ANESTHESIA OF ZOO-MANAGED AFRICAN PAINTED DOGS (LYCAON PICTUS) USING A DEXMEDETOMIDINE-KETAMINEBUTORPHANOL-MIDAZOLAM COMBINATION.

Zoo-managed adult African painted dogs (Lycaon pictus; n = 17) were anesthetized with mean dosages of dexmedetomidine 5 µg/kg, ketamine 1.93 mg/kg, butorphanol 0.2 mg/kg, and midazolam 0.15 mg/kg IM. Times to induction milestones (mean ± SD or median [range]) were initial effects at 2 min (1-4 min), recumbency at 3.2 ± 1.1 min, and intubation at 11.6 ± 1.3 min. Two dogs required isoflurane supplementation for intubation. Anesthesia was antagonized with mean dosages of atipamezole 0.05 mg/kg IM, naltrexone 0.2 mg/kg IM, and flumazenil 0.01 mg/kg IV. Times from antagonist administration to recovery milestones were extubation at 3.9 ± 1.5 min, control of head at 7.6 ± 2.5 min, sternal positioning at 8.8 ± 2.3 min, and standing at 12.1 ± 3.5 min. Animals were judged ready for reunification with conspecifics at 41.5 min (28-127 min), and reunification occurred at 62.1 ± 35.2 min. Paired arterial blood gas samples were obtained (n = 13). There was a significant decrease in temperature and blood pressure and increase in pO2, BEecf, and HCO3- (P < 0.05). Decreased respiratory rate with hypercapnia was occasionally observed. This protocol provided consistent anesthesia using a low dose α-2 agonist and permitted quick pack reunification.

DIAGNOSIS AND TREATMENT OF A NATURAL INFECTION WITH TRYPANOSOMA CRUZI (CHAGAS DISEASE) IN A SYMPTOMATIC DE BRAZZA'S MONKEY (CERCOPITHECUS NEGLECTUS) IN ALABAMA.

Trypanosoma cruzi, the causative agent of Chagas disease, is a zoonotic, vector-borne, protozoan hemoflagellate with a wide host range. An 11-yr-old, captive-bred male De Brazza's monkey (Cercopithecus neglecus) presented with weight loss despite normal appetite. Examination revealed hypoglycemia, nonregenerative anemia, and many trypanosomes on a blood smear. A whole blood sample was PCR-positive for T. cruzi discrete typing unit TcIV and the monkey seroconverted using two different methods. The monkey was treated with the standard human dose of benznidazole twice daily for 60 d; however, blood obtained over the next 1.5 yr posttreatment remained PCR-positive for T. cruzi. A second course of benznidazole at a higher dose but lower frequency for 26 wk was required for the monkey to convert to sustained PCR-negative status. The monkey recovered with no apparent lasting effects.

Fatal Sarcocystis calchasi-associated meningoencephalitis in 2 captive vulturine guineafowl.

Two captive vulturine guineafowl (Acryllium vulturinum) were presented with lethargy, hyporexia, weight loss, and progressive neurologic signs. One of the guineafowl was seropositive for Sarcocystis falcatula (1:50 dilution). Both guineafowl died within 5 d of presentation. Histologic examination revealed nonsuppurative meningoencephalitis with gliosis, associated with occasional schizonts in the neuropil. Using fresh-frozen brain tissue, PCR was performed to amplify the ITS1 RNA region and portions of the 18S ribosomal RNA gene (18S gene) and the 28S ribosomal RNA gene (28S gene). Analysis of nucleic acid sequences from the resulting amplicons indicated that Sarcocystis calchasi was the likely cause of disease. To our knowledge, S. calchasi-associated disease has not been reported previously in the order Galliformes.

Atypical Candidiasis of the Gnathotheca in a Lesser Flamingo (Phoeniconaias minor).

An approximately 38-year-old captive male lesser flamingo (Phoeniconaias minor) was presented with a mass involving the right ventral gnathotheca. The mass was surgically excised after which the flamingo was treated with parenteral nonsteroidal anti-inflammatory and antibiotic drugs. Histological analysis identified an abscess with intralesional fungal organisms. Culture and polymerase chain reaction sequencing identified the fungal organisms within the lesion as Candida albicans. Treatment with oral itraconazole was initiated 23 days after initial surgical excision; however, the flamingo continued to lose weight while being treated, and died after 10 days of antifungal therapy. Necropsy, histologic examination, and culture confirmed the persistence of a mycotic abscess that infiltrated the mandibular bone and was associated with C albicans.

Adjunctive Use of Intravenous Antibiotic Regional Limb Perfusion in Three Cranes with Distal Limb Infections.

