Knowledge about the normal metabolism and involvement of vitamin D in elephant calcium homeostasis is essential to understanding the possible role of vitamin D in Asian elephant (Elephas maximus) health, as well as to informing accurate diet formulation. This study provides an evaluation of analytes involved in vitamin D metabolism, in conjunction with dietary intake and ultraviolet light (UV) exposure, in Asian elephants managed in a northern temperate climate. Once monthly, for a total of 12 mo, serum from six adult Asian elephants was analyzed for 25-hydroxyvitamin D [25(OH)D], 24,25-dihydroxyvitamin D [24,25(OH)₂D], 1,25-dihydroxyvitamin D [1,25(OH)₂D], parathyroid hormone (PTH), total calcium (Ca), ionized calcium (iCa), phosphorus (P), and magnesium (Mg). The diet was analyzed monthly for vitamin D, Ca, and P. Monthly average vitamin D–weighted UV daily sums were determined to gauge average UV light exposure within the vitamin D action spectrum. No serum or diet parameters were affected by time or season. Average serum 25(OH)D₂ was 7.02 ± 0.85 ng/ml. 25(OH)D₃ levels were nondetectable in all samples despite supplementation of the diet with recommended levels of vitamin D₃, and UV exposure was at sufficient levels for cutaneous vitamin D synthesis for 6 mo of the year. Levels of 24,25(OH)₂D averaged 31.7% higher than 25(OH)D, and average 1,25(OH)₂D₂ was 11.24 ± 1.04 pg/ml. Values for PTH, Ca, iCa, P, and Mg were within expected ranges for Asian elephants. The information gained from this research expands the knowledge base for these analytes, evaluates 24,25-dihydroxyvitamin D for the first time, and provides new information regarding vitamin D metabolism and test interpretation in the Asian elephant.

Perturbations in serum prolactin secretion, both over- and underproduction, are observed in zoo African elephants (Loxodonta africana) that exhibit abnormal ovarian cycles. Similar prolactin problems are associated with infertility in other species. Pituitary prolactin is held under constant inhibition by a hypothalamic-derived neurotransmitter, dopamine; thus, regulation by exogenous treatment with agonists or antagonists may be capable of reinitiating normal ovarian cycles. This study tested the efficacy of oral administration of cabergoline (agonist) and domperidone (antagonist) as possible treatments for hyperprolactinemia or chronic low prolactin, respectively. Hyperprolactinemic (overall mean prolactin, >30 ng/ml), acyclic elephants were administered oral cabergoline (2 mg, n = 4) or placebo (dextrose capsule, n = 4) twice weekly. Overall mean prolactin concentration decreased in treated females compared with controls (32.22 ± 14.75 vs 77.53 ± 0.96 ng/ml; P = 0.01). Interestingly, overall mean progestagen concentrations also increased slightly (P < 0.05) in treated females (0.15 ± 0.01 ng/ml) compared with controls (0.07 ± 0.01 ng/ml), but no reinitation of normal cyclic patterns was observed. Chronic low prolactin (overall mean prolactin, <10 ng/ml), acyclic females were orally administered domperidone (2 g/day, n = 4) or placebo (dextrose capsule, n = 4) for 4 wk, followed by 8 wk of no treatment (four cycles) to simulate the prolactin pattern observed in normal cycling elephants. Overall mean prolactin concentrations increased (P = 0.005) during domperidone treatment (21.77 ± 3.69 ng/ml) compared with controls (5.77 ± 0.46 ng/ml), but progestagen concentrations were unaltered. Prolactin regulation by dopamine was confirmed by expected responses to dopamine agonist and antagonist treatment. Although prolactin concentrations were successfully reduced by cabergoline, and domperidone initiated the expected cyclic prolactin pattern, neither treatment induced normal ovarian activity.

The reference intervals of health parameters are valuable tools for veterinarians and conservationists to monitor the health status and viability of endangered species. Natural variation in the health of the long-lived Asian elephant (Elephas maximus) is poorly understood, particularly in relation to differences between males and females. Longitudinal health data were collected from clinical examination, hematology, and serum chemistry analyses over 3 yr from 227 healthy individually marked Asian elephants varying in age and sex. The study population was semicaptive and used in Myanmar's timber industry, but maintained natural feeding and breeding behavior. Body condition score (BCS) and blood pressure were investigated in clinical examinations. Hematological parameters included hematocrit, hemoglobin, total white blood cell count, and differential blood cell counts. Serum chemistry parameters included blood urea nitrogen, creatinine, total protein, albumin, globulins, aspartate aminotransferase, alkaline phosphatase, triglycerides, creatine kinase, glucose, calcium, potassium, sodium, and chloride. To the knowledge of the authors, this is the first description of BCS in an elephant population outside of zoos, and of blood pressure in this species using a novel adaptation of the Intelli Wrap Cuff pressure monitor. Several differences between the sexes were observed, with females generally having higher BCS and triglycerides, and males displaying higher alkaline phosphatase and glucose levels. This study provides important clinical tools that can be used to assess the health status and improve management in this endangered species.

