Decreases in Greater Sage Grouse (Centrocercus urophasianus) numbers throughout the western United States have been attributed to declining habitat quantity and quality. Improving our understanding of how interannual ecologic site variability affects nutritional status and fitness of different bird age classes will lead to improved land management and conservation strategies. Greater Sage Grouse were sampled from two Population Management Units located in northern Nevada, United States: Tuscarora (TU) and Lone Willow (LW) during 15 March–11 April 2004 and 14–20 March 2005. Twenty (16 yearlings, four adults) and 17 (7 yearlings, 10 adults) female Sage Grouse were captured and bled during 2004, and 12 (four yearlings, eight adults) and 14 (10 yearlings, four adults) were sampled during 2005 in TU and LW, respectively. Samples were evaluated to examine the effect of site, age, and year on specific hematologic and serum chemistry parameters. Several differences between age classes, sites, and years were detected for a number of fitness indicators; however, actual values fell within normal ranges of variation for Sage Grouse or other avian species. Differences were also detected for several parameters more closely related to reproductive fitness, including total plasma and serum proteins, and serum calcium and phosphorus. Yearlings had lower plasma protein (P<0.0001) and lower serum protein than did adults (P=0.0003). In 2004, TU yearlings had lower serum calcium levels than the adults, and in 2005, LW yearlings had lower levels than adults (P=0.008). Females on the TU site had lower serum phosphorus than the LW females (P<0.0001). Overall, adult females weighed more than yearlings (P=0.0004). Lower values found in yearlings, and on the TU management unit, indicate a lower production potential, particularly in unfavorable years. A lower intrinsic ability of yearlings to reproduce, combined with lower nutrition potentials and associated annual variations on certain types of habitat combinations, indicate that conservation measures must be flexible and based on local prescriptions. Fitness parameters of Sage Grouse should be used to assess effects of land management practices and conservation on Sage Grouse populations in order to provide more certainty of the outcome, whether positive, neutral, or deleterious.

Baseline blood chemistry data could be particularly valuable if reference values from free-ranging populations of rare or endangered species are not available. The Canada lynx (Lynx canadensis) is listed as threatened under the Endangered Species Act in the conterminous United States, even though the species is managed as a furbearer in Alaska and in most provinces of Canada. Body mass, blood chemistry, and hematologic data for free-ranging lynx were collected from 2003 to 2007 and for captive lynx from 1984 to 2007. Up to 2 yr of age, captive lynx were consistently heavier than free-ranging lynx. Body mass of adult free-ranging lynx was similar to body mass of captive adult lynx. Some differences in blood chemistry between free-ranging and captive lynx were statistically significant, but most measured values were within reference ranges for domestic cats. Free-ranging lynx had higher concentrations of aspartate aminotransferase, alanine aminotransferase, and blood urea nitrogen than did captive lynx, and these were outside the reference value ranges for domestic cats. Alkaline phosphatase and phosphorus were higher in juveniles (<12 mo when captured) as compared to adults. Free-ranging lynx maintained body mass between serial captures. Hematologic values, blood chemistry values, and body mass of free-ranging Canada lynx provide support for the hypothesis that Canada lynx in Minnesota, at the southern edge of their range, are in normal physical condition.

Alphaviruses (Togaviridae) infect wild birds, but clinical illness and death attributable to virus in naturally infected birds is rarely reported, particularly for small passerine species or nestlings. Buggy Creek virus is a unique alphavirus in the Western equine encephalomyelitis virus (WEEV) complex that is vectored by the cimicid swallow bug (Oeciacus vicarius), an ectoparasite of the colonially nesting Cliff Swallow (Petrochelidon pyrrhonota) and the introduced House Sparrow (Passer domesticus). While sampling birds for Buggy Creek virus (BCRV) during the summers of 2007 and 2008, we discovered large numbers of clinically ill or dead House Sparrow nestlings. Ill nestlings exhibited ataxia, torticollis, paresis, and lethargy. Histologic examination revealed that encephalitis was the most common finding, followed by myositis, myocarditis, and hepatic changes, but pathology was highly variable. We isolated BCRV from brain tissue in most of the ill or dead nestlings, and from blood, liver, kidney, spleen, lung, feather pulp, and skin in some birds. To our knowledge, this is the first report of clinical illness, gross pathology, and histopathology for a WEEV-complex alphavirus in a field-collected passerine species.

