis were found in large samples of other pre-Holocene (164 Oligocene, 397 Miocene, and 1,041 Plio-Pleistocene) North American mammals, including
114 antilocaprids. Given the unchanged frequency of bovid tubercular disease during the Pleistocene, it appears that most did not die from the disease but actually reached an
accommodation with it (as did the mastodon) (Rothschild and Laub 2006). Thus, they were sufficiently long-lived to assure greater spread of the disease. The relationships of the
proboscidean examples need further study, but present evidence suggests a Holarctic spread of tuberculosis during the Pleistocene, with bovids acting as vectors. While the role of other animals in the transmission of tuberculosis could be considered, the unique accommodation achieved by bovids and mastodons makes them the likely "culprits" in its spread.

   19.    Rothschild B.M. and Laub R. 2006. Hyperdisease in the late Pleistocene:validation of an early 20th century hypothesis.Naturwissenschaften 93: 557-564.

   20.    Bertelsen M.F., Bojesen M. and Olsen K.E.P. 2005. Fatal enterocolitis in two Asian elephants (Elephas maximus) caused by Clostridium difficile.  2005 Proceedings AAZV, AAWV, AZA Nutrition Advisory Group, pp. 66-67.
Abstract: Altered behavior, anorexia and listlessness were observed in four of five adult captive female Asian elephants (Elephas maximus). Two animals recovered, while two died after 2 days. The dead elephants were subjected to post mortem examination including histopathology, demonstrating fibrinonecrotic enteritis and colitis. Clostridium difficile was isolated from both dead elephants and from the feces of the two surviving affected animals, and identified by selective cultivation and PCR identification. All isolates had the tcdA and tcdB toxin genes and were positive in a toxigenic culture assay. C. difficile toxin from the intestinal content of one of the fatal cases was demonstrated using cell-culture based cytotoxin assays. Clostridium perfringens type A and Clostridium septicum were also isolated from both dead animals. Although C. perfringens has been associated with ulcerative enteritis in an elephant,¹ in this case these isolates likely are incidental, as C. perfringens enterotoxin was not demonstrated, and as C. septicum is well known for producing rapid post mortem overgrowth.  Amplified fragment length polymorphism typing, showed that the C. difficile isolates recovered from the outbreak, all had the same fingerprint profile, indicating that all four elephants were affected by the same bacterial clone. These findings appear to be the first to demonstrate that C. difficile may cause enterocolitis in elephants. The results emphasize the need to regard this organism as potentially dangerous for elephants. Although there was no prior exposure to antibiotic agents in this case, caution is recommended when treating elephants with antibiotics, as this may trigger C. difficile induced enterocolitis in other species, most notably humans and horses.²
LITERATURE CITED
1 Bacciarini, L.N., O. Pagan, J. Frey, and A. Grone. 2001. Clostridium perfringens beta2-toxin in an African elephant (Loxodonta africana) with ulcerative enteritis. Vet. Rec. 149: 618-20.
2 Songer, J.G. 1996. Clostridial enteric diseases of domestic animals. Clin. Microbiol. Rev. 9: 216-234.

   21.    Cousins D.V. and Florisson N. 2005. A review of tests available for use in the diagnosis of tuberculosis in non-bovine species.Rev.sci.tech.Off.int.Epiz. 24: 1039-1059.
Abstract: Bovine tuberculosis is an important disease that has impacts on regional and international trade. The disease can affect both social and economic stability and have a deleterious affect on species diversity. The intradermal tuberculin test has been in use for almost a century and, despite the technological advances of the last two decades, is still the only prescribed test for the
diagnosis of tuberculosis in cattle. Many other species of animal, including humans, can be infected with Mycobacterium bovis. This paper reviews the various tests that have been used by researchers for detecting infection with M. bovis in a variety of animal species, and attempts to prioritise or comment on the importance of having appropriately validated diagnostics for the different species. The difficulties of test validation using small numbers of animals, especially when tuberculosis occurs in only a few instances or the species of animal affected is rare and/or valuable, are discussed.

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

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

   24.    Lewerin S.S., Olsson S.L., Eld K. et al. 2005. Outbreak of Mycobacterium tuberculosis infection among captive Asian elephants in a Swedish zoo.Vet Rec. 156: 171-175.
Abstract: Between 2001 and 2003, there was an outbreak of tuberculosis in a Swedish zoo which involved elephants, giraffes, rhinoceroses and buffaloes. Cultures of
trunk lavages were used to detect infected elephants, tuberculin testing was used in the giraffes and buffaloes, and tracheal lavage and tuberculin testing
were used in the rhinoceroses. The bacteria isolated were investigated by spoligotyping and restriction fragment length polymorphism. Five elephants and
one giraffe were found to have been infected by four different strains of Mycobacterium tuberculosis. National Veterinary Institute, SE-751 89 Uppsala, Sweden.

