DETECTION AND DIFFERENTIATION OF TRICHURIS IN GIRAFFE (GIRAFFA CAMELOPARDALIS) UNDER HUMAN CARE.

Trichuris spp. are nematode parasites infecting many species, including domestic and wild ruminants in zoological and wildlife parks worldwide. These nematodes cause significant morbidity in giraffes (Giraffa camelopardalis) and other hoofstock. Parasite transmission between ruminant species is well reported; however, relative to domestic species, little is known about Trichuris infections in giraffes under human care. We hypothesized that Trichuris spp. differ between individual giraffes in different US regions, suggesting giraffes are susceptible to Trichuris from other ruminant hosts. The study sites used to assess this hypothesis included The Wilds in Cumberland, Ohio; Fossil Rim Wildlife Center in Glen Rose, Texas; White Oak Conservation in Yulee, Florida; and Binder Park Zoo in Battle Creek, Michigan. Trichuris eggs were collected from the feces of 14 individual giraffes located at the four different study sites and from soil samples from the enclosures where Trichuris-positive giraffes were housed. The eggs were isolated and their genes were amplified by PCR and compared at the molecular level. Trichuris samples from four giraffe hosts and one soil site were sequenced and portions of the cox1 and 18S genes compared. This study found that >12 eggs per gram of fecal-derived Trichuris eggs must be present to amplify Trichuris-specific DNA. The Trichuris spp. found in the majority of giraffes in this study were most similar to T. ovis and T. discolor, and one giraffe sample had greater similarity to T. skrjabini and T. leporis.

SUCCESSFUL TREATMENT OF BATRACHOCHYTRIUM DENDROBATIDIS IN EASTERN HELLBENDERS (CRYPTOBRANCHUS ALLEGANIENSIS ALLEGANIENSIS) WITH TERBINAFINE.

In 2019, two wild-caught adult female eastern hellbenders (Cryptobranchus alleganiensis alleganiensis) received a preshipment examination and were individually swabbed for chytrid testing via quantitative polymerase chain reaction (qPCR). Physical examination was unremarkable. Both females tested positive for Batrachochytrium dendrobatidis (Bd) and negative for B. salamandrivorans (Bsal). A course of terbinafine hydrochloride 1% in alcohol was administered in a 0.005% treatment bath for 5 min once daily for 5 d. Both animals were individually retested 1, 3, and 4 wk after treatment using qPCR. All post-treatment samples were negative for Bd and Bsal. This report represents the first successful treatment with terbinafine hydrochloride 1% in alcohol to eliminate subclinical Bd infection in eastern hellbenders and underlines the importance of preshipment testing for chytrid in all amphibians being transferred to new facilities or released into the wild as a means to minimize risk of disease introduction via subclinically infected individuals.

Detection of Trichuris eggs in feces and soil from giraffe (Giraffa camelopardalis) and other hoofstock enclosures under human care in the USA.

Trichuris spp. are nematode parasites infecting wild ruminants in zoological institutions worldwide. These helminths cause significant morbidity in giraffe (Giraffa camelopardalis) and other hoofstock located in zoological institutions throughout the United States. Historically, studies and institutions have used a variety of nematode detection methods with various flotation solutions. Optimization of Trichuris egg detection is necessary for monitoring collections. Fecal and soil optimized protocols were generated in this study using samples containing Trichuris eggs from multiple semi free-ranging zoological institutions. First, Sheather's sugar (specific gravity (SG) 1.27), sucrose (SG 1.40), magnesium sulfate (SG 1.26), and zinc sulfate (SG 1.18) were compared as flotation solutions by quantitative eggs per gram using a modified Stoll method. Then a soil recovery method was optimized comparing Tween 20, sodium hydroxide, Dawn™ (Procter and Gamble) detergent, and sodium chloride as liberating solutions to free eggs from the soil. We found that Sheather's sugar and sucrose solutions were the most effective for Trichuris egg detection, and either sodium hydroxide or sodium chloride liberated eggs from soil.

MORBIDITY AND MORTALITY OF TAKIN (BUDORCAS TAXICOLOR) IN NORTH AMERICAN ZOOLOGICAL INSTITUTIONS FROM 1997 TO 2017.