The pathogenic potential of deerpox virus was investigated via an experimental study utilizing seven black-tailed deer fawns (Odocoileus hemionus) between June and August of 2007. Successful transmission was achieved via intracutaneous and intravenous routes, and by commingling an uninoculated animal with experimentally infected fawns. One fawn became depressed and reluctant to eat but systemic clinical signs in the other fawns were confined to mild transient pyrexia. Typical multifocal poxviral cutaneous lesions of erythema, papules, pustules, ulceration, and crusting were observed. Two locally extensive ulcerative lesions also developed at inoculation sites. Oral lesions were seen in one commingled fawn, with some palatine epithelial cells containing intracytoplasmic inclusions. Microscopic cutaneous lesions included epithelial cell hyperplasia with hydropic change, intraepithelial pustules, erosions, folliculitis, and dense leukocytic dermal infiltrates. Transmission was confirmed by one or more of virus isolation, polymerase chain reaction or serum neutralization tests. Significant internal lesions were not seen at necropsy.

Three Blue Penguins (Eudyptula minor) were rescued between July 2006 and January 2007 off the east coast of Auckland, New Zealand. They were taken to a rehabilitation center, where they subsequently died in May 2007 and were submitted for necropsy. There was unilateral enlargement of the salt glands with disseminated small, pale, and firm foci in all birds. Histologic examination of the affected glands demonstrated the presence of multifocal granulomas and areas of severe squamous metaplasia of the collecting ducts. The remaining gland had areas of hyperplasia, dysplasia, and necrosis with a severe granulomatous inflammatory reaction. Intralesional gram-negative bacteria were detected, but, unfortunately, bacterial culture was unrewarding. No further cases were observed in penguins in the subsequent year, and the primary cause of the salt gland adenitis remains uncertain.

Mucor amphibiorum is the only pathogen known to cause significant morbidity and mortality in the free-living platypus (Ornithorhynchus anatinus) in Tasmania. Infection has also been reported in free-ranging cane toads (Bufo marinus) and green tree frogs (Litoria caerulea) from mainland Australia but has not been confirmed in platypuses from the mainland. To date, there has been little genotyping specifically conducted on M. amphibiorum. A collection of 21 Mucor isolates representing isolates from the platypus, frogs and toads, and environmental samples were obtained for genotypic analysis. Internal transcribed spacer (ITS) region sequencing and GenBank comparison confirmed the identity of most of the isolates. Representative isolates from infected platypuses formed a clade containing the reference isolates of M. amphibiorum from the Centraal Bureau voor Schimmelcultures repository. The M. amphibiorum isolates showed a close sequence identity with Mucor indicus and consisted of two haplotypes, differentiated by single nucleotide polymorphisms within the ITS1 and ITS2 regions. With the exception of isolate 96-4049, all isolates from platypuses were in one haplotype. Multilocus fingerprinting via the use of intersimple sequence repeats polymerase chain reaction identified 19 genotypes. Two major clusters were evident: 1) M. amphibiorum and Mucor racemosus; and 2) Mucor circinelloides, Mucor ramosissimus, and Mucor fragilis. Seven M. amphibiorum isolates from platypuses were present in two subclusters, with isolate 96-4053 appearing genetically distinct from all other isolates. Isolates classified as M. circinelloides by sequence analysis formed a separate subcluster, distinct from other Mucor spp. The combination of sequencing and multilocus fingerprinting has the potential to provide the tools for rapid identification of M. amphibiorum. Data presented on the diversity of the pathogen and further work in linking genetic diversity to functional diversity will provide critical information for its management in Tasmanian river systems.

The fungal pathogen Batrachochytrium dendrobatidis (Bd), the causative agent of chytridiomycosis, has been implicated in amphibian declines worldwide. It has been hypothesized that low inherent immunogenicity in Bd may be related to the high rates of morbidity and mortality that are associated with Bd-infected anuran populations. To test this idea, juvenile Rana muscosa (mountain yellow-legged frogs) were immunized with adjuvants in combination with a formalin-killed Bd culture to determine if it is possible to stimulate a protective immune response when challenged with a live inoculum of B. dendrobatidis. Three groups of juvenile R. muscosa (6 mo postmetamorphosis) were immunized with saline, Freunds Complete (FCA) and Incomplete Adjuvant (FIA), or the adjuvants in combination with a formalin-killed culture of B. dendrobatidis. The effects of immunization were modeled using survival analysis and a proportional hazards model. No significant differences were found between the groups in overall mortality, time to infection, infection prevalence, or intensity. While this study suggests that immunizing anurans against chytridiomycosis will not alter rates of infection or mortality among individuals, it does raise several questions regarding the attenuation and efficacy of anuran adaptive immune responses and whether they may be protective against this disease.