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

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

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

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

ACKNOWLEDGMENTS
The authors thank the zoos and individuals that have provided samples and assistance with this research, including Ray Ball, Carol Buckley, Jenifer Chatfield, Genny Dumonceaux, Javan Esfandiary, Rena Greenwald, Scott Larsen, Susan Mikota, Torsten Moller, Dick Montali, Mike Richards, Heidi Riddle, Mo Salman, Scott Terrell, and many others.  This research was supported by Chembio Diagnostics, Inc.
LITERATURE CITED
1 Larsen, R.S., M.D. Salman, S.K. Mikota, R. Isaza, R.J. Montali, and J. Triantis. 2000.  Evaluation of a multiple-antigen enzyme-linked immunosorbent assay for detection of Mycobacterium tuberculosis  
  infection in captive elephants.  J. Zoo Wildl. Med. 31:291-302.
2 Lyashchenko, K., et al.  2000. A multiantigen print immunoassay for the serological diagnosis of infectious diseases.  J. Immunol. Methods  242:91-100. 
3 Mikota, S.K., and J. Maslow.  2002.  Epidemiology and treatment of tuberculosis in elephants:  2002.  Proc. Am. Assoc. Zoo Vet. Annu. Meet.  Pp. 384-387.
4 Mikota, S.K., L. Peddie, J. Peddie, et al.  2001.  Epidemiology and diagnosis of Mycobacterium tuberculosis in captive Asian elephants (Elephas maximus).  J. Zoo Wildl. Med. 32:1-16.

Table 1.  Comparison of serodiagnostic results for tuberculosis in elephants.
Health status      # Elephants      # Rapid test positive    # MAPIA positive
Healthy                  63                                                                     1                                                                       0
TB infected            17                                                                     17                                                                    16</sup>

   26.    Maslow J.N., Mikota S.K., Zhu M. et al. 2005. Population pharmacokinetics of isoniazid in the treatment of Mycobacterium tuberculosis among Asian and African elephants (Elephas maximus and Loxodonta
africana).J Vet Pharmacol Ther. 28: 1-7.
Abstract: We recently described the clinical presentation and treatment of 18 elephants from six herds infected with TB. Treatment protocols and methods varied between herds to include both oral and rectal dosing using multiple drug doses and formulations. In this paper we present information regarding the pharmacokinetics (PK) of isoniazid (INH) in elephants and provide suggestions regarding initial treatment regimens. Forty-one elephants received INH daily by either oral or rectal administration with different formulations. Population PK analysis was performed using Non-linear Mixed Effect Modeling (NONMEM). Results of oral administration indicated that compared with premixed INH solution, the drug exposure was highest with a suspension prepared freshly with INH powder. When INH was concomitantly given as an admixture over food, Tmax was delayed and variability in drug absorption was significantly increased. Compared with oral administration, similar drug exposures were found when INH was dosed rectally. The data generated suggest that a starting dose of 7.5 mg/kg of INH is appropriate for initial TB treatment in elephants when premixed solution is administered directly into the oropharynx or rectal vault and 4 mg/kg are when INH is administered following immediate suspension from powdered form.  Section of Infectious Diseases, VA Medical Center, Division of Infectious Diseases, University of Pennsylvania, Philadelphia, PA, USA. jol.maslow@med.va.gov

   27.    Maslow J.N., Mikota S.K., Zhu M., Riddle H. and Peloquin C.A. 2005. Pharmacokinetics of ethambutol (EMB) in elephants.J Vet Pharmacol Ther 28: 321-323.

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

   29.    Pandey R. and Khuller G.K. 2005. Antitubercular inhaled therapy: opportunities, progress and challenges.Journal of Antimicrobial Therapy 55: 430-435.

   30.    Sanchez C.R., Murray S.Z., Isaza R. and Papich M.G. 2005. Pharmacokinetics of a single dose of enrofloxacin administered orally to captive Asian elephants (Elephas maximus).Am J Vet Res 66: 1948-1953.
Abstract: OBJECTIVE: To determine the pharmacokinetics of enrofloxacin after oral administration to captive elephants. ANIMALS: 6 clinically normal adult Asian elephants (Elephas maximus). PROCEDURE: Each elephant received a single dose of enrofloxacin (2.5 mg/kg, PO). Three elephants received their complete diet (pellets and grain) within 2 hours after enrofloxacin administration, whereas the other 3 elephants received only hay within 6 hours after enrofloxacin administration. Serum concentrations of enrofloxacin and ciprofloxacin were measured by use of high-performance liquid chromatography. RESULTS: Harmonic mean half-life after oral administration was 18.4 hours for all elephants. Mean +/- SD peak serum concentration of enrofloxacin was 1.31 +/- 0.40 microg/mL at 5.0 +/- 4.2 hours after administration. Mean area under the curve was 20.72 +/- 4.25 (microg x h)/mL. CONCLUSIONS AND CLINICAL RELEVANCE: Oral administration of enrofloxacin to Asian elephants has a prolonged elimination half-life, compared with the elimination half-life for adult horses. In addition, potentially therapeutic concentrations in elephants were obtained when enrofloxacin was administered orally at a dosage of 2.5 mg/kg. Analysis of these results suggests that enrofloxacin administered with feed in the manner described in this study could be a potentially useful antimicrobial for use in treatment of captive Asian elephants with infections attributable to organisms, such as Bordetella spp, Escherichia coli, Mycoplasma spp, Pasteurella spp, Haemophilus spp, Salmonella spp, and Staphylococcus spp.

