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

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

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

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

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

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

Successful management of hypertrophic cardiomyopathy in a Matschie's tree kangaroo (Dendrolagus matschiei).

A 3-yr-old, intact male Matschie's tree kangaroo (Dendrolagus matschiei) was examined for a 1-wk history of intermittent lethargy and tachypnea. An echocardiogram revealed concentric hypertrophy of the left ventricular free wall and interventricular septum. These findings were compared to measurements from healthy Matschie's tree kangaroos, supporting a diagnosis of hypertrophic cardiomyopathy. At the time of publication, the patient has been managed for over 11.5 yr, using a combination of enalapril, furosemide, diltiazem, and diet modifications. Hypertrophic cardiomyopathy should be considered as a differential diagnosis in tree kangaroos exhibiting signs of cardiovascular or respiratory distress. This case represents the first report of antemortem diagnosis and successful management of hypertrophic cardiomyopathy in a Matschie's tree kangaroo.

Non-lethal sampling of liver tissue for toxicologic evaluation of Florida cottonmouths snakes, Agkistrodon piscivorus conanti.

Due to their longevity, strong site tenure, poikilothermic metabolism, and low-energy specializations, reptiles might serve as excellent environmental sentinels. Cottonmouth snakes are generalist predators and scavengers, and as such, may have higher exposure to persistent environmental contaminants as a result of bioaccumulation. Traditionally, assessment and monitoring of contaminant exposure in reptiles have involved lethal sampling techniques. In this paper, we describe a non-destructive technique for sampling liver tissue in live anesthetized Florida cottonmouths. Wild-caught snakes (n = 21) were anesthetized with propofol, and a liver wedge biopsy was obtained by clamping the edge of the organ with two small hemostatic mosquito forceps via right-sided coeliotomy incision. A minimum required tissue sample weighing >100 mg was harvested from all except one of the animals. No mortalities occurred during the procedures or recovery from anesthesia, and all snakes were released back into the field after the animal had consumed prey and defecated, usually within 2 weeks following surgery. Hemorrhage was a minor complication in most snakes, especially those with friable discolored livers. The procedure appeared to have no short-term deleterious effects, and two biopsied individuals were captured after being released into the field and appeared to be normal and healthy. However, follow-up studies and recapture of more snakes are needed to assess long-term survivability. Our non-destructive liver sampling technique might be implemented in toxicological studies of other squamates and could help to minimize the lethal sampling of threatened species.

A mystery revealed: an update on eosinophil and other blood cell morphology of the Argentine black and white tegu (Salvator merianae).

Reptile white blood cell (WBC) morphological features are strikingly variable across species. In the Argentine black and white tegu (Salvator merianae), red tegu (Salvator rufescens), and Savannah monitor (Varanus exanthematicus), previous reports described a WBC type with a single distinct, clear, linear- to ovoid- to crescent-shaped inclusion of presumptive monocytic origin. The objective of this study was to further investigate the origin of this unique WBC type with crescent-shaped inclusions. Blood samples from two Argentine black and white tegus, tegu 1, a 4-year-old female, and tegu 2, a 2-year-old presumed male, were submitted for routine hematological evaluation. Additional blood films were prepared and stained with these cytochemical stains: alkaline phosphatase (ALP; naphthol AS-MX phosphate substrate), alpha-naphthyl butyrate esterase, alpha-chloroacetate esterase, myeloperoxidase, Periodic acid-Schiff, and Sudan black B. Blood films from tegu 1 were also stained with a second ALP stain (5-bromo-4-chloro-3-indoxyl-phosphate and nitroblue tetrazolium substrate), Luna, luxol fast blue, and toluidine blue. The blood from tegu 1 was cytocentrifuged to isolate and fix the buffy coat in glutaraldehyde 2.5% aqueous solution for transmission electron microscopy. Six morphologically distinct WBC types were identified from tegu 1, including heterophils, basophils, monocytes, azurophils, lymphocytes, and the unique WBC type, which were identified as eosinophils with inclusions. WBC types in tegu 2 were similar; however, eosinophils lacked a discernable inclusion. Proper WBC identification will be useful in obtaining accurate hemogram data for this species.

Use of Blood Lactate in Assessment of Manual Capture Techniques of Zoo-Housed Crocodilians.

