A wide range of systemic mycoses have been reported from captive and wild marine mammals from North America. Examples include regionally endemic pathogens such as Coccidioides and Blastomyces spp., and novel pathogens like Cryptococcus gattii, which appear may have been introduced to North America by humans. Stranding and necropsy data were analyzed from three marine mammal stranding and response facilities on the central California coast to assess the prevalence, host demographics, and lesion distribution of systemic mycoses affecting locally endemic marine mammals. Between 1 January 1998 and 30 June 2012, >7,000 stranded marine mammals were necropsied at the three facilities. Necropsy and histopathology records were reviewed to identify cases of locally invasive or systemic mycoses and determine the nature and distribution of fungal lesions. Forty-one animals (0.6%) exhibited cytological, culture- or histologically confirmed locally invasive or systemic mycoses: 36 had coccidioidomycosis, two had zygomycosis, two had cryptococcosis, and one was systemically infected with Scedosporium apiospermum (an Ascomycota). Infected animals included 18 California sea lions (Zalophus californianus), 20 southern sea otters (Enhydra lutris nereis), two Pacific harbor seals (Phoca vitulina richardsi), one Dall's porpoise (Phocoenoides dalli), and one northern elephant seal (Mirounga angustirostris). Coccidioidomycosis was reported from 15 sea lions, 20 sea otters, and one harbor seal, confirming that Coccidioides spp. is the most common pathogen causing systemic mycosis in marine mammals stranding along the central California coast. We also report the first confirmation of C. gattii infection in a wild marine mammal from California and the first report of coccidioidomycosis in a wild harbor seal. Awareness of these pathogenic fungi during clinical care and postmortem examination is an important part of marine mammal population health surveillance and human health protection. Temporal–spatial overlap may be observed for pathogenic mycoses infecting coastal marine mammals and adjacent human populations.

Chlamydia infection is known to impact the health of koalas (Phascolarctos cinereus) in New South Wales (NSW) and Queensland, but the clinical significance of Chlamydia infections in Victorian koalas is not well described. We examined the prevalence of Chlamydia infection and assessed associated health parameters in two Victorian koala populations known to be Chlamydia positive. The same testing regimen was applied to a third Victorian population in which Chlamydia had not been detected. We examined 288 koalas and collected samples from the urogenital sinus and conjunctival sacs. Detection and differentiation of Chlamydia species utilized real-time PCR and high-resolution melting curve analysis. Chlamydia pecorum was detected in two populations (prevalences: 25% and 41%, respectively) but only from urogenital sinus swabs. Chlamydia was not detected in the third population. Chlamydia pneumoniae was not detected. Chlamydia pecorum infection was positively associated with wet bottom (indicating chronic urinary tract disease) in one Chlamydia-positive population and with abnormal urogenital ultrasound findings in the other Chlamydia-positive population. The prevalence of wet bottom was similar in all populations (including the Chlamydia-free population), suggesting there is another significant cause (or causes) of wet bottom in Victorian koalas. Ocular disease was not observed. This is the largest study of Chlamydia infection in Victorian koalas, and the results suggest the potential for epidemiologic differences related to Chlamydia infections between Victorian koalas and koalas in Queensland and NSW and also between geographically distinct Victorian populations. Further studies to investigate the genotypes of C. pecorum present in Victorian koalas and to identify additional causes of wet bottom in koalas are indicated.

White-nose syndrome (WNS) has had a devastating effect on North American bat populations. The causal agent of WNS is the fungal pathogen, Pseudogymnoascus destructans (Pd), which has been shown to persist in caves after the eradication of host populations. As nonpathogenic Pseudogymnoascus spp. display saprophytic growth and are among the most commonly isolated fungi from caves, we examined whether Pd could grow in cave sediments and the contribution such growth could have to WNS disease progression. We inoculated a range of diverse cave sediments and demonstrated the growth of Pd in all sediments tested. These data indicate that environmental growth of Pd could lead to the accumulation of spores above the estimated infection threshold for WNS, allowing environment-to-bat infection. The obtained growth parameters were then used in a susceptible-infected-susceptible mathematic model to determine the possible contribution of environmental Pd growth to WNS disease progression in a colony of little brown bats (Myotis lucifugus). This model suggests that the environmental growth of Pd would increase WNS infection rates, particularly in colonies experiencing longer hibernation periods or in hibernacula with high levels of organic detritus. The model also suggests that once introduced, environmental Pd growth would allow the persistence of this pathogen within infected hibernacula for decades, greatly compromising the success of bat reintroduction strategies. Together these data suggest that Pd is not reliant on its host for survival and is capable of environmental growth and amplification that could contribute to the rapid progression and long-term persistence of WNS in the hibernacula of threatened North American bats.