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

   32.    Zhu M., Maslow J.N., Mikota S.K. et al. 2005. Population pharmacokinetics of pyrazinamide in elephants.Journal of Veterinary Pharmacology and Therapeutics 28: 403-409.
Abstract: This study was undertaken to characterize the population pharmacokinetics (PK),therapeutic dose, and preferred route of administration for pyrazinamide (PZA)in elephants. Twenty-three African (Loxodonta africana) and Asian (Elephas maximus) elephants infected with or in contact with others culture positive for Mycobacterium tuberculosis were dosed under treatment conditions. PZA was dosed daily at 20-30 mg/kg via oral (fasting or nonfasting state) or rectal (enema or suppository) administration. Blood samples were collected 0-24 h postdose. Population PK was estimated using nonlinear mixed effect modeling. Drug absorption was rapid with T(max) at or before 2 h regardless of the method of drug administration. C(max) at a mean dose of 25.6 (+/-4.6) mg/kg was 19.6 (+/-9.5 microg/mL) for PZA given orally under fasting conditions. Under
nonfasting conditions at a mean dose of 26.1 +/- 4.2 mg/kg, C(max) was 25% (4.87 +/- 4.89 microg/mL) and area under concentration curve (AUC) was 30% of the values observed under fasting conditions. Mean rectal dose of 32.6 +/- 15.2 mg/kg yielded C(max) of 12.3 +/- 6.3 microg/mL, but comparable AUC to PZA administered orally while fasting. Both oral and rectal administration of PZA appeared to be acceptable and oral dosing is preferred because of the higher C(max) and lower inter-subject variability. A starting dose of 30 mg/kg is recommended with drug monitoring between 1 and 2 h postdose. Higher doses may be required if the achieved C(max) values are below the recommended 20-50 microg/mL range.

   33.    Hirsch D.C. and Biberstein E.L. 2004. Mycobacterium. In: Hirsch DC, MacLachlan NJ and Walker RL (eds), Veterinary Microbiology pp. 223-234. Blackwell, Ames, Iowa.

   34.    Janssen D.L., Oosterhuis J.E., Fuller J. and Williams K. 2004. Field technique: A method for obtaining trunk wash mycobacterial cultures in anesthetized free-ranging African elephants (Loxodonta africana).  2004 PROCEEDINGS AAZV, AAWV, WDA JOINT CONFERENCE, pp. 582-583.
Abstract: The Guidelines for the Control of Tuberculosis in Elephants 2003 (Guidelines) of the National tuberculosis Working Group for Zoo and Wildlife Species were written to protect the health and safety of captive elephants together with their handlers and the viewing public.1 The Guidelines specifically address the display and transport of captive elephants but do not address the unique situation of free-living elephants being imported and subsequently displayed to the public.

Although the Guidelines describe a technique for collecting and handling a trunk wash in a trained, standing, non-anesthetized elephant, it does not describe a similar technique for anesthetized elephants in lateral recumbency. In an attempt to detect active mycobacterial infection in a group of 3 male and 8 female free-ranging African elephants scheduled for import into the United States, a technique was developed for collecting trunk washes in recumbent,  anesthetized elephants for mycobacterial culture.

A South African game-capture crew, experienced in translocating elephants, anesthetized elephants in groups via remote drug delivery and from a helicopter. The ground crew accomplished multiple simultaneous procedures including anesthesia maintenance and monitoring, physical and reproductive examinations, collection of general diagnostic and investigative samples, and trunk washes for mycobacterial cultures. This was accomplished while the capture crew was preparing animals for loading into specially designed trailers for transport to a holding boma. Little time was available for any one of procedure with multiple
animals being attended to at one time.

Once an elephant was stable in lateral recumbency, a 3-m foal stomach tube, prepackaged and sterilized, was inserted into the dependent side of the trunk tip. It was then gently fed up the trunk approximately 2.5 m. A 50-ml sample suction trap was attached to the end of the foal tube.The suction trap was then attached to a battery powered, portable aspirator pump designed for emergency medical care. The aspiration pump was activated to collect secretions from the most proximal portion of the trunk. If little or no secretions were collected by this means, the system was disconnected between the sample trap and the foal tube. Then, 100 ml of sterile saline was placed into raised end of the foal tube allowing it to drain toward the tip through gravity. The suction trap and aspiration pump were reattached to collect a sample in the sample trap. Then, the sample trap was replaced with a new trap, and the foal tube was inserted into the oral pharynx for collection of a separate oropharyngeal sample. This same procedure was repeated
with each elephant.

ACKNOWLEDGMENTS
So African veterinarians, Mike Bester, Larry Killmar, Janet Payeur, ARC/OVI, Thomas Hildebrant, Eric Zeehandelar, Kevin Reily, Denise SoFranko.

