Mycobacterial Airsacculitis Caused by Mycobacterium fortuitum in a Southern Rockhopper Penguin (Eudyptes chrysocome).

A 21-year-old male southern rockhopper penguin (Eudyptes chrysocome) was presented with a chronic history of intermittently decreased appetite, lethargy, and regurgitation. On the external physical examination, the bird was determined to be in fair-to-thin body condition with the complete blood count and plasma chemistry panel being largely unremarkable. Full-body radiographic images were considered normal, and gastroscopy showed only mild gastritis and duodenitis. The penguin was euthanatized shortly thereafter due to acute onset of respiratory distress. During the gross necropsy examination, the bird had severe airsacculitis with thick, yellow-to-tan, moist granular plaques adhering to the surface of many air sacs, as well as regional contiguous pneumonia. Intralesional acid-fast bacilli were observed in histologic sections of air sac tissue, and polymerase chain reaction of the affected air sacs was positive for Mycobacterium fortuitum. This clinical case study describes mycobacteriosis in a sub-Antarctic penguin and to the best of the authors' knowledge, the first reported isolation of M fortuitum from a penguin.

CLONAL SPREAD OF YERSINIA ENTEROCOLITICA 1B/O:8 IN MULTIPLE ZOO 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.

A RETROSPECTIVE STUDY OF MORBIDITY AND MORTALITY IN THE NORTH AMERICAN AMUR LEOPARD ( PANTHERA PARDUS ORIENTALIS) POPULATION IN ZOOLOGIC INSTITUTIONS FROM 1992 TO 2014.

The Amur leopard ( Panthera pardus orientalis) is one of the most critically endangered leopards on the International Union for the Conservation of Nature red list. The primary objective of this retrospective study was to identify common and significant causes of morbidity and mortality in the North American Amur leopard zoo population. This information provides insights that contribute to their improved care, health, and medical management and, ultimately, affects the sustainability of this leopard subspecies in the wild. Medical records and complete postmortem reports were requested from all North American zoologic institutions that held Amur leopards in their collections from 1992 to 2014. Information from 175 individuals, representing 93% of the population (188 animals), was received. Data were reviewed generally and by subdivision into the following age groups: neonate: 0 day-3 mo ( n = 45); juvenile-young adult: 3 mo-5 yr ( n = 21); adult: 5-10 yr ( n = 17); senior: 10-15 yr ( n = 31); or geriatric: >15 yr ( n = 61). The major causes of morbidity, listed by body system for the study population, excluding neonates, were musculoskeletal (40%), reproductive (24%), dental (19%), cardiopulmonary (13%), gastrointestinal (13%), and urogenital (12%). The two most common causes of death or euthanasia, primarily in senior and geriatric animals, were chronic renal disease and neoplasia; maternal neglect and maternal trauma were most common in neonates. The largest populations in this study were neonates and geriatrics, indicating that if animals survive the neonatal period, they often live into their late teens to early 20s. This is the first comprehensive study of the causes of morbidity and mortality in the Amur leopard zoo population in North America.

Pharmacokinetics of a Single Intramuscular Injection of Long-Acting Ceftiofur Crystalline-Free Acid in Cattle Egrets ( Bubulcus ibis).

We determined the pharmacokinetic properties of ceftiofur crystalline-free acid (CCFA), a long-acting antibiotic, after a single intramuscular injection in cattle egrets ( Bubulcus ibis). A dose of 20 mg/kg was administered intramuscularly to 18 birds and blood samples were collected via jugular venipuncture at 1, 2, 4, 8, 12, 24, 48, 72, 96, 120, 144, 168, 192, 216, and 240 hours after CCFA administration. Plasma concentrations of ceftiofur free acid equivalents (CFAEs) were measured via high-performance liquid chromatography. The minimum inhibitory concentration (MIC) of 1 µg/mL was reached by 1 hour after administration and remained higher than the MIC for at least 72 hours in all birds. This target concentration is effective for many bacterial infections in avian species. The area under the plasma concentration versus time curve was 451.3 h*µg/mL, maximum plasma concentration was 16.22 µg/mL, time to maximum plasma concentration was 3.2 hours, mean harmonic half-life was 37.92 hours, and time that the concentrations of CFAEs were higher than the target MIC was a minimum of 72 hours.

