In 2007, a National Park Service (NPS) biologist died from pneumonic plague after unprotected exposure to an infected mountain lion. This incident increased awareness of occupational zoonotic disease transmission and prompted an assessment of employees who handle wildlife. During April–June 2009, we conducted a national online survey of NPS biologists and other wildlife workers to assess in the preceding 12 mo: 1) potential work-related zoonotic disease exposures; 2) protective practices, including use of personal protective equipment (PPE); and 3) barriers and facilitators to PPE use. Summary protective measure scores were calculated and compared with sociodemographic and work-related factors. Surveys were completed by 238 employees from 131 parks in all NPS regions. Seventy-one percent were biologists or technicians, 16% natural resource specialists or managers, and 13% had other job titles. Among a majority of respondents, interactions with animals were infrequent and occurred approximately several times per year as follows: handling live (39%), sick (43%), or dead animals (46%), and drawing blood from animals (42%). The most frequently reported protective measures used were hand hygiene and gloves. Commonly agreed-upon measures that would facilitate PPE use included having PPE stocked and readily available (92%) and having specific PPE kits for use during necropsies (91%) and in remote field settings (91%). Significantly higher summary protective measure scores were found if respondents had either read or reviewed ‘‘NPS safe work practices for employees handling wildlife’’ with their supervisor, had zoonotic disease safety or PPE use included in their employee performance appraisal plans, or had conducted a job-hazard analysis for handling wildlife. Ninety (38%) respondents reported receiving zoonotic disease training. Our findings support the development and implementation of workplace interventions to increase zoonotic disease awareness and promote a culture of prevention among wildlife professionals.

The epizootiology of anatid herpesvirus 1 (AHV-1) infection in waterfowl is poorly understood but apparently involves persistence of the virus in latently infected birds. Epornitics have often occurred in captive waterfowl or semiwild ducks in parklike settings, and many wildlife professionals conclude that such ducks may be the source of infection for wild waterfowl. We assessed the prevalence of latent infection and viral shedding from four groups of waterfowl: naturally occurring populations of native waterfowl, captive-reared waterfowl released for shooting, introduced nonmigratory waterfowl (e.g., resident, wild Mallards; Anas platyrhynchos), and semiwild peridomestic waterfowl (e.g., park ducks) in North Carolina and Florida, USA from 2004 to 2009. A nested PCR assay was used to detect viral DNA in trigeminal ganglia and cloacal swabs. Detection of viral DNA in trigeminal ganglia, but not cloacal swabs, was assumed to indicate latent infection, whereas PCR-positive cloacal swabs indicated active shedding of the virus. We collected 2,045 samples from 23 species of native, wild waterfowl, and detected latent infections in nine species. Wild Northern Pintails (Anas acuta), a species reportedly resistant to the virus, had the highest prevalence (8.1%). However, low prevalences were identified in other waterfowl from various families. Cloacal shedding was rarely detected (0.1% prevalence) among native waterfowl and was observed in one Blue-winged Teal (Anas discors) and one Mottled Duck (Anas fulvigula). All captive-reared, released waterfowl (n=13) collected were Mallards and one was latently infected, suggesting that these birds could also serve as a source of AHV-1 for naive waterfowl. All nonmigratory waterfowl sampled (n=90) were also Mallards. None of the resident Mallards were shedding virus, but one was latently infected. The peridomestic waterfowl sampled included breeds of domestic Mallard (n=6) and Muscovy Ducks (Cairina moschata; n=73). One peridomestic Mallard and four Muscovy Ducks were shedding virus at the time they were sampled, but no latently infected, asymptomatic carriers were identified.

We conducted a serologic survey for selected infectious agents on two sympatric cormorants, the Imperial Cormorant (Phalacrocorax atriceps) and the Rock Shag (Phalacrocorax magellanicus). Blood was collected from 267 Imperial Cormorants and 106 Rock Shags at 17 colonies along the Patagonia Atlantic shore during nine breeding seasons (1994, 1999–2001, 2005– 2008, 2010). Antibodies to four pathogens were common to both species and frequently observed: avian paramyxovirus type 1 (56% of Imperial Cormorants and 56% of Rock Shags); avian adenovirus (67% of Imperial Cormorants and 40% of Rock Shags); infectious bronchitis virus serotypes IBV-41, IBV-46, IBV-99, and IBV-JMK (53% of Imperial Cormorants and 64% of Rock Shags); and Salmonella pullorum (18% of Imperial Cormorants and 7% of Rock Shags). Antibody prevalence for these pathogens varied significantly between species, except for avian paramyxovirus type 1. Exposure to avian paramyxovirus type 1 and all serotypes of infectious bronchitis virus varied significantly among seasons in both species. In contrast, the sporadic occurrence of positive titers suggest that cormorants had occasional exposure to Aspergillus spp. (3% of Rock Shags, only in 2000), avian paramyxovirus type 3 (5% of Rock Shags, only in 2008), Chlamydophila spp. (1% of Imperial Cormorants, only in 2010), and avian reovirus (1% of Rock Shags, only in 1999; 29% of Imperial Cormorants, in 2008 and 2010). Both species were antibody negative for avian encephalomyelitis virus, avian influenza virus, avian laryngotracheitis virus, avian paramyxovirus type 2, and infectious bursal disease virus. We provide the first information on pathogen exposure, indicated by detection of antibody in blood samples, for two sympatric species of South Atlantic cormorants. To determine major causes of morbidity and mortality in these birds future efforts should focus on necropsy surveys in cormorant colonies.

