Yersinia enterocolitica serotype O:9 has identical O-antigens to those of Brucella abortus and has apparently caused false-positive reactions in numerous brucellosis serologic tests in elk (Cervus canadensis) from southwest Montana. We investigated whether a similar phenomenon was occurring in brucellosis antibody–positive bison (Bison bison) using Y. enterocolitica culturing techniques and multiplex PCR of four diagnostic loci. Feces from 53 Yellowstone bison culled from the population and 113 free-roaming bison from throughout the Greater Yellowstone Ecosystem (GYE) were tested. Yersinia enterocolitica O:9 was not detected in any of 53 the bison samples collected at slaughter facilities or in any of the 113 fecal samples from free-ranging bison. One other Y. enterocolitica serotype was isolated; however, it is not known to cause cross-reaction on B. abortus serologic assays because it lacks the perosamine synthetase gene and thus the O-antigens. These findings suggest that Y. enterocolitica O:9 cross-reactivity with B. abortus antigens is unlikely to have been a cause of false-positive serology tests in GYE bison and that Y. enterocolitica prevalence was low in bison in the GYE during this study.

Wild yak (Bos mutus) are affected by a disorder known colloquially as “stiffness of extremities disease,” characterized by emaciation, lameness, stiffness in the gait, enlargement of the costochondral junctions, and abnormal curvature in the long bones. Results from preliminary epidemiologic and clinical observations suggested that this was a local, nutritional and metabolic disease associated with some mineral deficiency. Our objective was to determine the possible relationship between this disease and phosphorus (P) deficiency. We found that P concentrations in forage samples from affected areas were significantly lower than were those from unaffected areas, and the mean calcium:P ratio in the affected forage was 14:1. Phosphorus concentrations of blood, bone, teeth, and hair from affected yak were also significantly lower than were those from reference yak. Serum P levels of affected animals were much lower than were those of reference yak, whereas serum alkaline phosphatase levels were significantly higher than were those from reference yak. The P deficiency disease could be cured with supplement of disodium hydrogen phosphate (Na₂HPO₄). We conclude that the disease is mainly caused by P deficiency in forage.

Baseline reference ranges of serum chemistry and hematology data can be important indicators for the status of both individuals or populations of wild animals that are affected by emerging pathogens, toxicants, or other causes of disease. Frequently, reference ranges for these values are not available for wildlife species or subspecies. We present hematologic and serum chemistry reference ranges for moose (Alces alces) adults, yearlings, and calves in Norway sampled from 1992–2000. Additionally, we demonstrated that both induction time and chase time were correlated with initial rectal temperature, although they were not significantly correlated with cortisol, aspartate aminotransferase, glucose, or creatine kinase. Overall, the reference ranges given here are similar to those given for American moose, with a few differences that can be attributed to environment, testing methodology, or subspecies or species status. This is the first report, to our knowledge, of reference ranges for moose in Norway.

Hellbenders (n=97) were collected from the Little and Hiwassee Rivers in eastern Tennessee, USA, during 2009 and 2010. Location and morphometrics for each animal were recorded, and nonlethal tissue samples were collected to estimate the prevalence of infection with Batrachochytrium dendrobatidis (Bd) and Ranavirus in each watershed and year. Real-time polymerase chain reaction was performed on skin swabs for Bd and on tail clips for ranaviruses. Overall prevalences of DNA of Bd, Ranavirus, and coinfections (i.e., detectable DNA of both pathogens in the same individual) were 26%, 19%, and 5%, respectively. Differences in infection prevalence were detected between watersheds and years. Gross lesions were observed in 31 animals (32%), but the types of lesions were not consistent with chytridiomycosis or ranaviral disease. This is the first report of infection of eastern hellbenders with Bd and Ranavirus. Despite infection by both pathogens, it is unclear whether chytridiomycosis or ranaviral disease develops in wild populations of hellbenders. More research is needed to determine the susceptibility of hellbenders to Bd and ranaviruses and their role in the epidemiology of these pathogens.

The New Zealand Pigeon or kereru (Hemiphaga novaeseelandiae) frequently collides with windows and vehicles. In this study of 146 kereru collected from 1996 to 2009, we used 118 radiographs and 91 necropsies to determine skeletal and soft tissue injuries. Vehicle collisions resulted in more damage to the extremities (wing and femur), whereas collisions with windows resulted in trauma to the head, fractures/dislocations of the coracoids and clavicles, and ruptured internal organs. Soft tissue injuries included damage to the flight muscles and heart ruptures caused by fractured coracoid bones, as well as extensive bruising of pectoral muscles and hemorrhaging of the lungs. Rehabilitation time was not related to number of skeletal injuries sustained, nor was the time until death for those that did not survive. In general, kereru with greater numbers of injuries were less likely to survive rehabilitation. Flight speed and force calculations suggest that a 570-g kereru would collide with 3–70 times the force of smaller birds (5–180 g); this may explain the discrepancies between the injuries characterized here and those reported for North American passerines. The differences in injuries sustained from collisions with windows and cars can be used to inform rehabilitators about the possible nature of injuries if the source of impact is known.

