DIAGNOSTIC PERFORMANCE OF BLOOD ANALYTES FOR THE DIAGNOSIS OF RENAL DISEASE IN BLACK-FOOTED FERRETS (MUSTELA NIGRIPES) AT THE PHOENIX ZOO (2001-2020).

Renal disease is an important cause of morbidity and mortality in managed black-footed ferrets (BFF; Mustela nigripes).4,6,12 The objectives of this study were to establish reference intervals for blood analytes of clinically normal BFF (1-2 yr old), summarize the frequency of various renal histopathologic findings in a managed population of BFF, assess the diagnostic performance of blood analytes and urine specific gravity (USG) for the diagnosis of renal disease, and assess if comorbidities or age affects the performance of these analytes in diagnosing renal disease. Reference intervals were established using a cohort (n = 35) of clinically normal, young adult BFF. Postmortem records for all BFF at the Phoenix Zoo between 2001 and 2020 were reviewed, and those with available blood analyte data within 2 wk of death were included (n = 89). Ferrets were placed into one of three groups, based on the organ location of histopathologic abnormalities following necropsy: renal disease as the primary change; those with renal disease and at least one other affected major organ system; or absence of abnormalities in the kidneys. In ferrets with substantial renal changes, the primary diagnosis was amyloidosis (29 of 39; 74.4%). Creatinine, blood urea nitrogen, phosphorus (P), calcium (Ca), Ca:P ratio, USG, globulins, and cholesterol were the best-performing analytes for the diagnosis of renal disease, with an area under the curve of at least 0.90 (95% CI $ 0.80, 1.00). Serum renal markers were within reference intervals in BFF that died without histologic evidence of renal disease. Several blood analytes were significantly affected by age in animals that died of renal disease. This study provides reference intervals for blood analytes in young adult clinically normal BFF and illustrates the clinical utility for the diagnosis of renal disease in this species, particularly creatinine, USG, and P.

NOVEL SIMPLEXVIRUS (SIMPLEXVIRUS DOLICHOTINEALPHA1) ASSOCIATED WITH FATALITY IN FOUR PATAGONIAN MARA (DOLICHOTIS PATAGONUM).

Four of seven Patagonian maras (Dolichotis patagonum) at a zoological institution developed acute neurologic signs that progressed to tetraparesis and death. All affected were young adult females (10 mon-5 yr old) that presented over 11 d. Clinical signs were rapidly progressive and unresponsive to supportive therapies. Two of the four individuals were found deceased 4 d after hospitalization. Two individuals were euthanized due to poor prognosis and decline after 6 and 8 d, respectively. Simultaneously, an additional mara developed mild and self-resolving clinical signs, including a kyphotic gait and paraparesis. On gross examination, there were widespread petechiae and ecchymoses of the skeletal muscle, myocardium, skin, pericardium, urinary bladder mucosa, and spinal cord. On histopathology, all animals had necrotizing myelitis and rhombencephalitis, with intranuclear viral inclusions in three individuals. Electron microscopy confirmed herpesviral replication and assembly complexes in neurons and oligodendrocytes. Consensus PCR performed on spinal cord, brainstem, or cerebellum revealed a novel Simplexvirus most closely related to Simplexvirus leporidalpha 4. The virus was amplified and sequenced and is referred to as Simplexvirus dolichotinealpha1. It is unknown whether this virus is endemic in Patagonian mara or whether it represents an aberrant host species. Clinicians should be aware of this virus and its potential to cause severe, rapidly progressive, life-threatening disease in this species.

MAMMARY GLAND ADENOCARCINOMA IN FOUR INDIAN CRESTED PORCUPINES (HYSTRIX INDICA).

Neoplasia in porcupines is rarely reported in the literature, and the prevalence is unknown. A retrospective review of records from a private zoo diagnostic pathology service found four cases of mammary adenocarcinoma in Indian crested porcupines (Hystrix indica) from four separate zoological institutions. All cases presented in geriatric females (14-19 yr of age) as freely movable subcutaneous masses within the mammary chain. None of the individuals had additional clinical signs, radiographic, or hematologic changes at initial presentation. All cases were managed with surgical excision in the form of either an excisional biopsy or a partial mastectomy. Histologic examination diagnosed all tumors with anaplasia and moderate to high numbers of mitotic figures. Two cases required subsequent surgeries for management of local recurrence in the years following initial diagnosis. One case is 19 months postsurgical removal without evidence of metastasis or local recurrence. Two of the cases were euthanized after diagnosis of inoperable metastases to the lungs and spinal cord, including one previously treated with an oral nonsteroidal antiestrogen medication, tamoxifen. The third case was euthanized due to degenerative mobility changes and renal dysfunction and had no evidence of metastasis. The average survival time from initial surgical excision to euthanasia for the three applicable cases was 33 months. These cases suggest that surgical excision alone may result in temporary management of mammary adenocarcinoma in this species. Metastasis can occur, and routine screening with advanced imaging may aid in early detection of these lesions.

