COMPARISON OF SUBCUTANEOUS ALFAXALONE AND SUBCUTANEOUS ALFAXALONE-DEXMEDETOMIDINE FOR SEDATION IN THE HOUSTON TOAD (ANAXYRUS HOUSTONENSIS).

The Houston toad (Anaxyrus houstonensis), a primarily terrestrial amphibian of south-central Texas, has been listed as federally endangered since 1970. Sedation is an important tool for obtaining diagnostics and providing treatment in this species. This prospective, randomized, and blinded study compared the sedative effects of SC alfaxalone (Protocol A) at approximately 12 mg/kg (median [range] = 12.70 [12.09-13.95] mg/kg] to SC alfaxalone-dexmedetomidine (Protocol AD) at approximately 12 mg/kg (median [range] = 12.68 [12.16-13.56] mg/kg) and 0.1 mg/kg (median [range] = 0.1 [0.07-0.13] mg/kg), respectively, in adult Houston toads (n = 26). Toads from Protocol AD received atipamezole SC at approximately 1 mg/kg (median [range] = 0.96 [0.75-1.25] mg/kg) 45 min postinduction, whereas toads from Protocol A received the equivalent volume of SC sterile saline at the same time point. Heart rate, gular rate, and times to first effect, loss of righting reflex, ability to position for radiographs, loss of nociception, return of righting reflex, and full recovery were recorded. A significantly greater number of toads lost righting reflex, positioned for radiographs, and lost nociception with Protocol AD compared with Protocol A. Additionally, time to return of righting reflex and time to full recovery were significantly longer with Protocol AD than with Protocol A. The protocols did not differ significantly in time to first effect, time to radiographic positioning, or time to loss of nociception. Histologic examination of four toads euthanized during the study revealed acute injection site reactions from all administered drugs, including saline. No clinical adverse reactions were observed. This study demonstrates that the combination of SC alfaxalone and dexmedetomidine results in deeper sedation than SC alfaxalone alone, but also correlates with longer recovery times despite antagonist administration.

Neoplasia in an assurance population of Puerto Rican crested toads (Peltophryne lemur).

Disease monitoring of amphibian assurance populations is an important buffer against ongoing global extinctions. This study documents a high incidence of neoplasia in a zoo-managed assurance population of Puerto Rican crested toads (Peltophryne lemur; PRCTs). Over 5 years, neoplasia was diagnosed in 17/49 (35%) submitted adult PRCTs and was the cause of death or euthanasia in 13/17 (72%). Most toads were male (16/17; 94%) and 6 to 11-years-old (average 8.1 years). Notably, seven toads (41%) had multiple neoplasms. Of the 29 neoplasms identified, 17 (59%) were cutaneous or subcutaneous. The most common neoplasms included mast cell tumors (MCTs; 8/29; 28%), histiocytic sarcomas (6/29; 21%), lymphoma/leukemia (4/29; 14%), and squamous cell carcinomas (3/29; 10%). Distant metastases were documented in 6/8 (75%) toads with MCTs. Causes for neoplasia in this population were not determined though may include genetic or environmental factors. Continued investigations of managed endangered amphibians will help elucidate mechanisms of carcinogenesis.

METASTATIC MINERALIZATION IN A ZOOLOGIC COLLECTION OF SPOT-TAILED EARLESS LIZARDS (HOLBROOKIASPP.).

Vitamin and mineral supplements are commonly used in diets for zoologic and companion animals. Because specific nutrient requirements are often unknown, informed decisions are based on literature for related species. Over 18 mon beginning in November 2017, an entire population of spot-tailed earless lizards (Holbrookia lacerata and Holbrookia subcaudalis) died (N = 33). All but two lizards were submitted for histopathology (94%). All examined cases had mineralization in at least one tissue; 71% (22 of 31) had multisystemic mineral deposits consistent with metastatic mineralization. No underlying causes were detected histologically. The supplement used for dusting the food items fed five to six times per week was inadvertently switched for 2 to 4 mon, and the incorrect supplement was found to contain fourfold the intended vitamin D3 concentration. Thus, hypervitaminosis D was considered the most likely cause. Interestingly, eastern collared lizards (Crotaphytus collaris), also fed prey supplemented five to six times a week, and over 50 other insectivorous reptile and amphibian species possibly receiving the supplement one to seven times a week did not appear affected. During this time, only two other cases of metastatic mineralization were diagnosed in other herpetofauna at this institution. Prior to receiving the incorrect supplement, there were no cases of metastatic mineralization detected in the earless lizard population. These cases highlight species-specific sensitivities, and the deleterious effects of excessive or inappropriate supplementation. It is important to confirm product identification on arrival, regularly conduct chemical analysis of supplements, and educate keepers and owners about adverse effects of inappropriate supplementation.

