AIR SAC TREMATODES (CYCLOCOELIDAE STOSSICH, 1902) INFECTING BIRDS IN ZOOLOGICAL INSTITUTIONS IN THE UNITED STATES.

Air sac trematodes (Digenea: Cyclocoelidae) were detected in 23 avian species from eight aviaries in the United States. Most of the infected host species were passeriform birds, but a few species in other orders also were infected. Four species of adult flukes were encountered: Circumvitellatrema momota, Morishitium sp., Psophiatrema greineri, and Szidatitrema yamagutii. Findings from retrospective review of medical records, necropsy records, and author observations are presented. Potential terrestrial snail intermediate hosts were collected from three indoor aviaries. A high prevalence (47%) of larval trematode infections was demonstrated in one species of nonnative snail (Prosopeas achatinacea); one larva was isolated and matched to the adult species (C. momota) from birds using PCR. Problems with introducing potentially infected wild-caught birds into aviaries, and exchanging captive individuals between aviaries where they potentially may carry infections, are discussed.

MAST CELL NEOPLASIA IN THE GREEN IGUANA, IGUANA IGUANA.

This case series is the first description of mast cell neoplasia in green iguanas, Iguana iguana. All iguanas were adults. Six were females, and four were males. Lesions were associated with the oral cavity in 3 of 10 cases, and on the head in 3 of 10 cases. Three of the cases had masses over the trunk; a single case had a lesion on a limb. Single masses were most common involving the face or oral cavity (6 of 10), whereas four cases with neoplasia distributed along the trunk had multiple masses. One case developed peripheral blood mastocytosis. In most neoplasms, cells contained metachromatic granules in toluidine blue-stained sections. Polymerase chain reaction identified no internal tandem duplication mutation in exon 11 of c-kit, but a mutation in exon 8 in a single case. Compared with other reptiles, green iguanas may be predisposed to development of mast cell neoplasia.

Hepatic copper accumulates in ferrets with and without hepatobiliary disease.

OBJECTIVE: To determine hepatic copper concentrations and zonal distribution in ferrets with and without hepatobiliary disease, validate rhodanine-based qualitative copper scoring and digital copper quantification in ferret hepatic samples, and ascertain whether clinical features predicted copper accumulation. ANIMALS: 34 ferrets, including 7 with necroinflammatory disease, 5 with hepatocellular carcinoma, 13 with non-necroinflammatory disease, and 9 with no hepatobiliary disease. PROCEDURES: Rhodanine-based digital copper quantification was validated by use of liver dually measured by atomic absorption spectroscopy and digital scanning (R2 = 0.98). Clinical features and hepatic copper scores and concentrations (dry weight liver) were compared between groups. Zonal copper distribution was determined. RESULTS: Hepatic copper concentration was strongly correlated with copper scores (ρ = 0.88). Ferrets with hepatobiliary disease were significantly older and had significantly higher serum alkaline phosphatase and γ-glutamyltransferase activities and creatinine concentrations. Centrilobular copper accumulated in 23 of 34 (64%) ferrets with (n = 15) and without (8) hepatobiliary disease. Median copper concentrations were not significantly different between ferrets with and without hepatobiliary disease but were significantly higher within neoplastic hepatic tissue in ferrets with hepatocellular carcinoma. Hepatic copper concentrations exceeded feline (> 180 µg/g) and canine (> 400 µg/g) reference limits in 19 and 9 ferrets, respectively. Hepatic copper > 1,000 µg/g occurred in 5 ferrets with and 2 without hepatobiliary disease. Clinical features did not predict copper accumulation. CLINICAL RELEVANCE: Rhodanine-based digital copper quantification and qualitative copper scoring discerned liver copper accumulation in ferrets. Ferrets with and without hepatobiliary disease displayed a propensity for centrilobular hepatic copper accumulation of uncertain clinical importance. Clinical and clinicopathologic features could not exclusively implicate pathologic copper accumulation.

Clinical, surgical, and pathological findings in client-owned rabbits with histologically confirmed appendicitis: 19 cases (2015-2019).