LITERATURE CITED
1. National tuberculosis Working Group for Zoo and Wildlife Species. 2003. Guidelines for the Control of Tuberculosis in Elephants 2003. USDA-APHIS: http://www.aphis.usda.gov/ac/TBGuidelines2003.pdf

   35.    Stringfield C.E., Oh P., Granich R. et al. 2004. Epidemiologic investigation of a Mycobacterium tuberculosis infection of multiple animal species in a metropolitan zoo.  2004 PROCEEDINGS AAZV, AAWV, WDA JOINT CONFERENCE, pp. 46-48.
Abstract: From 1997 to 2000, six cases of Mycobacterium tuberculosis (TB) infection were diagnosed in three species of animals at, or recently originating from, the Los Angeles Zoo. Restriction fragment length polymorphism (RFLP) analysis showed that five of six animal isolates shared an identical IS6110 pattern, with the sixth differing only by one additional band. A multiinstitutional epidemiologic investigation was conducted to identify and interrupt possible transmission among the animal cases, and to screen personnel for active TB infection and TB skin-test conversion.
Animal Cases
In April and October of 1994, Asian elephant (Elephas maximus) #1 and Asian elephant #2 arrived at the Los Angeles Zoo from a private elephant facility where they had lived together. They were housed together at the zoo until November of 1996 when elephant #2 was returned to the facility for several months before transfer to another zoo. In the spring of 1997, Elephant #1 (30 yr old) died of salmonellosis, with M. tuberculosis found in granulomatous lymph node lesions from the thoracic and abdominal cavities, and Elephant #2 (30 yr old) was found to have a positive trunk wash culture for M. tuberculosis. In July of 1998, one of a closed herd of three Rocky Mountain goats (Oreamnos americanus) consisting of a sire and two offspring, died of pulmonary M. tuberculosis at 6 yr of age. The goat's asymptomatic herdmates were screened and had negative chest radiographs and tracheal wash cultures, but one of the two goats was positive on tuberculin skin-test. In October of 1998, a clinically normal Black rhinocerus (Diceros bicornis) was diagnosed with Mycobacerium tuberculosis after a positive skin test and nasal wash culture. In the winter of 1998, the two remaining goats were evaluated again with negative chest radiographs and tracheal wash cultures. However, 1 yr later, both were humanely euthanatized at 8 and 12 yr of age due to clinical evidence of tuberculosis on chest radiographs (both animals), and active clinical signs in one (neither were able to be orally treated). In January of 2001, a rhino was humanely euthanatized after a protracted illness that was nonresponsive to aggressive treatment. The rhino was found to have severe multifocal hemosiderosis and atypical mycobacterial infection in her lungs, with no M. tuberculosis cultured. This animal had been treated with oral Isoniazid and Rifampin for 1 yr, cultured routinely, and was never culture positive again.
Epidemiologic Investigation
Investigators examined medical and location histories of the affected animals, animal handling practices, health-care procedures, and performed an infection control assessment of the animal compounds and health-care facilities (including measuring air flow in the compounds by smoke testing). We conducted a review of zoo employee medical records for evidence of TB symptoms, tuberculin skin-test results, and chest radiograph information. A list of current and former employees was cross-matched with reported TB cases in the California state registry from 1985 to 2000. As part of the annual occupational health screening in June of 2000, zoo employees underwent questioning regarding TB symptoms, received tuberculin skin tests, and completed a questionnaire on medical history, job type, and history of contact with the infected animals.
Epidemiologic Findings
No common cross-species contact outside the animal compounds and no contact with an infectious human were found. The distance at which the public was kept from the animals and the distance of the compounds from each other (the elephant compound was 27 meters from the rhino compound and the goat compound was 90 m from both) suggests that direct transmission was unlikely. No active TB cases in humans were found, and no matches were found in the database of reporte d cases. The RFLP analysis of this strain of M. tuberculosis matched that of three elephants with which #1 and #2 were housed at a private elephant facility from September of 1993-February of 1994.1 We hypothesize that elephants #1 and #2 were infected at the private facility and were shipped with latent M. tuberculosis infection in 1994, subsequently infecting the black rhino and Mountain goats at the Los Angeles Zoo.
Of interest, animal caretaking and animal contact were not associated with a positive tuberculin skin-test, while groundskeepers were found to have an increased risk of tuberculin skin-test conversion compared with other job categories. Employees attending the elephant necropsy and employees who trained elephants were more likely to have tuberculin skin-test conversion than those who did not.
Conclusion
This is the first documented human and veterinary epidemiologic investigation of Mycobacterium tuberculosis affecting multiple species in a zoo. ² No evidence of transmission from humans to animals or active infections in humans were found. Genotyping evidence strongly suggests transmission from one species to another, although no evidence of transmission was discovered. Human tuberculin skin-test conversions associated with the elephants were most likely due to lack of respiratory protection for these employees when the risk of TB infection was not known. The finding that groundskeepers and not animal handlers were associated with a higher risk of tuberculin skin-test conversion was surprising, and we hypothesized that this may have to do with groundskeepers as a group being more likely to have
been born outside of the United States.
Control measures to eliminate the spread of disease to people and animals were undertaken immediately and throughout this outbreak, and no further cases of M. tuberculosis have been diagnosed at the zoo in the past 3 yr despite ongoing surveillance. Four elephants and three rhinos that had direct contact with the infected animals remain TB negative by trunk and nasal wash culture methods as outlined by the USDA for elephant TB surveillance. Methods of indirect transmission in mammalian zoo species and causes of variability in infection and morbidity within and among species warrant further investigation. Ongoing vigilance, occupational health programs and infection control measures in potentially exposed animals are recommended to prevent ongoing transmission of M. tuberculosis in zoo settings.
Acknowledgments
The authors thank the Animal Care and Animal Health staff of the Los Angeles Zoo who cared so well for these animals, and the veterinarians (including consulting pathologists), technicians, and medical records staff who collected, analyzed, and organized the clinical data. We could not have performed this evaluation without Sue Thisdell, Safety Officer at the Los Angeles Zoo; Jothan Staley and Donna Workman-Malcom of the City of Los Angeles Occupational Health Services Division; Lee Borenstein, Elenor Lehnkering, Patrick Ryan, Jeanne Soukup, and Annette Nita of the Los Angeles County Department of Health Services; and Diana Whipple for her RFLP expertise.
LITERATURE CITED
1. Mikota, S.K., L. Peddie, J. Peddie, R. Isaza, F. Dunker, G. West, W. Lindsay, R.S.Larsen, M. D. Salman, D. Chatterjee, J. Payeur, D. Whipple, C. Thoen, D. Davis, C. Sedgwick, R.J. Montali, M. Ziccardi, J. Maslow. 2001. Epidemiology and diagnosis of Mycobacterium tuberculosis in captive asian elephants (Elephas maximus). J. Zoo Wildl. Med. 32: 1-16.
2. Oh, P., R. Granich, J. Scott, B. Sun, M. Joseph, C. Stringfield, S. Thisdell, J. Staley, D. Workman-Malcolm, L. Borenstein, E. Lehnkering, P. Ryan, J. Soukup, A.Nitta, J. Flood. 2002. Human exposure following Mycobacterium tuberculosis infection of multiple animal species in a metropolitan zoo. Emerging Infectious Diseases. 8 (11): 1290-1293.orte