The meningeal worm (Parelaphostrongylus tenuis) is a nematode parasite that commonly infects white-tailed deer (Odocoileus virginianus; WTD) throughout the deciduous forest biome and deciduous-coniferous ecotone of eastern and central North America; the species is not known to occur west of the grassland biome of central North America. We used county-specific prevalence data to evaluate potential effects of landscape and climatologic factors on the spatial distribution of meningeal worm infection in South Dakota, US. Probability of infection increased 4-fold between eastern and western South Dakota and 1.3-fold for each 1-cm increase in summer precipitation. Sixty-three percent of WTD had only a single worm in the cranium. Expansion of meningeal worm infection across western South Dakota may be inherently low due to the combined effects of arid climate and potential attributes of the Missouri River that limit regional movements by infected WTD. Use of landscape genetic analyses to identify potential relationships between landscape features and population genetic structure of infected deer and parasites may contribute to a greater understanding of regional heterogeneity in meningeal worm infection rates across South Dakota, particularly in counties adjacent to the Missouri River. Future research evaluating heterogeneity in prevalence and intensity of infection between fawn and yearling deer, and the potential role of yearling male deer as dispersal agents of meningeal worms across the Missouri River, also is warranted.

We conducted surveillance for Riemerella anatipestifer (RA) in wild birds along the East Asian-Australasian flyway in South Korea. Detected RA were characterized by serotype, antibiotic susceptibility, and sequence analysis of the 16S rRNA gene. We collected 944 wild birds of 34 species from 19 of South Korea's major migratory wild bird habitats between 2011 and 2012. We identified RA by PCR and rRNA gene sequence in 71/102 (69.6%) pharyngeal swabs and 19/944 (2.0%) cloacal swabs of wild birds. Most RA positives (71/75 [95%] pharyngeal and 19/704 [(2.6%] cloacal) were from three duck species (family Anatidae): Mallard Duck (Anas platyrhynchos), Northern Pintail (Anas acuta), and Spot-billed Duck (Anas poecilorhyncha). Thirty-three RA isolates obtained and examined were highly resistant to aminoglycosides: kanamycin (100%), gentamicin (94%), amikacin (91%), neomycin (88%), and streptomycin (82%). Six isolates were identified as serotype 4 by agar gel precipitation. Serotypes 1 and 7, which are known virulent serotypes, were also identified in three isolates from wild duck species.

We investigated temporal and spatial trends in reporting of hemorrhagic disease (HD) in the midwestern and northeastern US using a 33-yr (1980–2012) questionnaire-based data set. This data set was supported by an additional 19 yr (1994–2012) of bluetongue virus (BTV) and epizootic hemorrhagic disease virus (EHDV) isolation results from clinically affected white-tailed deer (Odocoileus virginianus) in these regions. Both the number of counties that were reported positive for HD and the northern latitudinal range of reported HD increased with time. A similar increase was observed with both the number of states annually reporting HD and the number of counties where HD was reported. Large-scale outbreaks occurred in 1988, 1996, 2007, and 2012, and the scale of these individual outbreaks also increased with time. The predominant virus isolated from these regions was EHDV-2, but the prevalence of EHDV-6, which was first detected in 2006, appears to be increasing. Temporally, the extent of regional HD reporting was correlated with regional drought conditions. The significance of increases in reported HD and the incursions and establishment of new BTV and EHDV in the US currently are unknown.

The coccidian parasite Toxoplasma gondii infects humans and warm-blooded animals worldwide. The ecology of this parasite in marine systems is poorly understood, although many marine mammals are infected and susceptible to clinical toxoplasmosis. We summarized the lesions associated with T. gondii infection in the California sea lion (Zalophus californianus) population and investigated the prevalence of and risk factors associated with T. gondii exposure, as indicated by antibody. Five confirmed and four suspected cases of T. gondii infection were identified by analysis of 1,152 medical records of necropsied sea lions from 1975–2009. One suspected and two confirmed cases were identified in aborted fetuses from a sea lion rookery. Toxoplasmosis was the primary cause of death in five cases, including the two fetuses. Gross and histopathologic findings in T. gondii-infected sea lions were similar to those reported in other marine mammals. The most common lesions were encephalitis, meningitis, and myocarditis. The antibody prevalence in stranded, free-ranging sea lions for 1998–2009 was 2.5% (±0.03%; IgG titer 640). There was an increase in odds of exposure in sea lions with increasing age, suggesting cumulative risk of exposure and persistent antibody over time. The occurrence of disseminated T. gondii infection in aborted fetuses confirms vertical transmission in sea lions, and the increasing odds of exposure with age is consistent with additional opportunities for horizontal transmission in free-ranging sea lions over time. These data suggest that T. gondii may have two modes of transmission in the sea lion population. Overall, clinical disease was uncommon in our study which, along with low prevalence of T. gondii antibody, suggests substantially less-frequent exposure and lower susceptibility to clinical disease in California sea lions as compared to sympatric southern sea otters (Enhydra lutris nereis).

