PHARMACOKINETICS OF ORAL FLUNIXIN MEGLUMINE, MELOXICAM, OR GABAPENTIN IN THREE BLACK RHINOCEROS (DICEROS BICORNIS).

Pharmacokinetics of single, separate doses of IV flunixin meglumine (1 mg/kg), IV meloxicam (0.5 mg/kg), oral flunixin meglumine (1 mg/kg), oral meloxicam (1 mg/kg), and oral gabapentin (15 mg/kg) in three adult black rhinoceroses (Diceros bicornis) were determined from serial blood collection made over 72 h. The concentration versus time profiles were analyzed for each drug and route in each individual rhinoceros, and individual pharmacokinetic parameters were calculated for each medication administered. Meloxicam had near complete bioavailability in each trial, while flunixin meglumine was generally lower. Oral meloxicam was noted with similar half-life values between all animals (range 9.22-14.52 h) tested, while oral gabapentin had a larger range (range 10.25-24.85 h). Oral flunixin meglumine achieved a lower Cmax (range 170.67-664.38 ng/ml) in this study compared with the mean Cmax (1,207 ng/ml) reported in a similar study in white rhinoceroses (Ceratotherium simum), but some overlap in range of values was noted. Oral flunixin meglumine Tmax (range 1.05-10.78 h) and half-life (range 3.88-14.85 h) values in black rhinoceroses was similar to mean values reported in white rhinoceroses (3 and 8.3 h, respectively).

A 25-Year Retrospective Review of Mortality in Chimpanzees (Pan troglodytes) in Accredited U.S. Zoos from a Management and Welfare Perspective.

Understanding causes of death allows adjustment of health management strategies for animals in managed care. From 224 documented chimpanzee deaths occurring from 1995 to 2019 in 42 accredited U.S. zoos, post-mortem records and necropsy reports were analyzed for the primary cause of death, which were available for 214 individuals. In total, 37 cases of stillbirth and neonatal deaths were assessed (16.5%); however, the focus was otherwise placed on the remaining 177 cases in which the death occurred in individuals aged greater than 1 month. There were no sex-related differences in etiology; however, age variation in the cause of death was statistically significant (p < 0.001). Elderly (35 years and older) chimpanzees tended to die of intrinsic, often degenerative, etiologies, whereas infants, juveniles, and adolescents (less than 15 years) were more likely to be involved in fatal trauma. Overall, there were 27 deaths (15.3% of all post-neonatal deaths) related to trauma and 13 of these were directly or indirectly related to conspecific aggression. Understanding causes of mortality and the interrelation with management can benefit managed populations of chimpanzees.

HYPOCOAGULABILITY EFFECT OF ADEQUAN IN DOMESTIC CHICKENS (GALLUS GALLUS) AND CHILEAN FLAMINGOS (PHOENICOPTERUS CHILENSIS).

Effective management of articular injuries in avian species is a known and frequent challenge. Potential treatments include many domestic animal therapeutics, such as Adequan®, which is used widely in dogs and horses. However, clinical reports have described hemorrhagic diatheses in a variety of avian species treated with varying doses and administration frequency of Adequan. This study investigated the hypocoagulability associated with parenteral administration of Adequan in avian species. Following a pilot dosing study in domestic chickens (Gallus gallus), citrated plasma from Chilean flamingos (Phoenicopterus chilensis) (n = 42) was spiked with Adequan to represent three dosing regimens (1 mg/kg, 5 mg/kg, 10 mg/kg). The fibrinogen content of plasma samples was determined and thrombin-clotting times (TCTs) were compared for the untreated (control) and spiked flamingo samples. The TCT for control and 1-mg/kg spiked plasma were not significantly different; however, both 5 mg/kg and 10 mg/kg spiked samples demonstrated significantly prolonged TCT (P-value < 0.0001) indicating hypocoagulability. These results support that Adequan given parenterally at 1 mg/kg can be utilized safely in clinical case management as an adjunctive treatment for osteoarthritis in flamingos and potentially other avian species.

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.

Probable Enalapril Hepatotoxicity in a 13-Year-Old Male Western Lowland Gorilla: Pharmacologic Risks and Vigilance.

