Shell Lesions Associated With Emydomyces testavorans Infection in Freshwater Aquatic Turtles.

A newly described onygenalean fungus, Emydomyces testavorans, has been isolated from ulcerative shell and skin lesions of freshwater aquatic chelonians. To investigate the shell lesions associated with infection and determine if any lesional features were unique to E. testavorans, tissues from turtles housed in zoological institutions (n = 45) in the United States and free-living turtles (n = 5) submitted for diagnostic biopsy or necropsy were examined. Free-living turtles were from geographically distinct habitats in Florida (n = 1) and Washington (n = 4) at the time of sampling. Histologic shell sections were evaluated for the presence or absence of specific lesional features. Infection with E. testavorans was evaluated in all cases by screening GMS (Grocott-Gomori's methenamine silver)-stained histologic sections for the presence of morphologically consistent fungi and by quantitative PCR (polymerase chain reaction) on representative frozen tissue or formalin-fixed paraffin-embedded sections. Additionally, culture was performed for 15 cases with available fresh/frozen tissue. In total, there were 17 PCR-confirmed E. testavorans cases, 29 cases with morphologically consistent fungi on GMS-stained sections, and 21 cases of shell lesions without histologic or molecular evidence of E. testavorans infection. Epithelial inclusion cysts, defined as cystic structures within the dermis lined by keratinized stratified squamous epithelium and containing necrotic bone and keratin debris, were significantly (P < .01) associated with E. testavorans infection. Other significantly associated shell lesions included squamous metaplasia, hyperkeratosis, inflammation, and osteonecrosis (P < .05). This study identified characteristic shell lesions associated with E. testavorans infection. Further studies to prove causality are needed.

Novel Herpesvirus Associated With Oropharyngeal Squamous Cell Carcinoma in Smooth Green Snakes (Opheodrys vernalis).

During a 19-month period, 5 smooth green snakes (Opheodrys vernalis) maintained as an ex situ conservation colony presented with rapid clinical progression of locally invasive oropharyngeal squamous cell carcinoma. All 5 originated from the same wild source and were housed together or in close proximity. An infectious cause was considered likely, and nested conventional polymerase chain reaction (PCR) and in situ hybridization confirmed the presence of a novel alphaherpesvirus, Opheodrys herpesvirus 1, in the neoplastic tissue in 4 of the 5 snakes. Retrospective screening of previously submitted smooth green snakes by in situ hybridization did not detect virus in prior submissions from the colony. This report documents molecular characterization of an ophidian herpesvirus as well as colocalization of its viral nucleic acid with neoplastic transformation in snakes.

ORONASAL SQUAMOUS CELL CARCINOMAS IN FRANÇOIS' LANGURS (TRACHYPITHECUS FRANCOISI).

Squamous cell carcinomas (SCCs) are common oronasal tumors in nonhuman primates. In this study, 11 cases of oronasal SCC in François' langurs ( Trachypithecus francoisi ) are described. Five initial cases were discovered on review of the North American François' langur studbook, with a potential familial pattern observed. The studbook was used to identify related individuals, and records were requested for review. Six additional cases were documented, and samples from all cases were submitted for microscopic review, as well as polymerase chain reaction (PCR), immunohistochemistry (IHC), and in situ hybridization (ISH), for generic papillomaviruses and PCR for herpesviruses because either virus may cause SCC in humans and other nonhuman primates. Affected langurs commonly presented with facial swelling or ocular discharge but frequently did not have clinical signs, and carcinomas were diagnosed during routine examinations. Carcinomas were located in the oral or nasal cavities affecting the oral mucosa, tongue, hard palate, or oropharynx. Histologically, SCCs comprised anastomosing cords and nests of neoplastic epithelial cells that differentiated synchronously and asynchronously from peripheral basal type cells to central squamous-type cells and were occasionally oriented around accumulations of necrotic cell debris. Nuclear pleomorphism, anisokaryosis, prominent nucleoli, occasional mitoses, and a scirrhous response were common features. All animals tested negative for both viruses, except two langurs that were positive for generic papillomavirus by PCR, but no papillomavirus was detected by either IHC or ISH. In most cases, affected animals died within 5 mo of diagnosis.

Plasma Concentrations of Fentanyl Achieved With Transdermal Application in Chickens.