   36.    Guidelines for the control of tuberculosis in elephants.  2003.
Ref Type: Electronic Citation

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

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

   39.    Michel A.L., Venter L., Espie I.W. and Coetzee M.L. 2003. Mycobacterium tuberculosis infections in eight species at the National Zoological Gardens of South Africa, 1991-2001.Journal of Zoo and Wildlife Medicine 34: 364-370.
Abstract: Between 1991 and 2001 a total of 12 cases of Mycobacterium tuberculosis infection in eight different species were recorded in the National Zoological Gardens of South Africa in Pretoria (Tshwane). The genetic relatedness between seven of the M. tuberculosis isolates was determined by IS6110 restriction fragment length polymorphism analysis. For the majority of the isolates that were analyzed, a high degree of polymorphism suggested different sources of infection. Evidence of M. tuberculosis transmission between animals is reported in two chimpanzees (Pan troglodytes) housed together, from which samples were collected for analysis 29 mo apart.

   40.    Mikota S.K., Hammatt H. and Finnegan M. 2003. Occurrence and prevention of capture wounds in Sumatran elephants (Elephas maximus sumatranus).  Proc Amer Assoc Zoo Vet, pp. 291-293.
Abstract: The capturing of elephants in Indonesia began in 1986 as an attempted solution to human-elephant conflict.  The intent was to train "problem" elephants for use in agriculture, logging and tourism.  The initial captures were conducted under the guidance of Thai mahouts and Thai koonkie elephants (trained elephants used for capture).  A number of the Indonesians that were originally trained in capture techniques still work for the government forestry department (KSDA).  The younger pawangs (elephant handlers) that participate in captures have learned from their peers.  There is no formal training program. The actual mortality rate associated with elephant captures in Sumatra is unknown as official reports are lacking.  The age structure of the existing ~ 400 captive elephants is young (most under 25) which suggests that smaller, younger elephants are preferentially captured and / or that adult elephants do not survive the capture and training processes.  Our personal experiences (Mikota and Hammatt) in Sumatra show that mortality in newly captured elephants is high.In 2001, with endorsement from the World Wide Fund for Nature-Indonesia (WWF), the Wildlife Conservation Society (WCS), Fauna and Flora International (FFI), and the International Elephant Foundation (IEF), we requested a two-year Moratorium on elephant captures during which time capture techniques would be improved and alternative conflict mediation techniques evaluated.
A Moratorium against placing additional elephants into the Elephant Training Centers has been issued by the central government, however capture for translocation is still sanctioned.  Unfortunately, the provincial governments have increasingly acted in their own interests since the government of Indonesia began a de-centralization process a few years ago. Riau Province is thought to have the largest remaining populations of wild Sumatran elephants.Fifty-seven, human-elephant conflicts occurred in Riau between 1997-2000.  Although Riau is a hotbed of conflict, problems are occurring throughout Sumatra and we are aware of conflicts and captures in Bengkulu and North Sumatra. In October 2002, we were invited by KSDA (the provincial forestry department) to accompany their team into the field as they attempted to capture a large bull that had been raiding a palm oil plantation.  This opportunity was invaluable as we were able to observe first hand the techniques being used and where improvements were needed.  As a result of this and other experiences with newly captured elephants we observed: ·Equipment (Palmer) is old, poorly maintained, and used improperly. ·Essential supplies are lacking or homemade substitutes are used.
·The dose of xylazine is very high compared to wild elephant capture doses used in India and Malaysia.  The same dose is often used regardless of the size of the elephant. ·The needles are too short to reach muscle; open-ended needles are used which can become plugged with tissue, thus preventing injection. ·Neither the correct charge nor the correct load is selected.  We observed that many darts bounced making it difficult to ascertain the amount of drug injected or its depth of penetration.  Selection of an inappropriate charge results in unnecessary trauma. ·The preparation and use of darts, needles, and syringes lacks basic hygiene. ·Dart wounds are not treated and antibiotics are not administered.  ·There is no understanding of stress or capture myopathy. ·The capture team was not aware that sternal recumbency severely compromises respiration in elephants and that they can quickly die in this position. ·It is believed that elephant restraints must inflict pain to prevent wild elephants from escaping once captured.  ·There is no veterinarian on the capture team. The current capture techniques result in leg wounds from unprotected chains, neck wounds from "kahs" (neck yokes made of wood and wire), and abscesses from inappropriately administered darts.  Leg and neck wounds often become maggot infested.  Infections from dart wounds are, however, the primary cause of capture-related mortality.  These abscesses can drain for several months, even with treatment, and often progress to a necrotizing fasciitis, acute sepsis, and death. The Riau Province KSDA Team has been receptive to suggested changes to minimize wounds. Provision of heavier chains has alleviated the fear that elephants will escape.  Covering the chains with fire hose or heavy plastic minimizes injuries to legs and use of the kah has been discontinued.  A basic dart wound treatment protocol has been established. In June 2003, a comprehensive Elephant Immobilization and Translocation Workshop for Sumatra is planned to retrain all of Sumatra's field teams and to upgrade equipment. Sumatra's wild elephant population probably numbers fewer than 3000 and is under continued threat.  With so few elephants left, the preservation of as many viable herds as possible takes on increased urgency.  The Moratorium achieved in 2001 has set the groundwork for KSDA to choose translocation of wild elephants rather than capture and placement into already over-crowded and under-resourced Elephant Training Centers.  We cannot guarantee that Sumatra will capture elephants only for translocation, and it is inevitable that many more elephants will end up in captivity.  Regardless, all of the elephants that must suffer the interruption of their lives at the hand of man deserve, at the very least, humane treatment.  Translocations are neither simple nor a complete panacea.  Identifying suitable translocation areas and insuring that elephants remain there are significant challenges.  WWF-Indonesia is continuing its efforts to secure the lowland forest of Tesso Nilo in Riau Province as a "safe haven" for at least some of Sumatra's wild elephants (see WWF AREAS Program – Riau, Sumatra: http://www.worldwildlife.org/species/attachments/riau_profile.pdf).  The identification of interim release sites, together with improved capture techniques, offers the hope that fewer elephants will be removed from the wild.   ACKNOWLEDGMENTS: Our work in Sumatra has been supported by the Guggenheim Foundation, a CEF grant from the American Zoo and Aquarium Association, the International Elephant Foundation, Oregon Zoo, Columbus Zoo, Disney, Peace River Refuge, the Elephant Managers Association, the Riddles Elephant and Wildlife Sanctuary, Tulsa Zoo, Toronto Zoo, Niabi Zoo, San Antonio Zoo, Denver Zoo (AAZK Chapter), Milwaukee Zoo (AAZK Chapter), the Audubon Nature Institute (Youth Volunteers), Buttonwood Park Zoo, Melbourne Zoo, and private donors.  Special thanks to Harry Peachey, John Lehnhardt, Holly Reed, Kay Backues, Mike Keele, Steve Osofsky, and Heidi and Scott Riddle.

   41.    Nath I., Bose V.S.C., Panda S.K., Das B.C. and Singh L.A.K. 2003. A case of multiple abscesses in a baby elephant.Zoos' Print Journal 18: 1270.

   42.    Pavlik I., Ayele W.Y., Parmova I. et al. 2003. Mycobacterium tuberculosis in animal and human populations in six Central European countries during 1990-1999.Veterinarni Medicina 48: 83-89.
Abstract:  Results of Mycobacterium tuberculosis detection in animals from six Central European countries (Croatia, the Czech Republic, Hungary, Poland, Slovakia and Slovenia) spreading over 610402 km2 with a population of 11.8 million heads of cattle were analysed. In the monitoring period between 1990 and 1999, M. tuberculosis from animals was isolated only in two countries (Poland and Slovak Republic) from 16 animals with tuberculous lesions. These comprise 9 cattle (Bos taurus), 4 domestic pigs (Sus scrofa f. domestica) and three wild animals, an African elephant (Loxodonta africana), agouti (Dasyprocta aguti) and terrestrial tapir (Tapirus terrestris) from a zoological garden Gdansk in Poland. A steady decrease in the incidence of tuberculosis in humans was recorded during the monitoring period in all countries. The human population of the study countries was 68.03 million. In the period monitored, infection caused by M. tuberculosis was identified in a total of 241040 patients with a decreasing incidence of tuberculosis found in all countries. The lowest relative bacteriologically confirmed disease was found in the Czech Republic, Slovak Republic and Slovenia. Given the low number of infected domestic and wild animals, the epidemiological and epizootiological situation may be considered auspicious.