A 13-year-old male Western lowland gorilla presented acutely with a precipitous decline in health status from liver disease. Through diagnostic assessment, including serum chemistries and advanced imaging, it was diagnosed with probable hepatotoxicity resulting from its prescribed medication, enalapril. As one of several angiotensin converting enzyme inhibitors (ACE-I) available to zoo veterinarians, enalapril had been administered for treatment of mild ventricular hypertrophy diagnosed during routine examination 2.5 years prior to the presentation. The gorilla made a complete recovery with discontinuation of this medication, and provision of hepatoprotectants. Hepatotoxicity has been documented in humans receiving this product as an adverse drug reaction and is considered both rare and unpredictable in occurrence. In this event, an association was suspected with indulgent consumption of mulberry browse (Morus sp.) offered as nutritional enrichment immediately prior to clinical presentation and had potential impact on hepatic cytochrome P450 metabolism of the enalapril. Although ACE-I are important medications in this taxon due to its predisposition to cardiac disease, this event underscores the need for vigilance on the part of veterinarians and managers whenever pharmaceuticals are administered. Most drugs are modeled in a limited number of species but utilized in a wide variety, and unintended results are possible.

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.

SURVEY FOR EQUINE HERPESVIRUSES IN POLAR BEARS ( URSUS MARITIMUS) AND EXOTIC EQUIDS HOUSED IN US AZA INSTITUTIONS.

Infection by equine herpesvirus (EHV) strains (EHV-1, EHV-9) in ursid species, including polar bears ( Ursus maritimus), has been associated with neurological disease and death. A serosurvey of captive exotic equid and polar bear populations in US Association of Zoos and Aquaria institutions was performed to determine the prevalence of EHV strains using quantitative polymerase chain reaction (qPCR) and enzyme-linked immunosorbent assay (ELISA) tests. Equid species surveyed included zebra ( Equus spp.), Przewalski's wild horse ( Equus ferus przewalskii), Persian onager ( Equus hemionus), and Somali wild ass ( Equus africanus somaliensis). A questionnaire regarding husbandry and medical variables was distributed to institutions housing polar bears. No polar bears tested positive for EHVs on qPCR of blood or nasal swabs. No exotic equids tested positive for EHVs on qPCR of blood, but two exotic equids ( n = 2/22; 9%) tested positive for EHVs on qPCR of nasal swabs. On ELISA, polar bears infrequently were positive for EHV-1 ( n = 5/38; 13%). Exotic equids were positive for EHV-4 on ELISA more frequently ( n = 30/43; 70%) than for EHV-1 ( n = 8/43; 19%). Nine institutions submitted samples from both exotic equids and polar bears, two of which had both exotic equids and polar bears positive for EHVs by ELISA. Each of these institutions reported that the polar bear and exotic equid exhibits were within 80 m of each other and that risk factors for fomite transmission between exhibits based on husbandry practices were present. One institution that did not house exotic equids had a polar bear test positive for EHV-1 on ELISA, with no history of exposure to exotic equids. Further testing of captive polar bears and exotic equids is recommended, as is modification of husbandry practices to limit exposure of polar bears to exotic equids.

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 ADULT-ONSET CARDIAC DISEASE IN FRANÇOIS' LANGURS (TRACHYPITHECUS FRANCOISI) HOUSED IN U.S. ZOOS.

Cardiac disease is a common condition in captive primates, and multiple cases in François' langurs ( Trachypithecus francoisi ) were noted on review of the Species Survival Plan studbook. To determine the prevalence of cardiac disease in this species, surveys were distributed to current and previous holding institutions (n = 23) for the U.S. studbook population (n = 216). After exclusion of stillbirths (n = 48), animals less than 1 yr of age (n = 8), and animals housed internationally (n = 2), a study group (n = 158) was identified for this analysis. Robust data was received for 98.7% (n = 156) of the study group and antemortem and postmortem cardiac abnormalities were reported for 25.3% (n = 40) of these animals. Eight animals were reported as medically managed for clinical cardiac disease, and three of these were alive at the time of survey. Six of 11 animals with radiographic cardiac silhouette enlargement antemortem were noted with cardiomegaly on postmortem examination. Of 102 deceased animals in the study group, four were identified with dilated cardiomyopathy, and varying degrees of myocardial fibrosis was observed in 18 animals. Langurs with cardiac fibrosis were found to be significantly older than langurs without cardiac fibrosis (P = 0.003) and more commonly were male (P = 0.036). Screening tests for cardiac disease, such as thoracic radiographs and echocardiography, are recommended to diagnose affected animals earlier, to monitor progression of disease, and to guide treatment, although they should be interpreted with caution because of apparent insensitivity when compared with pathologic results.