Takin (Budorcus taxicolor) are classified as "Vulnerable" on the International Union for the Conservation of Nature Red List. Thus, ex situ conservation efforts provide assurance populations for future survival of this species. The objective of this study was to identify common causes of morbidity and mortality in takin populations in human care. Twenty North American institutions that housed takin from 1997 to 2017 completed a survey requesting medical and husbandry data. Data were examined broadly, by sex and age groups. There were 206 morbidity events (male = 133; female = 73) submitted across 102 takin (male = 62; female = 40). The most common causes of morbidity were infectious or inflammatory diseases (50%; 104/206), degenerative diseases (22%; 46/206), and traumatic events (17%; 34/206). Necropsy reports were provided for 42 takin that died during the study period. The most common causes of mortality were infectious or inflammatory diseases (26%; 11/42), traumatic events (24%; 10/42), and degenerative disease (12%; 5/42). Sixty-two percent of infectious or inflammatory diseases causing morbidity were associated with endoparasites (64/104). Degenerative joint diseases more commonly affected males (78%; 36/46) as well as forelimbs (48%; 22/46) when compared to hindlimbs (30%; 14/46) and unspecified limbs (22%; 10/46). The prevalence of trauma as a cause of morbidity and mortality was higher in neonate and juvenile takin groups combined (morbidity = 19%; mortality = 50%) as compared to adult and senior takin groups combined (morbidity = 15%; mortality = 11%). Older takin were euthanatized more often (57%; 16/28) than younger takin (29%; 4/14). Correlations between husbandry and health were difficult as a result of the inherent limitations of the survey. These data will inform takin-holding zoologic institutions and contribute to the successful management of takin in human care.

SAFETY OF AND HUMORAL IMMUNE RESPONSE TO THE MERIAL RECOMBITEK CANINE DISTEMPER VIRUS VACCINE IN MANED WOLVES (CHRYSOCYON BRACHYURUS).

This study evaluated the safety of and humoral response to the Merial Recombitek® recombinant canine distemper virus (rCDV) vaccine in maned wolves (n = 9, age 2-9 yr). All maned wolves had prior history of annual vaccination with the Merial Purevax® ferret rCDV vaccine. Serum neutralization (SN) to CDV was measured prior to initial vaccination with the rCDV Recombitek vaccine followed by a booster vaccination at 4-6 wk. Final SN titers were obtained at 13 wk post initial vaccination. The maned wolves developed no observable adverse side effects through the study. Pre-Recombitek vaccination SN titers ranged from negative to 1: 8. Postvaccination CDV titers ranged from negative to 1: 8, and were therefore below the range of that considered protective in domestic dogs.

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.

GASTRIC DILATATION VOLVULUS IN ADULT MANED WOLVES (CHRYSOCYON BRACHYURUS).

Gastric dilatation volvulus (GDV) was identified in six adult maned wolves ( Chrysocyon brachyurus ) housed at the Smithsonian Conservation Biology Institute in Front Royal, Virginia, and the Dickerson Park Zoo in Springfield, Missouri. Four individuals were found dead in their enclosures, and GDV was diagnosed postmortem based on radiographic and necropsy findings. Two individuals were diagnosed with GDV antemortem, with one wolf surviving following surgical intervention and supportive management. A concurrent splenic torsion was identified in three of six cases. Although GDV has been well documented in domestic dogs and is known to occur in maned wolves, objective, detailed case descriptions have been limited in this species. This report represents the first description of a surviving case of GDV in the maned wolf. Thorough species-specific documentation of any acutely fatal condition, such as GDV, is imperative for proper case recognition and medical management, with profound implications for species recovery efforts.

NOVEL RADIOGRAPHIC TECHNIQUE FOR PREGNANCY DETECTION IN THE MANED WOLF (CHRYSOCYON BRACHYURUS) WITHOUT ANESTHESIA.

Maned wolves ( Chrysocyon brachyurus ) maintained in ex situ populations challenge veterinarians and managers with high neonatal mortality and parental incompetence. These challenges led to the development of a novel diagnostic approach for pregnancy detection using radiographic imaging without anesthesia or sedation. To do this, a specialized crate was constructed to easily contain a single maned wolf, allowing the capture of lateral projection radiographic images of the abdomen prior to and throughout a 66-day pregnancy (days 20, 34, 48, and 55 of 66). Radiographs taken at days 48 and 55 postbreeding showed evidence of neonatal skeleton mineralization, confirming pregnancy with two pups. The dam gave birth at day 66 to two pups. This technical report describes a novel approach without anesthesia for successful radiographic pregnancy detection and determination of litter size in the maned wolf, a midsize carnivore, using a specially constructed crate.

ACUTE PHASE PROTEINS AS A MARKER OF RESPIRATORY INFLAMMATION IN PRZEWALSKI'S HORSE (EQUUS FERUS PRZEWALSKII).