Chytridiomycosis, an emerging infectious disease caused by the chytrid fungus Batrachochytrium dendrobatidis, threatens anuran populations worldwide. Effects of B. dendrobatidis on frog species are variable. Some species typically develop nonlethal infections and may function as carriers; others typically develop lethal infections that can lead to population declines. Nonlethal infections in the bullfrog (Lithobates catesbeianus) are well-documented. In contrast, recently metamorphosed wood frogs (L. sylvaticus) can die from chytridiomycosis. We conducted an ex-situ experiment between May and July 2010 to determine whether B. dendrobatidis-infected bullfrogs could transmit the fungus to wood frog tadpoles when the two species shared a body of water. We tested for B. dendrobatidis infections with quantitative polymerase chain reactions (qPCR) in a subsample of the wood frog tadpoles and in all metamorphosed wood frogs and compared risk of death of froglets exposed and unexposed to infected bullfrogs. We detected B. dendrobatidis sporadically in subsampled treatment tadpoles (nine of 90, 10%) and frequently in treatment froglets (112 of 113, 99.1%). Pooled risk of froglet death was higher (P<0.001) in treatment enclosures than in control enclosures. Our results indicate that, at the low infection loads bullfrogs tend to carry, swabbing for PCR analyses may underestimate prevalence of B. dendrobatidis in this species. We highlight bullfrog disease screening as a management challenge, especially in light of exotic bullfrog colonies on multiple continents and large-scale global trade in this species. We document the importance of quantifying lethal and sublethal effects of bullfrog vectors on B. dendrobatidis-susceptible species.

Hematologic and serum biochemical data collected must be interpreted by comparison with normal reference intervals generated from healthy animals, within a similar population, because many blood parameters are influenced by diet, environment, and stress. Species-specific reference intervals for the endangered island fox (Urocyon littoralis) are not available. We reviewed hematology and serum biochemistry panels from 280 island foxes sampled from 1999–2008 and established normal reference intervals from clinically healthy foxes using a nonparametric approach. Blood parameters were analyzed for differences in age, sex, island of origin, and captivity status. Alkaline phosphatase, alanine aminotransferase, aspartate aminotransferase, and creatine kinase activities, as well as calcium and phosphorus concentrations, were significantly higher in juveniles than in adults, but total protein and globulin concentration was lower for juveniles than for adults. Lymphocyte and eosinophil counts, and blood urea nitrogen (BUN) concentration, in foxes from the northern Channel islands of California, USA (Santa Cruz, Santa Rosa, and San Miguel) were higher when compared with foxes from Santa Catalina Island to the south. Higher lymphocyte and eosinophil numbers in the northern island foxes may be associated with increased levels of parasitism on the northern islands. Differences in BUN concentration in both free-ranging and captive foxes may reflect dietary differences among islands. Although aggressive conservation programs have been enacted, island foxes are still susceptible to infectious and neoplastic diseases and, potentially, to toxins. Island fox species-specific reference intervals will enable managers and veterinarians to better care for sick and injured foxes and will contribute to future population health monitoring.

We investigated the influence of detection probability (i.e., the probability of detecting the disease or organism of interest) on the repeatability of results reported from bacterial culture tests used to demonstrate the presence of species in the Pasteurellaceae family that infect bighorn sheep (Ovis canadensis). We also estimated occupancy probabilities (i.e., the probability an individual bighorn in a herd is infected) for each cultured biovariant and examined the effects of detection probability on the number of samples needed to detect the Pasteurellaceae biovariants from within an individual sheep as well as from within a herd. We collected 5–15 samples from free-ranging bighorns in Colorado, using oropharyngeal swabs or swabs of lungs, and submitted these swabs either immediately or after 2 days for bacterial culture. We saw significant variability in results for repeated samples from each of the sheep, and detection probabilities were ≤0.71 for all Pasteurellaceae biovariants cultured. The delayed (≥2 days) sample submission reduced both the microbial diversity detected and the detection probability for the biovariants characterized when compared to samples submitted immediately. Oropharyngeal sampling had higher detection probabilities of the individual biovariants than did lung swabs, and there was a difference in the biovariants detected between oropharyngeal and lung sampling. Depending on the biovariant of interest, estimates of occupancy probabilities ranged from 0.37–0.89, and we estimated that three to >30 swab samples were necessary to obtain a 95% confidence of detecting the cultured biovariants if they were present in an individual sheep. We estimated that the optimal sample sizes to detect the observed biovariants within a sheep herd with a 95% confidence ranged from sampling two bighorns twice to sampling 40 individuals once. Detection probability impacts the results reported from bacterial cultures for Pasteurellaceae in bighorn sheep, and confounding effects of the detection process should be addressed to improve the rigor of surveillance.

Klebsiella pneumoniae is a zoonotic, Gram-negative member of the family Enterobacteriaceae and is the causative agent of nosocomial septicemic, pneumonic, and urinary tract infections. Recently, pathogenic strains of K. pneumoniae sharing a hypermucoviscosity (HMV) phenotype have been attributed to multisystemic abscessation in both human and nonhuman primates. Although K. pneumoniae is a well-recognized zoonotic agent, there is a lack of general information including adequate diagnostic methods or treatments for nonhuman primates. In an effort to increase the body of knowledge of this enigmatic pathogen, K. pneumoniae isolates from African green monkeys (Chlorocebus aethiops sabaeus) on the island of St. Kitts, West Indies were genotypically and phenotypically characterized. Genetic fingerprints generated by PCR-mediated genomic fingerprinting, phenotypic characterization, and antimicrobial susceptibility all identified a high degree of similarity between the HMV and non-HMV K. pneumoniae isolates. The results obtained from this work will help establish a baseline for the development of efficacious diagnostic methods and treatment strategies for both human and nonhuman primates.

Porcine circovirus type 2 (PCV2) is a widespread, important pathogen of domestic swine and the causative agent of postweaning multisystemic wasting syndrome and other diseases and conditions referred to as “porcine circovirus diseases.” Specific antibodies and DNA to PCV2 have also been detected in European wild boars and North American feral pigs. We collected 312 tonsil samples from wild boars shot in 13 of 16 districts of Poland, and tested them for PCV2 DNA using a real-time PCR. We detected PCV2 DNA in 75.6% of tested tonsils, and in particular, in 60% of samples from the 2006–07 season, and 91% from 2007–08. The phylogenetic analysis that included 12 PCV2 sequences from wild boars revealed that they belonged to two genetic clusters, PCV2b and PCV2a. We present data on prevalence of PCV2 in Polish wild boars and for the first time report the PCV2a genotype in Poland.