OBJECTIVE: To report clinical, surgical, and pathological findings in client-owned rabbits with histologically confirmed appendicitis. ANIMALS: 19 rabbits. PROCEDURES: Medical records for client-owned rabbits that had a histologic diagnosis of appendicitis were reviewed. RESULTS: Median age of the rabbits at presentation was 24.0 months (range, 4 to 84 months). Seventeen cases occurred during the summer and fall seasons. Decreased appetite (17/19 rabbits), abnormal rectal temperature (hyperthermia, 9/16 rabbits; hypothermia, 4/16 rabbits), hypocalcemia (8/11 rabbits), and hypoglycemia (7/15 rabbits) were common signs. Abdominal ultrasonography and CT findings were suggestive of appendicitis in 6 of 8 rabbits and in 1 of 2 rabbits, respectively. Of the 6 rabbits that received medical treatment, 3 died at 48 hours, 1 died at 24 hours after hospitalization, and 1 died at 10 days after presentation; 1 rabbit was alive at 1,030 days after presentation. Of the 8 rabbits that underwent appendectomy, 3 died before discharge from the hospital and 1 died 113 days after surgery; 4 rabbits were alive at 315, 334, 1,433, and 1,473 days after presentation. The remaining 5 rabbits either died or were euthanized before treatment could be instituted. In each of the 19 rabbits, the appendix had evidence of severe inflammation with mucosal ulceration, heterophilic inflammation, and necrotic debris. CLINICAL RELEVANCE: For rabbits with decreased appetite and an apparently painful abdomen, hyperthermia, hypocalcemia, or hypoglycemia, appendicitis should be considered as a differential diagnosis. Further comparisons of medical and surgical treatments are required to establish treatment recommendations for rabbits with appendicitis.

Pathology of Diseases of Geriatric Exotic Mammals.

The review covers select disease conditions most frequently described in aging rodents (rats, mice, hamsters, guinea pigs), rabbits, and ferrets. The conditions are categorized by general organ systems, infectious diseases, and neoplasms. Two data systems, the Veterinary Medical Teaching Hospital and Comparative Pathology Laboratory at the University of California, Davis and Zoo/Exotic Pathology Service, Citrus Heights, California were used in the determining disease conditions to describe.

Development and validation of 2 probe-hybridization quantitative PCR assays for rapid detection of a pathogenic Coxiella species in captive psittacines.

Avian coxiellosis is an emerging cause of morbidity and mortality among captive psittacines, and the utility of a rapid detection test using easily obtained samples is paramount in a clinical setting. New sequences were obtained from 3 genes: groEL, dnaK, and rpoB. We developed probe-hybridization quantitative PCR (qPCR) assays using groEL and dnaK genes. Samples, including splenic aspirates, liver aspirates, whole blood, and choanal, conjunctival, and cloacal swabs, were collected from 4 psittacine species including 3 blue-and-gold macaws (Ara ararauna), 2 scarlet-chested parrots (Neophema splendida), 1 Timneh African grey parrot (Psittacus timneh), and 1 yellow-naped Amazon parrot (Amazona auropalliata). Retrospective review of postmortem findings from 3 of these psittacines included splenomegaly, hepatitis, and/or transmission electron microscopy confirmation consistent with previous reports of avian coxiellosis. There was 100% agreement between these assays and consensus PCR with sequencing. A Wilcoxon rank-sum test found a strong correlation between groEL and dnaK cycle threshold values (p < 0.001), validating these assays for detection of this avian Coxiella sp.

Mandibular squamous cell carcinoma in 2 psittacines.