Acute phase proteins are sensitive markers of inflammation, which are highly conserved across taxa. Although the utility of these proteins are becoming well defined in human and domestic animal medical fields, their role in nondomestic species remains unclear. In this communication, a 20-yr-old Przewalski's horse was presented for unresolving aspiration pneumonia, which cultured a unique Actinomyces-like bacteria. Despite waxing and waning clinical signs and minimal changes on baseline hematologic analysis, protein electrophoresis, serum amyloid A, and surfactant protein D serum concentrations showed changes that more accurately reflected the clinical severity of this case.

FATAL GASTRIC DILATION IN TWO ADULT BLACK-FOOTED FERRETS (MUSTELA NIGRIPES).

Acute gastric dilation resulting in death was identified in two adult black-footed ferrets (Mustela nigripes) housed at the Smithsonian Conservation Biology Institute in Front Royal, Virginia. Both individuals were adult males (3 and 5 yr) and previously clinically healthy prior to the event. The etiology of gastric dilation in both cases could not be definitively determined, and necropsy revealed severe cardiovascular compromise secondary to bloat. Limited literature is available regarding a syndrome of this type in adult black-footed ferrets. Differential diagnoses considered included gastric dilatation volvulus (GDV), severe gastric distention of unknown origin, and gastric outflow obstruction. Given the severity of this syndrome and the findings in these two cases, acute gastric dilation should be considered in black-footed ferrets presenting with acute abdominal distention, respiratory distress, and cardiovascular compromise.

Hidden in plain sight: Cryptic and endemic malaria parasites in North American white-tailed deer (Odocoileus virginianus).

Malaria parasites of the genus Plasmodium are diverse in mammal hosts, infecting five mammalian orders in the Old World, but were long considered absent from the diverse deer family (Cervidae) and from New World mammals. There was a description of a Plasmodium parasite infecting a single splenectomized white-tailed deer (WTD; Odocoileus virginianus) in 1967 but none have been reported since, which has proven a challenge to our understanding of malaria parasite biogeography. Using both microscopy and polymerase chain reaction, we screened a large sample of native and captive ungulate species from across the United States for malaria parasites. We found a surprisingly high prevalence (up to 25%) and extremely low parasitemia of Plasmodium parasites in WTD throughout the eastern United States. We did not detect infections in the other ungulate species nor in western WTD. We also isolated the parasites from the mosquito Anopheles punctipennis. Morphologically, the parasites resemble the parasite described in 1967, Plasmodium odocoilei. Our analysis of the cytochrome b gene revealed two divergent Plasmodium clades in WTD representative of species that likely diverged 2.3 to 6 million years ago, concurrent with the arrival of the WTD ancestor into North America across Beringia. Multigene phylogenetic analysis placed these clades within the larger malaria parasite clade. We document Plasmodium parasites to be common in WTD, endemic to the New World, and as the only known malaria parasites from deer (Cervidae). These findings reshape our knowledge of the phylogeography of the malaria parasites and suggest that other mammal taxa may harbor infection by endemic and occult malaria parasites.

Detection of a novel astrovirus from a black-naped monarch (Hypothymis azurea) in Cambodia.

BACKGROUND: Astroviruses are comprised of two genera with Avastrovirus infecting birds and Mamastrovirus infecting mammals. Avastroviruses have primarily been associated with infections of poultry, especially chicken, turkey, duck, and guineafowl production systems, but also infect wading birds and doves. Outcomes result in a spectrum of disease, ranging from asymptomatic shedding to gastroenteritis with diarrhea, stunting, failure to thrive and death. FINDINGS: Virological surveillance was conducted in birds from two sites in Cambodia in 2010. Samples were screened for influenza, astroviruses, coronaviruses, flaviviruses, and paramyxoviruses. A total of 199 birds were tested and an astrovirus was detected in a black-naped monarch (Hypothymis azurea). CONCLUSIONS: This is the first astrovirus detection in a passerine bird. Phylogenetic analysis and nucleotide distances suggest that this avastrovirus forms a distinct lineage and may constitute a fourth avastrovirus group.

A novel group of avian astroviruses in wild aquatic birds.

Using a pan-astrovirus reverse transcription-PCR assay, a great diversity of novel avastroviruses was detected from wild bird and poultry samples. Two groups of astroviruses detected from wild birds are genetically related or highly similar to previously known viruses in poultry. Most interestingly, a novel group of astroviruses was detected in wild aquatic birds. Our results also reveal that different groups of astroviruses might have difference host ranges. This study has expanded our understanding regarding avastrovirus ecology.