Mycoplasma conjunctivae, the causative agent of infectious keratoconjunctivitis (IKC), was recently detected in asymptomatic Alpine ibex (Capra ibex ibex). This suggested that an external source of infection may not be required for an IKC outbreak in wildlife but might be initiated by healthy carriers, which contradicted previous serologic investigations in chamois. Our aims were to 1) assess the prevalence of M. conjunctivae among asymptomatic ibex and Alpine chamois (Rupicapra rupicapra rupicapra) and its frequency in IKC-affected animals, 2) determine mycoplasma loads in different disease stages, and 3) characterize the M. conjunctivae strains involved. Eye swabs from 654 asymptomatic and 204 symptomatic animals were collected in diverse Swiss regions between 2008 and 2010, and tested by TaqMan real-time PCR. Data analysis was performed considering various patterns of IKC occurrence in the respective sampling regions. Strains from 24 animals were compared by cluster analysis. Prevalence of M. conjunctivae was 5.6% (95% confidence interval [CI]: 3.7–8.1%) in asymptomatic ibex and 5.8% (CI: 3.0–9.9%) in asymptomatic chamois, with significant differences between years and regions in both species. Detection frequency in symptomatic animals was significantly higher during IKC outbreaks than in nonepidemic situations (i.e., regular but low incidence or sporadic occurrence). Mycoplasma load was significantly lower in eyes from healthy carriers and animals with mild signs than from animals with moderate and severe signs. Although some strains were found in both asymptomatic and diseased animals of the same species, others apparently differed in their pathogenic potential depending on the infected species. Overall, we found a widespread occurrence of M. conjunctivae in wild Caprinae with and without IKC signs. Our results confirm the central role of M. conjunctivae in outbreaks but suggest that other infectious agents may be involved in IKC cases in nonepidemic situations. Additionally, presence and severity of signs are related to the quantity of M. conjunctivae in the eyes rather than to the strain. We propose that individual or environmental factors influence the clinical expression of the disease and that persistence of M. conjunctivae in populations of wild Caprinae cannot be excluded.

Information on health parameters, such as antibody prevalences and serum chemistry that can reveal exposure to pathogens, disease, and abnormal physiologic conditions, is scarce for Antarctic seal species. Serum samples from Antarctic fur seals (Arctocephalus gazella, n=88) from Bouvetøya (2000–2001 and 2001–2002), and from Weddell seals (Leptonychotes weddellii, n=20), Ross seals (Ommatophoca rossii, n=20), and crabeater seals (Lobodon carcinophagus, n=9) from the pack-ice off Queen Maud Land, Antarctica (2001) were analyzed for enzyme activity, and concentrations of protein, metabolites, minerals, and cortisol. Adult Antarctic fur seal males had elevated levels of total protein (range 64–99g/l) compared to adult females and pups (range 52–79 g/l). Antarctic fur seals had higher enzyme activities of creatine kinase, lactate dehydrogenase, and amylase, compared to Weddell, Ross, and crabeater seals. Antibodies against Brucella spp. were detected in Weddell seals (37%), Ross seals (5%), and crabeater seals (11%), but not in Antarctic fur seals. Antibodies against phocine herpesvirus 1 were detected in all species examined (Antarctic fur seals, 58%; Weddell seals, 100%; Ross seals, 15%; and crabeater seals, 44%). No antibodies against Trichinella spp., Toxoplasma, or phocine distemper virus (PDV) were detected (Antarctic fur seals were not tested for PDV antibodies). Antarctic seals are challenged by reduced sea ice and increasing temperatures due to climate change, and increased anthropogenic activity can introduce new pathogens to these vulnerable ecosystems and represent a threat for these animals. Our data provide a baseline for future monitoring of health parameters of these Antarctic seal species, for tracking the impact of environmental, climatic, and anthropogenic changes in Antarctica over time.

Surveillance is critical for identifying and monitoring pathogen activity in wildlife populations, but often is cost- and time-prohibitive and logistically challenging. We tested the hypothesis that wildlife rehabilitation centers are useful for monitoring pathogen activity using West Nile virus (WNV) as a case study. We hypothesized that birds submitted to wildlife rehabilitation centers would have a similar prevalence of antibody to WNV as free-ranging birds. From 2008 to 2010, we collected sera from peridomestic birds submitted to the Wildlife Care Clinic (WCC), a wildlife rehabilitation center in central Iowa, and tested them for antibodies to WNV. We also collected and tested sera from free-ranging peridomestic birds in the area from which approximately 50% of WCC submissions historically originated. Prevalences of WNV antibodies in free-ranging birds and in peridomestic WCC birds were 2.3% (44/1,936) and 2.8% (2/72), respectively. However, none of the birds submitted to the WCC from the area where we captured free-ranging birds had antibodies (0/29). Our results indicate that rehabilitation facilities are not likely to be useful for monitoring WNV activity at small spatial scales or over short-time periods due to the low endemic prevalence of WNV, and low and variable submission rates. However, at larger spatial scales (ca. nine Iowa counties) WNV antibody prevalence in peridomestic birds submitted to the WCC was similar to that of free-ranging birds. Although limitations to using rehabilitation birds to monitor WNV must be considered, testing these birds could be useful for monitoring WNV activity regionally, especially with many states limiting surveillance due to budgetary constraints.