While vector-borne diseases are known to be particularly influenced by environmental factors, the impact of land-cover change on vector-borne wildlife disease patterns is poorly understood, largely due to the paucity of data on disease occurrence at extensive spatial and temporal scales. Widespread and rapid anthropogenic land-cover change, especially urbanization, has transformed the US landscape during the last century. Epizootic hemorrhagic disease virus and blue tongue virus, vectored by Culicoides biting midges, are two RNA viruses in the Orbivirus genus that cause severe hemorrhagic disease (HD) in white-tailed deer (Odocoileus virginianus). We examine the spatial dynamics of HD affecting white-tailed deer in the contiguous United States in two periods covering 1980 to 2007 in connection with land-cover change over the same time. Using spatial statistical modeling, wetland cover emerges as a critical driver of HD morbidity, whereas the drivers of mortality patterns are more complex. Increasing wetland cover is positively associated with HD morbidity, which is consistent with the ecologic requirements of the Culicoides vector. Wetland cover is inherently dynamic due to its importance to biodiversity and water quality as well as its utility for other purposes when drained. Accordingly this analysis helps in understanding the consequences of changing wetlands on vector-borne disease patterns, to identify disease hotspots in a large landscape, and to forecast the spatial spread of HD and related diseases.

We investigated the exposure of jaguar (Panthera onca) populations and domestic carnivores to selected viral infections in the Cerrado, Amazon, and Pantanal biomes of Brazil. Between February 2000 and January 2010, we collected serum samples from 31 jaguars, 174 dogs (Canis lupus familiaris), and 35 domestic cats (Felis catus). Serologic analyses for antibodies to rabies virus, canine distemper virus (CDV), feline immunodeficiency virus (FIV), and for feline leukemia virus (FeLV) antigen were conducted. The jaguars from Cerrado and Pantantal were exposed to rabies virus, while the jaguars from the Pantanal and the dogs from all three areas were exposed to CDV. Two cats from the Amazonian site were antigen-positive for FeLV, but no jaguars had FeLV antigen or FIV antibody. Canine distemper and rabies viruses should be carefully monitored and considered potential threats to these jaguar populations. Currently FIV and FeLV do not appear to represent a health threat for jaguar populations in this area. Domestic dogs and cats in these areas should be vaccinated, and the movement of domestic animals around protected areas should be restricted.

Bovine brucellosis, caused by the bacterium Brucella abortus, is endemic in bison (Bison bison) and elk (Cervus elaphus nelsoni) populations in the area of Yellowstone National Park, USA. Two strategies have been proposed to reduce the risk of transmission of disease in bison: remote vaccination with the vaccine RB51, and the use of immunocontraception of bison to decrease shedding of organisms from infected females. The frequent occurrence of venereal transmission in bison would complicate either of these strategies, requiring vaccination of males as well as females, and rendering immunocontraception less effective in reducing transmission of B. abortus. To address the question of venereal transmission, we inoculated each of 18 bison cows with 4.5 × 10⁸ colony-forming units of B. abortus strain 19, as a surrogate of field strain, by three routes: intraconjunctival (IC), intravaginal (VI), and intracervical/intrauterine (AI). Bison semen was mixed with strain 19 inoculum for the latter route. Bison were monitored by serology and culture for 12 wk, at which time they were euthanized and specimens collected for culture. All IC-inoculated animals seroconverted on multiple tests and one was culture positive at 12 wk postexposure. Seven of eight VI bison developed suspect or positive serologic tests and four were positive at one or more time points. Weak transient serologic responses (suspect) were seen in four of five AI bison. Results showed that IC inoculation with strain 19 was a suitable surrogate for field strain to demonstrate exposure to the B. abortus. The seroconversion of four of eight VI bison indicated exposure of the immune system to the agent and the need for further studies on venereal transmission in bison.

We investigated the presence of Toxoplasma gondii infections in Eurasian lynx (Lynx lynx) in Finland by analyzing samples from 337 lynx that were legally hunted during the 2010–2011 season and by performing a retrospective nationwide database search of postmortem toxoplasmosis diagnoses in this species. We detected specific anti-T. gondii IgG antibodies in 290 (86.1%) of the 337 lynx. The method used was a direct agglutination test, and samples positive at the used dilution 1:40 were defined as antibody positive. Older lynx had 14.3 times higher odds of being antibody-positive than did lynx of the presumed age of 7–10 mo, and lynx weighing >15 kg had 16.7 times higher odds of being antibody positive than did those ≤15 kg. Lynx from the southwest were more often antibody positive, with an odds ratio 6.3, than lynx from the northeast. None of the 332 fecal samples available was positive for the presence of T. gondii-like oocysts with a quantitative MgSO₄ flotation technique, and none of the 167 free-ranging Eurasian lynx examined postmortem by veterinary pathologists from January 2000 to May 2010 had died from toxoplasmosis. Although Finnish lynx were confirmed to commonly encounter T. gondii, we found no evidence of an ongoing contribution to the environmental oocyst burden nor of the lynx dying from the infection.