SUPERFICIAL CERVICAL LYMPHADENITIS ASSOCIATED WITH STREPTOCOCCUS PHOCAE IN FIVE COMMON BOTTLENOSE DOLPHINS (TURSIOPS TRUNCATUS): A CASE SERIES.

Between 2009 and 2018, five common bottlenose dolphins (Tursiops truncatus) at the US Navy Marine Mammal Program presented with superficial cervical lymphadenitis. Clinical findings included ultrasonographic evidence of cervical lymph node enlargement, severe leukocytosis, elevated erythrocyte sedimentation rates, and reduced serum iron. Three of the dolphins presented with clinicopathologic changes without presence of clinical signs, and the other two cases additionally presented with partial to complete anorexia, lethargy, and refusal to participate in training sessions. Ultrasound-guided fine needle aspiration or biopsy of the affected lymph nodes yielded Streptococcus phocae by PCR in all cases, and the organism was cultured in one of five cases. Animals were treated with a combination of enteral, parenteral, intralesional antimicrobial, or a combination of those therapies and supportive care. Time to resolution of clinical disease ranged between 62 and 188 days. To the authors' knowledge, this is the first report of Streptococcus phocae cervical lymphadenitis in cetaceans. Streptococcus phocae lymphadenitis should be a differential for cervical lymphadenopathy in this species, especially when associated with pronounced systemic inflammation and a history of potential exposure.

RETROSPECTIVE ANALYSIS OF MORTALITY CAUSES IN MANAGED KIRK'S DIK-DIKS (MADOQUA KIRKII) IN NORTH AMERICA FROM 1988 to 2019.

Kirk's dik-diks (Madoqua kirkii) are the most common dik-dik species managed in North American zoological institutions, but their numbers are declining at a concerning rate, with less than 40 individuals currently housed in accredited institutions. This retrospective study reports the causes of mortality in Kirk's dik-diks in North American zoological institutions from 1988 to 2019. Out of 15 institutions accredited by the Association of Zoos and Aquariums (AZA) currently housing Kirk's dik-diks, nine contributed to this study (60% participation). Eighty-four necropsy records were reviewed to determine the primary affected body system and etiological cause of death across and within age categories. Neonatal death (prior to 1 mon of age) was most common (38.1%), followed by death in adults (29.8%), geriatric animals (19.0%), and juveniles (13.1%). As a whole population, causes of death by body system were multisystemic (47.6%), musculoskeletal (15.5%), respiratory (8.3%), and digestive (8.3%). Neonatal complications accounted for 50.0% of all deaths in animals prior to 1 mon of age. In juveniles, a nutritional cause of death was most common (27.3%) and significantly higher compared to measures of this cause within other age categories. In adults, metabolic etiologies and trauma each accounted for 28.0% of deaths. Degenerative etiologies were most common in geriatric individuals, representing 31.3% of the deaths. Death from infectious disease was found across all ages, representing 11.9% of all mortalities. Results from this study provide a baseline reference for this species and may aid clinicians in decision-making as it relates to the medical care and management of this species during different life stages.

COMPARISON OF FECAL CYTOLOGY AND PRESENCE OF CLOSTRIDIUM PERFRINGENS ENTEROTOXIN IN CAPTIVE BLACK-FOOTED FERRETS (MUSTELA NIGRIPES) BASED ON DIET AND FECAL QUALITY.