ASSESSMENT OF CAPILLARY ZONE ELECTROPHORESIS AND SERUM AMYLOID A QUANTITATION IN CLINICALLY NORMAL AND ABNORMAL SOUTHERN WHITE RHINOCEROS (CERATOTHERIUM SIMUM SIMUM) AND SOUTHERN BLACK RHINOCEROS (DICEROS BICORNIS MINOR).

Capillary zone electrophoresis (CZE) and an immunoassay for serum amyloid A (SAA) were used to examine serum samples from clinically normal and abnormal southern white rhinoceros (Ceratotherium simum simum) and southern black rhinoceros (Diceros bicornis minor) under managed care. CZE resolved seven fractions as well as subfractions for α1 globulins. Reference intervals were calculated for white rhinoceros (n = 33) and found to have some differences over previously reported intervals generated using agarose gel electrophoresis (AGE) methods in sera from free-ranging animals. In addition, the coefficient of variation related to fraction quantitation was found to be overlapping or superior to that reported for AGE. No significant differences were observed in CZE measurands and total protein between clinically normal and abnormal rhinoceros. In contrast to CZE, significant differences in SAA levels (P < 0.001) were observed in samples from the white rhinoceros between clinically normal and abnormal animals. In addition, in limited sample sets with repeated measures, SAA provided prognostic value. Future studies should generate more robust reference intervals and delineate the application of both SAA quantitation and CZE in routine health assessments and in prognostication.

CHRONIC PANCREATIC DISEASE IN THE LESSER KUDU (TRAGELAPHUS IMBERBIS): A REPORT OF 16 CASES IN THE UNITED STATES.

A review of archival cases at Northwest ZooPath from 1995-2018 identified 16/96 (17%; eight females, eight males) lesser kudus (Tragelaphus imberbis) with chronic pancreatic disease (CPD) from three institutions, all of which originated from an initial founder stock of 12 animals. Ages at time of death or euthanasia ranged from 2 to 132 mo (average age = 69 mo). All cases had varying degrees of pancreatic acinar cell depletion, ductular hyperplasia, and fibrosis. Serum fructosamine, glucose, insulin, and insulin to glucose ratio collected close to time of death from 13 affected animals were not significantly different from controls (n = 19). Of these analytes, receiver operating characteristic analysis identified fructosamine as the best-performing analyte with an area under the curve 0.671 (95% confidence interval = 0.480-0.828; sensitivity = 58.3%, specificity = 84.2%; cutoff point >352 µmol/L) in the diagnosis of CPD. With a 15% prevalence, there was a positive predictive value of 41% and a negative predictive value of 92%, indicating that the probability of false positives is high, but of false negatives is low. An etiologic agent was not identified by histology (n = 16), transmission electron microscopy (n = 1), or enterovirus PCR (n = 2). Serum zinc and hepatic heavy metal analyses were judged to be within normal limits. Chronic pancreatic disease is considered an important cause of morbidity and mortality in the lesser kudu; serum chemistry analysis warrants further investigation in its use for diagnosis. The etiopathogenesis is not understood, but the absence of obvious causes, the occurrence also in very young animals, and the inbred lineage of lesser kudus in the United States suggest a genetic basis for this disease.

HEMATOLOGY, PLASMA BIOCHEMISTRY, AND PLASMA PROTEIN ELECTROPHORESIS REFERENCE INTERVALS FOR BLUE IGUANAS (CYCLURA LEWISI) FROM GRAND CAYMAN ISLAND.

The blue iguana (Cyclura lewisi) is an endangered rock iguana species native to Grand Cayman, in the Cayman Islands. Health assessments were conducted on captive and free-roaming iguanas in 2001 and 2003-2014 and were performed in the summer wet season (June-July) of 2003-2004 and 2010-2014 and in the winter dry season (November-December) of 2001 and 2005-2009. Morphometric data were recorded from iguanas when blood samples were collected: 903 samples were collected and data from 890 samples from 775 iguanas were included. Samples were analyzed for hematology, plasma biochemistry, protein electrophoresis, mineral panels, 25-hydroxyvitamin D, and testosterone. Reference intervals were created for captive subadults, captive adults, and free-roaming adults when data were sufficient. Significant differences among these groups were described, as were differences on the basis of sex, season, and origin (captive vs free-roaming). In captive iguanas, most analytes were significantly different between subadults and adults, mature heterophils and copper were significantly higher in the dry season, zinc levels were significantly higher in the wet season, and cholesterol and triglycerides were significantly higher in adult females than adult males. Testosterone in adult males was significantly higher in the dry season. These results will aid in future health assessments and disease investigations in wild and captive populations of blue iguanas and are of comparative value for other Cyclura species that are free-roaming, captive, and, especially, in similar conservation release programs.