Wild Turkeys (Meleagris gallopavo) are susceptible to many of the same diseases as domestic turkeys. Before 2005, most Wild Turkeys in southern Georgia, US, had little or no exposure to commercial poultry operations. As part of a pathogen survey examining the effects of commercial poultry on Wild Turkeys, samples were collected from Wild Turkeys from March 2005 through May 2008. The turkeys were collected from 13 counties in southern Georgia and Madison County, Florida, and tested for antibodies to various pathogens of poultry. Three (13%) of the turkeys were positive for antibodies to Salmonella. Thirteen turkeys (54%) were positive for Newcastle disease virus antibodies, and 15 turkeys (63%) were positive for antibodies to reticuloendotheliosis virus. One turkey (4%) from Madison County was positive for avian encephalomyelitis virus antibodies.

The veterinary records of three species of free-living, endangered black cockatoos (Calyptorhynchus spp.; n = 565) admitted to the Perth Zoo Veterinary Hospital in Western Australia during a 10-yr period (2000–09) were analyzed to determine the effect of clinical presentation and treatment on survival to release. The most-common reason for admission was trauma (at least 76.7% of cases), and trauma was also the most-frequent finding on necropsy examination (80.1% of cases). Anemia and paralysis-paresis were significant factors determining the decreased likelihood of survival of cockatoos undergoing rehabilitation. Human activities, in particular vehicle strike, were significant causes of morbidity and mortality in free-living black cockatoo populations.

Aleutian mink disease virus (AMDV) causes a parvovirus infection, initially characterized in American mink (Neovison vison), that may have harmful effects on wild populations of susceptible animals. In North America, where American mink are native, the origin, host range, and prevalence of AMDV in wild species is not clear. We studied striped skunks (Mephitis mephitis) and raccoons (Procyon lotor) to determine whether species sympatric with mink are potential reservoirs in the transmission of AMDV to wild mink and mink farms. Antibodies to AMDV were detected in 41% of skunk serum samples (143/347) and AMDV nucleic acids were detected in 32% (14/40) of skunk spleen samples by PCR, indicating that AMDV exposure and infection were frequent in skunks. We detected no AMDV antibodies in 144 raccoon blood samples. Phylogenetic analysis revealed a newly identified AMDV haplogroup consisting of isolates from Ontario skunks and a free-ranging domestic mink from Ontario. Our findings of frequent AMDV infection in skunks, close genetic similarity between skunk and mink AMDV isolates, and evidence of AMDV transmission from skunks to mink support the hypothesis that skunks may be acting as alternative hosts and reservoirs of AMDV to wild mink through cross-species virus spillover.

Prairie dogs (Cynomys spp.) suffer high rates of mortality from plague. An oral sylvatic plague vaccine using the raccoon poxvirus vector (designated RCN-F1/V307) has been developed for prairie dogs. This vaccine is incorporated into palatable bait along with rhodamine B as a biomarker. We conducted trials in August and September 2012 to demonstrate uptake and apparent safety of the RCN-F1/V307 vaccine in two prairie dog species under field conditions. Free-ranging prairie dogs and other associated small rodents readily consumed vaccine-laden baits during field trials with no apparent adverse effects; most sampled prairie dogs (90%) and associated small rodents (78%) had consumed baits. Visual counts of prairie dogs and their burrows revealed no evidence of prairie dog decline after vaccine exposure. No vaccine-related morbidity, mortality, or gross or microscopic lesions were observed. Poxviruses were not isolated from any animal sampled prior to bait distribution or on sites that received placebo baits. We isolated RCN-F1/V307 from 17 prairie dogs and two deer mice (Peromyscus maniculatus) captured on sites where vaccine-laden baits were distributed. Based on these findings, studies examining the utility and effectiveness of oral vaccination to prevent plague-induced mortality in prairie dogs and associated species are underway.