   43.    Potters D., Seghers M., Muyldermans G. et al. 2003. Recovery of Mycobacterium elephantis from sputum of a patient in belgium.Journal of Clinical Microbiology 41: 1344.
Abstract: Mycobacterium elephantis was isolated from a human respiratory specimen in April 1999, demonstrating its presence in Europe. The biochemical reaction results, antimicrobial susceptibility pattern, and sequence data for this strain are all in agreement with those of M. elephantis strains isolated previously from other continents.

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

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

   46.    Schmitt D.L. 2003. Proboscidea (Elephants). In: Fowler ME and Miller RE (eds), Zoo and Wild Animal Medicine pp. 541-550. Elsevier Science USA.

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

   48.    Vodicka R. and Kral J. 2003. Purulent trunk dermatitis in a male Ceylon elephant (Elephas maximus).Verh.ber.Erkrg.Zootiere 41: 151-153.
Abstract: A report in given on the therapy of purulent trunk dermatitis in an aggressive male Ceylon elephant. Despite the non-standard steps we took (repeated anaesthesias during a short time, non-compliance with the recommendations as to the application of some drugs, etc.) and the difficult handling (an aggressive; incontrollable elephant, no restraint chute), it is possible even to treat a case like this.

   49.    Ziccardi M., Wong H.N., Tell L.A. et al. 2003. Further optimization and validation of the antigen 85 immunoassay for diagnosing mycobacteriosis in wildlife.  Proc Amer Assoc Zoo Vet, pp. 219-220.
Abstract: Mycobacteriosis caused by Mycobacterium bovis, M. tuberculosis and M. avium has been a well-documented health problem for zoological collections as long ago as the late 19^th century.  Prevalence estimation in these captive wildlife populations, however, has been hampered by diagnostic test methods that are oftentimes difficult or impossible to conduct and/or interpret (due to the requirement for multiple immobilizations for measurement of response), the occurrence of non-specific results with methods such as the intradermal skin test, and/or the near-total lack of validation, optimization and standardization of any of the available test methods in the species of interest.  Additionally, because intradermal skin testing is the primary screening method for many of these species, the ability to compare exposure in captive wildlife with exposure in free-ranging populations has been limited due to the difficulty with follow-up in free-ranging populations.  Lastly, unlike testing methods that use serological techniques, skin testing precludes retrospective studies of banked samples to determine onset of reactivity.

Recently, human tuberculosis researchers working with tuberculosis in humans have developed an immunoassay that detects a serum protein complex (the antigen 85, or Ag85, complex) produced by mycobacteria in the early stages of mycobacterial infections¹.  Previous work has shown that this method is a promising diagnostic tool in the evaluation of tuberculosis exposure in some primate (including orangutan (Pongo pygmaeus ), a species known for non-specific tuberculin responses)²  and captive hoofstock species³.  In order to determine the feasibility and applicability of a widespread use of this method for captive and free-ranging wildlife species, we have undertaken a number of pilot studies on different populations of interest, with the goals of optimizing and validating the immunoassay through analysis of serum from known infected and non-infected individuals and through comparisons with other diagnostic methods.  Thus far, we have begun evaluating the applicability of the antigen 85 immunoassay in various avian, primate, rhinoceros and hoofstock species for detecting tuberculosis and/or paratuberculosis (Johne's disease) infections.  Preliminary results, a summary of which will be presented, indicate that this method may be a valuable adjunct to other testing methods (including gamma interferon and multiple-antigen ELISA) to allow a better evaluation of true mycobacterial status in these species.

LITERATURE CITED

1.Bentley-Hibbert, S. I., X. Quan, T. G. Newman, K. Huygen and H. P. Godfrey. 1999.  Pathophysiology of Antigen 85 in patients with active tuberculosis. Infect Immun. 67(2):581-8.
2.Kilbourn, A. M., H. P. Godfrey, R. A. Cook, P. P. Calle, E. J. Bosi, S. I. Bentley-Hibbert, K. Huygen, M. Andau, M. Ziccardi and W. B. Karesh.  2001.  Serum Antigen 85 levels in adjunct testing for active mycobacterial infections in orangutans.  J. Wildl. Dis. 37(1): 65-71.
3.Mangold, B. J., R. A. Cook, M. R. Cranfield, K. Huygen, and H. P. Godfrey.  1999.  Detection of elevated levels of circulating antigen 85 by dot immunobinding assay in captive wild animals with tuberculosis.  J. Zoo Wildl. Med. 30(4): 477-483.

