NONDOMESTIC FELID ABC BLOOD PHENOTYPING, GENOTYPING, AND CROSSMATCHING.

Based upon previous clinical experience with domestic cats (Felis catus), the ability to assess ABC blood types and blood (in-)compatibilities of nondomestic felids, and adequately consider and plan for blood transfusions, may be important. Although nondomestic felids appear to have an ABC blood group system similar to domestic cats, typing with point-of-care kits and by CMAH genotyping for domestic cats have not been reported. In this study, 162 blood samples from 18 different nondomestic felid species (cheetah [Acinonyx jubatus, n = 42], lion [Panthera leo, n = 33], tiger [Panthera tigris, n = 23], Canada lynx [Lynx canadensis, n = 11], snow leopard [Uncia uncia, n = 10], puma [Puma concolor, n = 7], clouded leopard [Neofelis nebulosa, n = 6], serval [Leptailurus serval, n = 5], jaguar [Panthera onca, n = 5], fishing cat [Prionailurus viverrinus, n = 4], Pallas cat [Felis manul, n = 3], bobcat [Lynx rufus, n = 3], ocelot [Leopardus pardalis, n = 3], black footed cat [Felis nigripes, n = 2], leopard [Panthera pardus, n = 2], African wildcat [Felis lybica, n = 1], caracal [Caracal caracal, n = 1], and sand cat [Felis margarita, n = 1]) were ABC blood typed by laboratory and point-of-care tests, genotyped for four known CMAH variants for type B and type C (AB) phenotypes, and crossmatched with one another and domestic type A cats. Traditional tube typing identified blood type A (n = 106), type B (n = 8), type C (n = 43), and no discernible ABC type (n = 4). Several discrepancies were found between point-of-care and traditional typing test results. None of the tested felids possessed the four CMAH variants responsible for type B and C (AB) in domestic cats. Crossmatch incompatibilities (≥2+ agglutination) were identified within and between nondomestic felid species and beyond ABC incompatibilities. Of 26 crossmatches performed between domestic cats and various nondomestic felids, only 7 (27%) were compatible. In conclusion, point-of-care typing kits and CMAH genotyping, successfully used in domestic cats, may not identify the correct ABC blood type in nondomestic felids. Prior crossmatching is recommended to increase the likelihood of compatible transfusions between any nondomestic felids.

VITAMIN C DEFICIENCY-ASSOCIATED LESIONS IN A COLONY OF COMMON VAMPIRE BATS (DESMODUS ROTUNDUS) IN A ZOO FACILITY.

The Milwaukee County Zoo has housed common vampire bats (Desmodus rotundus) since 1973. The bats are fed defibrinated cow's blood supplemented with a liquid pediatric multivitamin. From July 2013 to May 2014, multiple deaths occurred in colony bats, including five juveniles with multiple bone fractures and failure of endochondral ossification, three adults with cerebellar necrosis, and one adult with subcutaneous hemorrhage. In November 2013, an adult bat developed a nonhealing left wing hematoma and eventually succumbed 9 mo later. A postmortem examination revealed multifocal extensive necrohemorrhagic and suppurative ulcerative dermatitis with no underlying cause determined. From July to December 2014, five of nine adult bats in the colony developed similar hematomas along with gingival bleeding. One euthanized bat had a serum ascorbic acid level of 0.08 mg/dl and marked generalized subcutaneous hemorrhage. A therapeutic trial was initiated in which two bats received defibrinated cow's blood supplemented only with oral vitamin C, 100 mg/kg PO q24h for 3 d, and then 50 mg/kg PO q24h. Two other bats received nonsupplemented defibrinated cow's blood and were given vitamin K 3.3 mg/kg SC q12h for 3 d, and then 3.3 mg/kg SC q24h for 7 d. The bats supplemented with vitamin C improved, supporting a diagnosis of vitamin C deficiency. All bats were subsequently supplemented with vitamin C leading to resolution of all lesions within 10 d to 2 mo. Vitamin C is necessary for collagen synthesis, which is required for proper wound healing, capillary and cartilage strength, osteoid production, and pial membrane formation of the cerebellum. Several bat species cannot synthesize vitamin C and require a dietary source. This is the first report of vitamin C deficiency in a colony of vampire bats leading to severe chronic subcutaneous hemorrhage, bone fragility, microfractures, cerebellar necrosis, and death.

OPHTHALMIC EXAMINATION FINDINGS AND INTRAOCULAR PRESSURE MEASUREMENTS IN SIX SPECIES OF ANURA.