Pallas' cat [Otocolobus (Felis) manul] experiences a high mortality rate from toxoplasmosis. During the period 2006–2016, the overall mortality rate for this species from all causes during the first year of life was 71.59% in European Association of Zoos and Aquaria institutions, with the most significant infectious cause from systemic toxoplasmosis (20.6%) as confirmed by postmortem examination and histopathology. Clindamycin was used starting in 2014 in two collections that had previously experienced 100% mortality rates by toxoplasmosis in kittens less than one year of age, covering key Toxoplasma gondii exposure periods for kittens (n = 17) as a prophylactic measure. This protocol resulted in a 67.03% (95% confidence interval 41.76–78.61%) reduction in the first year mortality rate over a two-year period to 5.88% in those animals treated.

Enrofloxacin is a fluoroquinolone widely used in animals including fish. Intramuscular (IM) injection of enrofloxacin is a feasible and efficacious option for drug delivery. In many species IM injection has been associated with injection site reactions and increases in serum muscle enzymes. Injection site reactions have not been well characterized in fish. Three groups of striped bass (Morone saxatilis) received an IM injection of enrofloxacin 2.27% in the right epaxial musculature 24, 48, or 96 hr prior to evaluation. Mean dose was 7.69 mg/ kg (6.14–9.69 mg/kg). The 24- and 48-hr groups received an injection of equal-volume 0.9% saline in the left epaxial musculature. A corresponding noninjected tissue sample was designated in the left epaxial musculature from each fish of the 96-hr group. Fish were euthanized and injection sites and noninjection control sites were evaluated grossly and histologically. Grades 1–4 were assigned to samples, with grade 1 corresponding to normal tissue and grades 2, 3, and 4 corresponding to mild, moderate, and severe inflammation and/or necrosis respectively. Externally, all control and injection sites appeared visually unremarkable. On cut surface, epaxial muscle of the enrofloxacin-injected tissue appeared moderately to severely hemorrhagic compared to saline and noninjected tissue, which was normal or mildly hemorrhagic. Histologically, eight of eight noninjected tissues were grade 1. For saline-injected tissues, 14 of 16 tissues were grade 2 and 2 samples were grade 3 when 24- and 48-hr groups were combined. For enrofloxacin-injected tissues, 8 of the 8 24-hr samples were grade 3 and 16 of the 16 48- and 96-hr samples were grade 4. These data show that IM injection of enrofloxacin 2.27% is associated with severe hemorrhage, necrosis, and inflammation in striped bass, and may negatively affect animal welfare.

Mavacoxib is a selective cyclooxygenase-2 nonsteroidal anti-inflammatory drug that has been used for management of osteoarthritis and other inflammatory conditions in dogs. The main advantage of mavacoxib over other nonsteroidal anti-inflammatory drugs is its longer plasma half-life, leading to decreased dosing frequency. This study determined the pharmacokinetics of mavacoxib in Caribbean flamingos (Phoenicopterus ruber ruber) after a single-dose oral administration of 6 mg/kg (n = 6). Plasma mavacoxib concentrations were determined using liquid chromatography with mass spectrometry, and pharmacokinetic analysis was performed using noncompartmental methods. Mean peak plasma concentration (C_max) was (mean; range) 2.97 (2.19––4.06) µg/ml; mean time to peak plasma concentration (T_max) was 18.68 (4.00–48.00) hr; mean area under the curve (AUC) was 455 (292–637) hr * µg/ml; and mean terminal half-life (T_1/2) was 74.47 (49.57–161.43) hr. Based on the results of this study, mavacoxib dosed at 6 mg/kg orally in Caribbean flamingos reaches plasma concentrations above the therapeutic concentration established for dogs, but further studies are needed to determine appropriate dosing recommendations in flamingos.

In this study, adult intact male and female (n = 10) naked mole-rats (Heterocephalus glaber) were anesthetized using a combination of dexmedetomidine (0.06 mg/kg intramuscularly [IM]), ketamine (20 mg/kg IM), and midazolam (1.0 mg/kg IM). Atipamezole (1.0 mg/kg IM) and flumazenil (0.1 mg/kg IM) were administered 40 min after induction. Induction and recovery times were monitored and recorded. Vital parameters, including heart rate, respiratory rate, and SpO₂, and reflexes were monitored every 5 min during the anesthetic period. Anesthetic induction was smooth and rapid. All monitored reflexes were lost within a median time of 60 sec (interquartile range, 15 sec). Heart rate and respiratory rate were significantly decreased from baseline, whereas there was no difference in SpO₂ over the anesthetic period. The mean time to recovery was 15 ± 7 min (mean ± SD). One animal was found dead 40 min after apparent recovery, which is suspected to be an anesthetic-related death. Based on these findings, dexmedetomidine-ketamine-midazolam anesthesia is an effective anesthetic protocol in naked mole rats that provides a consistent anesthetic plane but should be used with caution in animals with underlying conditions.

A review of anesthetic procedures used in Andean bears (Tremarctos ornatus), sloth bears (Melursus ursinus), and giant pandas (Ailuropoda melanoleuca) housed at the Smithsonian Institution's National Zoological Park (NZP) from 1995 to 2016 was performed. A total of 146 anesthetic procedures (55 procedures on 12 Andean bears, 38 procedures on nine sloth bears, and 53 procedures on five giant pandas) occurred at NZP during this time frame. Induction protocols involved some combination of ketamine (K; n = 121), tiletamine–zolazepam (TZ; n = 91), medetomidine (M; n = 67), xylazine (X; n = 42), midazolam (Mid; n = 9), and butorphanol (B; n = 1). The most commonly used protocols were TZKM for both Andean bears (n = 29) and sloth bears (n = 12), and KX in giant pandas (n = 17). Supplemental injections of K, TZ, Mid, or M were required to complete inductions in 66 cases (45%). Anesthetic maintenance was most often completed with isoflurane, ketamine, or propofol. The most commonly reported complications included perianesthetic emesis (n = 47), ptyalism (n = 16), and seizure-like activity (n = 11). The most frequent physiologic findings included low pulse oximetry values (n = 95), bradycardia (n = 95), hypothermia (n = 74), and hypertension (n = 55). Dose ranges, induction, and recovery times, supplementation and complication rates, and physiologic values are reported for each protocol by species.