Effective anthelmintics are valuable tools for biologists conducting manipulative field experiments to examine effects of parasites on wildlife. However, before such experiments are carried out the efficacy of these drugs must be determined. We conducted three field experiments (May 2010–September 2011) on free-ranging eastern grey kangaroos (Macropus giganteus) at a golf course in Victoria, Australia, treating animals with the anthelmintic drugs moxidectin (subcutaneous, 1 mg/kg, 2 mg/kg), ivermectin (subcutaneous, 200 µg/kg), and albendazole (oral, 3.8 mg/kg). After treatment we monitored strongylid fecal egg counts (FECs) over time and assessed anthelmintic efficacy using fecal egg count reduction tests (FECRTs). We also performed a larval development assay (LDA) to evaluate directly the efficacy in the nematode population. Unexpectedly, moxidectin and ivermectin had low efficacy with maximum FEC reductions of 82% and 28%, respectively. However, treatment with albendazole reduced FECs by 100% in all kangaroos and egg counts remained low for up to 3 mo. The results from the LDA supported the FECRTs, with low macrocyclic lactone efficacy and high albendazole efficacy. Macrocyclic lactones, at recommended dose rates, were much less effective against strongylid nematodes in kangaroos than has been reported for domestic herbivores. This may be partly due to pharmacokinetics in the host and partly due to low susceptibility in some of the nematodes infecting eastern grey kangaroos.

We tested 191 sera of red foxes (Vulpes vulpes; 78 females and 113 males) for antibodies to Toxoplasma gondii using an indirect immunofluorescent test. Tissue samples of myocardium, lymph nodes, and brains from antibody-positive animals were tested for T. gondii DNA using specific PCR and processed for histopathology and immunohistochemistry (IHC) to detect T. gondii antigen. Of 192 (53.4%) antibody-positive animals, eight were positive by PCR on myocardium and one on brain. All DNA extracts were genotyped. Histopathology showed lesions characteristic of protozoan encephalitis; IHC did not show T. gondii antigen in examined tissues. The high antibody prevalence found in our study, which is the first in Italy, and the occurrence of polymorphic strains (combination of different type I and III alleles) divergent from typical T. gondii strains, suggests red foxes may be a sentinel of T. gondii in the environment.

Backpack harnesses are commonly used to attach radio and satellite transmitters to a wide range of bird species for research and conservation management. They are an integral part of the conservation management of the New Zealand Takahē (Porphyrio hochstetteri), an endangered flightless rail. Radio transmitters mounted on backpack harnesses enable the birds to be tracked in their remaining native range of remote, mountainous Fiordland, New Zealand. We evaluated 26 Takahē retrospectively at necropsy by gross examination, radiography, and computed tomography to assess damage from the backpack harness. Ten birds that had never worn a harness had no evidence of wing injury. Of the 16 birds that had worn a harness, 10 (63%) had superficial soft tissue injury to skin or patagium or more severe injury, such as remodeling of the distal humerus at the harness cord-wing interface, or pathologic fractures. Such injuries are hypothesized to be associated with discomfort, increased risk of infection or fracture, and therefore reduced fitness. These findings have implications for all avian species deployed with backpack harnesses.

Using a prospective, randomized study design we demonstrate that midazolam sedation minimizes acidosis compared with physical restraint in captive juvenile estuarine crocodiles during handling or noninvasive procedures at preferred body temperature. A dose of midazolam (5.0 mg/kg) was administered intramuscularly into the forelimb of 20 male estuarine crocodiles weighing 2–3.5 kg. Their heart and respiratory rate and degree of sedation were monitored until recovery and then daily for 7 subsequent days. Blood samples were taken at 30, 60, 90, 180, and 360 min. We recorded lactate, partial pressure of carbon dioxide (CO₂), hematocrit, glucose, and blood pH. A second group (1.9–2.6 kg) was physically restrained for 5 min and the same parameters recorded. Physically restrained animals demonstrated elevated heart rate, respiratory rate, glucose, lactate, and anion gap compared with the midazolam-treated group. Physically restrained animals had lower pH, bicarbonate, and partial pressure of CO₂ compared with the midazolam-treated group. Behavior in the physically restrained group in the days following the study was disrupted, with reluctance to feed and bask, compared with midazolam-treated animals whose behavior was normal. We conclude that midazolam administered in the forelimb of captive estuarine crocodiles of 2–3.5 kg provides predictable onset and duration of sedation enabling physical examination, sample collection, and translocation of the animals with minimal disturbance to lactate, pH, and CO₂. Behavior following recovery appears normal.

Aflatoxin is a widely occurring and harmful mycotoxin produced by strains of Aspergillus spp. growing on vegetable matter. We investigated the concentration of aflatoxin needed to impair normal physiologic responses and induce acute morbidity and mortality in Northern Bobwhites (Colinus virginianus). Ten wild-caught adult bobwhites (five males and five females) from southern Texas were randomly assigned to each treatment group (0, 100, 500, 1,000, and 2,000 parts per billion (ppb) aflatoxin; n=50). We orally administered 100 μL of aflatoxin, derived from Aspergillus flavus, once per week for 4 wk and monitored bird mass, daily feed consumption, liver histology, and blood chemistries. An in vitro white blood cell proliferation test was conducted using spleen tissue to determine the effect of aflatoxin on the immune system. There was no mortality in the control groups, whereas mortalities occurred in all treatment groups except in the 100 ppb aflatoxin treatment. Immunosuppression, reduction in gamma-globulin, glucose, and gamma-glutamyltransferase blood levels, and abnormal liver histology were observed in aflatoxin-exposed quail. Blood chemistry indicated cellular damage to the liver and kidneys. We concluded that short-term, acute doses of aflatoxin as low as 100 ppb can be detrimental to the health of Northern Bobwhites.