A unilateral, malformed gnathotheca in a green-winged macaw (Ara chloropterus) was managed for 16 mo with periodic beak trims. Biopsy of the affected beak did not identify an underlying cause; however, at postmortem examination, a densely cellular neoplasm invaded the mandible from the beak, dissecting and effacing the cortex and trabecular bone and inciting periosteal proliferation. There was no evidence of metastasis. Neoplastic cells were strongly immunopositive for cytokeratin AE1/AE3, consistent with squamous cell carcinoma (SCC). A black-headed caique (Pionites melanocephalus) was presented with a similar clinical history, timeline, and gross appearance. Biopsy of the mandibular portion of the lesion resulted in a diagnosis of SCC. Medical management with toceranib at a palliative dose resulted in adverse side effects and was discontinued. Early diagnosis of mandibular SCC may allow for early intervention and development of treatment modalities. Our 2 cases suggest that the mandibular bone, not the beak, may be a preferred biopsy site in the diagnosis of mandibular SCC in psittacines.

Disease Overview of the Urinary Tract in Exotic Companion Mammals and Tips on Clinical Management.

Diseases of the urinary tract are reviewed, covering infectious (bacterial, viral, parasitic), degenerative, congenital, metabolic, nutritional, neoplastic, obstructive, and toxic causes. Some clinical presentations and diagnostic procedures are described for ferrets, rabbits, guinea pigs, hamsters, mice, rats, chinchillas, hedgehogs, and sugar gliders, as well as therapies.

Surgical Management of an Intrathoracic Branchial Cyst in a Yellow-crested Cockatoo (Cacatua sulphurea).

A 40-year-old, female lesser crested cockatoo (Cacatua sulphurea) was presented with a complaint of hyporexia and sudden onset of sneezing and wheezing. Physical examination revealed mild stertorous inhalation, and the apex of the heart was palpable caudoventral to the distal tip of the sternum. Projection radiographic images showed a soft tissue mass displacing the heart and the thoracic portion of the trachea. A subsequent computed tomography series revealed a single, large, and predominantly encapsulated soft-tissue mass. The mass was contained within the cranial thoracic region and occupied most of the anatomic location of the thoracic portion of the clavicular air sac, extending around a portion of the trachea. A surgical exploratory procedure was performed, with a thoracic inlet thoracotomy, and the mass was found to be cystic and deeply attached to surrounding tissues at its caudal-most aspect. Complete excision was not possible, and the mass was drained and an incomplete resection was accomplished with approximately one-half of the cystic structure removed and submitted for histopathology. The mass was found to be benign, epithelial-lined, dense, fibrous connective tissue that would be consistent with a branchial cyst.

Generation of red drum (Sciaenops ocellatus) hematology Reference Intervals with a focus on identified outliers.

BACKGROUND: The Deepwater Horizon (DWH) oil spill in 2010 released millions of barrels of crude oil into the northern Gulf of Mexico, exposing numerous species of animals to the toxic components of oil. A comprehensive assessment of morbidity and mortality caused by DWH oil exposure was undertaken by the DWH Natural Resource Damage Trustees to characterize ecosystem damages. OBJECTIVES: This study aimed to characterize normal hematologic RIs in red drum fish with blood cell descriptions, and to demonstrate the importance of identifying and removing outliers when generating RI. METHODS: Two years after the oil spill, 57 adult, red drum fish of mixed sexes were caught along the eastern Louisiana coastline. Eight different sites were chosen to catch the fish; 6 sites were contaminated with oil, and 2 sites were not contaminated at the time of the oil spill. Hematologic RIs were generated from heparinized whole blood samples of healthy red drum as determined by gross examination and histopathologic examination. Two methods were used to detect hematologic effects likely caused by oil contamination. RESULTS: Red drum PCVs (RI 42-62%) were higher than previously reported in cold water and bottom-dwelling fish species, while absolute WBC counts (RI 2.9-8.7  × 109 /L) were comparable to WBC counts previously reported in other fish species with heterophil and lymphocyte absolute concentrations frequently being equivalent. Anemic animals (PCV<42%) were only identified in oil-contaminated sites. CONCLUSION: RIs in many wild fish species are lacking, and therefore, this study provides valuable baseline data on healthy red drum fish. The outliers assessed using ASVCP RI guidelines can provide valuable clinical information regarding individuals in population health assessments, which may be more sensitive for the detection of abnormalities than for population statistics comparing the mean. The importance of removing outliers and rerunning RI statistics is highlighted by this field example.