Discrimination of individual tigers (Panthera tigris) from long distance roars.

This paper investigates the extent of tiger (Panthera tigris) vocal individuality through both qualitative and quantitative approaches using long distance roars from six individual tigers at Omaha's Henry Doorly Zoo in Omaha, NE. The framework for comparison across individuals includes statistical and discriminant function analysis across whole vocalization measures and statistical pattern classification using a hidden Markov model (HMM) with frame-based spectral features comprised of Greenwood frequency cepstral coefficients. Individual discrimination accuracy is evaluated as a function of spectral model complexity, represented by the number of mixtures in the underlying Gaussian mixture model (GMM), and temporal model complexity, represented by the number of sequential states in the HMM. Results indicate that the temporal pattern of the vocalization is the most significant factor in accurate discrimination. Overall baseline discrimination accuracy for this data set is about 70% using high level features without complex spectral or temporal models. Accuracy increases to about 80% when more complex spectral models (multiple mixture GMMs) are incorporated, and increases to a final accuracy of 90% when more detailed temporal models (10-state HMMs) are used. Classification accuracy is stable across a relatively wide range of configurations in terms of spectral and temporal model resolution.

Evaluating echocardiogram and indirect blood pressure results in male western lowland gorillas (Gorilla gorilla gorilla) during three phases of an anesthetic protocol.

Until the majority of the great ape population is trained for conscious cardiac evaluations, most individuals will require general anesthesia to perform echocardiograms. Within the veterinary community, concern exists that certain anesthetic protocols may exacerbate or artificially induce signs of cardiac disease. Because of potential cardiovascular effects, medetomidine has generally been used cautiously in patients with cardiac disease. The combination of ketamine and medetomidine is frequently used by many institutions because of its reversibility. To date, no published studies have obtained physiologic or echocardiographic parameters comparing different anesthetic protocols. In this study, with the use of seven adult male gorillas (Gorilla gorilla gorilla) with and without cardiac disease, echocardiographic and indirect blood pressure data during three phases of an anesthetic protocol were collected. The initial echocardiographic study was completed with ketamine/ medetomidine alone (5-7 mg/kg, i.m., and 0.05-0.07 mg/kg, i.m., respectively); the second study was completed after the addition of sevoflurane inhalant anesthesia to this procedure; and the third study was completed after reversal of medetomidine by administration of atipamezole (5:1 with the medetomidine dose given at induction). Without exception, ejection fractions were 15-25% lower under anesthesia with medetomidine as compared to ejection fractions after administration of atipamezole. Indirect blood pressures were higher on ketamine/ medetomidine, lower with addition of sevoflurane, and considerably lower after administration of atipamezole.

Serosurvey of free-ranging Amur tigers in the Russian Far East.

Wild Amur tigers (Panthera tigris altaica, n=44) from the Russian Far East were tested for antibodies to feline leukemia virus, feline corona virus (FCoV), feline immunodeficiency virus, feline parvovirus (FPV), canine distemper virus (CDV), Toxoplasma gondii, and Bartonella henselae. Antibodies to FCoV, CDV, FPV, and T. gondii were detected in 43, 15, 68, and 42% of tigers, respectively. No differences were detected in antibody prevalence estimates between tigers captured as part of a research program and those captured to mitigate human-tiger conflicts. Domestic dogs (Canis familiaris) were tested as a potential source for CDV; 16% were vaccinated against CDV and 58% of unvaccinated dogs were antibody positive for CDV. A high percentage of tigers were exposed to potential pathogens that could affect the survival of this species. We recommend continued monitoring of wild tigers throughout Asia, development of standardized sampling and postmortem examination procedures, and additional research to better understand potential domestic and wild animal sources for these pathogens.