Morbilliviruses pose a significant threat to marine mammal populations around the world and have been associated with multiple epizootics in pinnipeds and cetaceans. As part of a preventive veterinary medical program, five harbor seals (Phoca vitulina) in a managed collection were vaccinated with a recombinant canarypox-vectored canine distemper virus vaccine. The vaccine was evaluated for safety (by monitoring seals for local and systemic adverse effects and by testing for shedding of the canarypox vector) and efficacy (by testing for serum neutralizing antibodies). None of the seals showed signs of local or systemic adverse reactions to the vaccination. Three seals vaccinated once did not seroconvert, but the recombinant vaccine induced a persistent serum virus neutralizing titer (12 mo) in the two seals that were vaccinated twice, 1 mo apart.

Vector-borne and zoonotic pathogens have comprised a significant proportion of the emerging infectious diseases in humans in recent decades. The role of many wildlife species as reservoirs for arthropod-borne viral pathogens is poorly understood. We investigated the exposure history of various African wildlife species from the Congo basin to mosquito-borne flaviviruses and alphaviruses by testing archived serum samples. Sera from 24 African forest buffalo (Syncerus caffer nanus), 34 African elephants (Loxodonta africana), 40 duikers (Cephalophus and Philantomba spp.), 25 mandrills (Mandrillus sphinx), 32 mountain gorillas (Gorilla beringei beringei), five Grauer’s gorillas (Gorilla beringei graueri), two L’Hoest’s monkeys (Cercopithecus lhoesti), two golden monkeys (Cercopithecus kandti), and three chimpanzees (Pan troglodytes) sampled between 1991 and 2009 were tested for antibodies against chikungunya virus (CHIKV), o’nyong-nyong virus (ONNV), West Nile virus (WNV), dengue 2 virus (DENV-2), and yellow fever virus (YFV) by plaque reduction neutralization test. Specific neutralizing antibodies against ONNV were found in African forest buffalo in the Democratic Republic of the Congo (DRC) and Gabon, duikers in the DRC, and mandrills in Gabon, providing novel evidence of enzootic circulation of ONNV in these countries. African forest buffalo in the DRC and Gabon also demonstrated evidence of exposure to CHIKV, WNV, and DENV-2, while mandrills in Gabon were antibody positive for CHIKV, DENV-2, WNV, and YFV. All of the elephants tested had a strong neutralizing antibody response to WNV. We also document results from a survey of gorillas for arboviruses, of which 4/32 (13%) had antibody to an alphavirus or flavivirus. Overall, our results demonstrate a high prevalence of neutralizing antibodies against multiple arboviruses in wildlife in equatorial Africa.

The Yukon-Kuskokwim Delta (Y-K Delta) in western Alaska is an immense and important breeding ground for waterfowl. Migratory birds from the Pacific Americas, Central Pacific, and East Asian-Australasian flyways converge in this region, providing opportunities for intermixing of North American- and Eurasian-origin hosts and infectious agents, such as avian influenza virus (AIV). We characterized the genomes of 90 low pathogenic (LP) AIV isolates from 11 species of waterfowl sampled on the Y-K Delta between 2006 and 2009 as part of an interagency surveillance program for the detection of the H5N1 highly pathogenic (HP) strain of AIV. We found evidence for subtype and genetic differences between viruses from swans and geese, dabbling ducks, and sea ducks. At least one gene segment in 39% of all isolates was Eurasian in origin. Target species (those ranked as having a relatively high potential to introduce HP H5N1 AIV to North America) were no more likely than nontarget species to carry viruses with genes of Eurasian origin. These findings provide evidence that the frequency at which viral gene segments of Eurasian origin are detected does not result from a strong species effect, but rather we suspect it is linked to the geographic location of the Y-K Delta in western Alaska where flyways from different continents overlap. This study provides support for retaining the Y-K Delta as a high priority region for the surveillance of Asian avian pathogens such as HP H5N1 AIV.

Emerging infectious disease events are frequently caused by arthropod-borne viruses (arboviruses) that are maintained in a zoonotic cycle between arthropod vectors and vertebrate wildlife species, with spillover to humans in areas where human and wildlife populations interface. The greater Congo basin region, including Uganda, has historically been a hot spot for emergence of known and novel arboviruses. Surveillance of arthropod vectors is a critical activity in monitoring and predicting outbreaks of arboviral disease, and identification of blood meals in engorged arthropods collected during surveillance efforts provides insight into the ecology of arboviruses and their vectors. As part of an ongoing arbovirus surveillance project we analyzed blood meals from engorged mosquitoes collected at five sites in western Uganda November 2008–June 2010. We extracted DNA from the dissected and triturated abdomens of engorged mosquito specimens. Mitochondrial cytochrome c oxidase I gene sequence was amplified by PCR and sequenced to identify the source of the mosquito host blood. Blood meals were analyzed from 533 engorged mosquito specimens; 440 of these blood meals were successfully identified from 33 mosquito species. Species identifications were made for 285 of the 440 identified specimens with the remainder identified to genus, family, or order. When combined with published arbovirus isolation and serologic survey data, our results suggest possible vector-reservoir relationships for several arboviruses, including Rift Valley fever virus and West Nile virus.