Twenty-one free-ranging warthogs (Phacochoerus africanus) in the Kruger National Park, South Africa, were immobilized with a combination of medetomidine (0.07 ± 0.01 mg/kg), butorphanol (0.26 ± 0.04 mg/kg), tiletamine–zolazepam (0.69 ± 0.15 mg/kg), and ketamine (1.43 ± 0.21 mg/kg) administered intramuscularly by dart. Induction, immobilization, and recovery characteristics were evaluated using a standardized scoring system. In the immobilized warthogs, physiological variables were measured every 5 min and arterial blood gases were analyzed at 15-min intervals. At 45 min after initial drug administration, atipamezole (0.34 ± 0.050 mg/kg) and naltrexone (0.53 ± 0.079 mg/kg) were administered intravenously. Overall, induction quality after darting was scored as excellent and the mean time to safe handling was 5.9 ± 2.0 min. Based on muscle relaxation, and loss of palpebral and pedal reflexes, most subjects (17 out of 21) reached a plane of surgical anesthesia by 10 and 15 min; 20 out of 21 warthogs were in this plane for the duration of the monitoring period. In the immobilized warthogs the overall mean heart rate was 65 ± 15.3 beats per minute, mean respiratory rate was 14.7 ± 5.6 breaths per minute, and the mean rectal temperature was 37.9 ± 1.4°C during the 40 min. Arterial blood gas results showed hypoxemia (mean PaO₂ 62.1 ± 16.2 mmHg), hypercapnia (mean PaCO₂ 47.1 ± 5.1 mmHg), and acidemia (mean pH = 7.36 ± 0.04). Values for PaO₂ and pH improved over the immobilization period. After antagonist administration, overall recovery quality from immobilization was scored as good, with animals standing at a mean time of 7.3 ± 4.9 min. The drug combination proved to be effective in the immobilization of free-ranging warthogs with rapid induction, good anesthesia, and limited cardiorespiratory changes. This anesthetic protocol produces effective, safe, and partially reversible immobilization in warthogs.

A prospective clinical trial was performed to evaluate the efficacy of haloperidol premedication prior to xylazine–ketamine anesthesia with a goal of reducing capture stress in adult male captive spotted deer (Axis axis). On the morning of the study, deer were fed a banana either containing haloperidol tablets (1 mg/kg) (haloperidol group, n = 10) or without haloperidol (placebo group, n = 10). Six hours postadministration, xylazine (3 mg/kg) and ketamine (2 mg/kg) was administered intramuscularly via a dart. Rectal temperature, heart rate, respiratory rate, and SpO₂ (percent hemoglobin saturation) were recorded at 5-min intervals. Blood gas analysis was performed at time 0 (venous blood) and 10 and 20 min (arterial blood) postinduction. Serum cortisol was determined from venous blood (35 min postinduction), following which yohimbine was administered at a dose of 0.15 mg/kg intramuscular and 0.15 mg/kg intravenous. Statistical analysis of repeated measures data was performed with a two-way analysis of variance. Paired data were analyzed with a Wilcoxon rank-sum test (categorical data) or a paired t-test (continuous data). Significance was set at P ≤ 0.05, and results were expressed as mean ± SEM. There was no significant difference in induction time or recovery time between treatment groups. Rectal temperature and heart rate were significantly lower in the haloperidol group. Both groups demonstrated acidosis with venous pH being significantly lower in the placebo group when compared to the haloperidol group. Serum cortisol and arterial plasma lactate were lower in the haloperidol group indicative of reduced stress and physical exertion. Haloperidol premedication proved to be beneficial in reducing capture stress, when administered prior to xylazine–ketamine anesthesia, in spotted deer.

Alfaxalone is a neurosteroid anesthetic agent that has been extensively used in both human and veterinary medicine for more than 50 yr. Previous studies involving avian species demonstrated various dose ranges and multiple routes of administration. The aim of this study was to evaluate the short-term sedative, cardiorespiratory, and thermoregulatory effects of an intramuscular injection of alfaxalone on budgerigars (Melopsittacus undulatus). A crossover study was performed with a sample size of 10 male budgerigars, previously determined to be healthy based on physical examination. Alfaxalone was administered intramuscularly at two doses: 15 and 20 mg/kg. The lower dose resulted in mild to moderate sedation for 29 ± 5 min, whereas the higher dose resulted in moderate to profound sedation for 29 ± 7 min. A statistically significant decrease in heart rate was observed 2 min after administration of alfaxalone at 15 mg/kg; however, this finding was noted to be transient. A statistically significant decrease in respiratory rate was observed at 6 and 10 min after injection in both groups. Cloacal temperature measurement with a digital thermometer and eye temperature calculated from thermographic images demonstrated a decrease in body temperature over time but was not found to be statistically significant. Intramuscular use of alfaxalone proved to provide short-term sedation in budgerigars, with statistically significant but clinically mild cardiorespiratory effects. Due to a significant decrease in body temperature, active warming is recommended when using alfaxalone in budgerigars.