Species-specific welfare indicators are important in promoting positive welfare for zoo animals. Reptiles are a notoriously understudied group in regards to behavior, welfare needs, and husbandry requirements. Using opportunistically obtained samples, we evaluated how blood lactate is affected by variation in manual capture and restraint in crocodilians. Lactate is an indicator of anerobic metabolism in reptiles. It offers a relatively simple and indirect way to assess physiological stress. Determining the best way to reduce struggling during capture and associated stress is of great importance to these species' overall welfare. Blood samples (N = 53) were collected from 10 different species of crocodilians. It was found that age class was a significant predictor variable of lactate levels after capture, and longer handling time seemed to cause an increase in lactate. Finally, draining enclosure pools for a small number of the captures was associated with higher lactate levels compared to other capture factors that were recorded. This study showed that welfare of captive crocodilians could be improved by shortening the duration of physical restraint events when possible. Developing objective measures of welfare and establishing baseline recommendations for care and handling of crocodilians will ultimately promote and improve their wellbeing, along with that of other understudied reptiles in zoos.

AUSTWICKIA CHELONAE IN A WILD GOPHER TORTOISE (GOPHERUS POLYPHEMUS) AND EVIDENCE OF POSITIVE SELECTION ON THE DIPHTHERIA-LIKE TOXIN GENE.

Austwickia (Dermatophilus) chelonae is a filamentous, Gram-positive Actinobacteria in the Dermatophilaceae family. It has caused fatal granulomatous disease in diverse captive reptile species on three continents, but its presence in wild or free-ranging populations was unknown. An adult female gopher tortoise (Gopherus polyphemus) was presented euhydrated, but cachectic and infested with ticks, with two firm, encapsulated masses over the cranioventral neck and right stifle. The tortoise had moderate nonregenerative anemia and evidence of inflammation; plasma biochemistry data was within normal limits. Fine needle aspirate of the neck lesion revealed abundant necrosis and aggregates of cocci. Computed tomography delineated the masses and revealed an additional mass adjacent to the left zygomatic bone. After surgical excision, histology identified chronic granulomas with intralesional filamentous bacteria. Pan-bacterial 16S rRNA PCR and sequencing of the masses identified A. chelonae. Despite treatment with oxytetracycline and ceftazidime, the tortoise deteriorated and was euthanatized. An esophageal lesion consistent with A. chelonae was seen on postmortem examination, although it was determined that the tortoise ultimately succumbed to fungal pneumonia caused by Metarhizium robertsii, an entomopathogenic biotoxin sprayed as insect control. This case reveals A. chelonae is present in free-ranging chelonians in North America. This organism produces a toxin gene similar to diphtheria toxin, one of the most potent known biotoxins, which has not been previously identified outside the genus Corynebacterium. Novel PCR primers were designed for the toxin and rpoB genes, which were amplified and sequenced from two cases and compared with two available genomes. Selection analysis revealed that the toxin gene is under positive selection, which implies it interacts significantly with the immune system, making it a good candidate for immunodiagnostic test development.

Evaluation of parenteral drugs for anesthesia in the blue crab (Callinectes sapidus).

The objective of this study was to evaluate the efficacy and safety of several parenteral anesthetics in blue crabs (Callinectes sapidus). Thirty-one animals were administered one or more of the following drugs by injection into the hemolymph (i.v.) through an arthrodial membrane: etomidate, ketamine, lidocaine, pentobarbital, propofol, tiletamine-zolazepam, xylazine, and ketamine-xylazine. A subset of crabs received intracardiac ketamine. Etomidate had no effect. Lidocaine effects were ultrashort (<3 min). Pentobarbital had prolonged inductions (2 min) and often caused violent excitement and poor anesthesia. Propofol induced light anesthesia accompanied by distress and limb autotomy. Inductions with ketamine, xylazine, tiletamine-zolazepam, and ketamine-xylazine were usually fast (<60 sec). Their anesthetic effects were generally very short (5-10 min) but predictable, smooth, and with good muscle relaxation. The latter two protocols induced a deep plane of anesthesia more consistently but also more significant bradycardia. Intracardiac ketamine injection was fatal in four of five crabs. In conclusion, the anesthetic protocols were considered unsuitable for prolonged anesthesia. However, if very short-term anesthesia is desired, then tiletamine-zolazepam or ketamine-xylazine is recommended. Further studies are indicated to identify a safe anesthetic protocol of longer duration in C. sapidus as well as in other crab species.