Beginning in 1997 Michigan State University Mara Hyena Project investigators observed waxing and waning progression of oral and genital masses during long-term behavioral observations of a population of wild spotted hyenas (Crocuta crocuta) from the Masai Mara Game Reserve, Kenya. From 1999–2000, we darted adult spotted hyenas to obtain routine physiologic and hematologic data and collected small, raised, lobulated, pigmented masses from the oral or genital areas of eight animals. Microscopically, masses consisted of variably thickened epidermis with thick elongate rete pegs, prominent stratum spinosum, and few koilocytes, consistent with papillomavirus-induced lesions. Immunohistochemistry on formalin-fixed, paraffin-embedded papilloma tissue revealed positive intranuclear labeling for papillomavirus antigen in the superficial stratum granulosum and in sloughing keratin layers of multiple samples. Polymerase chain reaction on DNA extracts from tumor tissue amplified a papillomavirus-specific 418 base pair amplicon in the E1 ORF. Basic Local Alignment Search Tool analysis of the sequenced amplicon suggests a novel hyaenid papillomavirus. Confirmatory complete genomic sequencing was performed later by the Rega Institute in Belgium. To our knowledge, this is the first report of a papillomavirus in a Hyaenidae species. Spotted hyena social behavior might facilitate oral-genital transmission of papillomavirus in this population.

Brucella species infection in marine mammal species has been reported to have a global distribution. In 2007, the description of Brucella ceti was published and formally adopted for those isolates originating from cetaceans and pathologic lesions similar to those seen in terrestrial mammals infected with Brucella spp. have been associated with its isolation. Brucella ceti infection specific to the central nervous system has been described in two species of cetacean: striped dolphins (Stenella coeruleoalba) in Europe and Costa Rica and an Atlantic white-sided dolphin (Lagenorhynchus acutus) in the UK. We describe the first report, to our knowledge, of B. ceti–associated meningitis and arthritis in a third species, the short-beaked common dolphin (Delphinus delphis), in an animal that stranded in the UK.

We report the number of strandings caused by crude oil among loggerhead turtles (Caretta caretta) in the Canary Islands between 1998 and 2011 and analyze the impact of the designation of the Canary Islands as a Particularly Sensitive Sea Area (PSSA) in 2005. Among 1,679 stranded loggerhead turtles, 52 turtles stranded due to crude oil (3.1%). The survival rate of the turtles stranded by crude oil was 88%. All turtles that died because of crude oil stranding had signs of ingestion of crude oil and lesions, included esophageal impaction, necrotizing gastroenteritis, necrotizing hepatitis, and tubulonephrosis. The number of strandings caused by crude oil after 2005 was significantly lower than it was before 2006. We show that the designation of the Canary Islands as a PSSA in 2005 by the International Maritime Organization was associated with a reduction of sea turtle strandings caused by crude oil.

Leptospirosis is a zoonotic disease caused by the bacterium Leptospira interrogans. The organism is typically maintained within a geographic region by colonizing renal tubules of carrier animals and shed into the environment in urine. We assessed whether L. interrogans was present in fox squirrels (Sciurus niger) in Larimer County, Colorado, USA, and whether it is associated with disease. Twenty-two squirrels were trapped from 29 November 2011 to 15 December 2011 for use in an unrelated study. The squirrels were individually housed for 33–65 days and euthanized; no clinical disease was observed. On gross examination, significant renal lesions were observed in 6 of 22 animals (27%). Histologically, affected animals had severe neutrophilic tubulitis with interstitial nephritis. Immunohistochemistry was conducted on the kidneys of all animals and 10 of 22 (45%) were positive for L. interrogans, with varying severity of infection. The same 10 squirrels were serologically positive for antibodies specific to L. interrogans. These results suggest that L. interrogans is present in fox squirrels in Larimer County, Colorado, USA, and may be associated with varying degrees of renal disease. Further investigation into the role of wildlife in the ecology of leptospirosis within the region is warranted.

The Iriomote cat (IC; Prionailurus iriomotensis) and the Tsushima leopard cat (TLC; Prionailurus bengalensis euptilura) are endangered wild felids in Japan. As a part of ongoing conservation activities, we conducted a molecular, epidemiologic survey of Bartonella, Ehrlichia, and Anaplasma infections in wild IC and TLC populations. Blood samples (47 from 33 individual IC; 22 from 13 TLC) were collected between August 2002 and January 2011. Using PCR analysis, we confirmed the presence of Bartonella henselae in ICs and Bartonella clarridgeiae in TLCs, with prevalences of 6% and 8%, respectively. Using PCR and basic local alignment search tool analyses, we identified Ehrlichia canis in both cats and Anaplasma bovis in TLCs. The prevalence of E. canis was 12% in ICs and 8% in TLCs, and the prevalence of A. bovis was 15% in TLCs. This is the first report, to our knowledge, of B. henselae, B. clarridgeiae, E. canis, and A. bovis infections in these two endangered species. Continuous monitoring of these pathogens is needed for their conservation.

