The deaths of two Asian elephants (Elephas maximus) in August 1996 led the United States Department of Agriculture to require the testing and treatment of elephants for tuberculosis. From August 1996 to September 1999, Mycobacterium tuberculosis infection was confirmed by culture in 12 of 118 elephants in six herds. Eight diagnoses were made antemortem on the basis of isolation of M. tuberculosis by culture of trunk wash samples; the remainder (including the initial two) were diagnosed postmortem. We present the case histories, epidemiologic characteristics, diagnostic test results, and therapeutic plans from these six herds. The intradermal tuberculin test, enzyme-linked immunosorbent assay serology, the blood tuberculosis test, and nucleic acid amplification and culture are compared as methods to diagnose M. tuberculosis infection in elephants.

Humoral immune responses of black-footed penguins (Spheniscus demersus) to DNA-mediated immunization with a β-galactosidase reporter gene expression plasmid were evaluated. Six male and 6 female adult penguins received either test plasmid, pCMV-β, containing the β-galactosidase gene or control plasmid, pCI, lacking a gene for expression. Three birds from each group were used previously in a diluent control group and given one injection of sterile saline. All samples were screened for anti-β-galactosidase antibodies by indirect enzyme-linked immunosorbent assay with anti-chicken immunoglobulin G as secondary antibody. Antibodies to β-galactosidase were detected in the sera of pCMV-β–inoculated penguins, with a peak response on day 21. Antibody titers of the test plasmid group versus both control groups on days 21, 28, and 42 differed significantly. These results demonstrate that black-footed penguins can be safely transfected with the gene encoding β-galactosidase and will mount a humoral response against the in vivo–expressed protein. Knowledge from this initial study can be applied to the development of DNA-mediated vaccines against specific infectious diseases of penguins.

Feline coronavirus genetic elements were detected by polymerase chain reaction from blood, fecal samples, and effusive fluid collected from 33 cheetahs in the U.S.A. Feline coronavirus-specific serum antibodies were also measured by indirect immunofluorescence. Ten cheetahs were positive for viral shedding by polymerase chain reaction, whereas 13 were seropositive by immunofluorescence. Results of serology did not consistently correlate with shedding of virus, and the capture antigen used for detection of feline coronavirus-specific antibodies had a significant impact on results. Testing of samples from one population over a 1-yr period indicated chronic infection in some animals. These relatively healthy carrier animals were a source of virus for contact animals. Screening programs in cheetah populations for feline coronavirus infection may be most reliable if a combination of serologic analysis and viral detection by polymerase chain reaction is used.

Reproductive tracts or tissues from five male black rhinoceroses (Diceros bicornis), two male white rhinoceroses (Ceratotherium simum), two male one-horned Asian rhinoceroses (Rhinoceros unicornis), seven female black rhinoceroses, and six female white rhinoceroses from multiple institutions were examined to characterize their anatomy and histology. Some observations and measurements were obtained from in situ tracts of intact animals before or during necropsy. Formalin-fixed tissues were dissected and examined histologically. Retrospective reproductive data from each rhinoceros was obtained from the institutions of origin. Reproductive histology of these species was similar to that of other mammals. Male accessory gland structure varied among species, and the Asian rhinoceros epididymis was more loosely attached and had larger duct diameters than did the epididymides of the African species. Although histology was typically mammalian, rhinoceros reproductive morphology combined chacteristics of several different mammals. Defining this unique morphology of rhinoceroses may help in understanding their reproductive physiology and will effect the development of appropriate reproductive techniques.

To examine the waveforms of electrocardiograms, cardiac rhythm, heart rates at rest and during excitement, and the rate of increase of heart rate, electrocardiograms were recorded with standard bipolar limb leads from 79 free-living birds, including 19 species. The heart weights and heart-to-body weight ratios were obtained from an additional 96 free-living birds, including 20 species. In the majority of the electrocardiograms, lead I was of low amplitudes for all waves except the P wave, and leads II and III were very similar to each other with a predominant S wave and a very short or elevated ST segment. The P wave was often superimposed on the T wave when the heart rate increased to 330 beats/min. Four types of arrhythmia were observed in 50 of the 79 birds (63.3%): 48 sinus arrhythmias, four sinus arrests, two atrial premature contractions, and one ventricular premature contraction. The resting heart rate was negatively associated with the rate of increase, suggesting that a bird with a low resting rate might be able to maintain a greater capacity to increase its heart rate than one with a high resting rate. A negative correlation on a bilogarithmic scale was obtained between the heart weight and the resting heart rate, indicating that a bird with a high heart weight had lower resting heart rate than a bird with a low heart weight. When the heart-to-body weight ratios of free-living birds were compared according to their motility, the ratios of more active birds were greater than those of less active ones.