Marine mammals are at risk for infection by fecal-associated zoonotic pathogens when they swim and feed in polluted nearshore marine waters. Because of their tendency to consume 25–30% of their body weight per day in coastal filter-feeding invertebrates, southern sea otters (Enhydra lutris nereis) can act as sentinels of marine ecosystem health in California. Feces from domestic and wildlife species were tested to determine prevalence, potential virulence, and diversity of selected opportunistic enteric bacterial pathogens in the Monterey Bay region. We hypothesized that if sea otters are sentinels of coastal health, and fecal pollution flows from land to sea, then sea otters and terrestrial animals might share the same enteric bacterial species and strains. Twenty-eight percent of fecal samples tested during 2007–2010 were positive for one or more potential pathogens. Campylobacter spp. were isolated most frequently, with an overall prevalence of 11%, followed by Vibrio cholerae (9%), Salmonella spp. (6%), V. parahaemolyticus (5%), and V. alginolyticus (3%). Sea otters were found positive for all target bacteria, exhibiting similar prevalences for Campylobacter and Salmonella spp. but greater prevalences for Vibrio spp. when compared to terrestrial animals. Fifteen Salmonella serotypes were detected, 11 of which were isolated from opossums. This is the first report of sea otter infection by S. enterica Heidelberg, a serotype also associated with human clinical disease. Similar strains of S. enterica Typhimurium were identified in otters, opossums, and gulls, suggesting the possibility of land-sea transfer of enteric bacterial pathogens from terrestrial sources to sea otters.

Migratory (particularly aquatic) birds are the major natural reservoirs for type A influenza viruses. However, their role in transmitting highly pathogenic avian influenza (HPAI) viruses is unclear. Egypt is a “funnel” zone of wild bird migration pathways from Central Asia and Europe to Eastern and Central Africa ending in South Africa. We sought to detect and isolate avian influenza viruses in migratory birds in Egypt. During September 2003–February 2009, the US Naval Medical Research Unit Number 3, Cairo, Egypt, in collaboration with the Egyptian Ministry of Environment, obtained cloacal swabs from 7,894 migratory birds captured or shot by hunters in different geographic areas in Egypt. Samples were processed by real-time reverse transcriptase PCR for detection of the influenza A matrix gene. Positive samples were processed for virus isolation in specific-pathogen-free embryonated eggs and isolates were subtyped by PCR and partial sequencing. Ninety-five species of birds were collected. Predominant species were Green-Winged Teal (Anas carolinensis; 32.0%, n=2,528), Northern Shoveler (Anas clypeata; 21.4%, n=1,686), and Northern Pintail (Anas acuta; 11.1%, n=877). Of the 7,894 samples, 745 (9.4%) were positive for the influenza A matrix gene (mainly from the above predominant species). Thirteen of the 745 (1.7%) were H5-positive by PCR (11 were low-pathogenic avian influenza and two were HPAI H5N1). The prevalences of influenza A was among regions were 10–15%, except in Middle Egypt (4%). Thirty-nine influenza isolates were obtained from PCR-positive samples. Seventeen subtypes of avian influenza viruses (including H5N1 and H7N7) were classified from 39 isolates using PCR and partial sequencing. Only one HPAI H5N1 was isolated in February 2006, from a wild resident Great Egret (Ardea alba). No major die-offs or sick migratory birds were detected during the study. We identified avian influenza virus subtypes not previously reported in Egypt. The HPAI H5N1 isolated or detected indicates that migratory birds may play a role in the dispersal of HPAI virus, but a detailed mechanism of this role needs to be elucidated.

During the fall of 2006, in Israel, epizootic hemorrhagic disease virus (EHDV) serotype 7 caused an intense and widespread epizootic in domestic cattle that resulted in significant economic losses for the dairy industry. The susceptibility of potential North American vector and ruminant hosts to infection with EHDV-7 is not known but is essential to understanding the potential for establishment of this exotic orbivirus in North America if it were introduced. Our primary objective was to determine whether white-tailed deer (WTD; Odocoileus virginianus) are susceptible to infection with EHDV-7. Six, 8-mo-old WTD were experimentally infected with EHDV-7, and all became infected and exhibited varying degrees of clinical disease. Clinical signs, clinicopathologic abnormalities, and postmortem findings were consistent with previous reports of orbiviral hemorrhagic disease (HD) in this species. Four of six animals died or were euthanized because of the severity of disease, one on postinoculation day (PID) 5 and the remaining WTD on PID 7. All deer had detectable viremia on PID 3, which peaked on PID 5 or 6 and persisted for as long as PID 46 in one animal. Deer surviving the acute phase of the disease seroconverted by PID 10. Based on the 67% mortality rate we observed, this strain of EHDV-7 is virulent in WTD, reaffirming their role as a sentinel species for the detection of endemic and nonendemic EHDV. Further, the observed disease was indistinguishable from previous reports of disease caused by North American EHDV and bluetongue virus serotypes, highlighting the importance of serotype-specific diagnostics during suspected HD outbreaks.

We recently documented an epizootic of beak deformities in more than 2,000 Black-capped Chickadees (Poecile atricapillus) and other wild bird species in North America. This emerging avian disease, which has been termed avian keratin disorder, results in gross overgrowth of the rhamphotheca, the outer, keratinized layer of the beak. To test the hypothesis that the beak deformities characteristic of this disorder are associated with accelerated keratin production, we measured rates of beak growth and wear in affected Black-capped Chickadees (n=16) and a control sample of unaffected chickadees (n=14) collected from south-central (61°09′–61°38′N, 149°11′–149°48′W) and interior Alaska (64°51′–64°53′N, 147°49′–147°59′W). Rates of absolute growth were 50–100% higher in affected birds than they were in control birds and exceeded records from other passerine species. These results suggest that abnormally rapid epidermal growth is the primary physical mechanism by which beak deformities develop and are maintained in affected chickadees. Although beak overgrowth typically worsened over time, differential patterns of wear influenced the severity and morphology of deformities. In some cases, the effects of accelerated keratin growth were partially mitigated by frequent breakage of rhamphothecal tips. However, mortalities occurred in 9 of 16 birds (56%) with beak deformities during the study, suggesting that avian keratin disorder results in severe health consequences for affected birds. Additional study of factors that control beak keratin production is needed to understand the pathogenesis of this debilitating disease in wild birds.