Anesthesia is commonly employed in aquatic medicine to facilitate physical exams, diagnostics, and surgical interventions. Tricaine methanesulfonate (MS-222) is the most commonly used anesthetic for fish and is currently the only anesthetic approved by the US Food and Drug Administration Center for Veterinary Medicine for food-producing fish. Despite the frequency of anesthetic procedures in fish, anesthetic monitoring remains rudimentary in many facilities. This study evaluated the impact on blood gases, acid-base balance, and electrolytes in koi (Cyprinus carpio) anesthetized at concentrations of 100 mg/L and 150 mg/L MS-222. Blood samples from 25 fish per treatment were collected at 5 and 20 min of anesthetic immersion. Forty-nine of 50 fish recovered uneventfully from anesthesia; one fish did not recover and was euthanatized. Results showed significant increases in partial pressure of carbon dioxide (pCO₂) (P = 0.006) and hyperglycemia (P = <0.0001) with increasing anesthetic concentration and time under anesthesia and a significant decrease in partial pressure of oxygen (pO₂) with increased anesthetic time (P = 0.021). There were several electrolyte changes observed with both increasing anesthetic time and concentration. All electrolytes except potassium remained within published reference ranges for koi, while potassium showed a significant decrease in concentration associated with anesthetic time and concentration. The results of this study indicate that MS-222 at 100 mg/L and 150 mg/L represent safe anesthetic concentrations for koi undergoing minimally invasive diagnostics; however, koi anesthetized with MS-222 at a concentration of 150 mg/L experienced more significant changes in blood gases, acid-base balance, and electrolyte concentrations.

This study assessed the in vitro temporal changes that occur in blood pH and lactate concentrations for an elasmobranch species and a chelonian species, as well as blood gases (partial pressures of carbon dioxide [pCO₂] and oxygen [pO₂]) for a chelonian species, with a portable clinical point-of-care analyzer. Blood samples were collected from 10 cownose rays (Rhinoptera bonasus) and 10 red-eared sliders (Pseudemys scripta elegans), stored on ice, and serially analyzed at six time points up to 90 min postcollection. Results indicate that analysis should be conducted as soon as possible after blood collection for these species, with immediate analysis being preferred. However, if analysis must be delayed, syringes may be capped, placed on ice, and analyzed at a later time. Analysis within 90 min provided clinically acceptable results for pH and lactate in both species and for pCO₂ in red-eared sliders, whereas substantial artifactual increases of pO₂ were seen in red-eared sliders.

The processing of blood samples can be delayed during health assessments of wildlife populations in distant locations. The use of whole blood preservatives may be useful in these situations. However, there is scant information regarding their use in nonmammalian species. This study tested the efficacy of two cell preservatives on whole blood collected from 12 Attwater's prairie chickens (Tympanuchus cupido attwateri). The preservatives used were Streck Cell Preservative© (SCP), a proprietary proteinaceous stabilizer developed for human flow cytology and validated in other mammalian species, and formalin, which is commonplace in histopathology, but its use in whole blood has been limited to fish. Grouped blood samples were treated with heparin, SCP, or formalin and analyzed at 0, 1, 4, and 7 days after collection for packed cell volume (PCV), complete blood count (CBC), and cellular morphology. SCP effectively preserved most cell types in Attwater's prairie chicken blood samples over a period of 7 days, with the exception of monocyte cell counts, which were significantly reduced from day 0. Formalin maintained total white blood cell counts at baseline levels measured by hemocytometer, but irregular staining characteristics prevented accurate analysis of differential counts or cellular morphology. Both preservatives altered PCV compared with the heparin control, but these values remained constant over time, highlighting the need for method-specific reference intervals. The validation of SCP in Attwater's prairie chickens supports its potential for use in other avian species for the collection of accurate hematologic data when the processing of blood samples may be delayed.

Agarose gel electrophoresis (AGE) has been widely implemented throughout veterinary medicine and for analysis of plasma proteins of avian and reptile species. Capillary zone electrophoresis (CZE) is becoming a standard method in human clinical pathology laboratories but has not widely been used for the analysis of animal samples. The objective of the present study was to compare protein fractions derived from AGE and CZE methods using plasma from the green turtle (Chelonia mydas). Plasma samples were analyzed by AGE and CZE per manufacturer guidelines. The methods were assessed by CV analysis, Spearman's correlation, Passing-Bablok regression, and Bland Altman plots. CZE consistently resolved more fractions than AGE with three fractions observed in the prealbumin migrating region versus one for AGE and two fractions in the γ globulin region versus one for AGE. Compared with AGE, CZE showed a lower CV in intra-assay tests (1.0–4.9% vs 2.0–28.3%) and a lower or overlapping CV in interassay tests (1.0–10.6 vs 2.3–22.0). The prealbumin, α2 globulin, and β globulin fractions correlated the least between the methods (for all three fractions: r_s ≤ 0.28, P > 0.21). Moderate, significant correlations between AGE and CZE methods were observed for albumin (r_s = 0.78, P < 0.0001) and γ globulins (r_s = 0.78, P < 0.0001). CZE has a higher precision and ease of use over AGE and offers the opportunity to resolve additional protein fractions. This will necessitate the development of new conventions in placement of fraction delimits, definition of species-specific reference intervals, and evaluation of clinical utility in abnormal turtles.