The black-footed ferret (Mustela nigripes) is an endangered mustelid native to North America. Gastroenteritis is a documented cause of morbidity and mortality in managed individuals, particularly by infectious agents. Fecal cytology is an inexpensive and rapid test that can help guide clinical management strategies for animals with enteritis; however, normal parameters have not been established in black-footed ferrets. The objective of this study was to characterize fecal cytological findings of 50 fecal samples from 18 black-footed ferrets that received two different diet types (ground meat versus whole prey) and that were visibly judged to be normal or abnormal. This study also tested for the presence of Clostridium perfringens enterotoxin by enzyme-linked immunosorbent assay in all abnormal and a subset of normal fecal samples. Significantly higher spore-forming bacteria and yeast prevalence were present in normal feces from individuals following the meat-based compared with the whole-prey diet. Samples from individuals with abnormal feces had significantly more spore-forming bacteria than normal feces, regardless of diet. Normal feces had higher diplococci and spore-forming bacteria compared with domestic canine and feline standards. A single abnormal fecal sample was positive for enterotoxin and originated from the only animal requiring treatment. Results indicate that low numbers of spore-forming bacteria can be found in fecal samples from clinically normal black-footed ferrets. Fecal cytology shows significantly increased spore-formers in clinically abnormal ferrets and in clinically normal ferrets following a ground meat-based diet.

STREPTOCOCCUS EQUI SSP. ZOOEPIDEMICUS MENINGOENCEPHALOMYELITIS IN A SOUTHERN TAMANDUA (TAMANDUA TETRADACTYLA) ASSOCIATED WITH A RAW HORSEMEAT DIET.

A 1-y-old female southern tamandua (Tamandua tetradactyla) presented with vomiting, hyporexia, and neurologic signs. Magnetic resonance imaging revealed contrast-enhancing material within the lateral and fourth ventricles and a T2 hyperintense cerebellar lesion, consistent with meningoencephalitis. The tamandua rapidly declined and was euthanatized. On gross postmortem exam, the tamandua had diffusely injected leptomeninges, opaque fluid in the fourth ventricle, and subdural brainstem and spinal cord hemorrhage. Histologically, there was regionally hemorrhagic and multifocal fibrinosuppurative meningoencephalomyelitis, ventriculitis, choroid plexitis, cerebellar folia necrosis, ependymitis, radiculoneuritis, and abundant intralesional gram-positive cocci. Streptococcus equi ssp. zooepidemicus was cultured from brain, cardiac blood clot, and multiple samples of horsemeat collected from the animal's diet. This is the first report of streptococcal meningoencephalomyelitis in a southern tamandua. The route of infection was likely gastrointestinal inoculation, which may have implications for the routine practice of feeding diets containing raw meat to insectivores.

INTRAMUSCULAR EPINEPHRINE RESULTS IN REDUCED ANESTHETIC RECOVERY TIME IN AMERICAN ALLIGATORS (ALLIGATOR MISSISSIPPIENSIS) UNDERGOING ISOFLURANE ANESTHESIA.

Inhalants are commonly used to anesthetize reptiles, but volatile anesthetics have been associated with prolonged recovery times. The objective of this study was to determine the effects of intramuscular (IM) epinephrine on anesthetic recovery times following isoflurane anesthesia in a population of subadult American alligators ( Alligator mississippiensis ). In this prospective randomized crossover study, five clinically healthy alligators were anesthetized for 90 min with the use of isoflurane. Alligators were randomly assigned into one of two treatment groups: Group E received IM epinephrine (0.1 mg/kg), and Group S received an equal volume of 0.9% saline administered after isoflurane was discontinued. Time from the end of inhalant administration to return of spontaneous ventilation, return of the palpebral reflex, movement in response to a standardized toe pinch, and spontaneous movement was recorded. The time of extubation was noted and occurred following the return of spontaneous ventilation and movement. Pulse rate, surface body temperature, and airway gases including expiratory and inspiratory isoflurane concentrations and end-tidal carbon dioxide were measured every 5 min throughout the study. The time from the end of anesthesia to extubation was significantly faster in Group E (51.2 ± 16.7 min) compared to Group S (107.4 ± 43.7 min). Pulse rate was significantly higher within the first 15 min following epinephrine injection compared to the saline group at these time points. Therefore, IM epinephrine administered at the end of general anesthesia can significantly hasten anesthetic recovery from isoflurane in alligators.

GRANULOMATOUS ENCEPHALOMYELITIS IN A FALSE GHARIAL (TOMISTOMA SCHLEGELII) ASSOCIATED WITH A NOVEL CHLAMYDIA SPECIES.