We used 16S rRNA sequencing and leukotoxin gene (lktA) screening via PCR assay to clarify phylogenetic and epidemiologic relationships among Pasteurellaceae isolated from bighorn sheep (Ovis canadensis). Only six of 21 bighorn isolates identified as “Mannheimia haemolytica” in original laboratory reports appeared to be isolates of M. haemolytica sensu stricto based on 16S rRNA sequence comparisons; the remainder grouped with M. glucosida (n=8) or M. ruminalis (n=7). Similarly, 16S rRNA sequence comparisons grouped only 16 of 25 trehalose-fermenting bighorn isolates with reference strains of Bibersteinia trehalosi; nine other trehalose-fermenting bighorn isolates formed a clade divergent from B. trehalosi reference strains and may belong to another species. Of the 16 bighorn isolates identified as B. trehalosi by 16S rRNA sequences, only nine carried detectable lktA and thus seemed likely pathogens; none of the Bibersteinia clade isolates yielded detectable lktA despite reportedly showing β hemolysis in culture. Our findings suggest that traditional metabolism-based methods for identifying Pasteurellaceae isolates lack sufficient accuracy and resolution for reliably discerning bacterial causes of respiratory disease in bighorn sheep. Consequently, these traditional methods should minimally be augmented by molecular techniques to improve epidemiologic relevance. Streamlined surveillance approaches focused primarily on detecting pathogenic Pasteurellaceae (e.g., M. haemolytica sensu stricto and lktA-positive B. trehalosi) and other select pathogens may be most informative for investigating and managing bighorn respiratory disease.

We document the first associations of two recently described species of Pasteurellaceae bacteria with lesions in wild pinnipeds in rehabilitation. Samples were collected from nine lesions in four California sea lions (Zalophus californianus) and two Pacific harbor seals (Phoca vitulina) during necropsy or admission examinations at a rehabilitation facility in northern California. Seven Pasteurellaceae isolates were identified using phenotypic tests and partial rpoB gene sequencing. Six strains of Otariodibacter oris were isolated from California sea lions. Otariodibacter oris was isolated in pure culture from four abscesses, an affected lymph node, and a bone lesion consistent with osteomyelitis. Otariodibacter oris was also cultured with Arcanobacterium phocae and β-hemolytic streptococci. A pure culture of Bisgaardia genomospecies 1 was obtained from an abscess in a harbor seal. This is the first time, to our knowledge, that O. oris has been associated with infection. Isolation of these bacteria in pure culture from abscesses and osteomyelitis strongly indicates a pathogenic potential of this organism. Likewise, the isolation of Bisgaardia genomospecies 1 in pure culture from an abscess in a harbor seal implies causality.

Infection with Elaeophora schneideri, a filarial parasite, occurs commonly in mule deer (Odocoileus hemionus) and elk (Cervus elaphus nelsoni), but seemingly less so in moose (Alces alces). Of 109 carotid artery samples from moose harvested throughout Colorado, USA, in 2007, 14 (13%; 95% binomial confidence interval [bCI]=7–21%) showed gross and 91 (83%; 95% bCI=75–90%) showed histologic evidence of elaeophorosis. Although neither blindness nor other clinical signs associated with elaeophorosis were reported among the harvested moose we examined, the pervasiveness of this parasite may motivate further study of the potential effects of elaeophorosis on moose survival and population performance in the southern Rocky Mountains. Our data suggest histopathology may be more sensitive than gross examination in detecting elaeophorosis in harvested moose.

Coyotes (Canis latrans) are commonly infested with ticks, including Amblyomma americanum, the predominant vector of Ehrlichia chaffeensis and Ehrlichia ewingii; Dermacentor variabilis, an important vector of Rickettsia rickettsii; and Amblyomma maculatum, a major vector of Rickettsia parkeri, a spotted fever group (SFG) Rickettsia. To determine the degree to which coyotes are infected with or exposed to tick-borne bacterial disease agents, serum samples collected from coyotes in Oklahoma and Texas were tested for antibodies reactive to R. rickettsii, Ehrlichia canis, E. chaffeensis, E. ewingii, Borrelia burgdorferi, and Anaplasma phagocytophilum by indirect fluorescent antibody (IFA) testing or enzyme-linked immunosorbent assay (ELISA). Of the coyotes tested, 60% (46/77) and 64% (47/74) had antibodies reactive to R. rickettsii and E. chaffeensis, respectively, on IFA. Additionally, 5% (4/77) had antibodies reactive to E. canis, but not B. burgdorferi or A. phagocytophilum, on SNAP^® 4Dx^® ELISA; subsequent serologic analysis by plate ELISA using species-specific peptides revealed antibodies to E. ewingii, E. canis, and E. chaffeensis in 46% (23/50), 18% (9/50), and 4% (2/50) of serum samples, respectively. Taken together, these data indicate that coyotes in this region are commonly exposed to SFG Rickettsia and E. ewingii and that further consideration of coyotes as a component of the maintenance cycle for these pathogens may be warranted.

Seven grizzly (Ursus arctos; four male, three female) and three black (Ursus americanus; two male, one female) bears caught in culvert traps or leg snares were immobilized in northwestern Wyoming with carfentanil and xylazine at doses, respectively, of 0.011±0.001 and 0.12±0.01 mg/kg for grizzly bears and 0.014±0.002 and 0.15±0.04 mg/kg for black bears. These drugs were antagonized with 1 mg/ kg naltrexone and 2 mg/kg tolazoline. Induction and recovery times, respectively, were 4.3±0.5 and 7.1±0.8 min for grizzly bears and 5.2±0.4 and 9.1±2.2 min for black bears. Inductions were smooth and uneventful. Recoveries were characterized initially by increased respiration followed by raising of the head, which quickly led to a full recovery, with the bears recognizing and avoiding humans and moving away, maneuvering around obstacles. All bears experienced respiratory depression, which did not significantly improve with supplemental oxygen on the basis of pulse oximetry (P=0.56). Rectal temperatures were normothermic. Carfentanil-xylazine immobilization of bears provided significant advantages over other drug regimens, including small drug volumes, predictable inductions, quick and complete recoveries, and lower costs. On the basis of these data, both grizzly and black bears can be immobilized effectively with 0.01 mg/kg carfentanil and 0.1 mg/kg xylazine.