Sedation and Anesthesia of Galapagos (Chelonoidis nigra), Aldabra (Aldabrachelys gigantea), and African Spurred Tortoises (Centrochelys sulcata): A Retrospective Review (2009-2019).

Tortoises belong to the taxonomic family Testudinidae, which is considered one of the most imperiled families of the order Testudines. Anesthesia is often required for the medical and surgical management of large tortoises. The objectives of this retrospective study were to review drug regimens used to successfully anesthetize Galapagos (Chelonoidis nigra), Aldabra (Aldabrachelys gigantea) and African spurred (Centrochelys sulcata) tortoises, and to compare the times to effect and to extubation in tortoises administered different premedication protocols. Anesthetic records of giant tortoises admitted to the University of Florida College of Veterinary Medicine between January 2009 and December 2019 were reviewed. A total of 34 tortoises (six Aldabra, 23 Galapagos, and five African spurred) were included, resulting in 64 anesthetic events. Frequently used premedication protocols included an α2-adrenergic agonist and ketamine combined with either midazolam (group α2-adrenergic agonist, midazolam, ketamine, AMK; n = 34), a µ-opioid receptor agonist (group α2-adrenergic agonist, µ-opioid receptor agonist, ketamine, AOK; n = 13), or a µ-opioid receptor agonist and midazolam (group α2-adrenergic agonist, midazolam, µ-opioid receptor agonist, ketamine, AMOK; n = 10). Inhalant anesthetics (isoflurane, n = 21; sevoflurane, n = 23) were frequently used for maintenance of anesthesia following premedication. Out of the 34 total tortoises, 22 had only one anesthetic event, five had two anesthetic events, three had three anesthetic events, and four had four or more anesthetic events. Few adverse effects were observed and there was no mortality reported during the peri-anesthetic period. Sedation and general anesthesia of giant tortoises can be successfully performed with a combination of an α2-adrenergic agonist and ketamine in combination with midazolam and/or a µ-opioid receptor agonist.

Identification of a novel mortality-associated Helicobacter species in gopher tortoises (Gopherus polyphemus), qPCR test development and validation, and correlation with mortality in a wildlife rehabilitation population.

The genus Helicobacter includes spiral-shaped bacteria in the phylum Proteobacteria, class Epsilonproteobacteria, order Campylobacteriales, that have been associated with disease in animals, including reptiles. Three wild gopher tortoise (Gopherus polyphemus) index cases presented between 2012 and 2019 with nasal discharge, lethargy, and weight loss. Cytological examination of nasal discharge from all 3 tortoises identified marked heterophilic and mild histiocytic rhinitis with abundant extracellular and phagocytized spiral shaped bacteria that stained positive with Warthin-Starry stain. Polymerase chain reaction (PCR) and sequencing of the 16S rRNA gene revealed this to be a novel Helicobacter species. Two tortoises died despite treatment attempts, and the third was moribund and was euthanized. Histological examination of the nasal mucosa (n = 3) showed granulocytic to lymphocytic rhinitis with variable mucosal hyperplasia, erosion, and ulceration; Warthin-Starry staining highlighted the presence of spiral bacteria in the untreated tortoise. Genus-specific primers were designed, and the gyrA and groEL genes were amplified by PCR and sequenced. Phylogenetic analysis shows that this organism and other previously characterized Helicobacter from tortoises form a clade. Development and cross-validation of two qPCR diagnostic assays for the gyrA and groEL genes showed significant correlation of the results of two assays (P < 0.0001). These assays were used to survey nasal wash samples from 31 rehabilitating gopher tortoises. Mortality of tortoises significantly correlated with higher Helicobacter loads detected by qPCR (P = 0.028). Appropriate quarantine protocols for tortoises during rehabilitation should consider this organism. Upper respiratory disease in tortoises may involve complex microbial ecology; factors beyond Mycoplasmopsis (Mycoplasma) agassizii should be taken into account.