A 5-yr-old, captive, hatched, female false gharial (Tomistoma schlegelii) presented with a 1-mo history of cervical spinal curvature. Antemortem diagnostics, including blood work, electromyography, muscle biopsies, and advanced imaging tests, were either within reference ranges or did not identify any specific etiology. Necropsy revealed extensive, marked, chronic granulomatous encephalomyelitis along with neuronal necrosis, rarefaction, gliosis, and astrocytosis of the white and gray matter of the cerebrum, cerebellum, brainstem, and spinal cord. Pan-chlamydiae polymerase chain reaction protocols for the 16S ribosomal RNA and ompA genes were performed on samples of spinal cord and brain, and both resulted in amplicons. Sequencing of the products revealed that they were positive for a novel Chlamydia species. Infections by members of the phylum Chlamydiae have been reported in a diverse range of vertebrate hosts, including crocodilians. Chlamydia spp. infections are likely underdiagnosed because of a paucity of diagnostic techniques specific for detection. This is the first case report of a novel Chlamydia species associated with severe granulomatous encephalomyelitis in a false gharial.

EPINEPHRINE OR GV-26 ELECTRICAL STIMULATION REDUCES INHALANT ANESTHESTIC RECOVERY TIME IN COMMON SNAPPING TURTLES (CHELYDRA SERPENTINA).

Prolonged anesthetic recovery times are a common clinical problem in reptiles following inhalant anesthesia. Diving reptiles have numerous adaptations that allow them to submerge and remain apneic for extended periods. An ability to shunt blood away from pulmonary circulation, possibly due to changes in adrenergic tone, may contribute to their unpredictable inhalant anesthetic recovery times. Therefore, the use of epinephrine could antagonize this response and reduce recovery time. GV-26, an acupuncture point with reported ß-adrenergic and respiratory effects, has reduced anesthetic recovery times in other species. In this prospective randomized crossover study, six common snapping turtles (Chelydra serpentina) were anesthetized with inhalant isoflurane for 90 min. Turtles were assigned one of three treatments, given immediately following discontinuation of isoflurane: a control treatment (0.9% saline, at 0.1 ml/kg i.m.), epinephrine (0.1 mg/kg i.m.), or acupuncture with electrical stimulation at GV-26. Each turtle received all treatments, and treatments were separated by 48 hr. Return of spontaneous ventilation was 55% faster in turtles given epinephrine and 58% faster in the GV-26 group versus saline (P < 0.001). The times to movement and to complete recovery were also significantly faster for both treatments than for saline (P < 0.02). Treated turtles displayed increases in temperature not documented in the control (P < 0.001). Turtles administered epinephrine showed significantly increased heart rates and end-tidal CO(2) (P < 0.001). No adverse effects were noted in the study animals. The mechanisms of action were not elucidated in the present investigation. Nevertheless, the use of parenteral epinephrine or GV-26 stimulation in the immediate postanesthetic period produces clinically relevant reductions in anesthetic recovery time in common snapping turtle. Further research is necessary to evaluate the effects of concurrent GV-26 and epinephrine administration and to assess responses in other reptilian species.

BLASTOCYSTIS SP. AND BLASTOCYSTIS RATTI IN A BRAZILIAN PORCUPINE (COENDOU PREHENSILIS) WITH DIARRHEA.

A hand-raised, 5-mo-old, intact male Brazilian porcupine (Coendou prehensilis) was evaluated for chronic diarrhea, failure to thrive, and anorexia. On presentation the porcupette was dull, dehydrated, and passing yellow, malodourous, watery diarrhea. Cytologic examination of feces revealed a large number of organisms, morphologically consistent with Blastocystis. Blastocystis polymerase chain reaction (PCR) performed on feces was positive. Direct sequencing on two sequential samples confirmed the presence of Blastocystis ratti and a novel Blastocystis sequence. The porcupette was treated supportively, which included a 4-wk metronidazole course. Diarrhea resolved within 2 wk of treatment, and the animal's growth rate dramatically improved. Recheck PCR was negative for Blastocystis. Although an important and controversial cause of diarrhea in immunocompromised humans, this organism is not well recognized as a potential pathogen and zoonosis in zoo animals. Clinicians should be aware of the potential for disease associated with this organism, especially in immunocompromised animals.

MANAGEMENT OF A PATENT URACHUS AND YOLK COELOMITIS IN A PREHENSILE-TAILED SKINK (CORUCIA ZEBRATA).