Common Quail (Coturnix coturnix) were subjected to controlled and replicated experiments in the summer of 2008 to investigate the effects of short-term dehydration on cholinesterase activity in brain and plasma and the interaction between dehydration and exposure to the organophosphorus pesticide dicrotophos in these same tissues. Our objective was to determine if dehydration could confound the diagnosis of anticholinesterase exposure using inhibition of cholinesterase activity in quail tissues. The effect of dehydration was quantified using measures of plasma osmolality and hematocrit. Dicrotophos exposure caused significant inhibition of cholinesterase activity in brain, while the effects of dehydration and interaction were not significant. Dehydration caused significant duration-dependent increases in plasma osmolality and hematocrit. Dehydration also caused a significant increase in plasma cholinesterase activity. Variation in the change in plasma cholinesterase activity in response to dehydration was significantly and positively correlated with dehydration-induced variation in both the change in plasma osmolality and the change in hematocrit. These correlations suggest that plasma cholinesterase activity in quail is not limited to plasma but occupies some larger pool of the extracellular fluid volume, and we suggest lymph is part of that pool. The effects of dehydration on plasma cholinesterase activity masked the inhibitory effects of dicrotophos. Here, the combination of dehydration and dicrotophos exposure produced plasma cholinesterase activity that was not significantly different from reference and pre-exposure values, confounding the diagnosis of anticholinesterase exposure in dehydrated, dicrotophos-exposed quail. A method to adjust plasma cholinesterase activities for the confounding effects of dehydration and enable the diagnosis of anticholinesterase exposure in dehydrated, dicrotophos-exposed quail was developed. Clinicians and practitioners responsible for the diagnosis of anticholinesterase exposure in birds are cautioned that dehydration, commonly observed in sick wildlife, may mask the effect of anticholinesterases on plasma cholinesterase activity.

African savannah elephants (Loxodonta africana) are an ecologically and economically important species in many African habitats. However, despite the importance of elephants, research on their parasites is limited, especially in wild populations. Currently, we lack genetic tools to identify elephant parasites. We present genetic markers from ribosomal DNA (rDNA) and mitochondrial DNA (mtDNA) to identify five elephant-specific nematode parasites in the family Strongylidae: Murshidia linstowi, Murshidia longicaudata, Murshidia africana, Quilonia africana, and Khalilia sameera. We collected adult nematodes from feces deposited by wild elephants living in Amboseli National Park, Kenya. Using both morphologic and genetic techniques, we found that the internal transcribed spacer (ITS) region in rDNA provides a reliable marker to distinguish these species of strongyles. We found no evidence for cryptic genetic species within these morphologic species according to the cox-1 region of mtDNA. Levels of genetic diversity in strongyles from elephants were consistent with the genetic diversity seen within other strongyle species. We anticipate that these results will be a useful tool for identifying gastrointestinal nematode parasites in elephants.

Fecal analysis is commonly used to estimate prevalence and intensity of intestinal helminths in wild carnivores, but few studies have assessed the reliability of fecal flotation compared to analysis of intestinal tracts. We investigated sensitivity of the double centrifugation sugar fecal flotation and kappa agreement between fecal flotation and postmortem examination of intestines for helminths of coyotes (Canis latrans). We analyzed 57 coyote carcasses that were collected between October 2010 and March 2011 in the metropolitan area of Calgary and Edmonton, Alberta, Canada. Before analyses, intestines and feces were frozen at 280 C for 72 hr to inactivate Echinococcus eggs, protecting operators from potential exposure. Five species of helminths were found by postmortem examination, including Toxascaris leonina, Uncinaria stenocephala, Ancylostoma caninum, Taenia sp., and Echinococcus multilocularis. Sensitivity of fecal flotation was high (0.84) for detection of T. leonina but low for Taenia sp. (0.27), E. multilocularis (0.46), and U. stenocephala (0.00). Good kappa agreement between techniques was observed only for T. leonina (0.64), for which we detected also a significant correlation between adult female parasite intensity and fecal egg counts (R_s=0.53, P=0.01). Differences in sensitivity may be related to parasite characteristics that affect recovery of eggs on flotation. Fecal parasitologic analyses are highly applicable to study the disease ecology of urban carnivores, and they often provide important information on environmental contamination and potential of zoonotic risks. However, fecal-based parasitologic surveys should first assess the sensitivity of the techniques to understand their biases and limitations.

Dorsal-spined protostrongylid nematode larvae (Metastrongyloidea: Protostrongylidae) were recovered from the feces of the endangered pampas deer (Ozotocerus bezoarticus celer) in Campos del Tuyú Wildlife Reserve, Bahia Samborombón, Argentina. Partial DNA sequences from the large subunit ribosomal RNA (LSU rRNA) gene and from the second internal transcribed spacer region (ITS2) were amplified, cloned, sequenced, and compared to those of other nematodes. Nucleotide alignment and phylogenetic analysis of the sequences indicate that this protostrongylid nematode is most closely related to Parelaphostrongylus spp. as inferred from the LSU rRNA sequence analysis. Analysis of the ITS2 spacer indicated that the pampas deer protostrongylid is nested in a clade containing Parelaphostrongylus and Elaphostrongylus spp. These sequences differed considerably from those of other protostrongylid nematodes, and were most similar to those of Parelaphostrongylus spp. and Elaphostrongylus spp. in spite of clear variability from both genera. These results suggest that the protostrongylid from pampas deer is an undescribed nematode that likely belongs in the subfamily Elaphostrongylinae.