Clinical and pathologic characteristics of T-cell lymphoma with a leukemic phase in a raccoon dog (Nyctereutes procyonoides).

A 7.5-year-old raccoon dog (Nyctereutes procyonoides) from the Henry Doorly Zoo (Omaha, Nebraska) presented to the veterinary hospital for lethargy and weight loss. On physical examination, splenomegaly and hepatomegaly were noted on palpation and were confirmed by radiographic evaluation. Radiography also demonstrated a mass in the cranial mediastinum. A complete blood cell count revealed marked leukocytosis (115,200 cells/µl), with a predominance of lymphoid cells. The animal was euthanized due to a poor prognosis. Necropsy revealed splenomegaly, hepatomegaly, and a large multiloculated mass in the cranial mediastinum. The histologic and immunohistochemical diagnosis was multicentric T-cell lymphoma with a leukemic phase.

Comparison of carfentanil-xylazine and thiafentanil-medetomidine in electroejaculation of captive gaur (Bos gaurus).

Carfentanil citrate and thiafentanil oxalate have been used successfully to immobilize captive and free-ranging ungulates. The objective of this study was to compare the efficacy and certain physiologic parameters of protocols by using the 2 opioids in gaur (Bos gaurus). Eight adult gaur bulls were immobilized for electroejaculation at Omaha's Henry Doorly Zoo (Omaha, Nebraska, USA). All the animals were immobilized twice, by using each of the following protocols one time: 10 mg carfentanil combined with 100 mg xylazine (CX), reversed with 1,000 mg naltrexone and 24 mg yohimbine; and 12 mg thiafentanil combined with 20 mg medetomidine (TM), reversed with 120 mg naltrexone and 100 mg atipamezole. Immobilization drugs were delivered intramuscularly into the shoulder area via pole syringe. Electroejaculation was carried out by a standardized protocol to duplicate procedural stimulation on each animal. Induction and recovery times, initial rectal temperature, heart rate, respiratory rate, anesthetic depth, oxygen saturation, indirect blood pressure, and arterial blood gases were recorded at the time of initial handling, before ejaculation, and after ejaculation. Antagonists were administered 1/4 i.v. and 3/4 s.q. Both protocols require a small volume of drug for a large ungulate, provide smooth induction, and adequate anesthesia. Both protocols produced a significant hypoxemia, although the animals on CX showed slightly better blood gas values (based on lower partial pressure of carbon dioxide) and numerically lower blood pressure values. Animals on TM had better muscle relaxation and smoother recoveries, with no renarcotization noted. The results of the present study indicate the TM and CX protocols used for immobilizing gaur result in similar quality ejaculates that can be used for fertility examination as well as for assisted reproduction such as artificial insemination. Additional immobilizations need to take place to further compare these 2 combinations in this species.

Vocal power and pressure-flow relationships in excised tiger larynges.

Despite the functional importance of loud, low-pitched vocalizations in big cats of the genus Panthera, little is known about the physics and physiology of the mechanisms producing such calls. We investigated laryngeal sound production in the laboratory using an excised-larynx setup combined with sound-level measurements and pressure-flow instrumentation. The larynges of five tigers (three Siberian or Amur, one generic non-pedigreed tiger with Bengal ancestry and one Sumatran), which had died of natural causes, were provided by Omaha's Henry Doorly Zoo over a five-year period. Anatomical investigation indicated the presence of both a rigid cartilaginous plate in the arytenoid portion of the glottis, and a vocal fold fused with a ventricular fold. Both of these features have been confusingly termed 'vocal pads' in the previous literature. We successfully induced phonation in all of these larynges. Our results showed that aerodynamic power in the glottis was of the order of 1.0 W for all specimens, acoustic power radiated (without a vocal tract) was of the order of 0.1 mW, and fundamental frequency ranged between 20 and 100 Hz when a lung pressure in the range of 0-2.0 kPa was applied. The mean glottal airflow increased to the order of 1.0 l s(-1) per 1.0 kPa of pressure, which is predictable from scaling human and canine larynges by glottal length and vibrational amplitude. Phonation threshold pressure was remarkably low, on the order of 0.3 kPa, which is lower than for human and canine larynges phonated without a vocal tract. Our results indicate that a vocal fold length approximately three times greater than that of humans is predictive of the low fundamental frequency, and the extraordinarily flat and broad medial surface of the vocal folds is predictive of the low phonation threshold pressure.