Detection of West Nile virus (WNV) has been reported in a variety of wild ducks in the US, but little is known about the pathogenesis and outcome of exposure of the disease in these species. Previous experimental studies of WNV in ducks either have challenged a small number of ducks with WNV or have tested domesticated ducks. To determine susceptibility and immune response, we challenged 7-wk-old Wood Ducks (Aix sponsa) with a 1999 American Crow (Corvus brachyrhynchos) isolate of WNV. Wood Ducks were susceptible to infection with the virus, and, although clinical signs or mortality were not observed, microscopic lesions were noted, particularly in the heart and brain. West Nile virus viremia peaked on day 2 postinfection (pi) at 10^4.54 plaque-forming units (PFU) of virus/mL serum and WNV was shed orally (between 10² and 10^2.9 PFU per swab) and cloacally. Specific anti-WNV antibody response was rapid, with anti-WNV IgM detected on day 3 pi followed on day 5 pi by anti-WNV IgG. Neutralizing antibodies were detected by plaque-reduction neutralization assay in one duck on day 4 pi, and in all sampled ducks on day 5. These results indicate that Wood Ducks are susceptible to WNV, but it is unlikely that significant WNV mortality events occur in Wood Ducks or that they play a significant role in transmission. However, WNV viremia was sufficient, in theory, to infect mosquitoes, and oral and cloacal shedding of the virus may increase the risk of infection to other waterbirds.

Patterns of host–parasite association may vary across the landscape in part because of host and parasite diversity, divergence, local ecology, or interactions among these factors. In central coastal California, we quantified parasite prevalence, infection intensity, and diversity in two sister species of woodrats (Neotoma fuscipes and Neotoma macrotis) where the species co-occur (sympatry) and where each species exists alone (allopatry). In feces from 50 adults we identified seven taxa: the protozoans Eimeria, Giardia, and Cryptosporidium, the nematodes Trichuris, Aspicularis, and Eucoleus, and a cestode in the family Anoplocephalidae. Gastrointestinal parasite infection intensity and diversity were higher in males than in females, a difference that was most pronounced in the more aggressive N. fuscipes. Both species had lower infection intensity in sympatry than in allopatry and in sympatry the two species did not differ in infection intensity in total but did maintain distinct parasite communities. Taken together, our findings suggest that host evolutionary differences, including perhaps species-specific patterns of aggressive behavior, as well as local ecology, influence the likelihood of infection by these endoparasite taxa.

Giant liver fluke (Fascioloides magna) populations readily expand under suitable conditions. Although extirpated from the eastern slopes of the Canadian Rocky Mountains in the early 1960s, the fluke reappeared following natural spread through mountain passes from British Columbia. Herein, we assessed epizootiology of the fluke population two decades later. Between 1984 and 1991, 534 ungulates, including 381 elk (Cervus canadensis), 68 mule deer (Odocoileus hemionus hemionus), 54 white-tailed deer (Odocoileus virginianus), and 31 moose (Alces alces) from adjacent areas of Alberta (AB) and British Columbia (BC), Canada, were examined for giant liver flukes. Prevalence in elk increased from 53% to 79% (1984–91) in Banff National Park (BNP) in AB and 77% to 100% (1985–89) in Kootenay National Park (KNP) in BC. Super-infections (>100 flukes) were more common in later years. Generally, prevalence increased over time and with increasing age of elk. Intensity was lowest in young-of-year (BNP 8±5, KNP 3), but similar in yearlings (BNP 36±11, KNP 23±8) and adults (BNP 33±5, KNP 32±6). Prevalence was similar in male and female elk. Intensity was higher in males (BNP 47±7, KNP 46±12) than in females (BNP 28±6, KNP 22±4), although the maximum number of flukes (545) occurred in a female elk. Prevalence and intensity differed among other species of ungulates but patterns were similar in each park. Prevalence was lower in mule deer (BNP 6%, KNP 4%) than in white-tailed deer (BNP 44%, KNP 28%) and moose (BNP 52%, KNP 63%). Intensity differed among these species but never exceeded 30 flukes. Gravid flukes occurred only in elk and white-tailed deer. Transmission occurred primarily in late summer-fall and in wet habitats. At least seven elk died as a direct result of fluke infection. In this region, elk and white-tailed deer maintain the F. magna population with spillover into moose and, rarely, mule deer.