Providing appropriate analgesia is an important concern in any species. Fentanyl, a µ-receptor specific opioid, use is common in mammalian species but has been incompletely evaluated for this purpose in avian species. Transdermal fentanyl patches were applied to domestic chickens (n = 10) of varying breeds for 72 hours. Repeated blood samples were collected from the birds to assess time-concentration of fentanyl and norfentanyl in plasma, as assayed by liquid chromatography-mass spectrometry, throughout patch application and for 48 hours after patch removal. Compartmental modeling was used to characterize the elimination profiles. Evaluation as a large bolus, followed by slower elimination rates over the remaining time, best fit the data as a one-compartment open model. Although maximum plasma fentanyl concentrations varied substantially by individual birds, chickens trended into 2 general groups of maximum plasma concentration, clearance, and volume of distribution, which was attributed to absorption variability. For all birds, harmonic mean of elimination half-life was 7.2 ± 3.7 hours and showed less individual variation than the other pharmacokinetic parameters. Because the application of transdermal fentanyl patches in the chickens achieved plasma fentanyl concentrations considered therapeutic in people, this approach could provide an additional analgesic option for avian patients.

PROSPECTIVE POPULATION MANAGEMENT FOR HEPATITIS B IN THE CHIMPANZEE (PAN TROGLODYTES) SSP® POPULATION.

Hepatitis B virus causes horizontally transmitted infectious hepatopathy of primates and may progress to hepatocellular carcinoma. Historically, a small number of chimpanzees ( Pan troglodytes ) living in accredited North American zoos have been confirmed with positive hepatitis B serology consistent with exposure. However, the overall status for this population and the interpretation of these individual test results have not been established previously. The current U.S. zoo-housed population (n = 259) was assessed serologically for hepatitis B by surface protein antigen (HbsAg) and surface antibodies (anti-Hbs). Signalment, origin, current health status, history of liver disease, and hepatitis B vaccination history were obtained for each animal. Serologic status was measured directly in 86.5% (n = 224) of these individuals, with 2.2% (n = 5) of the study population determined to be chronically infected by positive HbsAg and negative anti-Hbs status. Additionally, 11.6% (n = 26) of the directly measured population tested were HbsAg negative and anti-Hbs positive, which was indicative of viral exposure. No animals were determined to be acutely infected as HbsAg and anti-Hbs positive. Although these results demonstrated a relatively low prevalence of hepatitis B infection among these chimpanzees, the varied serologic results between institutions underscored the importance of routine serologic testing, especially at times of proposed transfers, and consideration of species vaccination protocols.

RETROSPECTIVE ANALYSIS OF MORTALITY IN THE NORTH AMERICAN CAPTIVE RED PANDA (AILURUS FULGENS) POPULATION, 1992-2012.

Red pandas ( Ailurus fulgens ) are managed as captive populations in both North America and Europe. Regular review of pathology reports is a useful tool for developing veterinary care and husbandry strategies for such populations. Though thorough pathology reviews have been conducted for the European studbook, the North American population has not been reviewed similarly until now. Complete gross and histopathology reports were requested from institutions holding red pandas that died during 1992 through 2012 (n = 530), and reports were received for 95.8% of the individuals, including full necropsy records for 366 red pandas. These reports were classified by subspecies, gender, and age, then reviewed for primary cause of death and secondary pathological findings. A substantial portion of the deaths (40.2%) were neonates (<30 days of age). In both neonatal and juvenile (age = 31-365 days) animals, pneumonia was the most common cause of death. In adult (age = 366 days-10 yr) and geriatric red pandas (age >10 yr), cardiovascular disease was the most common cause of death. Renal disease and gastrointestinal disease also were common pathologic findings in adult and geriatric animals. These findings suggest that stress associated with captivity and husbandry practices, including those associated with social, environmental, and nutritional conditions, may contribute to immune and cardiovascular pathologies, and other common necropsy findings.

Detection and Management of Air Sac Trematodes (Szidatitrema Species) in Captive Multispecies Avian Exhibits.