Dermatitis in captive Wyoming toads (Bufo baxteri) associated with Fusarium spp.

From May 2007 to June 2008, 30 of 49 Wyoming toads (Bufo baxteri) kept at Omaha's Henry Doorly Zoo (Nebraska, USA) died showing clinical signs of ventral erythema, inappetance, lethargy, and delayed righting reflex. Treatment with antifungals and antibiotics was unsuccessful in all cases. Histopathologic analyses revealed dermatitis as the primary problem in 20 of 21 toads in which skin was examined. Fungal dermatitis was present in 17 toads, with hyphae approximately 1-3 µm in diameter, and parallel cell walls and frequent septations. In 14 animals, the fungal dermatitis was the main pathologic lesion. Several species of bacteria were associated with all cases. A few animals tested positive for Ranavirus using polymerase chain reaction. Fusarium sp. was consistently cultured from skin, feces, kidneys, and from powdered food provided to crickets. Four isolates were identified as Fusarium proliferatum, Fusarium oxysporum, Fusarium solani, and Fusarium verticillioides, which suggested a secondary role of fungi. A specific underlying cause of disease could not be found, although the roles of humidity and Ranavirus infection are discussed, along with the well-known susceptibility of bufonids to fungal dermatitis.

Morbillivirus infection in a wild siberian tiger in the Russian Far East.

We report the first documented case of morbillivirus infection in a wild, free-ranging Siberian tiger (Panthera tigris altaica). The tigress entered a small village in the Russian Far East in an ambulatory but stuporous state with no apparent recognition or fear of humans. Her condition progressed rapidly with neurological signs, anorexia, and ultimately death. Histologic lesions included vacuolated to malacic white matter in the brain stem, cerebellum, and thalamus, with associated lymphocytic meningoencephalitis. Large, intranuclear, eosinophilic inclusions were within regional astrocytes, and the brain lesions were immunohistochemically positive when stained for canine distemper viral antigen. Hematologic and blood chemistry results were consistent with overwhelming systemic infection and starvation. The animal also was antibody-positive for canine distemper virus, feline panleukopenia, and feline coronavirus.

Scoliosis in a tiger salamander (Ambystoma tigrinum) associated with encysted digenetic trematodes of the genus Clinostomum.

A group of 202 tiger salamanders (Ambystoma tigrinum) was brought into captivity due to habitat destruction. Half of these animals died, over two mo, showing generalized hemorrhages compatible with an infectious disease, but an etiologic agent was not determined. Encysted metacercarial stages of trematodes within the skeletal musculature, coelomic cavity, and subcutaneous space were additional necropsy findings. One salamander had scoliosis and multiple skin nodules. A radiograph showed no skeletal abnormality to explain the scoliosis; however, numerous round nodules were more radiodense than the surrounding tissue. A presumptive diagnosis of generalized trematodiasis was made, yet the salamander did not improve after a course of praziquantel and subsequently died. Necropsy revealed massive, encysted trematode infection. Histologic examination revealed marked multifocal intramuscular, subcutaneous, and coelomic trematodiasis with associated necrosis and inflammation. Based on gross morphology, the trematode was identified as a member of the genus Clinostomum.

Mixed infection with reovirus and Chlamydophila in a flock of budgerigars (Melopsittacus undulatus).