Novel Paranannizziopsis species in a Wagler's viper (Tropidolaemus wagleri), tentacled snakes (Erpeton tentaculatum), and a rhinoceros snake (Rhynchophis boulengeri) in a zoological collection.

We report several cases of fungal infections in snakes associated with a new species within the genus Paranannizziopsis. Three juvenile Wagler's vipers (Tropidolaemus wagleri) presented with skin abnormalities or ulcerative dermatitis, and two snakes died. Histologic examination of skin from the living viper revealed hyperplastic, hyperkeratotic, and crusting epidermitis with intralesional fungal elements. The terrestrial Wagler's vipers were housed in a room with fully aquatic tentacled snakes (Erpeton tentaculatum), among which there had been a history of intermittent skin lesions. Approximately 2 months after the biopsy of the viper, a skin sample was collected from one tentacled snake (TS1) with skin abnormalities and revealed a fungal infection with a similar histologic appearance. Fungal isolates were obtained via culture from the Wagler's viper and TS1 and revealed a novel species, Paranannizziopsis tardicrescens, based on phenotypic characterization and molecular analysis. P. tardicrescens was cultured and identified by DNA sequence analysis 8 months later from a dead tentacled snake in an exhibit in an adjacent hallway and 13 months later from a living rhinoceros snake (Rhynchophis boulengeri) with two focal skin lesions. Antifungal susceptibility testing on three of four cultured isolates demonstrated potent in vitro activity for terbinafine and voriconazole.

HEALTH SURVEY OF FREE-RANGING RACCOONS (PROCYON LOTOR) IN CENTRAL PARK, NEW YORK, NEW YORK, USA: IMPLICATIONS FOR HUMAN AND DOMESTIC ANIMAL HEALTH.

We conducted health assessments on 113 free-ranging raccoons ( Procyon lotor ) in Central Park, New York City, US, in February 2010, September 2010, and November 2011 in conjunction with a trap-vaccinate-release program to control a raccoon rabies epizootic. Five individuals were sampled at two time points for 118 raccoon examinations in total. We tested 13 of 13 and 8 of 13 euthanized raccoons for rabies and canine distemper virus (CDV), respectively, by antigen testing on brain tissue; all were negative for both viruses. Endoparasitism was the most common necropsy finding, with definitive identification of Baylisascaris procyonis in six of eight (75%) necropsied raccoons. Multiple intestinal parasites were detected in feces of living raccoons, including ascarid-type ova in 25 of 80 (31%) raccoons, with B. procyonis confirmed in one sample. Median blood lead level was 7.3 µg/dL (n=104). Rabies virus neutralizing antibody titer was ≥0.5 IU/mL in 9 of 88 (10%) raccoons naive to rabies vaccination and in 13 of 20 (65%) previously vaccinated raccoons. The majority of raccoons we tested were seropositive for canine parvovirus-2 (54/59, 92%) and Toxoplasma gondii (39/60, 65%). Fewer were seropositive for Rickettsia rickettsii (3/30, 10%). None were seropositive for CDV (n=108), canine adenovirus-1 (n=60), or Borrelia burgdorferi (n=30). Ectoparasites found during 16 of 118 (13.6%) physical examinations included Ixodes texanus ticks (15/118, 12.7%) and Trichodectes octomaculatus lice (1/118, 0.8%). We detected Campylobacter jejuni in 5 of 79 (6%) fecal samples. We detected 11 Salmonella enterica serotypes in 70 of 111 (63.1%) enteric cultures, the most common of which were Salmonella Newport (20/70, 29%) and Salmonella Oranienburg (20/70, 29%). These results indicate that raccoons in Central Park likely are involved in the environmental occurrence and potential disease transmission of a variety of infectious and noninfectious diseases of concern for human, wildlife, and domestic animal health.

RETROSPECTIVE INVESTIGATION OF CATARACT MANAGEMENT IN AVIAN SPECIES IN A ZOOLOGIC COLLECTION.