The objective of this study was to establish a standardized protocol to monitor Besnoitia tarandi prevalence and intensity in barren-ground caribou (Rangifer tarandus) herds by: 1) calculating the relative sensitivity and specificity of the gross visual assessment of four anatomical sites compared with microscopic evaluation, and 2) determining which of four anatomical sampling sites was the most sensitive for detecting B. tarandi cysts by microscopy. Sampled tissues consisted of the conjunctiva of the left eye and skin sections from the rostrum, metatarsus, and thigh from 312 harvested caribou. Diagnosis of infection with B. tarandi was based on observation of at least one cyst by microscopic examination. For each tissue, the maximal density of cysts (number of B. tarandi cysts/mm² in the section examined) was calculated for a measured area consisting of the dermis extending from the epidermis of the skin to the base of the hair follicles and adnexal structures. For the conjunctiva, the entire submucosa was evaluated. Gross visual evaluation markedly underestimated B. tarandi prevalence in caribou with a relative sensitivity ranging from 0.29 in the conjunctiva to 0.13 in the skin section from the thigh, whereas relative specificities ranged from 0.98 to 1.00. The metatarsus and rostrum skin sections had the highest probabilities of cyst detection of all four anatomical sampling sites. The metatarsus harbored significantly higher densities of B. tarandi cysts than the rostrum, thigh, or conjunctiva. In conclusion, microscopic evaluation of a skin section from the anterior aspect of the mid-third portion of the metatarsal region could be used as a standardized comparative indicator of density of B. tarandi infection in Rangifer.

Field immobilization of captive antelope may be required for medical examination, blood sample collection, and animal identification. The aim of this study was to evaluate the effects of a combination of butorphanol, detomidine, and midazolam (BDM) and its partial reversibility in Nile lechwe antelope (Kobus megaceros). Nine captive lechwes, weighing 28–64 kg, were immobilized, in February 2011, with butorphanol 0.20±0.05 (mean±SD) mg/kg, detomidine 0.20±0.05 mg/kg, and midazolam 0.31±0.08 mg/kg administered intramuscularly (IM) with a blowpipe. Physiologic parameters and depth of anesthesia were recorded when the animals became recumbent at 19.55±8.36 min after darting (T0) and after 10 (T10), 20 (T20), and 30 (T30) min. An arterial blood sample was collected at T20. At the end of the procedures, immobilization was partially reversed with atipamezole 0.25 mg/kg IM. Quality of induction, immobilization, and recovery was scored. The BDM combination induced immobilization and lateral recumbency in 13.44±5.61 min. Median induction score (scored 1 [excellent] to 4 [poor]) was 1 (range 1–2). Heart rate varied 40–104 beats/min, respiratory rate 16–108 breaths/min, and rectal temperature 36.5–40.3 C. Hyperthermia was observed and rapidly treated in three animals that demonstrated insufficient immobilization after darting. Arterial blood gas analyses revealed a mean pH of 7.43±0.07, partial arterial pressure of CO₂ of 44.1±6.0 mmHg, partial arterial pressureof O₂ of 74.0±13.5 mmHg, and an arterial O₂ saturation of 94.77±3.96%. Recovery was smooth and animals were walking in 13.44±7.85 min. Median recovery score (1 = excellent to 4 = poor) was 1 (range 1–2). The BDM was effective in immobilizing captive healthy lechwes with minimal cardiorespiratory changes.

Following field observations of wild Agassiz's desert tortoises (Gopherus agassizii) with oral lesions similar to those seen in captive tortoises with herpesvirus infection, we measured the prevalence of antibodies to Testudinid herpesvirus (TeHV) 3 in wild populations of desert tortoises in California. The survey revealed 30.9% antibody prevalence. In 2009 and 2010, two wild adult male desert tortoises, with gross lesions consistent with trauma and puncture wounds, respectively, were necropsied. Tortoise 1 was from the central Mojave Desert and tortoise 2 was from the northeastern Mojave Desert. We extracted DNA from the tongue of tortoise 1 and from the tongue and nasal mucosa of tortoise 2. Sequencing of polymerase chain reaction products of the herpesviral DNA-dependent DNA polymerase gene and the UL39 gene respectively showed 100% nucleotide identity with TeHV2, which was previously detected in an ill captive desert tortoise in California. Although several cases of herpesvirus infection have been described in captive desert tortoises, our findings represent the first conclusive molecular evidence of TeHV2 infection in wild desert tortoises. The serologic findings support cross-reactivity between TeHV2 and TeHV3. Further studies to determine the ecology, prevalence, and clinical significance of this virus in tortoise populations are needed.

While it has been demonstrated that persistent bovine viral diarrhea virus (BVDV) infections can be established in white-tailed deer (Odocoileus virginianus) following in utero exposure in the first trimester of gestation, there is little to no information regarding the outcome of infection in later stages of pregnancy in deer. Our goal was to observe the impact of infection of white-tailed deer in the second and third trimesters of pregnancy. Five white-tailed deer in the second trimester of pregnancy and four in the third trimester were infected with a BVDV type 2 virus previously isolated from a BVDV-infected deer harvested from the wild. Infection of deer in the second trimester of pregnancy resulted in loss of the pregnancy in three of five deer. Fawns born to the two remaining deer appeared normal and were born BVDV antigen–negative with neutralizing serum antibodies against BVDV. Infection of does in the third trimester of pregnancy did not result in fetal death or persistent infection and all does gave birth to live, healthy fawns that were BVDV antigen-negative and born with antibodies against BVDV. These results, combined with those previously reported regarding BVDV infection in the first trimester of pregnancy, suggest that the impact of BVDV infection of pregnant white-tailed deer is very similar to that observed in pregnant cattle.