Application of the laryngeal mask airway for anesthesia in three chimpanzees and one gibbon.

Three pediatric chimpanzees and one pediatric gibbon were anesthetized for routine physical examination. Anesthesia was maintained with inhalant delivered via a laryngeal mask airway (LMA). The LMA was easy to insert, provided adequate control of the airway for ventilation, and caused no tracheal stimulation. No complications were observed. As compared with a face mask, the LMA has the advantage of a more secure airway; the ability to effectively ventilate the patient; less dead space, which leads to lower rebreathing of carbon dioxide; and less exposure of personnel to waste gases. As compared with an endotracheal tube, the LMA causes less airway trauma, is easier to place, and is less stimulating to the patient. The LMA should be considered for use in fasted non-human primates presented for procedures lasting less than 60 min where high peak inspiratory pressures are not needed.

Preliminary evaluation of deslorelin, a GnRH agonist for contraception of the captive variable flying fox Pteropus hypomelanus.

BACKGROUND: This study was conducted to study the effects of a gonadotropin-releasing hormone (GnRH) agonist, deslorelin, on luteinizing hormone (LH), testosterone (males), semen characteristics and pregnancy in the variable flying fox Pteropus hypomelanus. STUDY DESIGN: Male (n = 3) and female (n = 5) bats received a 4.7-mg implant and were housed with untreated bats (eight females and three males, respectively). Plasma was collected twice monthly and analyzed for hormone concentrations, and semen was collected from untreated and treated males 1 month preimplantation, 3 months postimplantation and 4 months postimplantation. RESULTS: Administration of a GnRH challenge 1 month postimplantation showed an attenuated response in treated (n = 4), but not in untreated (n = 4), male and female bats. Plasma LH was lower in treated versus untreated males (p = .04), but not in females. Testosterone was lower in treated versus untreated males (p < .001). Spermic ejaculates were obtained from treated males, although no untreated females became pregnant during the 8-month study. One treated female became pregnant 6 months after implantation. CONCLUSION: Deslorelin is a useful and reversible contraceptive for P. hypomelanus.

A blood survey of elements, viral antibodies, and hemoparasites in wintering Harlequin Ducks (Histrionicus histrionicus) and Barrow's Goldeneyes (Bucephala islandica).

Twenty-eight Harlequin Ducks (Histrionicus histrionicus) and 26 Barrow's Goldeneyes (Bucephala islandica) were captured in Prince William Sound, Alaska, between 1 and 15 March 2005. Blood was collected for quantification of element concentrations, prevalence of antibodies to several viruses, and hemoparasite prevalence and identification. Although we found selenium concentrations that have been associated with selenosis in some birds (>or=2.0 ppm ww), our findings contribute to a growing literature describing relatively high selenium in apparently healthy birds in marine environments. Avian influenza virus antibodies were detected in the plasma of 28% of the ducks. No antibodies against adenovirus, reovirus, or paramyxovirus 1 were detected. Several hemo-parasite species were identified in 7% of ducks. Our findings are similar to those in other free-living marine waterfowl and do not indicate unusual concerns for the health of these species in this area in late winter.

Hyperplastic dermatitis associated with acariasis in a siamang (Symphalangus syndactylus).

A 36-yr-old male captive siamang (Symphalangus syndactylus) was evaluated for mange in the form of generalized alopecia, flaky skin, and pruritus of 1 mo duration. Multiple skin scrapings and biopsies revealed high numbers of trombidiiform mites identified as Psorobia (formerly Psorergates) sp. near cercopitheci (Acarina: Psorergatidae) based on morphologic characteristics. Prolonged repetitive treatment with ivermectin killed the mites and resolved the clinical signs. Psorergatid mites should be considered as a cause of dry flaky skin in even long-term captive siamangs or other primates. This is the first record of psorergatid mites from a primate host in the family Hylobatidae.

Radiographic evaluation of cardiac size in flying fox species (Pteropus rodricensis, P. hypomelanus, and P. vampyrus).