A complete ophthalmic exam, including intraocular pressure (IOP) measurement, is key to diagnosing ocular diseases such as uveitis and glaucoma in frogs. We performed complete ophthalmic anterior segment examinations and IOP rebound tonometry measurements using two different settings (other "p" and canine "d") for six anuran species. The objectives were to describe common ocular abnormalities found in these species, to compare IOP values between different tonometer settings, and to compare IOPs between species. Examinations revealed abnormalities including cataracts (11/98 total eyes), lenticular sclerosis (10/98) and lipid keratopathy (9/98). IOP was measured with the TonoVet® and the ranges (oculus uterque, OU mm Hg other "p" setting, canine "d" setting) were giant waxy monkey tree frogs (Phyllomedusa bicolor) (3.5-7.6; 6.5-11.7; n = 5), mission golden-eyed tree frogs (Trachycephalus resinifictrix) (7.0-9.7, 13.2-15.7; n = 6), boreal toads (Bufo boreas boreas) (0.8-5.5, 5.7-10.5; n = 13), Mexican giant tree frogs (Pachymedusa dacnicolor) (3.8-5.0, 8.3-11.8; n = 3), Lake Titicaca frogs (Telmatobius culeus) (8.8-10.5, 14.0-17.2; n = 8), and mossy tree frogs (Theloderma corticale) (9.7-11.0, 15.7-17.0; n = 5). The TonoVet canine "d" setting IOP measurements were statistically higher (P = 0.01) than the other "p" setting measurements for all species except the giant waxy monkey tree frogs. IOP was significantly lower for giant waxy monkey tree frog eyes with cataracts (P < 0.05) with the other "p" setting. IOP did not statistically differ in eyes with lenticular sclerosis. IOP can be measured by rebound tonometer in anurans, but more research is needed for species-specific references using consistent settings.

INTRAOCULAR PRESSURE AND EXAMINATION FINDINGS IN THREE SPECIES OF CENTRAL AND SOUTH AMERICAN TREE FROGS (CRUZIOHYLA CRASPEDOPUS, CRUZIOHYLA CALCARIFER, AND ANOTHECA SPINOSA).

The purpose of this prospective study was to describe intraocular pressure (IOP) and examination findings in three tree frog species (Cruziohyla craspedopus [fringe leaf frog], Cruziohyla calcarifer [splendid leaf frog], and Anotheca spinosa [spiny-headed or coronated tree frog]). Thirty-one C. craspedopus, four C. calcarifer, and five A. spinosa were weighed, sexed based on phenotype where possible, and examined using slit-lamp biomicroscopy and indirect ophthalmoscopy. IOP was measured using the TonoVet and TonoLab rebound tonometers while the frogs were held two ways (unrestrained, then restrained). Statistical differences were determined using one-way analysis of variance (ANOVA) and t-tests. Mean ± SD IOP (TonoVet and TonoLab, respectively) was 15.1 ± 2.5 mmHg and 15.6 ± 4.1 mmHg in C. craspedopus; 14.8 ± 1.5 mmHg and 18.8 ± 3.1 mmHg in C. calcarifer; and 9.1 ± 2.1 mmHg and 10.8 ± 1.4 mmHg in A. spinosa. There was no significant difference in IOP in C. craspedopus by eye (Right vs Left), tonometer, or restraint method. IOP in female C. craspedopus was 1-3 mm Hg higher than in males with both devices (P < 0.05). IOP was statistically significantly different between all species for the TonoLab and between Cruziohyla genus frogs and A. spinosa for the TonoVet (P < 0.05). There was no difference in IOP measurements between the TonoVet and TonoLab in C. craspedopus. IOP varied by gender in C. craspedopus and between species, but not by tonometer. Ocular abnormalities were minimal in this group of captive bred frogs.

MEASURING INTRAOCULAR PRESSURE IN WHITE'S TREE FROGS (LITORIA CAERULEA) BY REBOUND TONOMETRY: COMPARING DEVICE, TIME OF DAY, AND MANUAL VERSUS CHEMICAL RESTRAINT METHODS.