From 2 exhibits in a zoological collection, 2 juvenile fairy bluebirds ( Irena puella ) and 1 adult blue-grey tanager (Thraupis episcopus) died within 3 months of one another. The cause of death was attributed to air sac trematodes, which were identified as Szidatitrema species based on morphology of adult trematodes and miracidia isolated from a snail intermediate host. To determine the extent of trematodiasis in the collection, individual exhibits within the same building as the original presenting cases were assessed, with birds representing 27 avian species from 9 orders. Sampling consisted of individual (n = 244) and pooled same-species group (n = 193) fecal examinations, and for some individuals, and tracheal swab (n = 106), resulting in a total of 543 samples. In addition, tracheal swabs were performed on 14 birds for comparative cytology, but no parasites were found. Flukes were positively identified in 4 tracheal swab samples (4%), 37 individual fecal samples (15%), and 52 of the group fecal samples (27%). When results of the swab method were compared with those of fecal examination, fecal testing was significantly associated (P < .001) with positive results. Based on these results, a screening process was instituted of 3- or 4-day combined fecal samples evaluated by both sedimentation and flotation techniques 2 weeks before outgoing shipments or intrazoo transfers of birds housed in exhibits known to have snail populations. Snail control methods also were initiated in all exhibits. Treatment with praziquantel was carried out on a case-by-case basis, and included oral, parenteral, and nebulized administration. Although control measures were expected to manage the infection and reduce distribution of the parasite to other collections, complete eradication of trematodes in the population is unlikely.

Esophageal squamous cell carcinoma in six harbor seals (Phoca vitulina spp.).

Six cases of esophageal squamous cell carcinoma were identified in six captive adult Pacific (Phoca vitulina richardsii; n = 2) and Atlantic (Phoca vitulina concolor; n = 4) harbor seals. These seals presented with intermittent dysphagia, regurgitation, inappetence, and abnormal posturing. Common clinical pathology findings in these seals included azotemia, hyperproteinemia, hyperglobulinemia, and leukocytosis. Gastrointestinal endoscopy commonly revealed an ulcerated mass near the gastroesophageal junction. Each seal was euthanized (n = 3) due to poor prognosis, subsequently died while undergoing an anesthetic procedure (n = 2), or found dead (n = 1). The diagnosis of squamous cell carcinoma was confirmed via biopsy of esophageal mucosa during endoscopy or histopathologic examination of affected tissues after necropsy. On the basis of clinical and postmortem findings, esophageal squamous cell carcinoma should be considered as a differential diagnosis in aged harbor seals exhibiting clinical signs of regurgitation, decreased appetite or anorexia, vomiting, and/or abnormal posturing.

Pharmacokinetics of a single dose of intravenous and oral meloxicam in red-tailed hawks (Buteo jamaicensis) and great horned owls (Bubo virginianus).

Pharmacokinetic data were determined after a single dose of meloxicam in red-tailed hawks (RTH; Buteo jamaicensis) and great horned owls (GHO; Bubo virginianus). In a nonrandomized crossover design, individual birds of each species received 1 dose of intravenous meloxicam (0.5 mg/kg i.v.; n = 7 for each species) followed by a 2-week washout period, and then each received 1 dose of oral meloxicam (0.5 mg/kg PO; n = 5 for each species). Blood samples were collected intermittently after administration, and meloxicam was detected in plasma by high-performance liquid chromatography. Time versus plasma concentration data were subjected to noncompartmental analysis. Red-tailed hawks were determined to have the shortest elimination half-life for meloxicam (0.49 +/- 0.5 hours) of any species documented. Great horned owls also eliminated meloxicam very rapidly (0.78 +/- 0.52 hours). Great horned owls achieved higher plasma concentrations (368 +/- 87 ng/mL) of meloxicam than RTH (182 +/- 167 ng/mL) after oral administration, although RTH had a markedly higher volume of distribution (832 +/- 711 mL/kg) than GHO (137.6 +/- 62.7 mL/kg). The differences in meloxicam pharmacokinetics between these 2 raptor species supports the need for species-dependent studies and underlines the challenges of extrapolating drug dosages between species. Results of this study suggest that the current recommended once-daily dosing interval of oral meloxicam is unlikely to maintain plasma concentrations anticipated to be therapeutic in either RTH or GHO, and practical dosing options are questionable for this nonsteriodal anti-inflammatory drug in these raptor species.

Calcium carbonate obstructive urolithiasis in a red kangaroo (Macropus rufus).

A 6-yr-old male red kangaroo (Macropus rufus) presented for a history of inappetance, abnormal behavior, and unconfirmed elimination for 6 hr prior to presentation. Based on abdominal ultrasound, abdominocentesis, and cystocentesis, a presumptive diagnosis of urinary tract obstruction with uroabdomen and hydronephrosis was reached. Abdominal radiographs did not assist in reaching an antemortem diagnosis. Postmortem examination confirmed a urinary bladder rupture secondary to urethral obstruction by a single urethrolith. Bilateral hydronephrosis and hydroureter were identified and determined to be a result of bilateral ureteroliths. Urolith analysis revealed a composition of 100% calcium carbonate. A dietary analysis was performed, implicating an increased Ca:P ratio from a food preparation miscommunication as a contributing factor. Appropriate husbandry changes were made, and mob surveillance procedures were performed, which resolved the urolithiasis risk for the remaining five animals.