An 11-day-old, captive-born, male prehensile-tailed skink (Corucia zebrata) was evaluated for a chronically swollen umbilicus. On presentation, the skink appeared dehydrated and weak. The umbilical stump was sensitive, edematous, and erythematous. Yellow fluid was readily expressed during palpation of the surrounding area, suggestive of urine. Following several days of supportive care, a positive contrast cloacagram supported the diagnosis of a patent urachus. An exploratory celiotomy was performed, confirming yolk coelomitis and a patent urachus. Both were corrected surgically, and the skink improved steadily thereafter. This report confirms the presence of a urinary bladder in C. zebrata and is the first report of a patent urachus in a reptile. Surgical intervention and medical management of concurrent infectious coelomitis were curative in this case.

Meningoencephalitis with secondary obstructive hydrocephalus caused by probable coccidioides species in a buff-cheeked gibbon (Nomascus gabriellae).

An 8-yr-old male buff-cheeked gibbon (Nomascus gabriellae) acutely developed abnormal behavior, decreased appetite, and dull mentation. Mild generalized muscle wasting and weight loss were the only other abnormalities noted on examination. Routine immunodiffusion serology for Coccidioides spp. were IgG and IgM positive. Magnetic resonance imaging of the brain was suggestive of an infectious meningoencephalitis with secondary obstructive hydrocephalus. A ventriculoperitoneal shunt was placed in standard fashion to reduce the imminent risk of mortality from increased intracranial pressure. Postoperative treatment included oral fluconazole, a tapered course of prednisolone, and physical therapy. Clinical signs improved steadily and the gibbon was fit to return to exhibit 8 wk post-shunt placement. This case of coccidioidomycosis demonstrates the complications that can occur with dissemination to the central nervous system and its management. It is the first published report describing the use of ventriculoperitoneal shunt placement in this species.

Septicaemic Listeriosis in a White-Faced Saki (Pithecia pithecia).

A 27-year-old female white-faced saki (Pithecia pithecia) died following an onset of vomiting and ptyalism. Necropsy revealed lesions of suppurative ventriculitis, choroid plexitis, periventricular encephalitis and meningitis with intralesional gram-positive coccobacilli and paired rods. The saki also had suppurative to mononuclear hepatitis, mild intestinal crypt necrosis, proliferative glomerulonephritis, aortic arteriosclerosis, pulmonary interstitial fibrosis, chronic mild epicarditis, ovarian medullary arteriopathy and a focal superficial cerebral fibrotic nodule with surrounding chronic mixed cell inflammation. Listeria monocytogenes was cultured from liver and spinal cord. Intralesional Listeria bacteria were immunolabelled in brain sections and real-time polymerase chain reaction of brain tissue detected L. monocytogenes. Whole genome multilocus sequence typing characterized the cultured bacterial isolates as sequence type 6 and clonal complex 6. A database search for related clinical and food listerial outbreaks identified genetically related isolates but, because these isolates were more than 20 alleles distant from the saki isolates, they were not a related cluster. Reports of listeriosis in non-human primates are infrequent, and when infections do occur, they tend to be haematogenous with the propensity to cause meningoencephalitis. This saki likely ingested environmental L. monocytogenes, which resulted in disease that may have been facilitated by pre-existing co-morbidities and age.

Eumycetoma and disseminated phaeohyphomycosis in a Sumatran tiger.

A 16-y-old female Sumatran tiger (Panthera tigris sumatrae) was evaluated for hyporexia. Examination revealed chronic kidney disease and a large subcutaneous axillary mass with draining tracts that contained numerous small black grains. Histologic examination revealed the presence of intralesional fungal hyphae. Persistent hyporexia and pyogranulomatous disease, as well as progressive cachexia and azotemia occurred despite treatment, and euthanasia was performed. Disseminated phaeohyphomycosis was diagnosed on postmortem examination, additionally affecting various lymph nodes, the nasal cavity, mesenteric adipose tissue, abdominal aorta, pericardium, and kidney. Fungal culture from a deep-tissue sample isolated a pure growth of Curvularia sp., a dematiaceous opportunistic fungus able to cause eumycetomas and/or phaeohyphomycosis. Phaeohyphomycosis is a rare but emerging condition, not previously reported as disseminated disease in an exotic carnivore, to our knowledge. Aggressive systemic antifungal treatment was unsuccessful, likely complicated by diagnostic challenges and concurrent renal disease. The presence of a swelling with abundant grains exiting draining tracts should direct clinicians to the diagnosis of a mycetoma, warranting early and aggressive treatment.