The southern sea otter (Enhydra lutris nereis) is a threatened marine sentinel. During postmortem investigations of stranded sea otters from 2004 to 2013 in California, US, papillomas were detected in the oral cavity of at least seven otters via necropsy and histopathology. Next-generation sequencing of viral particles purified from a single papilloma revealed a novel papillomavirus, Enhydra lutris papillomavirus 1 (ElPV-1). The genome of ElPV-1 was obtained, representing the first fully sequenced viral genome from southern sea otters. Phylogenetic analysis of the entire L1 gene, as well as a concatenated protein identities plot of all papillomaviral genes revealed that ElPV-1 is a λ-papillomavirus, related to a raccoon papillomavirus (Procyon lotor papillomavirus type 1) and a canine oral papillomavirus. Immunohistochemical staining, using a cross-reactive bovine papillomavirus antibody, suggested that ElPV-1 is present in intranuclear inclusions and intracytoplasmic keratin granules. Virus-infected cells were scattered throughout the stratum granulosum and stratum spinosum of the gingival and buccal papillomas. Using ElPV-1–specific PCR, we confirmed viral DNA in oral papillomas from all seven stranded sea otters, with identical L1 sequences. This virus is associated with the development of oral papillomatosis in southern sea otters.

Worldwide, stranded marine mammals and the network personnel who respond to marine mammal mortality have provided much of the information regarding marine morbillivirus infections. An assay to determine the amount of virus present in tissue samples would be useful to assist in routine surveying of animal health and for monitoring large-scale die-off events. False negatives from poor-quality samples prevent determination of the true extent of infection, while only small amounts of tissue samples or archived RNA may be available at the time of collection for future retrospective analysis. We developed a one-step duplex real-time reverse transcriptase-quantitative-PCR assay (RT-qPCR) based on Taqman probe technology to quantify phocine distemper virus (PDV) isolated from an outbreak in harbor (Phoca vitulina concolor) and gray seals (Halichoerus grypus) along the northeast US coast in 2006. The glyceraldehyde-3-phosphate-dehydrogenase (GAPDH) gene was selected to assess RNA quality. This duplex assay is specific for PDV and sensitive through a range of 10⁰ to 10⁹ copies ds-plasmid DNA. For the GAPDH target, the reaction in duplex amplified 10⁰ to 10⁹ copies of ds-plasmid DNA and was detectable in multiple seal species. This assay reduced the likelihood of false negative results due to degradation of tissues and well-to-well variability while providing sensitive and specific detection of PDV, which would be applicable in molecular epidemiologic studies and pathogen detection in field and laboratory investigations involving a variety of seal species.

Mycoplasma infections can cause significant morbidity and mortality in captive and wild chelonians. As part of a health assessment of endangered bog turtles (Glyptemys muhlenbergii) in the northeastern US, choanal and cloacal swabs from these and other sympatric species, including spotted turtles (Clemmys guttata), eastern box turtles (Terrapene carolina carolina), wood turtles (Glyptemys insculpta), and common snapping turtles (Chelydra serpentina) from 10 sampling sites in the states (US) of Delaware, New Jersey, and Pennsylvania, were tested by PCR for Mycoplasma. Of 108 turtles tested, 63 (58.3%) were PCR positive for Mycoplasma including 58 of 83 bog turtles (70%), three of three (100%) eastern box turtles, and two of 11 (18%) spotted turtles; all snapping turtles (n = 7) and wood turtles (n = 4) were negative. Sequence analysis of portions of the 16S–23S intergenic spacer region and the 16S ribosomal RNA gene revealed a single, unclassified species of Mycoplasma that has been previously reported in eastern box turtles, ornate box turtles (Terrapene ornata ornata), western pond turtles (Emys marmorata), and red-eared sliders (Trachemys scripta elegans). We document a high incidence of Mycoplasma, in the absence of clinical disease, in wild emydid turtles. These findings, along with wide distribution of the identified Mycoplasma sp. across a broad geographic region, suggest this bacterium is likely a commensal inhabitant of bog turtles, and possibly other species of emydid turtles, in the northeastern US.

Three cases of lethal sheep-associated malignant catarrhal fever (SA-MCF) in free-ranging moose (Alces alces) were diagnosed in Lesja, Norway, December 2008–February 2010. The diagnosis was based on PCR identification of ovine herpesvirus 2 (OvHV-2) DNA (n = 3) and typical histopathologic lesions (n = 1). To study the possibility of subclinical or latent MCF virus (MCFV) infection in this moose population and in red deer (Cervus elaphus), we examined clinically normal animals sampled during hunting in Lesja 2010 by serology and PCR. Sera from 63 moose and 33 red deer were tested for antibodies against MCFV by competitive-inhibition enzyme-linked immunosorbent assay. To test for MCFVs, a consensus PCR for herpesviral DNA was run on spleen samples from 23 moose and 17 red deer. All samples were antibody and PCR negative. Thus, there is no evidence of previous exposure, subclinical infection, or latent infection in this sample. This seasonal cluster of SA-MCF cases (2008–10) may be attributable to exposure of moose to lambs when OvHV-2 shedding is presumed to be maximal, compounded by an unusual extended grazing period by sheep in the autumn.