Eight capuchin monkeys (Cebus apella) were vaccinated against rabies with an inactivated suckling mouse brain vaccine (SMBV). Three 1-ml doses of 2% brain tissue suspension were given by i.m. injection at 0, 30, and 60 days. Blood samples were collected at 0, 30, 60, 90, 150, 210, 240, 300, and 365 days and were tested by simplified fluorescence inhibition to titer-neutralizing antibodies. All of the animals developed neutralizing antibodies with titers >0.5 IU/ml after vaccination, but the immune response persisted for only 122.3 ± 32.6 days. The SMBV was able to induce immune response in the capuchin monkeys, but protection was short-lived.

Spondyloarthropathy was observed in 25 (2.8%) of 895 preserved canid museum specimens and was catalogued by species. The associated skeletal alterations in canids are indistinguishable grossly and physiologically from those in humans with spondyloarthropathy of the reactive type. Rate of affliction was independent of captive or wild-caught status or gender. In canids, spondyloarthropathy was much more common than osteoarthritis (0.3%), which predominantly is limited to captive animals. Animal well-being may be enhanced by recognition of the condition and initiation of specific treatment.

Eimeria gruis and Eimeria reichenowi are common coccidial parasites of a number of species of cranes. Until recently, little was known about either the site for invasion or the dynamics of early development of the crane coccidia because of the difficulty of identifying sporozoites and early developmental stages of these parasites by conventional staining methods. In the present study, monoclonal antibodies (MAbs) elicited against Eimeria spp. of chickens and turkeys were found to cross-react with sporozoites and developmental stages of E. reichenowi in the tissues of Florida sandhill cranes (Grus canadensis). With these Mabs, E. reichenowi sporozoites were found in specimens taken at 6 hr postinoculation (PI) from just proximal to Meckel's diverticulum in the jejunum to the ileocecal juncture. Fewer were found in the ceca and rectum and none in the duodenal loop. At 24 hr PI, there were markedly fewer sporozoites and their location had shifted to the duodenum. No stages were seen in intestinal cells at 5 days PI (DPI), but trophozoites had developed in the liver and spleen. At 10 DPI, sexual stages were detected in the intestine from the duodenal loop through Meckel's diverticulum but not in other organs. By 14 DPI, numerous developmental stages were detected in the intestine (ceca and jejunum), liver, and lungs but not in the heart, kidney, or brain. The number, location, and maturity of the stages in the ceca differed markedly from those in the jejunum.

Approximately 350 Amazon parrots were destined for relocation in Peten province, northeastern Guatemala. In random sampling of the parrots, 95 blood and 75 fecal samples were examined individually for parasites. Coccidia were present in 6.0% (3/50) of Amazona autumnalis autumnalis, and they were the only parasites detected. There were no blood parasites observed in 64 A. a. autumnalis, four Amazona pionus senilis, 16 Amazona ferinosa guatemala, 10 Amazona albifronsus albifronsus, and one Amazona xantholora. No fecal parasites were observed in four A. p. senilis, 12 A. f. guatemala, eight A. a. albifronsus, and one A. xantholora.

Penicillin G and antipyrine, which served as model drugs to assess the relative capacities of renal and hepatic elimination pathways, respectively, were each administered intravenously to six ostriches (Struthio camelus) and to six emus (Dromaius novaehollandiae). Drug concentrations in blood samples collected over a period of 12 hr after administration were assayed, and elimination half-life, mean residence time, clearance, and steady-state volume of distribution were calculated. Mean values for elimination half-life and mean residence time of penicillin G were significantly higher in emus than in ostriches; no significant differences in antipyrine pharmacokinetics between species were demonstrated.