In 1996, the Hook Lake Wood Bison Recovery Project was initiated to establish a small, disease-free, captive, bison-breeding herd. Founders originated from wild bison herds in the Slave River Lowlands in northern Canada, which, like other bison herds in and around Wood Buffalo National Park, are endemically infected with bovine tuberculosis (caused by Mycobacterium bovis) and brucellosis (caused by Brucella abortus). After 9 yr of apparent disease freedom, tuberculosis was detected within the captive herd, leading to complete depopulation. This study examined the performance of antemortem tuberculosis diagnostic tests used during the project. Performances of the caudal-fold test, fluorescent polarization assay, multiantigen print immunoassay (MAPIA), and the rapid test (RT) were assessed by estimating sensitivity, specificity, positive predictive value, and negative predictive value for each test. Kappa values measuring agreement between tests were calculated. Overall, the tests did not differ with respect to sensitivities and specificities, which ranged from 50% to 92% and from 34% to 100%, respectively. The MAPIA tended to show high sensitivity, and there was significant agreement only between the MAPIA and RT. Serum collected from infected animals at slaughter produced highly variable results on the different assays, and one infected bison was negative on all antemortem tests. The results of this analysis suggest use of multiple antemortem tests in parallel, particularly those incorporating multiple antigens, to optimize sensitivity in detecting bovine tuberculosis in bison. However, as demonstrated in this herd, even a seemingly optimal antemortem testing regimen can fail to detect M. bovis–infected individuals.

Brucella abortus has been an important wildlife disease issue for most of the last century, especially because wildlife species are considered to be important disease reservoirs for cattle. Diagnostic uncertainty, caused in part by cross-reactions of antibodies to environmental pathogens such as Yersinia enterocolitica O:9 on standard Brucella serology, has exacerbated the challenges of managing the disease and has highlighted the need for test validation in wildlife species. The western immunoblot was evaluated for use in detecting B. abortus exposure in elk (Cervus elaphus) and for ruling out exposure to cross-reacting bacteria. Samples collected from 2003 to 2006, including 54 female and immature elk from four different elk herds, were tested using standard Brucella serologic methods (card, rapid automated presumptive [RAP], and rivanol tests), as well as the western immunoblot. Samples (n=28) from animals known to be naturally infected with B. abortus biovar 1 served as positive controls. For presumed negative samples, sera (n=26) were collected from two elk herds in which negative serologic tests, and the absence of clinical signs of disease such as abortions, supported Brucella-negative classification. In addition to these study samples, serologic data from 12 tule elk (Cervus elaphus nannodes) were provided from the California Department of Fish and Game in order to illustrate a field application of the western blot. The western immunoblot had the highest sensitivity (1.0;% 0.899–1.0) and specificity (1.0; 0.891–1.0) among all tests used in the study. The Kappa statistic for agreement between the western blot and the card, rivanol, and RAP tests were 0.701, 0.808, and 0.921, respectively, showing good to excellent agreement with the standard diagnostic tests currently in use. Although the western immunoblot is more expensive and time intensive than other tests, in this limited study, it was shown to be reliable for establishing and confirming B. abortus disease status in elk. In addition to this study, subsequent applications of the western blot assay have been successful in detecting Yersinia sp. exposure in elk after their antibodies cross-reacted on standard Brucella serology.

Based on previous necropsy results, Microcystis blooms in constructed water impoundments in the Kruger National Park (KNP) have been identified as a cause of wildlife mortality. In response to wildlife mortality during 2007, water samples, containing algal bloom material, were collected during February 2007 and July 2007 from four dams (Nhlanganzwani, Mpanamana, Makhohlola, and Sunset) in the southeastern part of the KNP as part of the follow-up investigation. The toxicity of the Microcystis blooms was determined using the enzyme-linked immunosorbent assay (ELISA), protein phosphatase inhibition (PPI) assay, mouse bioassay, and African sharptooth catfish (Clarias gariepinus) primary hepatocytes. Both the ELISA and PPI assays indicated that the water sample collected during February 2007 from the Nhlanganzwani Dam, and samples collected from the Nhlanganzwani and Sunset dams in June 2007, were toxic. These dams, exhibiting the toxic Microcystis blooms, were also associated with the wildlife mortality. Mice injected intraperitoneally with water samples from Nhlanganzwani Dam (February 2007) induced hepatotoxicity and mortality within 1 hr. Primary hepatocytes from the sharptooth catfish exposed to samples from these dams gave similar results. This laboratory investigation and results strongly incriminate the toxic Microcystis blooms as the cause of the wildlife mortality. Eutrophication and bloom formation appear to have been the consequence of the high numbers of hippopotami (Hippopotamus amphibius) in specific dams.