Pododermatitis and wing lesions are commonly reported issues in captive crane species. Regional limb perfusion has been used as a treatment for distal limb infections in several avian species, as systemic antibiotic therapy is often prolonged and unrewarding. A black-necked crane (Grus nigricollis), Siberian crane (Leucogeranus leucogeranus), and wattled crane (Bugeranus carunculatus) were treated with amikacin (5-10 mg/kg IV infusion) regional limb perfusion for cellulitis and osteomyelitis of hind limb digits and alular osteomyelitis and septic arthritis of the carpus, respectively, with a range of 1-3 treatments per case. Clinical signs of infection resolved within 10-40 days following regional limb perfusion combined with oral or parental antibiotic therapy. No side effects were observed following regional limb perfusion. This is the first report of regional limb perfusion in cranes and the first report of intravenous regional limb perfusion in a wing of an avian species.

EMYDID HERPESVIRUS 1 INFECTIONS IN WESTERN POND TURTLES (ACTINEMYS MARMORATA) AND A RED-EARED SLIDER (TRACHEMYS SCRIPTA ELEGANS) WITH FATAL AND NONFATAL OUTCOMES.

Herpesviruses are important pathogens in zoologic chelonian collections and have been associated with fatal disease in turtles of the family Emydidae. In this report, three western pond turtles (Actinemys marmorata), living in a mixed-species freshwater turtle exhibit, presented with varying degrees of lethargy, pallor, generalized edema, and cloacal hemorrhage before death within a 2-wk period. Postmortem findings included necrohemorrhagic enterocolitis, necrotizing splenitis, hepatic necrosis, esophagitis, thymic necrosis, and pneumonia with epithelial necrosis and degeneration of the trachea and kidneys. Intraepithelial, intranuclear, amphophilic to eosinophilic, Cowdry type A viral inclusion bodies were identified in the intestinal tract, liver, spleen, kidney, trachea, lung, and thymus. PCR amplification and sequencing of liver tissue produced amplicons that were 100% homologous with emydid herpesvirus 1 (EmyHV-1). Molecular screening of cohoused emydid turtles revealed that a red-eared slider (Trachemys scripta elegans) and a western pond turtle, both asymptomatic, were PCR-positive for EmyHV-1 on combined oral-cloacal swabs. This report describes, for the first time, EmyHV-1-associated disease in western pond turtles and molecularly identifies EmyHV-1 in an asymptomatic red-eared slider.

MYONECROSIS AND DEATH DUE TO PRESUMED MICROCYSTIN TOXICOSIS IN AMERICAN WHITE PELICANS (PELECANUS ERYTHRORHYNCOS).

Over a period of 5 mo, seven out of eight American white pelicans (Pelecanus erythrorhynchos) housed on a spring-fed pond at a zoo died or were euthanized. Clinical signs included inability to stand, anorexia, and weight loss. Clinicopathologic findings included heterophilic leukocytosis and elevated creatine kinase and aspartate aminotransferase. Histopathologic findings on all pelicans demonstrated severe, chronic, diffuse rhabdomyofiber degeneration and necrosis, making vitamin E deficiency a differential diagnosis despite routine supplementation. Based on tissue and pond water assays for the cyanobacterial toxin, microcystin, toxicosis is suspected as the inciting cause of death in these cases. We hypothesize that vitamin E exhaustion and resultant rhabdomyodegeneration and cardiomyopathy were sequelae to this toxicosis.

NOVEL PREGNANCY DIAGNOSIS AND ALTRENOGEST SUPPLEMENTATION LEAD TO SUCCESSFUL PREGNANCY IN A BLACK-FOOTED CAT (FELIS NIGRIPES).

This is the first time fecal prostaglandin F2α metabolite (PGFM) analysis and altrenogest were used to determine and maintain a pregnancy in a zoo-housed black-footed cat (BFC; Felis nigripes). The established pair had not produced offspring during the year since their arrival at Birmingham Zoo. Fecal samples were collected daily and analyzed by enzyme immunoassay for progestagen, estrogen metabolite, and PGFM concentrations. After a 4-mo separation, the pair was reintroduced, and breeding was reinitiated. Two pregnancies were suggested by sustained, elevated progestagen concentrations (mean >3 × baseline) but were confirmed by elevated PGFM concentrations (mean 4-25 × baseline) beginning about 1 mo after presumed or observed breeding. The first pregnancy was lost after ∼51 days, and altrenogest was administered (0.088 mg/kg/day) in order to help sustain the second pregnancy, which went to term, but the kitten did not survive. Ultrasonographic and radiographic evaluations of pregnancy were utilized during the second pregnancy to provide information on litter size.

PHARMACOKINETICS OF A SUSTAINED-RELEASE FORMULATION OF MELOXICAM AFTER SUBCUTANEOUS ADMINISTRATION TO AMERICAN FLAMINGOS ( PHOENICOPTERUS RUBER).