A review of avian cataracts at the Wildlife Conservation Society's Bronx Zoo between 1992 and 2011 was conducted. Ninety cataracts in 54 birds from 42 species were identified. Cataracts were found primarily during examination for ocular abnormalities (29/54, 53.7%) or opportunistically (13/54, 24.1%) and were most commonly diagnosed as mature (22/90, 24.4%). Systemic medical conditions diagnosed in these birds included West Nile virus (4/54, 7.4%), head trauma (3/54, 5.6%), plumbism and Salmonella Pullorum (1/54, 1.9%), Marek's disease (1/54, 1.9%), leukocytosis (1/54, 1.9%), and hyperglycemia (1/54, 1.9%). Cataracts were progressive in seven birds of four species. Unilateral enucleation was performed in 2/54 (3.7%) birds, and 12/54 (22.2%) underwent cataract removal (phacoemulsification in 16 eyes and standard extracapsular cataract extraction in 2 eyes). Concurrent ocular abnormalities, such as corneal scarring and lens-induced uveitis, were seen in 2/18 (11.1%) eyes preoperatively in the group undergoing cataract removal, 2/2 (100%) eyes preoperatively in the group undergoing enucleation, and 33/70 (47.1%) of eyes that did not undergo surgery. For birds undergoing cataract removal, complications included successfully treated cardiorespiratory arrest intraoperatively (1/12, 8.3%) as well as postanesthetic complications of acute respiratory distress and tracheal stricture (2/12, 16.7%). The most common postoperative ocular abnormalities included posterior capsular opacity (4/18 eyes, 22.2%) and corneal scarring (2/18 eyes, 11.1%). Lens cortical regrowth and marked posterior lens capsular opacity occurred in one eye of one bird after phacoemulsification, necessitating a second ocular surgery. A successful outcome, as determined by improved postoperative visual acuity, was seen in 10/12 (83.3%) birds undergoing cataract removal, and 5/12 (41.7%) of these birds were alive >3 yr after surgery. The results of this review will aid clinicians in identifying common stages of cataracts, determining eligibility for cataract surgery, and managing postoperative complications in avian patients.

Comparison of two injectable immobilization combinations in Himalayan tahr (Hemitragus jemlahicus): medetomidine, ketamine, and atipamezole versus carfentanil, xylazine, naltrexone, and atipamezole.

A crossover study was performed in 12 Himalayan tahr (Hemitragus jemlahicus) undergoing preshipment examinations to compare a partially reversible to a fully reversible intramuscular chemical immobilization protocol. The partially reversible protocol (MKA) consisted of induction with medetomidine (0.06 +/- 0.009 mg/kg) and ketamine (2.03 +/- 0.315 mg/kg) and antagonism by atipamezole (0.30 +/- 0.044 mg/kg). The fully reversible protocol (CXNA) consisted of induction with carfentanil (0.009 +/- 0.003 mg/kg) and xylazine (0.08 +/- 0.019 mg/kg) and antagonism by naltrexone (0.867 +/- 0.332 mg/kg) and atipamezole (0.105 +/- 0.023 mg/ kg). Animals were monitored for quality and length of induction and recovery, depth of immobilization, heart rate, respiratory rate, rectal temperature, indirect mean blood pressure (MBP), oxygen saturation, and end-tidal carbon dioxide concentration. Blood was collected for serum cortisol measurement. Significant (P < 0.05) differences included a higher MBP, higher serum cortisol, and longer recovery time with MKA compared to CXNA. In addition, the quality of induction and recovery were different. With MKA, animals ambulated less during induction, remained recumbent longer during recovery, and demonstrated more ataxia on rising. Despite differences between the protocols, both provided an acceptable level of immobilization for pre-shipment testing to be done and appeared to be safe in the study population. These differences should be taken into consideration when selecting the anesthetic protocol because either regimen may be more or less desirable for different medical or immobilization settings.

Early life stress and novelty seeking behavior in adolescent monkeys.

Recent evidence suggests that early exposure to mild stress promotes the development of novelty seeking behavior. Here we test this hypothesis in squirrel monkeys and investigate whether novelty seeking behavior is associated with differences in cerebrospinal fluid (CSF) levels of the serotonin metabolite 5-hydroxyindoleacetic acid (5HIAA), the dopamine metabolite homovanillic acid (HVA), the norepinephrine metabolite 3-methoxy-4-hydroxyphenylethylene glycol (MHPG), and the neuropeptide corticotrophin-releasing factor (CRF). Monkeys were randomized early in life to either mild intermittent stress (IS) or no stress (NS) conditions, and subsequently presented with opportunities to interact with a familiar or novel object in a test box that was connected to each monkey's home cage. To further minimize the potentially stressful nature of the test situation, monkeys were acclimated to the test procedures prior to study initiation. Post-test plasma levels of cortisol in IS and NS monkeys did not differ significantly from baseline levels measured in undisturbed conditions. During testing, more IS than NS monkeys voluntarily left the home cage, and IS monkeys spent more time in the test box compared to NS monkeys. More IS than NS monkeys engaged in object exploration in the test box, and IS monkeys preferred to interact with the novel vs. familiar object. Novelty seeking was not associated with differences in 5HIAA, HVA, MHPG, or CRF, but correlated with differences in object exploration observed in a different test situation at an earlier age. These trait-like differences in novelty seeking appear to reflect mild early stress-induced adaptations that enhance curiosity and resilience.