Ocular diseases reported in frogs include uveitis and glaucoma, which are associated with changes in intraocular pressure (IOP). The objectives of this study were to characterize the normal IOP for White's tree frogs ( Litoria caerulea ) using two types of rebound tonometers, and to assess whether time of day or method of restraint affected IOP. Eighteen conscious, unrestrained, ophthalmologically normal frogs were used to measure IOP using TonoVet® and TonoLab® tonometers, at three time points during the day. In a subset of 12 frogs, IOP was measured while under manual restraint using the TonoVet. Anesthesia was induced in 9 frogs using two different concentrations of MS-222 (0.5 g/L and 2 g/L) in order to evaluate for changes in IOP with the TonoVet. Mean (± SD) IOP values for the TonoLab (16.8 ± 3.9 mm Hg) were significantly higher than TonoVet values (14.7 ± 1.6 mm Hg; P < 0.01). TonoVet IOP values did not significantly change with time of day. TonoLab values were significantly lower in the evening (1600-1800; 14.5 ± 3.1 mm Hg), compared with morning and midday measurements (0800-1000 and 1200-1400; 18.0 ± 3.8 mm Hg; P < 0.01). Manually restrained frogs had significantly lower IOP (13.4 ± 1.5 mm Hg) compared with unrestrained frogs (15.3 ± 1.2 mm Hg; P < 0.01). Chemical restraint did not cause significant changes in IOP. Intraocular pressure can be measured with both types of rebound tonometers in White's tree frogs, but time of day and manual restraint can affect IOP values.

CLINICAL MANAGEMENT OF THIRD PHALANX FRACTURES IN LESSER (TRAGELAPHUS IMBERBIS) AND GREATER KUDU (TRAGELAPHUS STREPSICEROS).

Two greater kudu ( Tragelaphus strepsiceros ) and one lesser kudu ( T. imberbis ) from two zoological institutions presented with overgrown front hooves, and were diagnosed with fractures of the third phalanges in the affected digits. Both greater kudu had milder lamenesses at diagnosis, and were managed conservatively with hoof trims, stall rest, and nonsteroidal anti-inflammatory medications. Ongoing management through regular hoof trims led to improvement and eventual resolution of lameness. The more severely lame lesser kudu received hoof blocks on the front claws not associated with fractured phalanges. This therapy was well tolerated and resulted in resolution of lameness immediately after application. Radiographic evidence of healing was present 8 wk posttherapy. Diagnosis of these fractures was greatly aided by radiographic views obtained at a 45° oblique angle with the claws distracted.

MYOSITIS OSSIFICANS TRAUMATICA IN A VAMPIRE BAT (DESMODUS ROTUNDUS).

A 15-yr-old sexually intact female vampire bat ( Desmodus rotundus ) was diagnosed with myositis ossificans traumatica of the abdominal wall. The bat presented with a large ulcerated firm mass along the abdomen. Radiographs and cytology were performed, followed by surgical exploration. The mass was determined to be nonresectable and the bat was euthanized. Histopathology showed severe necrotizing, degenerative, and pyogranulomatous myositis with osseous and cartilaginous metaplasia, fibrosis, and ulceration, which were consistent with myositis ossificans traumatica. Myositis ossificans traumatica is commonly associated with previous trauma to skeletal muscle. Two years prior, this bat had an emergency Caesarian section at this site, which was postulated to elicit a marked tissue response leading to this condition. Myositis ossificans traumatica is infrequently reported in humans, dogs, cats, pigs, and horses. To the author's knowledge, this is the first report of this condition in a bat.

Bilateral Uveitis and Hyphema in a Catalina Macaw (Ara ararauna × Ara macao ) With Multicentric Lymphoma.

A 20-year-old, female Catalina macaw (Ara ararauna × Ara macao ) was presented with bilateral uveitis and hyphema. The hyphema initially improved with 0.12% prednisolone acetate ophthalmic drops (1 drop OU q4h for 7 days), but the hyphema recurred after the drops were tapered. The bird subsequently developed inappetance, weight loss, regurgitation, and lethargy and was euthanatized 24 days after initial presentation. Necropsy revealed marked splenomegaly and hepatomegaly, with significant mucosal ulcerations of the proventriculus and petechiation associated with both kidneys. Histopathologic examination revealed multicentric lymphoma, with neoplastic cells observed in ocular, splenic, hepatic, renal, proventricular, intestinal, pancreatic, and choanal tissue. Neoplastic lymphocytes effaced the iris, ciliary body, and the choroid of the eyes, and neoplastic lymphocytes were attached to the corneal endothelium and infiltrated the sclera, episclera, and conjunctivae. Immunohistochemical results indicated that the neoplastic lymphocytes were CD3(+) and CD79a(-), which is consistent with T-cell lymphoma.

EXPERIMENTAL CHALLENGE STUDY OF FV3-LIKE RANAVIRUS INFECTION IN PREVIOUSLY FV3-LIKE RANAVIRUS INFECTED EASTERN BOX TURTLES (TERRAPENE CAROLINA CAROLINA) TO ASSESS INFECTION AND SURVIVAL.