Amphibian pathogens in Southeast Asian frog trade.

Amphibian trade is known to facilitate the geographic spread of pathogens. Here we assess the health of amphibians traded in Southeast Asia for food or as pets, focusing on Batrachochytrium dendrobatidis (Bd), ranavirus and general clinical condition. Samples were collected from 2,389 individual animals at 51 sites in Lao PDR, Cambodia, Vietnam and Singapore for Bd screening, and 74 animals in Cambodia and Vietnam for ranavirus screening. Bd was found in one frog (n = 347) in Cambodia and 13 in Singapore (n = 419). No Bd was found in Lao PDR (n = 1,126) or Vietnam (n = 497), and no ranavirus was found in Cambodia (n = 70) or Vietnam (n = 4). Mild to severe dermatological lesions were observed in all East Asian bullfrogs Hoplobatrachus rugolosus (n = 497) sampled in farms in Vietnam. Histologic lesions consistent with sepsis were found within the lesions of three frogs and bacterial sepsis in two (n = 4); one had Gram-negative bacilli and one had acid-fast organisms consistent with mycobacterium sp. These results confirm that Bd is currently rare in amphibian trade in Southeast Asia. The presence of Mycobacterium-associated disease in farmed H. rugolosus is a cause for concern, as it may have public health implications and indicates the need for improved biosecurity in amphibian farming and trade.

Avian coronavirus in wild aquatic birds.

We detected a high prevalence (12.5%) of novel avian coronaviruses in aquatic wild birds. Phylogenetic analyses of these coronaviruses suggest that there is a diversity of gammacoronaviruses and deltacoronaviruses circulating in birds. Gammacoronaviruses were found predominantly in Anseriformes birds, whereas deltacoronaviruses could be detected in Ciconiiformes, Pelecaniformes, and Anseriformes birds in this study. We observed that there are frequent interspecies transmissions of gammacoronaviruses between duck species. In contrast, deltacoronaviruses may have more stringent host specificities. Our analysis of these avian viral and host mitochondrial DNA sequences also suggests that some, but not all, coronaviruses may have coevolved with birds from the same order.

Seroprevalence of retrovirus in North American captive macropodidae.

Laboratory records of serology results from captive macropodidae sampled between 1997 and 2005 were reviewed to assess the seroprevalence of retrovirus exposure. Serum samples from 269 individuals (136 males, 133 females) representing 10 species of macropods housed in 31 North American captive collections were analyzed for retrovirus antibody using an indirect immunofluorescent assay. The prevalence of positive antibody titers comparing male versus female, between species, between age groups, and among animals with identified parentage was examined by nonparametric statistical analyses. Median age of animals at time of sample collection was 36 mo (range 2-201 mo). Total percentage seropositive was 20.4%. Serum antibody was detected in 31 of 47 (66.0%) tammar wallaby (Macropus eugenii), nine of 24 (37.5%) yellow-footed rock wallaby (Petrogale xanthopus), four of 11 (36.4%) swamp wallaby (Wallabia bicolor), 10 of 80 (12.5%) red-necked wallaby (Macropus rufogriseus), and one of 54 (1.9%) parma wallaby (Macropus parma). No individuals of western gray kangaroo (n=3) (Macropus fuliginosus), eastern gray kangaroo (n=19) (Macropus giganteus), common wallaroo (n=6) (Macropus robustus), red kangaroo (n=11) (Macropus rufus), or Matschie's tree kangaroo (n=14) (Dendrolagus matschiei) were positive for retrovirus antibody. These results demonstrate that five species of captive macropods have a history of exposure to retrovirus, with the highest percentage seropositive and highest statistical correlation in M. eugenii (pair-wise Fisher's exact test, alpha = 0.05). Additionally, one wild-caught M. eugenii was confirmed seropositive during quarantine period, indicating that retrovirus exposure may exist in wild populations.

Inability to induce tympanic squamous metaplasia using organochlorine compounds in vitamin A-deficient red-eared sliders (Trachemys scripta elegans).