There are limited reports of the genetic characterization of Toxoplasma gondii infecting captive macropods in North America. A novel genotype, ToxoDB PCR-RFLP genotype 263, was reported from six wallabies at a zoological facility in Virginia, USA, prompting an investigation into the genotypes from T. gondii strains infecting macropods at a zoological park in Florida, USA. Cardiac muscle and/or lung samples from an agile wallaby (Macropus agilis, n = 1), red kangaroos (Macropus rufus, n = 8), red-necked wallaby (Macropus rufogriseus, n = 1), and a tammar wallaby (Macropus eugenii, n = 1) that died between 2014 and 2018 were collected. All 11 cases were confirmed to have died from systemic toxoplasmosis by histopathology and immunohistochemical staining. Multilocus PCR-RFLP genotyping of T. gondii was performed directly on tissue samples or on parasites isolated from myocardium by mouse bioassay. Two cases of toxoplasmosis were identified as the reported novel genotype, ToxoDB PCR-RFLP genotype 263, but no common source of exposure could be identified. Five cases were identified as genotype 2 (type III strain, haplogroup 3), and four cases were identified as genotype 216, which has been previously reported in North American wildlife. There were no overt differences in lesion severity or distribution related to genotype. These results suggest that the premise was contaminated with at least three genotypes of T. gondii causing systemic toxoplasmosis in macropods. The largest cluster of fatal toxoplasmosis in macropods in the study period occurred following severe rainfall flooding of the exhibit, suggesting the transmission of T. gondii by water and pointing out the importance of this transmission mechanism. In summary, our study revealed three T. gondii outbreaks that caused significant loss of macropods within 5 yr in a zoological facility in Florida. More studies are needed to understand transmission and prevention of toxoplasmosis in sensitive zoo animals.

Vector-borne Plasmodium spp. infect a wide range of bird species. Although infections may be asymptomatic, certain genera, especially those that evolved in regions without endemic malaria, appear particularly susceptible to symptomatic disease, leading to morbidity and mortality. High mortalities associated with malaria infections have been documented in captive species of Sphenisciformes, Somateria, and Larosterna, all genera that evolved in climates with low mosquito exposure. To better characterize trends in Plasmodium-related mortality in a zoological collection in New York, necropsy reports for birds of all three genera that died between 1998 and February 2018 were analyzed; comparisons were made between birds that died with or without evidence of malaria infection. A seasonal peak in deaths was observed in birds regardless of their malaria status. There was no significant difference in the age of birds at death between malaria-positive and malaria-negative animals. These results suggest that age and season of death were not associated with malaria status. To investigate an association between parasite lineage and clinical outcome, polymerase chain reaction was used to identify parasite lineage in necropsied birds as well as healthy birds sampled as part of surveillance studies. Twelve different Plasmodium lineages were identified. The relative prevalence of parasite lineages was compared between necropsy and surveillance samples. A single parasite lineage, SGS1 (species: Plasmodium relictum), was significantly more likely to be found in surveillance samples; it was detected in a plurality of surveillance data but found in only one necropsy case. Other parasite lineages were more likely to be found in necropsies than in surveillance samples, most notably SEIAUR01 (species: Plasmodium cathemerium). These data may be consistent with a difference in virulence between parasite lineages. This investigation has implications for the monitoring and care of vulnerable avian species.

The clouded leopard (Neofelis nebulosa) is classified as vulnerable on the International Union for the Conservation of Nature Red List of Threatened Species. However, diseases affecting this species across zoo populations are not well documented. The primary objective of this retrospective study was to identify common and significant causes of morbidity and mortality in captive-bred clouded leopards from European, Asian, and Australian institutions. Medical records from 44 zoological parks that held 271 clouded leopards from 1934 to 2017 were reviewed. Major causes of mortality in the dead leopards (n = 141) were respiratory disease (17%), maternal neglect and starvation (12%), generalized infectious disease (10%), digestive disease (10%), and trauma (10%). Six animals lived more than 20 yr and two were older than 22 yr. Diseases were recorded 344 times (average of two per leopard) in 166 living leopards. The body systems most frequently affected by disease in these 166 individuals were, in order of frequency, integumentary (prevalence = 21%), digestive (21%), respiratory (16%), musculoskeletal (12%), and urinary (10%) systems. Neoplasia (7%) was less frequent, followed by cardiovascular (5%), genital (3%), and viral (3%) disorders. Extensive, self-induced alopecia on the tail and dorsum was the most frequently reported dermatological disease, which is proposed to be called the “clouded leopard alopecia syndrome.” The most common neoplasm was pheochromocytoma (1%), followed by squamous cell carcinoma of the paw pads, pleural mesothelioma and multicentric lymphomas (<1% each). Dilated cardiomyopathy (2%) was the most common cardiovascular disease. Bronchopneumonia (7%), enteritis (4%), and nephritis (4%) were the most frequently reported respiratory, digestive, and renal diseases, respectively. Diagnosed disease incidence was significantly higher in Europe. This paper reports the results of a comprehensive study of the causes of morbidity and mortality in European, Asian, and Australian clouded leopard zoo populations.