   50.    Alexander K.A., Pleydell E., Williams M.C. et al. 2002. Mycobacterium tuberculosis: An Emerging Disease of Free-Ranging Wildlife.Emerging Infectious Diseases 8: 598-601.
Abstract: Expansion of ecotourism-based industries, changes in land-use practices, and escalating competition for resources have increased contact between free-ranging wildlife and humans. Although human presence in wildlife areas may provide an important economic benefit through ecotourism, exposure to human pathogens
may represent a health risk for wildlife. This report is the first to document introduction of a primary human pathogen into free-ranging wildlife. We describe outbreaks of Mycobacterium tuberculosis, a human pathogen, in free-ranging banded mongooses (Mungos mungo) in Botswana and suricates (Suricata suricatta) in South Africa. Wildlife managers and scientists must address the potential threat that humans pose to the health of free-ranging wildlife.

   51.    Auclair B., Mikota S., Peloquin C.A., Aguilar R. and Maslow J.N. 2002. Population pharmacokinetics of antituberculous drugs and treatment of Mycobacterium bovis  infection in Bongo Antelope (Tragelaphus eurycrus isaaci ).Journal of Zoo and Wildlife Medicine 33: 193-203.

   52.    Benkirane A. and de Alwis M.C.L. 2002. Haemorrhagic septicaemia, its significance, prevention and control in Asia.Vet.Med.-Czech 47: 234-240.
Abstract: Haemorrhagic septicaemia (HS) is an endemic disease in most countries of Asia and sub Saharan Africa. Within the Asian Region, countries can be classified into three categories, on the basis of incidence and distribution of the disease; these are respectively countries where the disease is endemic or sporadic, clinically suspected but not confirmed, or free. Economic losses due to HS are not only confined to losses to the animal industry, but also rice production on account of its high prevalence among draught animals used in rice fields. Only a few attempts have been made to estimate economic losses, the methodologies used in different countries have varied, and many are not based on active surveillance, and a consideration of all components of direct and indirect losses. Most Asian countries rank HS as the most important contagious disease or the most important bacterial disease in cattle and buffaloes. Resource allocation for prevention and control of HS nationally or internationally will evidently depend on a correct estimate of its economic impact. The key factors in prevention and control would be timely and correct reporting, accurate and rapid diagnosis, strategic use of vaccines with the attainment of a high coverage where necessary with a high quality vaccine. National level activities geared towards attainment of these objectives may be with advantage supported and strengthened by international organisations involved in animal health. ?e present paper attempts to review aspects related to the epidemiology, control and containment of HS in Asia and, proposes some key issues on which a regional programme for HS control in this continent should be centred.

   53.    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.

   54.    Gavier-Widen D., Hard Af Segerstad C., Roken B. et al. 2002. Mycobacterium tuberculosis infection in Asian elephants (Elephas maximus) in Sweden.  European Association of Zoo and Wildlife Veterinarians 4th Scientific Meeting, Heidelberg, Germany.

   55.    Mikota S.K. and Maslow J. 2002. Epidemiology and Treatment of Tuberculosis in Elephants: 2002. In: Baer CK (ed), American Association of Zoo Veterinarians Annual Conference, 2002, pp. 384-387.

   56.    Nayar K.N.M., Chandrasekharan K. and Radhakrishnan K. 2002. Management of surgical affections in captive elephants.Journal of Indian Veterinary Association Kerala 7: 55-59.

   57.    Oh P., Granich R., Scott J. et al. 2002. Human exposure following Mycobacterium tuberculosis infection of multiple animal species in a Metropolitan Zoo.Emerg Infect Dis 8: 1290-1293.
Abstract: From 1997 to 2000, Mycobacterium tuberculosis was diagnosed in two Asian elephants (Elephas maximus), three Rocky Mountain goats (Oreamnos americanus), and one black rhinoceros (Diceros bicornis) in the Los Angeles Zoo. DNA fingerprint patterns suggested recent transmission. An investigation found no active cases of tuberculosis in humans; however, tuberculin skin-test conversions in humans were associated with training elephants and attending an elephant necropsy.

   58.    Payeur J.B., Jarnagin J.L., Marquardt J.G. and Whipple D.L. 2002. Mycobacterial isolations in captive elephants in the United States.Ann N Y Acad Sci 969: 256-258.
Abstract: Interest in tuberculosis in elephants has been increasing over the past several years in the United States. Several techniques have been used to diagnose mammalian tuberculosis. Currently, the test considered most reliable for diagnosis of TB in elephants is based on the culture of respiratory secretions obtained by trunk washes.

   59.    Peloquin CA. 2002. Therapeutic drug monitoring in the treatment of tuberculosis.Drugs 62: 2169-2183.

   60.    Singh V.N. 2002. Symptomatic study of haemorrhagic septicaemia in elephant in Mudumalai Wildlife Sanctuary, Tamil Nadu.Indian Forester 128: 1089-1100.
Abstract: Symptoms of haemorrhagic septicaemia, a dreaded disease in elephant is recorded in this study which reveals systematic spread of Oedema from jowl to throat, neck, brisket, abdomen and perenical regions. It also records the changes in character/colour of dung, urine, eye, tongue, trunk, body temperature, feeding habit and body condition alo