We measured the abundance of sarcoplasmic masses within skeletal muscle myocytes of an adult female stranded pigmy sperm whale (Kogia breviceps). The presence of these masses in other species has been reported in association with myopathies, including myotonic dystrophy, the most frequently related pathology. Other histopathologic muscle changes included a high number of internal nuclei, variations in fiber size and shape, and the predominance of type I fibers.

We captured 140 bats of seven species in Merida City in the Yucatan Peninsula of Mexico in 2010. Serum was collected from each bat and assayed by plaque reduction neutralization test (PRNT) using six flaviviruses: West Nile virus, St. Louis encephalitis virus, and dengue viruses 1–4. Flavivirus-specific antibodies were detected in 26 bats (19%). The antibody-positive bats belonged to three species: the Pallas’s long-tongued bat (Glossophaga soricina), Jamaican fruit bat (Artibeus jamaicensis), and great fruit-eating bat (Artibeus lituratus), and their flavivirus antibody prevalences were 33%, 24%, and 9%, respectively. The PRNT titers were usually highest for dengue virus 2 or dengue virus 4, but none of the titers exceeded 80. These data could indicate that most of the antibody-positive bats had been infected with dengue virus. However, because all titers were low, it is possible that the bats had been infected with another (perhaps unrecognized) flavivirus not included in the PRNT analysis, possibly a virus more closely related to dengue virus than to other flaviviruses. Each serum sample was assayed for flavivirus RNA by reverse transcription PCR, but all were negative.

Surveillance for evidence of West Nile virus (WNV) infection in Morelet’s crocodiles (Crocodylus moreletii) was conducted in Campeche State, Mexico, in 2007. Sera from 62 crocodiles (32 free-ranging and 30 captive) were assayed for antibodies to WNV by epitope-blocking enzyme-linked immunosorbent assay. Antibodies to WNV were detected in 13 (41%) wild and nine (30%) captive crocodiles, and the overall antibody prevalence was 35%. Although evidence of WNV infection in captive crocodiles has been reported in Mexico, we provide the first evidence of WNV exposure in wild crocodiles in Mexico.

Chronic wasting disease (CWD) adversely affects both wild and captive cervid populations. A vaccine to prevent CWD would be a highly desirable tool to aid in disease management. To this end, we tested in mule deer a combination of CWD vaccines consisting of cervid prion peptide sequences 168-VDQYNNQNTFVHDC-182 and 145-NDYEDRYYRENMYRYPNQ-164 that had previously been shown to delay onset of clinical disease and increase survival in a mouse-adapted scrapie model. Thirteen captive mule deer (Odocoileus hemionus) were divided into vaccine (n=7) and control groups (n=6), and given prime and boost vaccinations intramuscularly 5 wk apart. Eight weeks postprime (3 wk postboost), all animals were challenged via natural exposure to an environment contaminated with infective CWD prions. Deer were monitored intermittently for prion infection by rectal and tonsil biopsies beginning 275 days postchallenge. All vaccinates responded to both peptide conjugates present in the combination vaccine as measured by enzyme-linked immunosorbent assay. However, all deer eventually became infected regardless of vaccine status.

Anthrax outbreaks in white-tailed deer, Odocoileus virginianus, are frequent in west Texas, USA, particularly across the Edwards Plateau. However, the outbreak severity varies among years. We summarize the outbreak history in white-tailed deer at a ranch north of Del Rio, Texas, from 2001 to 2010 and compare mortality rates to remotely sensed vegetation indices derived from Moderate Resolution Imaging Spectroradiometer satellite data. It has long been posited that the occurrence of mid- to high-latitude epizootics is associated with hot, dry summer conditions preceded by a wet spring, with cases occurring after summer rain events. Here we employed vegetation green-up indices as a proxy for such environmental conditions. Annual trajectories of vegetation indices identified a clear pattern of early green springs with dry summers in severe outbreak years. In contrast, later, less intense spring green-up with greener summers were associated with enzootic mortality years, when few cases occurred. There was a statistically significant difference in the annual timing and intensity of spring green-up from vegetation indices for epizootic and enzootic years. Years with epizootics have early, intense spring conditions, whereas enzootic years have low-intensity green-up. These results suggest that early green-up signatures may be useful in identifying epizootic climatic conditions ahead of the summer anthrax period. Such analyses are required to ultimately develop an early warning system for wildlife managers and veterinary public health officials.

Mycoplasma agassizii causes upper respiratory tract disease (URTD) in Texas tortoises (Gopherus berlandieri). To determine exposure to and shedding of M. agassizii, we collected blood samples and nasal swabs from 40 free-ranging Texas tortoises on public and private lands in Texas, USA, from May to October 2009. We used an enzyme-linked immunosorbent assay (ELISA) to detect M. agassizii–specific antibodies. Eleven (28%) tortoises were antibody positive, three (8%) were suspect, and the remaining 26 (65%) were negative. Nasal lavage samples were collected from 35 of the 40 tortoises for M. agassizii culture and PCR to detect shedding of M. agassizii. Current infection with M. agassizii was confirmed in one tortoise that had mild clinical signs of URTD and was positive by ELISA (antibody titer >512), PCR, and culture. The clinical isolate was confirmed as M. agassizii by restriction fragment length polymorphism and immunobinding.