Nobuto filter paper strips are widely used for storing blood-serum samples, but the recovery of proteins from these strips following rehydration is unknown. Poor recovery of proteins could reduce the concentration of antibodies and antigens and reduce the sensitivity of diagnostic assays. We compared the protein concentration, and its association with test sensitivity, of eluted Nobuto strip samples with paired sera. We collected and froze serum from five gray wolves (Canis lupus) for 8 mo. When thawed, we used a spectrophotometer (absorbance 280 nm) to determine the serum protein concentration for paired sera and Nobuto eluates for each animal in 2-fold serial dilutions. Total protein concentration was similar for both sample storage methods (Nobuto eluates and control sera), except for the undiluted samples in which Nobuto eluates had higher total protein concentrations. Both sample storage methods appear to produce similar results using the SNAP® 4Dx® Test to detect antibodies against pathogens causing Lyme disease, anaplasmosis, and ehrlichiosis as well as antigen for canine heartworm disease.

Chlamydia pecorum, a recognized pathogen of domesticated ruminants and koalas (Phascolarctos cinereus), has been recently reported in a broad range of other wildlife species including water buffalo (Bubalus bubalis), ibex (Capra ibex), chamois (Rupicapra rupicapra), red deer (Cervus elaphus), and birds. This identification raises questions as to whether cross-host transmission may be a factor in the epidemiology of infections in these species. To begin to address this question, we employed a C. pecorum species-specific multi-locus sequence typing (MLST) scheme to characterize a small collection of C. pecorum-positive samples from wild, free-range ibex, a chamois, and a red deer from Grison, Switzerland, a canton where domesticated and wild ruminants graze in close proximity during the summer. Screening by PCR confirmed low to moderate levels of Chlamydia pecorum DNA in the eyes of healthy ibex (n = 4) and in the deer fecal sample (n = 1). The MLST analysis revealed three novel sequence types (STs; 88, 90, and 89) in these samples. On phylogenetic analysis, the ibex and deer sequences clustered by host species in their own well-supported clades and away from C. pecorum STs found in other hosts. Even though the analyzed sample size was small, the identification of unique C. pecorum STs infecting free-ranging Alpine ibex and red deer provides useful information for further C. pecorum epidemiologic studies.

We evaluated impact of the needle length, sex, and body condition on chemical immobilization induction time in 50 (29 males and 21 females) free-ranging fallow deer (Dama dama) in Sweden, 2006–11. Induction time is probably the single most important factor when immobilizing free-ranging wildlife with the use of a remote drug-delivery system. Induction times should be short to minimize stress and risk of injury, and to ensure that immobilized animals can be found and clinically monitored as soon as possible. We measured the distance between the darting location and where we recovered the immobilized animal and also the time occurring between the two events. We used two types of needles: 2.0 × 30– or 2.0 × 40–mm barbed needles with side ports. The most important result is that a 10-mm-longer dart needle can reduce the retrieval time substantially (>20 min) until an animal is under monitoring. On average after the darting, the retrieval time decreased from 51 to 29 min and the distance decreased from 519 m from the darting location to 294 m. We suggest that a needle length of 40 mm is preferable for immobilization of wild fallow deer, especially for animals in over-average–to–fat body condition.

We assessed risk factors for Toxoplasma gondii exposure in semiaquatic mammals in east-central Illinois, US. This agricultural region has extensive drainage systems that could potentially transport T. gondii oocysts into the watershed. We used muskrats (Ondatra zibethicus) and American mink (Neovison vison) as sentinels of watershed contamination. We predicted individuals from larger subwatersheds would more likely be antibody-positive for T. gondii, as they were exposed to drainage from larger areas. We also evaluated amount of urban land cover within the subwatershed, proximity to farmsteads, and age of individuals in competing models of T. gondii infection. Antibodies to T. gondii were assayed in animal sera by modified agglutination tests (titer 25 or higher) and detected in 18 (60%) of 30 muskrats and 20 (77%) of 26 mink. Infection rates were ≥1.7 times higher than those typical for mammals in upland habitats in this region. Subwatershed size and age class were important predictors of T. gondii infection in muskrats (R² = 0.35). Models incorporating urban land cover and proximity to farmsteads had little support. None of our models of antibody prevalence in mink were well supported, possibly because mink are less strictly associated with riparian habitats. Because ~91% of our study area is devoted to agricultural production and urbanization, transport of T. gondii into freshwater ecosystems is likely facilitated by modified drainage practices common in these areas.