The Maryland Zoo in Baltimore experienced an outbreak of Frog virus-3 (FV3)-like ranavirus during the summer of 2011, during which 14 of 27 (52%) of its captive eastern box turtles (Terrapene carolina carolina) survived. To assess survival, immunity, and viral shedding, an experimental challenge study was performed in which the surviving, previously infected turtles were reinfected with the outbreak strain of FV3-like ranavirus. Seven turtles were inoculated with virus intramuscularly and four control turtles received saline intramuscularly. The turtles were monitored for 8 wk with blood and oral swabs collected for quantitative polymerase chain reaction (qPCR). During that time, one of seven (14%) inoculated turtles and none of the controls (0%) died; there was no significant difference in survival. Clinical signs of the inoculated turtles, except for the turtle that died, were mild compared to the original outbreak. Quantitative PCR for FV3-like ranavirus on blood and oral swabs was positive for all inoculated turtles and negative for all controls. The turtle that died had intracytoplasmic inclusion bodies in multiple organs. Three inoculated and two control turtles were euthanized at the end of the study. No inclusion bodies were present in any of the organs. Quantitative PCR detected FV3-like ranavirus in the spleen of a control turtle, which suggested persistence of the virus. The surviving five turtles were qPCR-negative for FV3-like ranavirus from blood and oral swabs after brumation. Quantitative PCR for Terrapene herpesvirus 1 found no association between ranavirus infection and herpesvirus loads. In conclusion, previously infected eastern box turtles can be reinfected with the same strain of FV3-like ranavirus and show mild to no clinical signs but can shed the virus from the oral cavity.

Comparison of Serum Protein Electrophoresis Values in Wild and Captive Whooping Cranes ( Grus americana ).

Protein electrophoresis of serum samples from endangered, wild whooping cranes ( Grus americana ) was performed to help assess the health of the only self-sustaining, migratory population in North America. Serum samples from wild adult cranes (n = 22) were taken at Aransas National Wildlife Refuge, Texas, USA during winter. Wild juvenile cranes (n = 26) were sampled at Wood Buffalo National Park, Northwest Territories, Canada, in midsummer. All captive crane samples were acquired from the International Crane Foundation, Baraboo, WI, USA. Captive adult cranes (n = 30) were sampled during annual examinations, and archived serum samples from captive juvenile cranes (n = 19) were selected to match the estimated age of wild juveniles. Wild juveniles had significantly lower concentrations of all protein fractions than wild adults, except for prealbumin and γ globulins. All protein fraction concentrations for wild juveniles were significantly lower compared with captive juveniles, except for prealbumin and γ globulins, which were higher. Wild adults had significantly greater γ globulin concentrations than captive adults. Captive juveniles had significantly lower prealbumin and albumin concentrations and albumin : globulin ratios than captive adults. The higher γ globulin concentrations in wild versus captive cranes are likely because of increased antigenic exposure and immune stimulation. Protein fraction concentrations vary significantly with age and natural history in this species. Reference intervals for serum protein electrophoresis results from captive adult whooping cranes are provided in this study.

Gut microbiota and age shape susceptibility to clostridial enteritis in lorikeets under human care.

BACKGROUND: Enteritis is a common cause of morbidity and mortality in lorikeets that can be challenging to diagnose and treat. In this study, we examine gut microbiota in two lorikeet flocks with enteritis (Columbus Zoo and Aquarium-CZA; Denver Zoo-DZ). Since 2012, the CZA flock has experienced repeated outbreaks of enteritis despite extensive diet, husbandry, and clinical modifications. In 2018, both CZA and DZ observed a spike in enteritis. Recent research has revealed that the gut microbiota can influence susceptibility to enteropathogens. We hypothesized that a dysbiosis, or alteration in the gut microbial community, was making some lorikeets more susceptible to enteritis, and our goal was to characterize this dysbiosis and determine the features that predicted susceptibility. RESULTS: We employed 16S rRNA sequencing to characterize the cloacal microbiota in lorikeets (CZA n = 67, DZ n = 24) over time. We compared the microbiota of healthy lorikeets, to lorikeets with enteritis, and lorikeets susceptible to enteritis, with "susceptible" being defined as healthy birds that subsequently developed enteritis. Based on sequencing data, culture, and toxin gene detection in intestinal contents, we identified Clostridium perfringens type A (CZA and DZ) and C. colinum (CZA only) at increased relative abundances in birds with enteritis. Histopathology and immunohistochemistry further identified the presence of gram-positive bacilli and C. perfringens, respectively, in the necrotizing intestinal lesions. Finally, using Random Forests and LASSO models, we identified several features (young age and the presence of Rhodococcus fascians and Pseudomonas umsongensis) associated with susceptibility to clostridial enteritis. CONCLUSIONS: We identified C. perfringens type A and C. colinum associated with lorikeet necrohemorrhagic enteritis at CZA and DZ. Susceptibility testing of isolates lead to an updated clinical treatment plan which ultimately resolved the outbreaks at both institutions. This work provides a foundation for understanding gut microbiota features that are permissive to clostridial colonization and host factors (e.g. age, prior infection) that shape responses to infection.