Four cases of iridociliary tumors in domestic rabbits (Oryctolagus Cuniculus).

Spontaneously occurring ocular neoplasia is rarely reported in rabbits. This case series presents four cases of rabbits diagnosed with iridociliary tumors, which have not been previously reported in this species. Major pathological findings include epithelial tumors affecting the anterior uvea with variable pigmentation and basement membrane formation. Follow-up information was only available for two cases, but neither showed evidence of metastasis, suggesting that the prognosis for these tumors in rabbits, as in other species, may be very good.

Organ weights and histopathology of double-crested cormorants (Phalacrocorax auritus) dosed orally or dermally with artificially weathered Mississippi Canyon 252 crude oil.

A series of toxicity tests were conducted to assess the effects of low to moderate exposure to artificially weathered Deepwater Horizon Mississippi Canyon 252 crude oil on representative avian species as part of the Natural Resource Damage Assessment. The present report summarizes effects of oral exposure (n=26) of double-crested cormorants (DCCO; Phalacrocorax auritus) to 5 or 10ml oil kg-1 day-1 for up to 21 days or dermal application (n=25) of 13ml oil to breast and back feathers every three days totaling 6 applications in 21 days on organ weights and histopathology. Absolute and relative kidney and liver weights were increased in birds exposed to oil. Additionally, gross and/or histopathologic lesions occurred in the kidney, heart, pancreas and thyroid. Clinically significant renal lesions in the orally dosed birds included squamous metaplasia and increased epithelial hypertrophy of the collecting ducts and renal tubules and mineralization in comparison to controls. Gross cardiac lesions including thin walls and flaccid musculature were documented in both orally and dermally dosed birds and myocardial fibrosis was found in low numbers of dermally dosed birds only. Cytoplasmic vacuolation of the exocrine pancreas was noted in orally dosed birds only. Thyroid follicular hyperplasia was increased in dermally dosed birds only possibly due to increased metabolism required to compensate damaged feather integrity and thermoregulate. Gastrointestinal ulceration was found in orally dosed birds only. There were no significant hepatic histopathologic lesions induced by either exposure route. Therefore, hepatic histopathology is likely not a good representation of oil-induced damage. Taken together, the results suggest that oral or dermal exposure of DCCOs to artificially weathered MC252 crude oil induced organ damage that could potentially affect survivability.

Coinfection with a novel fibropapilloma-associated herpesvirus and a novel Spirorchis sp. in an eastern box turtle (Terrapene carolina) in Florida.

Herpesviruses are important pathogens of chelonians, and include Chelonid herpesvirus 5, which is associated with fibropapillomatosis in sea turtles. Spirorchid trematodes are blood flukes that reside within the cardiovascular system of marine turtles and may be associated with severe disease. An eastern box turtle (Terrapene carolina) at the South Florida Wildlife Care Center (Fort Lauderdale, Florida) was presented to the facility with papillomatous growths behind both rear legs. Surgical removal resulted in remission for 8 months; however, lesions recurred, prompting a second surgery and acyclovir therapy. Surgical biopsies revealed subacute superficial inflammation associated with the supporting stroma of the cutaneous papillomas and granulomas within the superficial dermis containing fragmented and collapsed brown trematode eggs surrounded by multinucleated giant cells and epithelioid macrophages. Pan-herpesviral and pan-trematode consensus polymerase chain reaction and sequencing were run on tissue samples. Comparative sequence analysis revealed a novel alphaherpesvirus and a novel trematode in the genus Spirorchis. The animal became anorexic and was euthanized due to poor quality of life. While we do not yet have a complete understanding of the effects of herpesvirus and trematode infections in eastern box turtles, the findings thus presented provide initial insights into the disease relationships among these chelonians.

Widespread recombination, reassortment, and transmission of unbalanced compound viral genotypes in natural arenavirus infections.