This study evaluated the immune response of 47 (22 males, 25 females) captive maned wolves (Chrysocyon brachyurus) to modified-live canine parvovirus and canine distemper virus (Onderstepoort and Rockborn strains) vaccines. Sera were collected from 33 adults and 14 pups, including five free-ranging pups captured at 1 yr of age or younger. All the adults and four captive-born pups had been vaccinated prior to this first blood collection. Virus neutralization and hemagglutination-inhibition assays were performed for quantitating antibodies against canine distemper and canine parvovirus, respectively. Distemper antibody titers ≥100 were present in 57% of adults and 14% of pups. All adults and 29% of pups had parvovirus antibody titers ≥80. After vaccination, 72% of the wolves developed antibody titers ≥100 against distemper and 98% developed titers ≥80 against parvovirus. Both vaccines used were safe and immunogenic to juvenile and adult maned wolves, regardless of prior vaccination history.

With the use of a crossover study design, we investigated the respiratory and cardiovascular effects of naloxone administration in eight healthy Rocky Mountain wapiti (Cervus elaphus nelsoni) anesthetized with carfentanil (10 μg/kg i.m.) and xylazine (0.1 mg/kg). Anesthetized animals showed profound hypoxemia with mild hypercapnia, tachycardia, hypertension, and acidosis prior to naloxone administration. After monitoring equipment was placed, animals were administered either naloxone (2 μg/μg carfentanil i.v.) or an equivalent volume of normal saline. Mean values for P_aO₂, P_aCO₂, heart rate, and respiratory rate were significantly different between naloxone- and saline-treated groups, but mean blood pressure, hematocrit, and serum electrolyte concentrations were not. Mean P_aO₂ was 23.0 ± 4.1 mm Hg prior to administration of naloxone or saline and increased to 50.2 ± 7.3 mm Hg after naloxone administration. Mean P_aO₂ of saline-treated animals did not change significantly. Electrocardiograms of three saline-treated animals suggested myocardial hypoxia. Hypoxemia appeared to be caused by respiratory depression, hemodynamic alterations, and lateral recumbency. All but one animal remained anesthetized after naloxone administration. Anesthesia in all animals was reversed in ≤4 min with naltrexone (100 mg/mg carfentanil i.v. s.c.) and yohimbine (0.1 mg/kg i.v.). One bolus of naloxone improved oxygenation in carfentanil–xylazine-anesthetized wapiti.

A 3-yr-old bearded dragon (Acanthodraco vitticeps) presented with lethargy, a swollen right elbow joint, inability to move its rear limbs normally, and marked leukocytosis. The majority of leukocytes were an abnormal mononuclear lymphoid-type cell with a high nuclear to cytoplasmic ratio, a slightly blue cytoplasm, nuclei with coarsely granular chromatin, and some nuclear clefts. Acute leukemia of lymphoid or myeloid origin was tentatively diagnosed. The abnormal mononuclear leukocyte cell population stained positively for the myeloid cytochemical stains: peroxidase, chloroacetate esterase, and L1-calprotectin. The abnormal cell population of the peripheral blood did not stain with the lymphoid cytochemical stains: α-naphthyl butyrate esterase, CD3, and CD79a.

Acid-fast organisms were identified by histopathology of granulomatous lesions in an ostrich (Struthio camelus). The organisms were grown in Herrold's egg media with and without mycobactin and identified as Mycobacterium avium. An agar gel immunodiffusion (AGID) test for Mycobacterium avium paratuberculosis was performed for detection of antibody for M. avium in this infected ostrich and seven other ostriches that were in contact. The results of the AGID were consistent with the pathologic diagnosis of mycobacteriosis and the isolation of M. avium in the affected ostrich.

An epizootic of severe Cryptosporidium sp.–associated enteritis occurred in a group of 15 wild-caught juvenile rough green snakes (Opheodrys aestivus) at the Baltimore Zoo quarantine facility. All of the animals died with no premonitory signs. Histopathologic examination of the small and proximal large intestine of eight of the green snakes showed moderate to severe Cryptosporidium sp. infection and enteritis characterized by dense heterophilic and lymphocytic inflammatory infiltrates throughout the lamina propria with epithelial necrosis. Cryptosporidium sp. was also found in feces of an adult common garter snake (Thamnophis sirtalis) that was wild caught on zoo grounds and held in quarantine during the epizootic. After euthanasia, histologic examination of the garter snake showed a severe small intestinal Cryptosporidium sp. infection with only mild enteritis consisting of sparse heterophilic and lymphocytic infiltrates. There was no gross or histologic evidence of Cryptosporidium sp. gastritis in the nine snakes evaluated, and this is the first report of Cryptosporidium sp.–associated enteritis in snakes without gastric lesions.