Meloxicam is commonly used in avian medicine to relieve pain and inflammation, but the recommended dosing frequency can be multiple times per day, which can contribute to stress during convalescence. In this study, the pharmacokinetics of a sustained-release formulation of meloxicam were determined after subcutaneous administration of a single 3-mg/kg dose to eight healthy adult American flamingos ( Phoenicopterus ruber). Blood samples were collected before (time 0) and at 0.5, 1, 4, 8, 12, 24, 48, 96, and 120 hr after drug administration. Analysis of meloxicam in plasma samples was conducted with the use of reversed-phase high-performance liquid chromatography, and pharmacokinetic parameters were calculated by noncompartmental analysis. Plasma concentrations reached a mean maximum (±standard deviation) of 7.65 (±2.39) µg/ml at 0.56 (±0.18) hr with a terminal half-life of 1.76 (±1.41) hr. Based on these findings, this sustained-release formulation of meloxicam does not extend the interval between treatments as compared to the regular formulation, so it is not recommended in American flamingos at this time.

USE OF COMPOSITE MATERIALS AS A COMPONENT OF TUSK FRACTURE MANAGEMENT IN AN ASIAN ELEPHANT (ELEPHAS MAXIMUS) AND AN AFRICAN ELEPHANT (LOXODONTA AFRICANA).

Tusk fractures in Asian (Elephas maximus) and African elephants (Loxodonta africana) can result in damage to the distal end or to longitudinal cracks, potentially progressing to pulpitis. With pulp exposure, endodontic therapy is the treatment of choice, but conservative therapy has sufficed for some elephants. This manuscript describes the use of composite materials as a component of tusk fracture management. A 7-yr-old male Asian elephant fractured the distal end of both tusks with pulp exposure in one. Capping of each tusk with a Kevlar/fiberglass composite prevented further damage, and a modification allowed care of the exposed pulp tissue. A 34-yr-old male African elephant with a longitudinal crack received a carbon fiber/fiberglass composite circumferential wrap to potentially stabilize the crack. Compression of the crack was achieved, but follow-up was truncated due to bacterial pulpitis. Both cases show that composite material allows for lightweight, durable management of tusk fractures with continued radiographic monitoring.

SURVEY OF ANAPLASMA PHAGOCYTOPHILUM ANTIBODIES IN CAPTIVE PRZEWALSKI'S HORSES (EQUUS FERUS PRZEWALSKII).

Anaplasma phagocytophilum (formerly Ehrlichia equi ) is a tickborne pathogen of domestic horses and the causative agent of equine granulocytic anaplasmosis. After the occurrence of clinical anaplasmosis in a Przewalski's horse ( Equus ferus przewalskii) housed at the Smithsonian Conservation Biology Institute in 2008, opportunistic serosurveillance of the herd was initiated. From 2008 to 2014, 57 serum samples were collected from 27 individuals (10 males; 17 females). Using indirect immunofluorescent antibody assays for anti- Anaplasma phagocytophilum antibodies, it was determined that prevalence was 53%. No significant sex differences were identified. A statistical association between increasing age and seropositive status suggests cumulative risk of exposure to Anaplasma phagocytophilum . After exclusion of four clinical cases of anaplasmosis, it was found that 22-57% of those sampled each year were seropositive and clinically normal, suggesting that the majority of Przewalski's horses develop subclinical or self-limiting anaplasmosis after exposure to A. phagocytophilum .

CLINICAL DISEASE ASSOCIATED WITH ANAPLASMA PHAGOCYTOPHILUM INFECTION IN CAPTIVE PRZEWALSKI'S HORSES (EQUUS FERUS PRZEWALSKII).

Anaplasma phagocytophilum is a tick-borne pathogen of domestic horses and the causative agent of equine granulocytic anaplasmosis. This case series describes three confirmed cases of clinical anaplasmosis, and a fourth case of presumptive anaplasmosis in Przewalski's horses ( Equus ferus przewalskii) housed at the Smithsonian Conservation Biology Institute from 2008 to 2014. Clinical signs varied among individuals with affected horses exhibiting lethargy, weakness, pyrexia, hypophagia, reluctance to move, or ataxia. Anaplasmosis cases were confirmed with a combination of identification of neutrophilic inclusions (morulae) on peripheral blood smear, positive polymerase chain reaction (PCR) testing of whole blood, or convalescent titers. All animals recovered after antimicrobial therapy with oxytetracycline. Diagnosis should be made by a combination of clinical signs plus identification of morulae or positive A. phagocytophilum PCR. Treatment is curative using oxytetracyline intramuscularly or intravenously followed by daily therapy with oxytetracyline or minocycline for 14-30 days. The authors recommend that A. phagocytophilum infection be included on any differential list for Przewalski's horses presenting with fever or ataxia within or near an enzootic area.