The Mauritian pink pigeon (Nesoenas mayeri) is vulnerable, with only 400 individuals remaining in the free-living population. A European captive population was established in 1977 and a European Endangered Species Program (EEP) in 1992. The EEP long-term management plan recommends integrating the EEP and free-living Mauritius populations through pigeon transfers. A retrospective mortality review of the captive population was performed as part of a disease risk assessment process and to inform infectious disease screening prior to exporting captive birds to Mauritius. Six hundred pink pigeons from 34 institutions died from 1977 to 2018. Each individual was categorized according to age at time of death. Records from 404 individuals were categorized according to cause of death. Neonatal mortality (39%) and juvenile mortality (10.8%) were most commonly caused by noninfectious diseases (52% and 54.4%, respectively), including parental neglect and failure to thrive in neonates and nutritional secondary hyperparathyroidism in juveniles. Trauma (43.1%) was the most common cause of mortality in adults, with significantly higher mortality in males from interspecific aggression and in females due to intraspecific aggression. Yersinia pseudotuberculosis, Mycobacterium avium, and Escherichia coli were the most common infectious causes of adult mortality, and E. coli was the most common infectious cause in neonates. The following infectious diseases were identified as priorities for pre-export disease risk analysis, though not all caused mortality: Y. pseudotuberculosis, M. avium, Trichomonas spp., Chlamydia psittaci, and Coccidia spp. Husbandry changes have been made over the years to mitigate many of the noninfectious causes of mortality. These include alterations to nest sites to reduce neonatal trauma and abandonment, ultraviolet light supplementation and diet optimization to reduce metabolic disorders, improving enclosure design to reduce impact trauma, allowing females rest periods during breeding season, and avoiding housing with certain species.

Yersinia enterocolitica (YE) bioserotype 1B/O:8 (YE 1B/O:8) was identified in routine culture of a variety of zoo species housed at Omaha's Henry Doorly Zoo and Aquarium (OHDZA) from April to July 2011. Animal cases representing 12 species had YE detected from 34 cases during routine fecal monitoring and/or during postmortem examination: Coquerel's sifakas (Propithecus coquereli, two cases), black & white (BW) ruffed lemurs (Varecia variegata variegata, six cases), red ruffed lemurs (Varecia rubra, seven cases), white handed gibbon (Hylobates lar albimana, one case), black lemurs (Eulemur macaco, three cases), mongoose lemurs (Eulemur mongoz, two cases), African hunting dogs (Lycaon pictus, five cases), agile gibbons (Hylobates agilis, three cases), siamangs (Hylobates syndactylus, two cases), colobus monkey (Colobus angolensis palliates, one case), argus pheasant (Argusianus argus, one case), and orangutan (Pongo pygmaeus, one case). Most species were not symptomatic; however, three symptomatic cases in Coquerel's sifakas (two) and a white handed gibbon (one) showed clinical signs of diarrhea and lethargy that resulted in death for the Coquerel's sifakas. One unexpected death also occurred in a BW ruffed lemur. To the authors' knowledge, this is the first report of YE 1B/O:8 in such a large variety of zoo species. The source of the YE could not be identified, prompting the initiation of a diseases surveillance program to prevent further cases for the species that are sensitive to YE. To date, no additional cases have been identified, thus suggesting a single introduction of the YE 1B/O:8 strain into the zoo environment.

This case series includes a single case of disseminated tuberculous disease due to Mycobacterium pinnipedii in a New Zealand fur seal (Arctocephalus forsteri), which was being cared for by a zoo in New Zealand. The remaining five pinnipeds in the colony underwent extensive mycobacterial disease surveillance over the following 4 yr, involving a total of 26 anesthetic procedures and numerous diagnostic tests that included comparative intradermal tuberculin skin tests, mycobacterial antibody serology, respiratory and gastric lavages, and computed tomography (CT) scans. An additional case of chronic sinusitis due to Mycobacterium marinum and Pseudomonas aeruginosa was identified in a California sea lion (Zalophus californianus). Results from CT and the respiratory lavages were the most helpful antemortem diagnostic tests for active mycobacterial disease in this case series. Of the remaining four animals, two were euthanatized and two remain alive, and none of them had evidence of active mycobacterial disease. Further mycobacterial disease surveillance in staff and animals was performed, and no other case was identified. There are no validated mycobacterial surveillance tests available for pinnipeds and so it remains unknown whether the two surviving pinnipeds are truly negative or whether they have latent mycobacterial infection that could develop into active mycobacterial disease in the future. For this reason, increased levels of biosecurity and quarantine remain permanently in place for the pinniped colony.

Two geriatric red pandas (Ailurus fulgens) over a 4-yr period presented with vague clinical signs including anorexia, lethargy, and difficulty ambulating. Treatment protocols using enrofloxacin, steroids, and clindamycin were unsuccessful. Necropsy examination confirmed disseminated toxoplasmosis infection in these cases, and a modified agglutination test had been positive for a prolonged period of time before one panda showed signs of disease. A review of the Red Panda Species Survival Plan pathology database revealed two additional cases of disseminated toxoplasmosis in geriatric red pandas. Many organ systems were affected, but dissemination to the brain, lungs, and liver predominated. Immunohistochemistry or polymerase chain reaction was required to confirm a diagnosis in serologically positive animals, as well as in animals in which a histological diagnosis was suspected. This case series describes the clinical and pathological features of toxoplasmosis in geriatric red pandas.