A male Cope's grey tree frog (Hyla chrysoscelis) died spontaneously with ventral subcutaneous edema and was necropsied. Thickening of the intestinal mucosa was observed histopathologically, with villous atrophy and intraepithelial nematodes present. Adult female Strongyloides sp. nematodes were isolated from the fixed intestinal tract, the first time this nematode genus has been recovered from this frog genus. Intestinal strongyloidiasis should be considered as a cause for protein-losing enteropathy and death in frogs.

Preductal aortic coarctation and patent ductus arteriosus are described in a neonatal Sumatran tiger, Panthera tigris sumatrae. Eight days postpartum, the cub appeared weak, and it was separated from the dam for hand rearing. On examination it was dehydrated and hypothermic. Despite treatment, the animal's condition worsened and the cub died 12 days postpartum. Gross postmortem and histologic examinations revealed a preductal aortic coarctation and patent ductus arteriosus with a patent foramen ovale and moderate dilatation of the right ventricle of the heart. Focal pneumonia and mild hepatitis were also present; however, diffuse pulmonary congestion and edema were considered to be the proximate cause of death.

Both male and female Foleyella furcata were found in the subcutaneous tissue and abdominal cavity of an adult male wild-caught Senegalese chameleon (Chamaeleo senegalensis) This nematode species is endemic to Madagascar but has never been recorded from the continent of Africa. Prior to the chameleon's death, a migrating worm was seen under the skin in the abdominal and thoracic region. Huge numbers of small, sheathed microfilariae were detected in the blood smears. The chameleon was treated with a single dose (0.2 mg/kg s.c.) of ivermectin. Serious adverse reactions (complete inertia) developed within 24 hr after injection and lasted for 7 days, indicating either ivermectin toxicity or a systemic reaction involving the release of endotoxins from the microfilariae dying in the bloodstream as a result of parasiticide therapy. Therefore, ivermectin treatment of chameleons infected with Foleyella should be avoided.

A 10-day-old female southern white rhinoceros calf (Ceratotherium simum simum) was diagnosed with a patent urachus after urine was observed dribbling from the umbilicus. After being separated from its mother, the animal was sedated with i.m. butorphanol and anesthetized with isoflurane in oxygen for surgical correction of the patent urachus. Mild postoperative complications involved seroma formation and partial skin incision dehiscence, which necessitated three follow-up immobilizations for reevaluation and treatment of the surgical site. Histopathology did not reveal an infectious etiology as the cause for the complications or for the patent urachus. The etiology of the patent urachus in this animal remains undetermined. This report represents the first documented case of a patent urachus in a white rhinoceros.

On 27 May 1999, a big brown bat (Eptesicus fuscus) was discovered on an island exhibit at the Denver Zoo that contained a troop of 15 hooded capuchin monkeys (Cebus apella cay). The monkeys were attacking the bat when it was discovered. The bat was collected and humanely euthanatized without direct handling and submitted to the Colorado Department of Public Health and Environment Virology Laboratory for rabies evaluation. The monkeys had not been vaccinated against rabies virus. The next day, the laboratory confirmed that the bat was positive for rabies. The recommendations from the Colorado Department of Public Health and Environment and the Centers for Disease Control and Prevention were to euthanatize the monkeys or quarantine them and comply with the human nonvaccinated postexposure protocol. A 1-ml dose of a killed rabies vaccine was administered i.m. in the hip on each of days 2, 7, 12, 19, and 33 postexposure, and a single dose of human rabies immune globulin was administered i.m. 5 days postexposure. Blood was collected under anesthesia in order to evaluate the immune response after rabies vaccination from six monkeys 5 days postexposure, six monkeys 19 days postexposure (five of the six monkeys were the same monkeys bled 5 days postexposure), 15 monkeys 67 days postexposure, and 13 monkeys approximately 1 yr postexposure. All of the monkeys developed and maintained levels of rabies virus neutralizing antibody above 0.05 IU/ml by 67 days postexposure. Although a serologic titer of 0.05 IU/ml indicates an adequate human response after rabies vaccination, no similar information is available for nonhuman primates. To date, none of the monkeys has succumbed to rabies.