Diagnosis of tuberculosis in wildlife often relies on postmortem samples because of logistical challenges and lack of field-friendly techniques for live animal testing. Confirmation of infection through detection of infectious organisms is essential for studying the pathogenesis and epidemiology of disease. We describe the application of a technique to obtain respiratory samples from free-ranging living lions to facilitate detection of viable Mycobacterium bovis under field conditions. We identified M. bovis by mycobacterial culture and PCR in tracheobronchial lavage samples from 8/134 (6.0%) lions tested in Kruger National Park, South Africa. This confirms the respiratory shedding of viable M. bovis in living lions. The implications of these results are that infected lions have the potential to transmit this disease and serve as maintenance hosts.

If a parasite is not detected during a survey, one of two explanations is possible: the parasite was truly absent or it was present but not detected. We fit occupancy models to account for imperfect detection when combing fleas (Siphonaptera) from black-tailed prairie dogs (Cynomys ludovicianus) during June–August 2012 in the Vermejo Park Ranch, New Mexico, USA. With the use of detection histories from combing events during monthly trapping sessions, we fit occupancy models for two flea species: Oropsylla hirusta (a prairie dog specialist) and Pulex simulans (a generalist). Detection probability was <100% for both species and about 21% lower for P. simulans. Pulex simulans may be especially difficult to detect because it is about half the size of O. hirusta. Monthly occupancy (prevalence) for P. simulans was estimated at 24% (June, 95% confidence interval = 19–30), 39% (July, 32–47), and 56% (August, 49–64) in new prairie dog colonies, and 43% (32–54), 61% (49–71), and 79% (70–87) in old colonies. These results suggest P. simulans can attain high prevalence on prairie dogs, especially in old colonies. If P. simulans is highly prevalent on prairie dogs, it may serve as a “bridge vector” between Cynomys and other mammalian hosts of the plague bacterium Yersinia pestis, and even function as a reservoir of Y. pestis between outbreaks.

The bacterium Listeria monocytogenes causes disease in a wide variety of mammals including rabbits and hares. We describe naturally acquired metritis and septicemic listeriosis in wild female hares from Saskatchewan, Canada. Between April 2012 and July 2013, two white-tailed jackrabbits (Lepus townsendii) and a snowshoe hare (Lepus americanus) were presented to the Veterinary Medical Centre at the Western College of Veterinary Medicine, Saskatoon, Saskatchewan, Canada with nonspecific neurologic signs. The hares were euthanized and autopsied. Necrotizing fibrinosuppurative metritis was present in all. Additional findings in individual hares included fetal maceration, multifocal necrotizing myocarditis, multifocal hepatic necrosis, and nonsuppurative encephalitis. Listeria monocytogenes was cultured from multiple tissues in each hare. Although listeriosis in pregnant domestic rabbits has been studied, this is the first detailed description in wild North American hares. The epidemiology of listeriosis, including prevalence and the role of environmental sources and coprophagy in transmission among hares, requires further investigation.

Data on reptile analgesia are scarce for nonsteroidal anti-inflammatory drugs (NSAIDs) and opioids and almost completely lacking in sea turtles, even though emergencies requiring correct pain management are very frequent in their rehabilitative medicine; therefore, dosage regimens extrapolated from other species involve the risk of clinical failure and damage to the animals. We describe the pharmacokinetic behavior of meloxicam in the loggerhead sea turtle (Caretta caretta). We chose meloxicam because of its selective anti-cyclooxygenase-2 activity and lesser adverse side effects. No data are available on the capacity of turtles to tolerate NSAIDs, so we chose a dose of 0.1 mg/kg of meloxicam. Plasma concentrations of meloxicam were unexpectedly low both for intravenous (IV; maximum concentration [C_max] = 0.04±0.02 µg/mL) and intramuscular (IM; C_max = 0.07±0.09 µg/mL) administration. A double-peak phenomenon occurred after both IV (time for second peak concentration T_max2 = 10.33±10.89 h) and IM (T_max2 = 1.17±0.75 h). The second peak after IM injection was premature, so some difficulty and delay in absorption appears to be an appropriate explanation. Furthermore, the area under the curve, and therefore systemic bioavailability (F = 31.82±28.24%), after both IV (0.30±0.29) and IM (0.10±0.03) injection appeared particularly limited. Terminal elimination slope and mean residence time indicated fast elimination after IM dosing; as a consequence, plasma concentrations dropped below analytic limits in 8 h. Considering that IM is the favored route of administration of drugs in rescue centers, it is unlikely that meloxicam at 0.1 mg/kg is an appropriate choice, particularly in long-term pain management protocols.