Arenaviruses are one of the largest families of human hemorrhagic fever viruses and are known to infect both mammals and snakes. Arenaviruses package a large (L) and small (S) genome segment in their virions. For segmented RNA viruses like these, novel genotypes can be generated through mutation, recombination, and reassortment. Although it is believed that an ancient recombination event led to the emergence of a new lineage of mammalian arenaviruses, neither recombination nor reassortment has been definitively documented in natural arenavirus infections. Here, we used metagenomic sequencing to survey the viral diversity present in captive arenavirus-infected snakes. From 48 infected animals, we determined the complete or near complete sequence of 210 genome segments that grouped into 23 L and 11 S genotypes. The majority of snakes were multiply infected, with up to 4 distinct S and 11 distinct L segment genotypes in individual animals. This S/L imbalance was typical: in all cases intrahost L segment genotypes outnumbered S genotypes, and a particular S segment genotype dominated in individual animals and at a population level. We corroborated sequencing results by qRT-PCR and virus isolation, and isolates replicated as ensembles in culture. Numerous instances of recombination and reassortment were detected, including recombinant segments with unusual organizations featuring 2 intergenic regions and superfluous content, which were capable of stable replication and transmission despite their atypical structures. Overall, this represents intrahost diversity of an extent and form that goes well beyond what has been observed for arenaviruses or for viruses in general. This diversity can be plausibly attributed to the captive intermingling of sub-clinically infected wild-caught snakes. Thus, beyond providing a unique opportunity to study arenavirus evolution and adaptation, these findings allow the investigation of unintended anthropogenic impacts on viral ecology, diversity, and disease potential.

Isolation of a Bohle-like iridovirus from boreal toads housed within a cosmopolitan aquarium collection.

A captive 'survival assurance' population of 56 endangered boreal toads Anaxyrus boreas boreas, housed within a cosmopolitan collection of amphibians originating from Southeast Asia and other locations, experienced high mortality (91%) in April to July 2010. Histological examination demonstrated lesions consistent with ranaviral disease, including multicentric necrosis of skin, kidney, liver, spleen, and hematopoietic tissue, vasculitis, and myriad basophilic intracytoplasmic inclusion bodies. Initial confirmation of ranavirus infection was made by Taqman real-time PCR analysis of a portion of the major capsid protein (MCP) gene and detection of iridovirus-like particles by transmission electron microscopy. Preliminary DNA sequence analysis of the MCP, DNA polymerase, and neurofilament protein (NFP) genes demonstrated highest identity with Bohle iridovirus (BIV). A virus, tentatively designated zoo ranavirus (ZRV), was subsequently isolated, and viral protein profiles, restriction fragment length polymorphism analysis, and next generation DNA sequencing were performed. Comparison of a concatenated set of 4 ZRV genes, for which BIV sequence data are available, with sequence data from representative ranaviruses confirmed that ZRV was most similar to BIV. This is the first report of a BIV-like agent outside of Australia. However, it is not clear whether ZRV is a novel North American variant of BIV or whether it was acquired by exposure to amphibians co-inhabiting the same facility and originating from different geographic locations. Lastly, several surviving toads remained PCR-positive 10 wk after the conclusion of the outbreak. This finding has implications for the management of amphibians destined for use in reintroduction programs, as their release may inadvertently lead to viral dissemination.

Gastrointestinal cryptosporidiosis in captive psittacine birds in the United States: a case review.

Histopathologic analysis performed on tissue samples from 34 captive psittacine birds over a 14-year period demonstrated gastrointestinal cryptosporidiosis. Although 8 birds had died unexpectedly, at least 10 showed gastrointestinal signs before death. Most birds had other significant pathologic lesions, unrelated to cryptosporidiosis, in which case Cryptosporidium was likely a secondary pathogen. However, in 11 birds the only significant histopathologic finding was gastrointestinal cryptosporidiosis. In 2 lovebird (Agapornis species) cases, direct sequencing of proventricular samples confirmed the presence of Cryptosporidium avian genotype III. A description and analysis of the pathologic findings in these 34 birds, as well as a review of avian gastrointestinal cryptosporidiosis, is provided.