Short-acting neuroleptic drugs are used to prevent adverse effects of stress in wildlife. We compared the effect of aceproma-zine and haloperidol in Iberian ibex (Capra pyrenaica) captured with box-traps. We captured 23 male Iberian ibex at the National Game Reserve of Ports de Tortosa i Beseit, northeastern Spain, March 2003–June 2005. Seven animals received 0.1 mg/kg of acepromazine maleate, eight received 0.33 mg/kg of haloperidol and eight animals acted as controls. Clinical, hematologic, and serum biochemical parameters were analyzed. Both treatments decreased rectal temperature, white blood cells, lymphocytes, and concentrations of creatinine, creatine kinase, and lactate dehydrogenase. Acepromazine also decreased red blood cells, packed cell volume, hemoglobin concentration, neutrophils, and concentrations of glucose and cholesterol. Haloperidol also decreased heart rate and concentrations of urea and potassium. Our results demonstrate the suitability of using acepromazine and haloperidol in capture operations to reduce stress and prevent its adverse effects.

Wild waterbirds sampled July 2006–September 2009 in Mongolia were tested for antibodies to avian influenza (AI) virus with the use of a commercially available blocking enzyme-linked immunosorbent assay. Antibodies were detected in 25% (572/2,282) of tested birds representing 26 species, and all antibody-positive samples were from 12 species in the orders Anseriformes and Charadriiformes. The highest antibody prevalence was in Ruddy Shelducks (Tadorna ferruginea; 61.7%; n=261; 95% confidence interval [CI] 55.8–67.6%), Whooper Swans (Cygnus cygnus; 38.4%; n=242; 95% CI 32.3–44.5%), Swan Geese (Anser cygnoides; 15%; n=127; 95% CI 8.6–21.4%), Bar-headed Geese (Anser indicus; 13%; n=738; 95% CI 10.3–15.1%), and Mongolian Gulls (Larus mongolicus; 3.9%; n=255; 95% CI 1.3–6.5%). There was no significant temporal or spatial variation in the presence of antibodies in the sampled species. However, Bar-headed Geese and Mongolian Gulls showed spatial variation in antibody prevalence in 2007 and 2008, respectively. Our study provides insights into the hatch year waterbirds' exposure to AI virus at their natal and molting sites in Mongolia.

A free-ranging, adult, male Persian leopard (Panthera pardus ciscaucasica) was found at Geloul-Sarani protected zone, province of North-Khorasan, Iran and transported to the Ferdowsi University of Mashhad Veterinary Teaching Hospital. The leopard had normal temperature and respiratory and cardiac frequency, but was significantly dehydrated and had elevated capillary perfusion. The animal also was cachectic, with pale mucus membranes, third-eyelid protrusion, and bilaterally enlarged submandibular lymph nodes. The leopard was stabilized by intensive fluid and electrolyte therapy and hospitalized. In 2 days, the leopard had improved clinically but had severe ataxia and head pressing. Blood smears revealed gamonts of Hepatozoon sp. within some neutrophils. Hematologic and plasma chemistry abnormalities included moderate anemia, leukocytosis, hypocholestrolemia, and hypophosphatemia. In radiographic evaluations, no sign of periosteal reactions or new bone formation was seen on the skull, spine, long bones, pelvis, or vertebrae. The leopard was treated successfully with Tazocin and clindamycin for 1 mo. This is the first detection of a Hepatozoon sp. in wild Felidae in Iran. Because most Iranian wild felids and canids are endangered, knowing whether Hepatozoon infection represents a threat for these animals is important.

We administered a suite of treatments to a herd of Rocky Mountain bighorn sheep (Ovis canadensis canadensis) that was experiencing poor lamb recruitment and showing signs of respiratory disease. Despite 3 yr of treatment with various combinations of anthelmentics, antibiotics, vaccines, and hyperimmune serum products, recruitment was not improved.

Wild animal meat represents an important source of protein for many people in central Africa. Also known as bushmeat, this meat commodity is derived from wild animals hunted under uncontrolled conditions, transported to distant markets under rudimentary or no hygienic methods, and often eviscerated >24 hr after death. Considering the plausible role of wildlife as a reservoir for bacterial zoonotic pathogens, bushmeat may be an important public health risk in Central Africa. This cross-sectional survey served to evaluate the presence of Campylobacter, Salmonella, and Shigella in the muscle tissue of 128 wild animal carcasses from several hunted wildlife species (guenons [Cercopithecus spp.], collared mangabeys [Cercocebus torquatus], gray-cheeked mangabeys [Lophocebus albigena], African crested porcupines [Atherurus africanus], duikers [Cephalophus spp.], and red river hogs [Potamocherus porcus]) sold in two markets of Port-Gentil, Gabon, in July and August 2010. Salmonella was detected from one carcass; no Campylobacter or Shigella was detected. If Campylobacter and Shigella were present, the maximum expected prevalence was estimated at 6% and 1%, respectively. In light of such very low apparent muscle contamination levels, bushmeat likely does not represent a health risk per se with respect to Campylobacter, Salmonella, or Shigella. However, because carcass evisceration and skinning can take place within households prior to consumption, consumers should follow strict hygiene and food safety practices to avoid potential health hazards associated with the handling, preparation, or consumption of bushmeat.