Dilated cardiomyopathy is a relatively common pathology in captive flying foxes (Pteropus spp.). The goal of this study was to establish quantitative reference range measurements that could be used to support a diagnosis of cardiac disease in these animals. Lateral and ventrodorsal thoracic radiographs from apparently healthy flying foxes (n = 66) of three species (Rodriguez island flying fox, P. rodricensis, n = 18; small island flying fox, P. hypomelanus, n = 16; and Malaysian flying fox, P. vampyrus, n = 32) were evaluated objectively to describe the cardiac appearance. Absolute and relative cardiac dimensions also were measured. The same methods were used to evaluate radiographs from flying foxes (n = 9) with known dilated or acute cardiomyopathy. The following ratios were most appropriate for categorizing normal cardiac silhouette size. In the ventrodorsal projection, heart width to thoracic width and heart width to clavicle length were the preferred measurements. In the lateral projection, heart width compared with thoracic height was the preferred measurement. From radiographs of the bats with known dilated and acute cardiomyopathy, the apicobasilar heart length compared with thoracic height and heart width compared with thoracic height on lateral films were the most sensitive ratios for diagnosing cardiomegaly.

Gastric Intussusceptions in a Red Corn Snake (Pantherophis guttatus) Associated with Cryptosporidiosis.

A 3-year-old female red corn snake (Pantherophis guttatus) was presented for a three-week history of anorexia and decreased defecations. On physical examination, a soft midbody intracoelomic swelling was palpated. Transcutaneous coelomic ultrasound revealed a target-like mass on a transverse section of the stomach, suggesting the presence of a gastrointestinal intussusception. On exploratory coeliotomy, a double compounded esophagogastric and gastroduodenal intussusception was diagnosed and reduced surgically. A gastropexy was also performed to prevent recurrence. On histopathology, the gastric glandular mucosa showed moderate to marked proliferation. Diffusely lining the luminal surface of glandular epithelium and free within the lumen were a myriad of protozoa consistent with Cryptosporidium sp. A diagnosis of chronic proliferative gastritis due to Cryptosporidium sp. was made based on these findings. Intussusceptions are rare in reptiles and are infrequently reported in snakes. This is the first report of a double compounded intussusception in a nonmammalian species and the first report of an intussusception involving the stomach in a snake with gastritis due to Cryptosporidium sp.

Comparison of serum and plasma for determination of blood biochemical values in Malaysian flying foxes (Pteropus vampyrus).

The effects of the anticoagulant sodium heparin and time of centrifugation on 20 biochemical analytes in the blood of Malaysian flying foxes (Pteropus vampyrus) were evaluated. Paired plasma and serum samples were centrifuged at 1 hr and 6 hr postcollection. Heparinization and time of centrifugation did not significantly affect albumin, cholesterol, triglycerides, amylase, blood urea nitrogen, creatinine, alanine aminotransferase, aspartate aminotransaminase, alkaline phosphatase, calcium, sodium, and total carbon dioxide levels. Plasma was associated with higher globulin and lower potassium values. Glucose and chloride levels decreased significantly over time, whereas phosphorus levels increased. Serum creatine kinase activity at 6 hr postcollection was significantly higher than the other creatine kinase means. Sodium levels were not significantly affected by sodium heparin as used as an anticoagulant in this study.

Medical management of Toxoplasma meningitis in a white-throated capuchin (Cebus capucinus).

A 32-yr-old white-throated capuchin monkey (Cebus capucinus) was presented for paraparesis. A diagnosis of protozoal meningitis was made on the basis of a lymphocytic pleocytosis with elevated protein in cerebrospinal fluid and a positive serum titer to Toxoplasma gondii of 131,072 using an indirect hemagglutination test. Treatment with clindamycin and trimethaprim-sulfamethoxazole resulted in improvement within 3 days and complete clinical resolution within 2 wk.

Use of a nerve locator to facilitate administration of mandibular nerve blocks in crocodilians.

As part of a clinical workup of dental problems in a large crocodilian collection, mandibular nerve blocks were performed in the animals. A nerve locator was used to facilitate placement of the nerve blocks in American alligators (Alligator mississippiensis), Yacare caiman (Caiman yacare), and a dwarf crocodile (Osteolaemus tetraspis). Provision of analgesia is a frequently underused aspect of patient care in reptiles. Use of a nerve stimulator provides an objective measurement of nerve conduction blockade and may be useful in exotic species in which anatomic landmarks for nerve block placement are not well established.