EVALUATION OF THYROID HORMONES AND AS INFLUENCED BY TREATMENT WITH DESLORELIN IN PALLAS' CATS (OTOCOLOBUS (FELIS) MANUL).

Thyroid hormones regulate a variety of physiologic functions including metabolism, growth, and reproductive cycling, and these other hormones can impact the thyroid function via the hypothalamic-pituitary axis. For instance, the gonadotropin-releasing hormone agonist, deslorelin, used in nondomestic carnivores for contraception and behavioral control, down-regulates reproductive hormones through this mechanism and so may impact thyroid function. Due to clinical concerns of hypothyroidism in a bachelor group of adult male Pallas' cats (Otocolobus (Felis) manul) which also had deslorelin implants, serum samples from treated captive (n = 8) individuals, untreated captive (n = 25), and free-ranging (n = 9) individuals were analyzed for thyroid hormone concentrations. Total and free thyroxine (TT4 and FT4), total and free tri-iodothyronine (TT3 and FT3), and thyroid stimulating hormone (TSH) were measured although, due to sample volume limitations, not every hormone could be analyzed for every sample. Of these hormones, only FT4 was found statistically different between the deslorelin-treated and untreated groups. As samples were unevenly distributed across season, true comparison between seasons could not be made. The values reported for the untreated captive and free-ranging group, while representing a small sample size, can serve as a baseline assessment when evaluating the thyroid status of captive Pallas' cats.

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.

EXTRACORPOREAL SHOCK WAVE LITHOTRIPSY AND ENDOSCOPIC URETERAL STENT PLACEMENT IN AN ASIAN SMALL-CLAWED OTTER (AONYX CINEREA) WITH NEPHROLITHIASIS.

Urolithiasis is a significant disease concern in Asian small-clawed otters (Aonyx cinerea), with over 60% of captive animals affected. Bilateral ureteral stent placement, using endoscopic and fluoroscopic guidance, and extracorporeal shock wave lithotripsy (ESWL) were performed as salvage procedures in a 13-yr-old intact female Asian small-clawed otter following a 7-yr history of nephrolithiasis and progressive renal insufficiency. Following the procedure, radiographs revealed a slight shifting of urolith position, although a decrease in urolith mass was not observed. As a result of declining quality of life related to severe osteoarthritis, the otter was euthanized 5 wk after the procedure. While this treatment approach was unsuccessful in this case, the technique was clinically feasible, so ESWL and ureteral stent placement may remain a consideration for other individuals of this species presented earlier in the course of this disease.

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.

Human metapneumovirus infection in chimpanzees, United States.

Zoonotic disease transmission and infections are of particular concern for humans and closely related great apes. In 2009, an outbreak of human metapneumovirus infection was associated with the death of a captive chimpanzee in Chicago, Illinois, USA. Biosecurity and surveillance for this virus in captive great ape populations should be considered.

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.

Molecular detection of Plasmodium in free-ranging birds and captive flamingos (Phoenicopterus chilensis) in Chicago.

Frozen blood samples from 13 species of free-ranging birds (n = 65) and captive Chilean flamingos (Phoenicopterus chilensis) (n = 46) housed outdoors in the Chicago area were screened for Plasmodium. With the use of a modified polymerase chain reaction, 20/65 (30.8%) of free-ranging birds and 26/46 (56.5%) of flamingos were classified as positive for this parasite genus. DNA sequencing of the parasite cytochrome b gene in positive samples demonstrated that eight species of free-ranging birds were infected with five different Plasmodium spp. cytochrome b lineages, and all positive Chilean flamingos were infected with Plasmodium spp. cytochrome b lineages most closely related to organisms in the Novyella subgenus. These results show that Chilean flamingos may harbor subclinical malaria infections more frequently than previously estimated, and that they may have increased susceptibility to some Plasmodium species.

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.