Previously, we reported that wild eastern box turtles (Terrapene carolina carolina) with aural abscesses contained higher body burdens of organochlorine (OC) compounds than those without the lesion. This lesion in captive chelonians is associated with turtles that are fed diets deficient in vitamin A. To examine the pathophysiology of this lesion and evaluate the relationship between OC burdens and vitamin A metabolism, we maintained red-eared sliders (Trachemys scripta elegans) under different conditions of OC exposure and dietary vitamin A concentrations from August 2005 to February 2006. Dietary vitamin A concentration (0 or 5 international units/g in the diet) and OC exposure (no OC compound or the mixture of 2 mg/kg chlordane, 0.25 mg/kg aroclor, and 1 mg/kg lindane) did not affect histologic score based on degree of squamous metaplasia of the tympanic epithelium or levels of plasma or liver vitamin A among the study groups. The results of this study suggest that 6 mo of exposure to the selected OC compounds, or similar duration of reduced dietary vitamin A concentrations do not influence the formation of squamous metaplasia and aural abscesses in red-eared sliders. Further studies are required to determine whether the duration of the experiment was insufficient, the OC compounds selected were inappropriate, the dosing was incorrect, and whether there are other unknown mechanisms causing the reported association between OC exposure and aural abscesses seen in eastern box turtles.

Induction and recovery characteristics and cardiopulmonary effects of sevoflurane and isoflurane in bald eagles.

OBJECTIVE: To compare induction and recovery characteristics and cardiopulmonary effects of isoflurane and sevoflurane in bald eagles. Animals-17 healthy adult bald eagles. PROCEDURES: Anesthesia was induced with isoflurane or sevoflurane delivered in oxygen via a facemask in a crossover design with 4 weeks between treatments. Eagles were intubated, allowed to breathe spontaneously, and instrumented for cardiopulmonary measurements. Time to induction, extubation, and recovery, as well as smoothness of recovery, were recorded. RESULTS: Administration of sevoflurane resulted in a significantly quicker recovery, compared with isoflurane. Temperature, heart rate, and respiratory rate significantly decreased over time, whereas systolic (SAP), diastolic (DAP), and mean arterial blood pressure (MAP) significantly increased over time with each treatment. Temperature, heart rate, SAP, DAP, and MAP were significantly higher with isoflurane. Blood pH significantly decreased, whereas PaCO(2) significantly increased over time with each treatment. Bicarbonate and total carbon dioxide concentrations significantly increased over time with each treatment; however, there was a significant time-treatment interaction. The PaO(2) and arterial oxygen saturation increased over time with isoflurane and decreased over time with sevoflurane with a significant time-treatment interaction. Six eagles developed cardiac arrhythmias with isoflurane, as did 4 with sevoflurane anesthesia. CONCLUSIONS AND CLINICAL RELEVANCE: Isoflurane and sevoflurane administration resulted in smooth, rapid induction of and recovery from anesthesia similar to other species. Isoflurane administration resulted in tachycardia, hypertension, and more arrhythmias, compared with sevoflurane. Sevoflurane was associated with fewer adverse effects and may be particularly beneficial in compromised bald eagles.

Disseminated histoplasmosis in an African pygmy hedgehog.

CASE DESCRIPTION: A 2-year-old captive-bred sexually intact female African pygmy hedgehog (Atelerix albiventris) was evaluated because of vague signs of illness including inappetence, weakness, lethargy, and weight loss over a 20-day period. CLINICAL FINDINGS: Abnormalities detected via initial clinicopathologic analyses included anemia, thrombocytopenia, leukopenia, hypoproteinemia, and hypoglycemia. Results of a fecal flotation test were negative. Three weeks after the initial evaluation, splenomegaly was detected via palpation and ultrasonography. TREATMENT AND OUTCOME: The hedgehog was treated with broad-spectrum antibacterial agents, resulting in an initially favorable response. Fenbendazole was also administered against possible occult parasitic infestation. After 3 weeks of illness, the hedgehog's condition had worsened and supportive care and administration of additional antibacterial agents were instituted. The hedgehog died, and pathologic examinations revealed severe splenomegaly; granulomatous infiltrates were evident in multiple organs, and Histoplasma capsulatum yeasts were detected intralesionally. CLINICAL RELEVANCE: Histoplasmosis can develop in a wide range of mammalian species. African pygmy hedgehogs are becoming increasingly popular as exotic pets, and vague signs of illness and splenomegaly are often attributed to hemolymphatic malignancies, which are somewhat common in this species. Practitioners should be aware that similar clinical signs may be associated with histoplasmosis in these animals. Although the hedgehog of this report was confined indoors, it originated from an area where histoplasmosis was endemic; this indicates that the disease should be included as a differential diagnosis for hedgehogs that develop vague signs of illness and are known to originate from such geographic regions.