This article describes the urinogenital condition of three female Iberian ibexes (Capra pyrenaica—one infertile 3-yr-old adult and two prepubertal animals aged 1 (PP1) and 2 (PP2) yr, respectively, all raised in captivity. All showed constant urinal dribbling, leading to ulcerative dermatitis in the vulvar area. Housed in a stable with other females, the adult did not become pregnant after male contact in either of two consecutive mating seasons. Vaginoscopy and laparoscopic exploration performed on the prepubertal females revealed abnormalities of the vagina and urinary bladder. Ultrasound examination revealed atrophy of the left kidney in the adult female and PP1, and of the right kidney in PP2, with degeneration of the renal pelvis. A paraovarian cyst with hydrosalpinx was also detected in the left oviduct of the adult female. Postmortem analysis of the adult and PP2, which shared a mother, confirmed an extramural single ectopic ureter with vaginal insertion associated with atrophy of the ipsilateral kidney. Though PP1 was officially unrelated to the latter animals, all three might have had a common ancestor in their lineages.

Nine cases of amyloidosis in caracals (Caracal caracal) from three different institutions in Europe were reviewed and evaluated histopathologically. The six males and three females died between 2008 and 2018 at an age of 6 yr ± 2.5 mo (median ± interquartile range). In two out of nine (2/9) animals, amyloidosis was an incidental postmortem finding; the animals died of bronchopneumonia and gastric ulceration due to Helicobacter spp., respectively. Seven (7/9) animals suffered from acute renal failure due to amyloidosis, one of them additionally of cardiac decompensation. The predominant clinical signs were weight loss, lethargy, dys- or anorexia, dehydration, increased BUN and creatinine, and azotemia. The main gross lesion was a pale renal cortex on cut surface; in two animals, the kidneys appeared enlarged. Histologically, glomerular amyloid was present in every animal (9/9), and was the predominant renal manifestation of amyloidosis. Additional findings included splenic amyloid (8/8), amyloid in the lamina propria of the intestine (5/5), and amyloid in the lingual submucosa (4/4). Gastric mineralization was present in four animals suffering from renal failure. In the animal dying from bronchopneumonia, severe pancreatic amyloid deposits mainly affecting the exocrine pancreas (1/5) were identified. Immunohistochemistry was employed to identify amyloid AA in eight cases; only in the caracal dying from bronchopneumonia AA was amyloid confirmed. In several organs, especially in those where only small amyloid deposits were detected, a Congo red stain was often necessary to confirm the deposition. The etiology of the amyloidosis remains unknown. Three caracals were related within two generations, another three within four generations, so one might hypothesize a familial trait. In conclusion, amyloidosis should be considered as a significant disease in the caracal. Particularly in cases with renal disease, it should be included as a major differential diagnosis.

Cheetahs (Acinonyx jubatus) are particularly susceptible to feline herpesvirus-1 (FHV-1). Recommendations for preventive health care in cheetahs include vaccination against FHV-1 using killed and modified live virus (MLV) vaccines. Although MLV vaccines tend to induce a more robust immune response than killed vaccines, they can induce disease. This case series details an FHV-1 outbreak in four adult cheetahs following the use of MLV vaccine in one of them. All four cheetahs developed severe FHV-1 clinical signs and were euthanized. Clinical signs included depression, anorexia, nasal discharge, ocular discharge, sneezing, and ulcerative dermatitis. Herpesvirus infection was diagnosed using history, clinical signs, polymerase chain reaction, and histologic evaluation. The timeline of events suggests the MLV vaccine was the inciting cause, although this was not conclusively proven. Outcome of this case suggests that when considering MLV vaccines for cheetahs, careful risk and benefit discussions are merited.

An anorexic 5-yr-old female giant anteater (Myrmecophaga tridactyla) developed multifocal ulcerative and vesicular lesions affecting the rostrum, oral cavity, and tongue. Disseminated skin lesions were also found on the body, affecting the feet, flanks, and genital area. Polymerase chain reaction confirmed a systemic viremic orthopoxvirus infection. Cowpox virus was considered to be the only likely etiological agent. Intensive supportive treatment, including daily fluid therapy, force-feeding, and anti-inflammatory administration achieved a successful outcome after 3 wk. To the authors' knowledge, this is the first time a giant anteater with severe orthopoxvirus lesions has survived the disease. This unique case discusses current and possible future therapeutic and prophylactic options for the treatment of orthopoxvirus infections in giant anteaters and other nondomestic animal species.

Leptospirosis is the most common zoonotic disease worldwide and is considered endemic in countries with tropical climates. It is caused by 10 species of the Leptospira genus and by more than 275 serovars which can affect a wide range of vertebrates. In the Americas, 122 species of four classes of vertebrates have been reported to be infected or exposed to many Leptospira species. Many of these reports are from zoos and rehabilitation centers. Mexico has one single study that reported antibody titers against Leptospira in zoo animals. The purpose of this research was to identify the degree of exposure of some captive mammals and reptiles in Veracruz, a Mexican state with endemic leptospirosis, through microagglutination using 14 live strains of five Leptospira species. Sera samples were collected from 55 animals of 11 species from two classes (Mammalia and Reptilia), four orders (Primates, Artiodactyla, Carnivora, Crocodilia), and nine genera. The more prevalent serovars were Icterohaemorrhagiae and Tarassovi and the highest titers were reactive to the serovar Icterohaemorrhagiae with a value of 1: 51,200.