Lesions of the avian pancreas.

Although not well described, occasional reports of avian exocrine and endocrine pancreatic disease are available. This article describes the lesions associated with common diseases of the avian pancreas reported in the literature and/or seen by the authors.

Cutaneous neoplasia in ferrets, rabbits, and guinea pigs.

The frequency of common cutaneous tumors in ferrets, rabbits, and guinea pigs is summarized, with descriptions of their appearance and behavior, prognosis, and treatment options. This information assists clinicians when talking to clients, and helps owners make an informed decision whether or not to pursue aggressive treatment or to palliate.

Prevalence of and risk factors associated with atherosclerosis in psittacine birds.

OBJECTIVE: To estimate the prevalence of clinically relevant atherosclerotic lesions in birds and identify epidemiological variables and illness types associated with development of atherosclerosis. DESIGN: Retrospective case-control study. SAMPLE: Records of 7683 psittacine birds, including 525 with advanced atherosclerosis. PROCEDURES: 5 pathology centers provided databases and access to histopathology slides. Age and sex were collected for all birds of the Amazona, Ara, Cacatua, Nymphicus, and Psittacus genera. Databases were searched for atherosclerosis cases, and slides were reviewed for the presence of type IV through VI atherosclerotic lesions. Results were used to build several multiple logistic models to define the association between advanced atherosclerosis and age, sex, genus, illness type, and specific lesions. Prevalence was reported as a function of age, sex, and genus. RESULTS: In the first model including 7683 birds, age, female sex, and the genera Psittacus, Amazona, and Nymphicus were significantly associated with clinically relevant atherosclerosis detected via necropsy. Subsequent models of 1,050 cases revealed further associations with reproductive disease, hepatic disease, and myocardial fibrosis, controlling for age, sex, and genus. CONCLUSIONS AND CLINICAL RELEVANCE: Age, female sex, and 3 genera appeared to be positively associated with the presence of advanced atherosclerotic lesions in psittacine birds. This information may be useful in clinical assessment of the cardiovascular system and patient management. Reproductive diseases were the only potentially modifiable risk factor identified and could be a target for prevention in captive psittacine birds.

Diagnosis and treatment of an insulinoma in a guinea pig (Cavia porcellus).

CASE DESCRIPTION: A 5-year-old male guinea pig (Cavia porcellus) was examined because of lethargy, weight loss, and episodic neurologic signs, including paddling in lateral recumbency, head tilt, and circling. Prior to initial examination, the animal was treated with corn syrup whenever it appeared lethargic, plus an unknown dosage of sulfadimethoxazole. CLINICAL FINDINGS: The animal was thin, with abdominal distention and subtle torticollis. Chemistry panel results documented hypoglycemia (45 mg/dL). Corn syrup was discontinued in favor of a high-fiber formula fed via a syringe. Measurement of blood insulin concentration demonstrated hyperinsulinemia (> 1,440 pmol/L [> 201 µU/L]), with concurrent hypoglycemia (0.6 mmol/L [11 mg/dL]). TREATMENT AND OUTCOME: Diazoxide treatment for presumptive insulinoma was started at a dosage of 5 mg/kg (2.3 mg/lb), p.o., every 12 hours. A blood glucose curve demonstrated persistent hypoglycemia, and the diazoxide dosage was gradually increased to 25 mg/kg (11.4 mg/lb), p.o., every 12 hours. A second glucose curve measurement 12 days later confirmed adequate euglycemic control. Three weeks after the initial diazoxide dosage increase, the animal was reexamined for constipation and abdominal distension and died the following day. Histologic analysis confirmed a pancreatic beta-cell tumor (insulinoma). CLINICAL RELEVANCE: To the authors' knowledge, this is the first report of premortem diagnosis and treatment of an insulinoma in a guinea pig. This case demonstrates that diazoxide treatment can help achieve euglycemia in hypoglycemic guinea pigs and is a potential treatment option for guinea pigs with insulinoma.