Medicinal baits are distributed to manage zoonotic diseases, including raccoon (Procyon lotor) rabies, but efficient distribution strategies are needed for suburban environments. We developed an automated dispenser that transfers fishmeal polymer baits at user-specified intervals from a magazine to a receptacle fitted with a filter that exploits raccoon dexterity to limit consumption by nontarget species. We introduce the concept of automated dispensers and describe bait removal success rates for raccoons versus nontarget species. We monitored visitation with remote cameras after deploying a dispenser, programmed to present two baits per night, in three disjunct forest patches in northwest Indiana. Raccoons removed 72% of baits; nontarget, white-footed mice (Peromyscus leucopus) removed 11%; Virginia opossums (Didelphis virginiana) removed 9%. Bait removal success varied significantly between raccoons (76%) and opossums (21%), improving bait delivery specificity relative to hand baiting. Accumulation of baits in receptacles resulted in excess (more than one) bait consumption (39% of baits consumed by raccoons were excess), suggesting design improvements are needed to present additional baits only after previous baits have been consumed. Automated dispensers successfully sustained bait availability throughout the operational period. Subsequent research is needed to determine whether a sustained availability of baits achieved with automated dispensers is more effective for the treatment of raccoons in suburban environments than traditional distribution methods.

Pseudogymnoascus destructans, the causal agent of white-nose syndrome (WNS), is responsible for widespread mortality of hibernating bats across eastern North America. To document P. destructans exposure and infections on bats active during winter in the southeastern US, we collected epidermal swabs from bats captured during winters 2012–13 and 2013–14 in mist nets set outside of hibernacula in Tennessee. Epidermal swab samples were collected from eight Rafinesque's big-eared bats (Corynorhinus rafinesquii), six eastern red bats (Lasiurus borealis), and three silver-hair bats (Lasionycteris noctivagans). Using real-time PCR methods, we identified DNA sequences of P. destructans from skin swabs of two Rafinesque's big-eared bats, two eastern red bats, and one silver-haired bat. This is the first detection of the WNS fungus on Rafinesque's big-eared bats and eastern red bats and the second record of the presence of the fungus on silver-haired bats.

The use of serologic assays for influenza A virus (IAV) surveillance in wild birds has increased because of the availability of commercial enzyme-linked immunosorbent assays (ELISAs). Recently, an H5-specific blocking ELISA (bELISA) was shown to reliably detect H5-specific antibodies to low- and high-pathogenic H5 viruses in experimentally infected waterfowl. Mute Swans (Cygnus olor) were frequently associated with highly pathogenic H5N1 outbreaks in Europe and may have a similar role if highly pathogenic H5N1 is introduced into North America. We measured the prevalence of antibodies to the nucleoprotein and H5 protein in Mute Swans using three serologic assays. We collected 340 serum samples from Mute Swans in Michigan, New Jersey, New York, and Rhode Island, US. We detected antibodies to the IAV nucleoprotein in 66.2% (225/340) of the samples. We detected H5-specific antibodies in 62.9% (214/340) and 18.8% (64/340) using a modified H5 bELISA protocol and hemagglutination inhibition (HI) assay, respectively. The modified H5 bELISA protocol detected significantly more positive samples than did the manufacturer's protocol. We also tested 46 samples using virus neutralization. Neutralization results had high agreement with the modified H5 bELISA protocol and detected a higher prevalence than did the HI assay. These results indicate that North American Mute Swans have high nucleoprotein and H5 antibody prevalences.

We harvested 21 fallow deer (Dama dama) and 17 axis deer (Axis axis) in northern Mexico. Two fallow deer were positive for Babesia bigemina and one for Babesia bovis. Amplicons had the expected 170 and 291 base pairs and were identical to B. bigemina (S45366) and B. bovis (M38218), respectively.

We report infection by Ichthyophonus sp. in a South American amphibian, the hylid frog Hypsiboas pulchellus in Uruguay. This frog had a large subcutaneous mass over the urostyle and dorsal musculature comprised of parasitic cysts with mild granulomatous inflammation but otherwise appeared healthy.

A dead Intermediate Egret (Egretta intermedia) was found on the shore of a stream in South Korea in January 2013. Salmonella Typhimurium was isolated from purulent exudates in the foot joints, demonstrating bacterial arthritis. The isolate was similar to a poultry isolate determined by pulsed-field gel electrophoresis.

We describe gross, histopathologic, and immunonocytologic features of a primary uterine T-cell lymphoma with multiple metastases in an adult female Atlantic spotted dolphin (Stenella frontalis). Neoplastic T-cell lymphocytes (CD3 ) partially effaced the uterus, ovaries, mesosalpinx, and urinary bladder and variably infiltrated abdominal and thoracic organs. Herpesvirus and morbillivirus PCR were negative.