Eleven budgerigars (Melopsittacus undulatus) from a zoological collection presented at necropsy with emaciation and splenomegaly or hepatomegaly or both. Polymerase chain reaction assays performed on liver and spleen samples were positive for Chlamydophila psittaci in 2 of 3 birds tested, and histologic findings in 2 additional birds were compatible with chlamydiosis. The aviary was subsequently closed to the public, and a 45-day treatment regimen with doxycycline in the seeds was initiated. No further deaths of birds with hepatomegaly or splenomegaly were observed after the first day of treatment. Further investigation of birds that died during the outbreak with emaciation and with hepatic and splenic enlargement revealed severe necrosis of the spleen and liver suggestive of reovirus infection, which was supported with polymerase chain reaction analysis from paraffin-embedded tissue. This reovirus was sequenced and had 100% homology with a reovirus previously identified in an African grey parrot (Psittacus erithacus). The outbreak did not affect cockatiels (Nymphicus hollandicus) and blue quail (Coturnix chinensis) kept in the same aviary. A group of budgerigars added to the collection soon before the opening of the aviary may have introduced reovirus and Chlamydophila into the collection.

Herpesvirus infections in rock hyraxes (Procavia capensis).

Seven juveniles and 3 adults from a closed group of 19 rock hyraxes (Procavia capensis) housed in a zoo's indoor rock exhibit died or were euthanized after developing blepharoconjunctivitis and orofacial ulcers over a 2-week period. Histopathologic examination of dermal ulcers and ulcerated tongues revealed amphophilic to basophilic intranuclear inclusion bodies in epithelial cells bordering ulcers. Epithelial cells with inclusion bodies were often characterized by cytomegaly and karyomegaly, and many cells had formed syncytia. Examination of inclusion bodies in tongue epithelium by transmission electron microscopy revealed icosahedral nucleocapsids, approximately 80-95 nm in diameter, with morphologic features consistent with herpesvirus. Cytopathic effect (CPE) typical of alphaherpesvirus infection was seen in bovine turbinate, equine dermal, and Vero cell monolayers after inoculation with homogenates of the skin lesions, but CPE was not seen after inoculation onto Madin-Darby canine kidney or swine testicle cell monolayers. Polymerase chain reaction analysis using degenerate primers that targeted a portion of the herpesvirus polymerase gene generated a product of approximately 227 base pairs. The product was cloned, sequenced, and then analyzed using BLAST. At the nucleotide level, there was 86%, 77%, and 76% shared identity with Eidolon herpesvirus 1, Human herpesviruses 1 and 2, and Cercopithecine herpesvirus 2, respectively. Herpesvirus infections in rock hyraxes have not been characterized. The data presented in the current study suggest that a novel alphaherpesvirus caused the lesions seen in these rock hyraxes. The molecular characteristics of this virus would tentatively support its inclusion in the genus Simplexvirus.

Safety and immunogenicity of the Mycobacterium tuberculosis DeltalysA DeltapanCD vaccine in domestic cats infected with feline immunodeficiency virus.

Feline immunodeficiency virus (FIV)-positive and FIV-negative cats (n=4/group) received 2 x 10(6) CFU Mycobacterium tuberculosis DeltalysA DeltapanCD intramuscularly. Vaccination elicited antibody responses, albeit at lower levels in FIV-positive cats than in FIV-negative cats. Delayed-type hypersensitivity responses were minimal in both groups. No adverse reactions were found.

An outbreak of avian mycobacteriosis caused by Mycobacterium intracellulare in little blue penguins (Eudyptula minor).

Mycobacterium intracellulare (MIT) was diagnosed postmortem by culture and supporting histopathology in seven birds from a flock of little blue penguins (Eudyptula minor) at the Henry Doorly Zoo (HDZ). These birds represented 20% of the deaths in the population over a 4 yr period. Clinical signs in affected birds included severe respiratory distress characterized by open-mouth breathing with chronic debilitation. On exam, plaques were noted in the larynx, trachea, and soft tissue of the caudal oropharynx. Index cases were identified on necropsy in two birds on loan to another institution in 2003. Following a case confirmed antemortem at the HDZ, a three-drug protocol of rifampin (15 mg/kg p.o. s.i.d.), ethambutol (15 mg/kg p.o. s.i.d.), and clarithromycin (10 mg/kg p.o. s.i.d.) was started on this bird in 2004 and extended to the entire flock in 2005. Gastric wash, fecal samples, and throat plaques were obtained antemortem on five birds within the flock, selected because of the presence of oral plaques, and tested by culture followed by a polymerase chain reaction assay. MIT was detected in gastric washes from four birds and in throat plaques from all five. Three more birds died during treatment. After the seventh bird died, antimicrobial susceptibility testing performed in July 2007 indicated that the MIT was now resistant to most antibiotics tested, including rifampin and ethambutol. The treatment regimen was changed to minocycline (10 mg/kg p.o. b.i.d.) and clarithromycin (10 mg/kg p.o. s.i.d.). Oral plaques were not seen on monthly rechecks of the flock through November 2008. The proposed mechanism of transmission is exposure to wild birds but the source has not been determined. These cases of avian mycobacteriosis caused by MIT are the first known cases reported in little blue penguins.