RANAVIRUS EPIZOOTIC IN CAPTIVE EASTERN BOX TURTLES (TERRAPENE CAROLINA CAROLINA) WITH CONCURRENT HERPESVIRUS AND MYCOPLASMA INFECTION: MANAGEMENT AND MONITORING.

Frog virus 3 (FV3) and FV3-like viruses are members of the genus Ranavirus (family Iridoviridae) and are becoming recognized as significant pathogens of eastern box turtles (Terrapene carolina carolina) in North America. In July 2011, 5 turtles from a group of 27 in Maryland, USA, presented dead or lethargic with what was later diagnosed as fibrinonecrotic stomatitis and cloacitis. The presence of FV3-like virus and herpesvirus was detected by polymerase chain reaction (PCR) in the tested index cases. The remaining 22 animals were isolated, segregated by severity of clinical signs, and treated with nutritional support, fluid therapy, ambient temperature management, antibiotics, and antiviral therapy. Oral swabs were tested serially for FV3-like virus by quantitative real-time PCR (qPCR) and tested at day 0 for herpesvirus and Mycoplasma sp. by conventional PCR. With oral swabs, 77% of the 22 turtles were FV3-like virus positive; however, qPCR on tissues taken during necropsy revealed the true prevalence was 86%. FV3-like virus prevalence and the median number of viral copies being shed significantly declined during the outbreak. The prevalence of herpesvirus and Mycoplasma sp. by PCR of oral swabs at day 0 was 55% and 68%, respectively. The 58% survival rate was higher than previously reported in captive eastern box turtles for a ranavirus epizootic. All surviving turtles brumated normally and emerged the following year with no clinical signs during subsequent monitoring. The immediate initiation of treatment and intensive supportive care were considered the most important contributing factors to the successful outcome in this outbreak.

Identification of a novel herpesvirus in captive Eastern box turtles (Terrapene carolina carolina).

Herpesviruses are significant pathogens of chelonians which most commonly cause upper respiratory tract disease and necrotizing stomatitis. Herpesvirus infection was identified in two populations of captive Eastern box turtles (Terrapene carolina carolina) using histopathology and polymerase chain reaction (PCR) with DNA sequencing. Necrotizing lesions with eosinophilic to amphophilic intranuclear inclusion bodies were identified in the tissues of one hatch-year individual in January 2013, which was herpesvirus positive by PCR. A separate captive group of adults had an observed herpesvirus prevalence of 58% using PCR in July 2011. In these cases, a novel herpesvirus, Terrapene herpesvirus 1 (TerHV1), was identified and serves as the first herpesvirus sequenced in the genus Terrapene. Similar to the other herpesviruses of the Order Testudines, TerHV1 clusters with the genus Scutavirus of the subfamily Alphaherpesvirinae.

A comparison of oral and topical vitamin A supplementation in African foam-nesting frogs (Chiromantis xerampelina).

Vitamin A is essential for a variety of functions, including cellular differentiation, morphogenesis, growth, vision, immune response, and reproduction. A captive population of African foam-nesting frogs (Chiromantis xerampelina) with a known history of vitamin A deficiency had higher than expected incidence of sudden death, bacterial osteomyelitis, and stunted growth. Due to the high prevalence and untreatable nature of the diseases in the population, euthanasia of the population was recommended. Before euthanasia, the population was entered into a study to compare oral dietary supplementation of vitamin A to topical treatment with water-miscible vitamin A palmitate (AQUASOL A Parenteral, Mayne Pharma Inc., Paramus, New Jersey 07652, U.S.A.). Eighty-four frogs, weighing 2-7 g, were divided into a control and three treatment groups of 21 frogs per group, with normalized weight distribution. The control group received standard daily nutrition of crickets dusted with a supplement containing 342,000 international units (IU) vitamin A/kg. The treatment groups consisted of oral supplementation with crickets dusted with a fortified supplement containing 822,510 IU vitamin A/kg; topical vitamin A palmitate 50 IU every other day; and topical vitamin A palmitate 50 IU once a week. After 30 days, all frogs were euthanized, and 12 frogs from each group were analyzed for whole-body vitamin A levels. The control and treatment groups 1, 2, and 3 had average whole-body vitamin A levels of 1371.4 IU/kg (SE 284.4), 908.7 IU/kg (SE 186.5), 6385.9 IU/kg (SE 675.9), and 3521.8 IU/kg (SE 575.1), respectively. These results suggest that oral supplementation using a product high in vitamin A may be ineffective at raising whole-body vitamin A levels above those achieved with standard nutrition. Topical administration of vitamin A on an every other day and once a week dosing schedule achieved levels 4.5- and 2.5-fold higher than standard nutrition, respectively.