Fourteen captive and five free-ranging Minnesota gray wolves (Canis lupus) were tested for the presence of rabies virus neutralizing antibodies (RVNA) after vaccination with an inactivated canine rabies vaccine. Blood was collected from all wolves prior to vaccination and at 1 mo postvaccination (PV) and from all captive and three wild wolves at 3 mo PV. In addition, one free-ranging wolf was sampled at 4 mo PV, and two free-ranging wolves were sampled at 6 mo PV. All wolves were seronegative prior to vaccination. RVNA were detected in 14 (100%) captive wolves and in four of five (80%) free-ranging wolves. The geometric mean titer of the captive wolves at 1 mo PV was significantly higher (P = 0.023) than in the free-ranging wolves. Five of 13 (38.5%) captive wolves and none of the three (0%) free-ranging wolves had measurable RVNA at 3 mo PV. No measurable RVNA were detected in the serum samples collected from the free-ranging wolves at 4 and 6 mo PV. These results should be interpreted with caution because of the small number of free-ranging wolves tested. Further research is needed to properly assess immune function and antibody response to vaccination in captive wolves in comparison with their free-ranging counterparts.

A neonate male owl monkey (Aotus sp.) was identified cytogenetically as a hybrid after it failed to nurse and died. Phenotypically, the male parent possessed characteristics of the “gray-neck group,” and G-banded karyotypes identified him as Aotus lemurinus griseimembra (2n = 53), heterozygous for the centric fusion of chromosomes 13 and 14. The female parent belonged to the “red-neck group” and was identified cytogenetically as Aotus nancymaae (2n = 54). The neonate hybrid had 2n = 54 chromosomes with 13 homologous pairs of autosomes, 26 nonhomologous autosomes, and XY sex chromosomes. Thirteen of the nonhomologous chromosomes represented the paternal complement, and 13 were from the maternal complement. Chromosomal rearrangements occurring between the karyotypes of A. l. griseimembra and A. nancymaae were believed to include two paracentric inversions, a reciprocal translocation, and two complex rearrangements involving pericentric inversion, telocentromeric fusion, and centromeric adjustment. Cytogenetic analyses are necessary to identify most Aotus taxa and thus should be utilized to pair chromosomally compatible animals and avoid interspecies hybridization.

Two 6-yr-old male sibling Amur leopards (Panthera pardus orientalis) housed together at the Pittsburgh Zoo presented for acute onset of diarrhea with no changes in appetite or behavior. Heat-fixed modified Wright-stained and Gram-stained fecal smears revealed a mixed bacterial population with a large number of gram-positive Clostridium perfringens-like spores (>20 per high-power oil immersion field). In addition, C. perfringens enterotoxin was isolated from one leopard at 1:256, confirming the presence of C. perfringens enterotoxicosis. Treatment with oral metronidazole, tylosin tartrate, and psyllium fiber was prescribed, with return of more normal stool by the third day of treatment. Fecal consistency steadily improved and was considered normal by the time all prescribed treatments were complete. Diarrhea has not recurred. Partially thawed meat in the leopards' diet may have precipitated the production of an endogenous clostridial enterotoxicosis by disrupting digestive tract flora with resultant clostridial overgrowth and sporulation.

Thirty randomly selected game bird feeders were sampled at 25–33-day intervals from November 1996 to March 1997 to quantitate ochratoxin A concentrations in supplemental feed. Monthly mean ochratoxin A concentration of grain in feeders was 8.3 ± 0.8 ppb (n = 167). Ochratoxin A concentrations from individual feeders ranged from <5 to 109.9 ppb, levels that have not been demonstrated to negatively affect game birds in a laboratory environment. Stress may increase the chance of ochratoxin-induced mortality or morbidity for wild game birds. Only mean relative humidity was significantly correlated with monthly mean ochratoxin A concentration.