Pharmacokinetics of florfenicol after a single intramuscular dose in white-spotted bamboo sharks (Chiloscyllium plagiosum).

This study evaluated the pharmacokinetics of florfenicol in the white-spotted bamboo shark (Chiloscyllium plagiosum). In addition to the pharmacokinetics, the potential application for treatment of bacterial meningitis was explored. A pilot study was used to compare doses of 30, 40, and 50 mg/kg i.m. Following that study, 10 adult sharks were administered a single i.m. dose of florfenicol at 40 mg/kg. Plasma and cerebrospinal fluid were collected and analyzed for florfenicol by a sensitive and specific high-pressure liquid chromatographic method. Pharmacokinetic analysis was performed using both non-compartmental and compartmental techniques. The absorption produced an average peak at 54 (+/-19) hr from the i.m. site of administration, and the half-life was prolonged, averaging 269.79 hr (+/-135.87). Florfenicol plasma concentrations peaked at an average of 11.85 microg/ml (+/-1.45) and were maintained above our target minimum inhibitory concentration of 4-8 microg/ml for at least 120 hr. Cerebrospinal fluid concentrations peaked at an estimated 9 microg/ml around 48 hr, surpassing the target minimum inhibitory concentration for at least 72 hr.

Xylazine-midazolam-ketamine versus medetomidine-midazolam-ketamine anesthesia in captive Siberian tigers (Panthera tigris altaica).

Two alpha2-adrenoceptor agents, xylazine and medetomidine, in combination with midazolam and ketamine safely and effectively immobilized Siberian tigers (Panthera tigris altaica). The medetomidine protocol used smaller drug volumes, and induction and recovery times were shorter. Although cardiopulmonary abnormalities were noted, none were likely to be life threatening.

Efficacy of porcine gonadotropins for repeated stimulation of ovarian activity for oocyte retrieval and in vitro embryo production and cryopreservation in Siberian tigers (Panthera tigris altaica).

A comparison of the amino acid sequences demonstrated that Siberian tiger gonadotropins are more homologous with those of porcine than any other commercially available preparation. The present study measured the efficacy of repeated ovarian stimulation with purified porcine gonadotropins on the follicular, hormonal, and immunogenic responses in Siberian tigers as well as on the ability of oocytes retrieved by laparoscopic follicular aspiration to fertilize and cleave in vitro. Controlled rate and vitrification cryopreservation methods were also compared for their ability to support ongoing cleavage following thawing of presumptive 2- to 4-cell tiger embryos generated in vitro. Vitrification supported continued embryonic cleavage in vitro while controlled rate freezing did not. Stereological microscopy indicated an excellent ovarian response with the recovery of quality cumulus-oocyte complexes that apparently fertilized and cleaved in vitro. However, ultrastructural and physiological examination revealed abnormal and unnatural responses such as the failure of some cumulus-oocyte complexes to reach maturity and progestagen levels to approach normalcy. At the same time, analyses of blood for antibodies failed to detect an immune reaction to these foreign gonadotropins in an assay that tested positive for the chorionic gonadotropin-stimulated domestic cat. Together, these observations suggest that porcine gonadotropins may be effective for the ovarian stimulation of tigers but that some modifications to administration protocols are needed to produce a more natural response.