Effects of ultracentrifugation on plasma biochemical values of prefledged wild peregrine falcons (Falco peregrinus) in northeastern Illinois.

Centrifugation is performed on whole blood samples to obtain serum or plasma for biochemical analysis. Although blood samples centrifuged in a microhematocrit tube may maximize recovery of plasma from small-volume samples, plasma biochemical values from such samples have been implicated as causing erroneous results. To compare blood biochemical values obtained by microhematocrit centrifugation and centrifugation with a commercial tilt-rotor machine, blood samples were collected from peregrine falcon (Falco peregrinus) eyases aged 32-40 days (n=51). The samples were separated into 2 equal aliquots with 1 aliquot centrifuged in a tilt-rotor machine and the other aliquot ultracentrifuged in microhematocrit tubes. Separated plasma from both processes was sent to a commercial veterinary reference laboratory for routine clinical biochemical analysis. No significant differences were found in the biochemical results of the paired samples by the 2 centrifugation methods. These results show that the centrifugation method has no effect on the plasma quality for biochemical analysis in young peregrine falcons.

Acute pancreatitis in slender-tailed meerkats (Suricata suricatta).

Four adult, full-sibling slender-tailed meerkats (Suricata suricatta) were diagnosed with acute pancreatitis. The incident case presented with lethargy, anorexia, abdominal guarding, and a cranial abdominal mass. Serum was grossly lipemic, with elevated cholesterol and triglyceride concentrations and increased amylase and lipase activity. An exploratory laparotomy confirmed chylous peritonitis and included excision of a saponified spleno-duodenal mass, a partial pancreatectomy, and a splenectomy. Histopathology revealed severe, multifocal, subacute necrotizing and granulomatous pancreatitis. Within 13 days of the incident case, the second meerkat was identified with essentially identical clinical, surgical, and histologic findings. During subsequent physical examinations of apparently unaffected cohorts (n=12), physical and hematologic findings suggestive of pancreatitis were identified in the two remaining siblings of the first two cases. The definitive cause for these four cases is undetermined; however, common risk factors identified were obesity and hyperlipidemia, a change to a higher-fat diet, and genetic predisposition. To assess its usefulness in the diagnosis of meerkat pancreatitis, serum canine and feline pancreatic lipase immunoreactivity (cPLI and fPLI) concentrations were measured in serum samples (n=61) from two unrelated meerkat populations. Although these assays are highly sensitive and specific for the diagnosis of pancreatitis in domestic carnivores, similar correlation was not apparent for meerkats. In addition, hyperlipidemia was inconsistently present in many meerkats, with no apparent correlation to the development of clinical illness. Based on these observations, sensitive and specific diagnostic tests for pancreatitis in meerkats are currently unavailable.

Internal anatomy of the hornbill casque described by radiography, contrast radiography, and computed tomography.

Hornbills are distinguished from most other avian taxa by the presence of a casque on the dorsal maxillary beak, which, in all but 1 of the 54 extant hornbill species, is described as essentially an air-filled cavity enclosed by minimal cancellous bone. The external casque has been described in detail, but little has been described about its internal anatomy and the communications between the casque and the paranasal sinuses. In this study, 10 intact casque and skull specimens of 7 hornbill species were collected opportunistically at necropsy. The anatomy of the casque and the skull for each of the specimens was examined by radiography, contrast radiography, and computed tomography. After imaging, 8 specimens were submitted for osteologic preparation to directly visualize the casque and the skull interior. Through this standardized review, the baseline anatomy of the internal casque was described, including identification of a novel casque sinus within the paranasal sinus system. These observations will assist clinicians in the diagnosis and treatment of diseases of the casque in hornbill species.

Two cases of atypical mycobacteriosis caused by Mycobacterium szulgai associated with mortality in captive African elephants (Loxodonta africana).

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.

Cestode cysts in two African giant pouched rats (Cricetomys gambianus).

Multiple cestode cysts identified as Taenia serialis were present in the bodies of two wild-caught African giant pouched rats (Cricetomys gambianus) at necropsy. This rodent species can be an intermediate host for this parasite. Exotic rodents kept as pets in the United States may be affected.