Avian pox is an enveloped double-stranded DNA virus that is mechanically transmitted via arthropod vectors or mucosal membrane contact with infectious particles or birds. Magellanic Penguins (Spheniscus magellanicus) from two colonies (Punta Tombo and Cabo Dos Bahías) in Argentina showed sporadic, nonepidemic signs of avian pox during five and two of 29 breeding seasons (1982– 2010), respectively. In Magellanic Penguins, avian pox expresses externally as wart-like lesions around the beak, flippers, cloaca, feet, and eyes. Fleas (Parapsyllus longicornis) are the most likely arthropod vectors at these colonies. Three chicks with cutaneous pox-like lesions were positive for Avipoxvirus and revealed phylogenetic proximity with an Avipoxvirus found in Black-browed Albatross (Thalassarche melanophrys) from the Falkland Islands in 1987. This proximity suggests a long-term circulation of seabird Avipoxviruses in the southwest Atlantic. Avian pox outbreaks in these colonies primarily affected chicks, often resulted in death, and were not associated with handling, rainfall, or temperature.

Leiomyosarcoma with associated retrovirus were found in North America for the first time in adult Atlantic salmon (Salmo salar) held in a quarantine facility at the North Attleboro National Fish Hatchery (NANFH), Massachusetts, USA. The fish had been collected as age 1–2 yr animals from the Pleasant River, Maine, and were to be used as brood stock in a population augmentation program for that river. Neoplastic disease was observed at NANFH initially in older (age 4 yr) fish, followed by age 3 yr fish. Disease was not observed in age 2 yr fish. The mortality pattern was chronic.

Eastern box turtle (Terrapene carolina carolina) populations are in decline, likely due to anthropogenic forces and disease, necessitating hematologic and biochemical data from healthy individuals for evaluation of wild populations. We repeatedly sampled 21 free-ranging eastern box turtles from May to September 2009 in the spring, summer, and fall to establish temporal hematologic and biochemical reference intervals. Packed cell volume, aspartate aminotransferase, and potassium levels declined significantly as the active season progressed. High levels of albumin, globulin, and calcium coincided with the presence of eggs in females. These reference intervals should provide baseline data for the clinical evaluation of wild box turtles presented for veterinary care or for studies of wild populations.

We isolated Corynebacterium pseudotuberculosis from an abscess of the head of a captive elk submitted for necropsy and from a similar abscess of a living herd mate. To our knowledge, this is the first documented case in elk and should be considered a potential cause of subcutaneous abscesses in wild elk.

Limited information exists pertaining to the occurrence of uterine prolapse in free-ranging white-tailed deer and mule deer. We report a case of uterine prolapse in a free-ranging white-tailed deer (Odocoileus virginianus) in northeastern South Dakota and a free-ranging mule deer (Odocoileus hemionus) in central South Dakota.

Cathaemasia hians is a known parasite of storks (Ciconiformes), though reports on it are infrequent. We describe an adult, injured, wild Black Stork (Ciconia nigra) infected with Cathaemasia hians in Hungary. We also report an ectopic occurrence of Cathaemasia hians in the submucosa of the small intestine.

Surveillance has revealed low prevalence of avian influenza viruses (AIV) in shorebirds except Ruddy Turnstones (Arenaria interpres) on the North American Atlantic coast. Similarly, of five species of shorebirds surveyed in Alaska in 2010, Ruddy Turnstones had the highest AIV antibody prevalence; prevalence of AIV RNA was low or zero.

Rhodococcus equi was isolated from the submaxillary lymph nodes of wild boars (Sus scrofa) in Wakayama and Hyogo, Japan, with a high prevalence. Rhodococcus equi isolation rates between locations, sexes, or body weights were not different, except in the prevalence of vapB-positive R. equi between locations.

This study documented the influence of age, sex, and body condition on blood hemoglobin concentrations (Hb) in wild meerkats (Suricata suricatta). Peripheral Hb increased with age, leveling off in adulthood, but was not influenced by sex or body condition.

We analyzed seven bot fly larvae extracted from six howler monkeys (Alouatta palliata) in Los Tuxtlas, Mexico. The spine distribution and structure of the larvae were consistent with those reported for Cuterebra baeri from Panama. This report sets the northern limit for its distribution in the Americas.

Multiple, nodular, pigmented masses protruding from the cornea and adjacent sclera of the left eye of a white-tailed deer (Odocoileus virginianus) were diagnosed as choristomas (dermoids). Microscopically, the masses contained well-differentiated skin, cartilage, and bone. This appears to be the first report of a corneoscleral choristoma in a cervid.

Prevalence of anti-Toxoplasma antibody in free-ranging ocelots (Leopardus pardalis) in northeastern Mexico was 69% (n=26). Grouping by age and sex, there were significant differences between sexes in subadults (P=0.03) and between age classes in males (P=0.01). Antibody titer increased in two recaptured ocelots and decreased in two others.

A Peregrine Falcon (Falco peregrinus) died of verminous and bacterial pneumonia and aerosacculitis associated with capillarid nematodes of the genus Eucoleus. This represents an aberrant location for these helminths in avian hosts. Although positive identification was not possible, the worms resembled Eucoleus aerophilus, a species not reported previously from birds.

Road-killed common toads (Bufo bufo; n=1,740) from Flanders, Belgium, were tested for ranavirus and Batrachochytrium dendrobatidis (Bd) using polymerase chain reaction. Both infections were present at a very low prevalence (<0.2% with a confidence interval of 95% for ranavirus and 0.63% for Bd).

Hypoglycemia (glucose <65 mg/dl) was detected in 21 of 28 immobilized free-ranging black rhinoceroses (Diceros bicornis). At repeat sampling 25 min later, only 6 of 28 were hypoglycemic (P<0.05). Lactate was significantly higher (P<0.0001) early in immobilization. Hypoglycemia and elevated lactate may increase risk of anesthetic complications and postrecovery problems.