Baseline health parameters are limited in the primary literature for gray seals (Halichoerus grypus) in the northwest Atlantic. Accurate normal physiologic reference ranges for both species and specific geographic populations are vital tools for assessing the health of individuals and understanding the health of the entire population. This study developed comprehensive reference intervals for biochemical and hematologic parameters of recently weaned gray seal pups on Cape Cod, Massachusetts from samples collected in 2013, 2016, and 2017. Reference ranges were developed using methodology outlined by the American Society of Clinical Veterinary Pathology. By establishing more comprehensive biochemical and hematologic reference ranges for this population based on a robust sample size, this study provides a new tool for clinicians, researchers, and rehabilitation organizations to improve individual patient care and population research.

This study presents the gross and histopathological findings of adenoviral hemorrhagic disease (AHD) in two yearling and one adult mule deer (Odocoileus hemionus). These cases represent the first known outbreak of deer adenovirus (Odocoileus adenovirus 1) in Arizona. Over the span of a month, three female captive mule deer were submitted to Midwestern University's Animal Health Institute for postmortem examination. All of these deer were from the same deer farm and historical findings were similar, consisting of acute presentation of hemorrhagic diarrhea and sudden death. Grossly and histopathologically, all cases had severe pulmonary edema and hemorrhagic enteritis. Additionally, two of the three cases had low numbers of large amphophilic intranuclear inclusions expanding endothelial cells within the small intestine and lungs. Viral PCR of pooled small intestine, lung, and spleen from each of the three cases were positive for deer adenovirus and negative for blue tongue and epizootic hemorrhagic disease.

A group of eight Wagler's pit vipers (Tropidolaemus wagleri) from a private collection died with respiratory signs within 6 mo of one another. The group consisted of an adult breeding pair that was wild caught and six offspring from this pair. Four of the dead snakes were submitted for gross and histopathology. Signs of bacterial pneumonia were detected in all four examined snakes. No inclusion bodies suggestive of viral infection were found in any of the examined tissues. Polymerase chain reactions for the detection of ferla-, adeno-, reo-, and nidoviruses were all negative, but reptarenaviruses closely related to viruses previously described in boa constrictors (Boa constrictor) with inclusion body disease were detected in two of the four snakes. This is the first description of reptarenaviruses in viperid snakes. The pathogenic role of the virus in illness is unknown.

A 0.5-kg, 9-yr-old, male central bearded dragon (Pogona vitticeps) presented with a proliferative mass (0.4 × 0.2 inches) on the left rostral aspect of the lower lip. Physical examination, blood work, and whole-body radiography did not reveal any other abnormalities. Histopathology confirmed squamous cell carcinoma. Considering the small size of the tumor, absence of deep tissue infiltration, and its radioresponsive characteristics, iridium 192 high dose rate brachytherapy was attempted. The dragon initially received three doses of 4 Gy/site at days 0, 7, and 17. Recurrence developed 3 mo later. Three more fractions of 6 Gy/site at days 0, 7, and 14 were delivered according to the same procedure. A second recurrence appeared after 2 mo. Surgical excision was then performed, followed by four fractions of 6 Gy/site on the surgical site at 2-wk intervals. Sixteen months posttreatment, no recurrence of the mass was observed.

Hypervitaminosis D was diagnosed in a giant anteater (Myromecophaga tridactyla) and a large hairy armadillo (Chaetophractus villosus) being fed a commercial insectivore diet. Clinical findings included weight loss, reduced appetite, vomiting, and suspected abdominal discomfort. Hypercalcemia (3.68 and 2.04 mmol/L total and ionized calcium, respectively) was detected in the anteater, and plasma 25(OH)D levels were measured and found to be 808.7 and 379.4 nmol/L for the anteater and armadillo, respectively. Dietary change resulted in a reduction of 25(OH)D levels in both animals and resolution of hypercalcemia in the giant anteater. Dietary analysis of the commercial insectivore food revealed levels of vitamin D₃ higher than the data-sheet values. This case report demonstrates that hypervitaminosis D in Xenarthra can be associated with significant clinical signs.

An adult female spotted eagle ray (Aetobatus narinari) presented for medical evaluation due to a swelling located on the dorsal head. Ultrasound revealed that the swelling originated from a large pocket of fluid in the cranial vault. The swelling was aspirated, and purulent discharge was obtained; Enterococcus faecalis was cultured. An incision was made over the swelling in an attempt to drain fluid but was unsuccessful. Multiple aspirates were performed to drain the abscess, and the animal was treated with oxytetracycline injections. The initial incision sloughed and resulted in a large defect in the cranium that allowed exhibit water to come into the cranial vault and come in contact with the protective membrane of the brain. Forty-two days after initial presentation, the defect in the cranium was healed; fluid from the cranial vault was sampled and appeared normal. During and after treatment, the ray exhibited no abnormal neurologic signs.

Acinetobacter baumannii is a major cause of illness in hospitalized patients and the most important and common pathogen in nosocomial outbreaks worldwide. In animals, A. baumannii has been associated with respiratory infections in a group of minks, leading to pneumonia and acute mortality. This report documents a case of aspiration bronchopneumonia in a wild European hare caused by A. baumannii. A free-ranging, adult male European hare was submitted to necropsy after acute trauma due to being hit by a car. Its lungs showed consolidation with abscess in the middle and cranial lobes. Histopathologic evaluation revealed liquefactive necrosis associated with neutrophilic infiltration, cellular debris, plant material, and bacterial myriads surrounded by moderate neutrophils, macrophages, multinucleated giant cells, lymphocytes, and plasma cell inflammation. Acinetobacter baumannii was isolated from lung tissue.