Although pseudorabies virus can affect a wide range of mammalian and avian hosts, swine are the only natural hosts of the virus. The US commercial swine industry obtained pseudorabies-free status in 2004, which was important because of the economic value of domestic swine production; however, feral swine remain competent hosts and represent a constant threat for reintroducing the virus into the commercial industry. To better assess feral swine infection status, we collected 8,498 serum samples from feral swine across the United States between 1 October 2009 and 30 September 2012. Of these, 18% were antibody positive in 25 of 35 states where samples were collected, indicating that transmission risk is widespread.

To determine if bison (Bison bison) bulls from Yellowstone National Park (YNP), Montana, USA, shed an infective dose of Brucella abortus in semen, 50 YNP bulls were captured on public lands in Montana during the winter and early spring (April–May) of 2010 and 2011. The bulls were immobilized, and blood and semen samples were collected for serology and Brucella culture. Thirty-five bulls (70%) were antibody-positive, and B. abortus was cultured from semen in three (9%) of the 35 antibody-positive or suspect bulls, though not at concentrations considered an infective dose. Eight bulls (six antibody-positive, two negative) had palpable lesions of the testes, epididymides, or seminal vesicles consistent with B. abortus infection. Breeding soundness exams and semen analysis suggested that antibody-positive bulls were more likely to have nonviable ejaculate (8/35; 23%) than bulls without detectable antibody (2/15; 13%).

Feline immunodeficiency virus (FIV) classically infects felid species with highly divergent species-specific FIVs. However, recent studies have detected an FIV strain infecting both bobcats (Lynx rufus) and pumas (Puma concolor) in California and Florida. To further investigate this observation, we evaluated FIV from bobcats in Florida (n=25) and Colorado (n=80) between 2008 and 2011. Partial viral sequences from five Florida bobcats cluster with previously published sequences from Florida panthers. We did not detect FIV in Colorado bobcats.

In live animals, lead poisoning can be diagnosed by analyzing blood samples. For postmortem testing, blood samples are not available and analysis of liver or kidney is often used for diagnosis. Liver and kidney analysis is relatively expensive and results might not be quickly available. We examined an inexpensive, rapid method to screen animals for lead toxicosis postmortem by testing the mixture of body fluids (termed ‘‘tissue fluids’’) that pool in the body cavity at necropsy for lead. At necropsy we collected body fluid and liver samples from Common Loon (Gavia immer) and Bald Eagle (Haliaeetus leucocephalus) carcasses and determined concentrations of lead in tissue fluid using a desk-top blood lead analyzer. Concentrations of lead in liver were determined by inductively coupled plasma mass spectroscopy. There was strong correlation between tissue fluid and liver tissue lead concentrations, and receiver-operating characteristic analysis gave an area under the curve of 0.91, indicating that postmortem measurements of lead in tissue fluids can be utilized as a screening method for lead toxicosis.

Hemotropic mycoplasmas are bacteria that infect erythrocytes and cause sub-clinical infections to life-threatening disease. We describe hemotropic mycoplasma infection in a free-ranging black howler monkey (Alouatta caraya). This is the first molecular detection of a hemotropic mycoplasma in a nonhuman primate from Brazil.

We describe the first reported case of mycobacterial infection in a free-ranging pinniped in the Northern Hemisphere. Acid-fast bacteria were demonstrated histologically in the liver of an adult female common seal (Phoca vitulina), and Mycobacterium avium subsp. avium was cultured from the liver.

X-rays of the deformed femur of a least shrew (Cryptotis parvus) revealed an ossified callus enclosing a greatly displaced, spiral fracture. Based on models of fracture healing, this shrew lived for at least 28 days postfracture. Survival of major fractures in the wild may not be uncommon.

During 2007–09, we necropsied striped skunks (Mephitis mephitis) from Colorado, USA. Eight of 51 (16%) had severe infections with the subcutaneous filarid nematode Filaria taxideae, and four of the infected skunks (50%) had dermatitis that was histologically associated with parasite ova in the skin.

We detected Cryptosporidium sp. by direct immunofluorescence in fecal samples from greater bamboo lemurs (Prolemur simus) and eastern rufous mouse lemurs (Microcebus rufus) inhabiting the Ranomafana National Park, Madagascar. This is the first report of an occurrence of these potentially zoonotic parasites in free-ranging lemurs in the rain forest of Madagascar.

We screened 18 endangered Chilean huemul (Hippocamelus bisulcus) for antibodies to infectious agents. We detected no antibody to bovine herpesvirus-1 (BHV-1) or Brucella abortus (BA); two huemul had antibody to bovine viral diarrhea virus (BVDV). Cattle (n=35) had antibody to BVDV and BHV-1 but not BA.

Rodents often live in close association with anthropogenic environments, offering rodent-associated pathogens numerous opportunities for transmission to humans. We screened for zoonotic gastrointestinal pathogens in rodents caught during autumn/winter in human dwellings in rural areas surrounding Uppsala, Sweden. We detected Campylobacter jejuni, Giardia sp., and Cryptosporidium sp. in rodent intestines.

We report blood parameters for 18 adult and 12 juvenile, free-ranging, female Nile monitors (Varanus niloticus) in Egypt. We report differences between adult and young monitors and compare parameters to those of other reptile species.