Analgesic Efficacy of Tramadol and Morphine in White's Tree Frogs (Litoria caerulea).

Published data are sparse regarding the recognition of clinically relevant pain and appropriate analgesia in amphibians. The amphibian analgesia literature has primarily focused on nociceptive pathways in a single species, the northern leopard frog (Rana pipiens). The objective of the current study was to assess the analgesic efficacy and safety of oral tramadol and subcutaneous morphine in a commonly maintained zoo and pet species, White's tree frog (Litoria caerulea). We hypothesized that tramadol and morphine would provide dose-dependent antinociception, as measured by significant increases in hindlimb withdrawal latency after exposure to a noxious thermal stimulus. Two randomized, placebo-controlled, complete crossover studies were performed, with tramadol (n = 12) administered at 15, 25, and 40 mg/kg PO and morphine (n = 12) administered at 5 and 10 mg/kg SC. Hindlimb withdrawal latency was measured for a maximum of 72 h. No adverse side effects or signs of sedation were observed with any dose or drug evaluated. No significant difference in withdrawal latency was detected between the control and either tramadol or morphine. These negative results were surprising, suggesting that the thermal nociceptive model may not be biologically relevant in amphibian species.

Urethral diverticulum and urolithiasis in a female guinea pig (Cavia porcellus).

CASE DESCRIPTION A 5-year-old sexually intact female guinea pig was evaluated because of mild dysuria and a subcutaneous mass located cranioventral to the urogenital openings. CLINICAL FINDINGS Non-contrast-enhanced CT and surgical exploration of the distal aspect of the urethra revealed a urethral diverticulum with an intraluminal urolith. Analysis revealed that the urolith was composed of calcium carbonate and struvite. TREATMENT AND OUTCOME The urolith was surgically removed and ablation of the urethral diverticulum was attempted. Approximately 3 months later, the guinea pig was reevaluated for masses in the perineal region, and positive-contrast urethrocystography revealed 2 uroliths present in the same diverticulum. Uroliths were manually expressed with the patient under general anesthesia. Approximately 2 weeks later, urethroplasty was performed to create an enlarged stoma with the diverticulum, thereby preventing urine from pooling in the diverticulum and potentially reducing the risk of future urolith formation. The urethroplasty site healed well with no reported complications or evidence of urolith recurrence 6 months after surgery. CLINICAL RELEVANCE Urolithiasis is common in guinea pigs, and urethral diverticulum and intraluminal urolith formation should be considered as a potential differential diagnosis for a subcutaneous mass along the distal aspect of the urethra. Creation of a urethral stoma from a urethral diverticulum via urethroplasty achieved a successful outcome in this patient.

Distribution and outcome of ocular lesions in snakes examined at a veterinary teaching hospital: 67 cases (1985-2010).

OBJECTIVE: To determine the distribution and clinical outcome of ocular lesions in snakes. DESIGN: Retrospective case series. ANIMALS: 67 snakes with ocular lesions. PROCEDURES: Signalment, lesion duration, diagnosis, treatment, and clinical outcome were recorded for all snakes with ocular lesions that were examined at a veterinary teaching hospital from 1985 to 2010. RESULTS: 71 ocular lesions were detected in 67 of 508 (13%) snakes examined. Affected snakes were of the families Boidae, Pythonidae, Colubridae, and Viperidae. The distribution of ocular lesions did not vary by taxonomic family, age, or sex; however, snakes from the genus Epicrates with ocular lesions were overrepresented in the population. The most commonly diagnosed ocular lesions were retained spectacle (n = 41), pseudobuphthalmos or subspectacular abscess (13), trauma (8), and cataracts (4). Pseudobuphthalmos or subspectacular abscess developed more frequently in Colubridae than in non-Colubridae snakes. Of the 16 snakes with retained spectacles for which data were available, the lesion recurred once in 4 snakes and multiple times in 5 snakes. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that retained spectacle was the most common ocular lesion diagnosed in snakes. Compared with other snakes with ocular lesions, snakes of the genus Epicrates had a higher than expected frequency of ocular lesions in general and snakes of the family Colubridae had a higher than expected frequency of pseudobuphthalmos or subspectacular abscess.