Environmental contamination by petroleum hydrocarbons from anthropogenic sources can be a cause of stress for free-ranging wildlife. The response of wildlife to chemical contaminants requires that the hypothalamic–pituitary–adrenal (HPA) axis be precisely regulated to allow for proper glucocorticoid-mediated adaptive responses. Chronic oral exposure to low concentrations of bunker C fuel oil causes the development of adrenal hypertrophy in male ranch mink (Mustela vison) without increasing serum or fecal glucocorticoid concentrations. This hypertrophy is an adaptive response to fuel oil-induced adrenal insufficiency. To determine if the same phenomenon occurs in female mink or male mink exposed to artificially weathered fuel oil, female mink were fed 0 ppm (mineral oil) or 420 ppm fuel oil and male mink were exposed to 0 ppm, 420 ppm fuel oil, or 480 ppm artificially weathered fuel oil in the diet for 60–62 days. At the end of the exposure, serum glucocorticoid concentrations were assayed along with body and organ weight measurements. Fecal glucocorticoid concentrations were assayed at time points throughout the exposure. Male mink fed fuel oil or weathered fuel oil and female mink fed fuel oil had adrenal enlargement without any significant increases in the serum or fecal concentration of glucocorticoids, which is consistent with fuel oil–induced adrenal insufficiency. To address the physiological consequences of adrenal insufficiency, fuel oil–exposed male mink were administered an adrenocorticotropic hormone (ACTH) stimulation test. Fuel oil–exposed animals had a smaller incremental increase in serum glucocorticoid concentration after ACTH challenge compared to control animals. Our findings provide further evidence that the HPA axis of fuel oil–exposed animals is compromised and, therefore, not able to respond appropriately to the diverse stressors found in the environment.

An increase in the incidence of debilitated loggerhead sea turtle (Caretta caretta) strandings in the southeastern United States has been observed in recent years. These turtles are characterized by emaciation and heavy burdens of external and internal parasites, and bacterial infections, but the underlying cause of their condition is unknown. To investigate further the causes of these strandings, a health assessment was performed on stranded, debilitated loggerhead turtles, and contaminant concentrations in various tissues were compared to those from healthy turtles. This portion of the study investigated the potential role of mercury (Hg) toxicity in the debilitated condition described above. Hematocrit, total protein, albumin, globulin, glucose, calcium, lymphocyte counts, heterophil:lymphocyte ratios, aspartate aminotransferase, uric acid, sodium, and chloride were altered in debilitated loggerheads relative to healthy animals. However, none of the aforementioned health indicators correlated with Hg concentrations in either red blood cells (RBCs) or plasma. The Hg concentration in RBCs was 129±72 (mean±standard deviation) times higher than in plasma, causing a significant dilution of Hg in whole blood due to extreme anemia. Mercury concentrations in RBCs (73.7±21.2 ng/g) and scutes (455±57 ng/g) from debilitated turtles were similar to or lower than those reported for healthy animals, indicating no elevation in Hg exposure before and during the progression of this condition. These findings suggest that Hg toxicity does not play a role in the debilitated loggerhead condition observed in the southeastern United States.

A key component of wildlife disease surveillance is determining the spread and geographic extent of pathogens by monitoring for infected individuals in regions where cases have not been previously detected. A practical challenge of such surveillance is developing reliable, yet cost-effective, approaches that remain sustainable when monitoring needs are prolonged or continuous, or when resources to support these efforts are limited. In order to improve the efficiency of chronic wasting disease (CWD) surveillance in Colorado, United States, we developed a weighted surveillance system exploiting observed differences in CWD prevalence across demographic strata within infected mule deer (Odocoileus hemionus) populations. We used field data to estimate sampling weights for individuals from eight demographic strata distinguished by differences in apparent health, sex, and age. In this system, individuals from a sample source with high prevalence and low inclusion probability (e.g., clinical CWD ‘‘suspects'') received ≥10.3 times more weight than those from a source with low prevalence and high inclusion probability (e.g., apparently healthy, hunter-harvested individuals). We simulated use of this alternative surveillance system for a deer management unit in Colorado and evaluated the potential effects of using biased weights on the probability of failing to detect CWD and on relative surveillance costs. We found that this system should be transparent, cost-effective, and reasonably robust to the inadvertent use of biased weights. By implementing this, or a similar, weighted surveillance system, wildlife agencies should be able to maintain or improve current surveillance standards while, perhaps, collecting and examining fewer samples, thereby increasing the efficiency and cost-effectiveness of ongoing CWD surveillance programs.

Genital epithelial tumors of Atlantic bottlenose dolphins (Tursiops truncatus [Tt]) and Burmeister's porpoises (Phocoena spinipinnis) were formerly shown to be associated with papillomavirus (PV) infection. Papillomaviruses are highly prevalent viruses involved in the development of various tumor types in a wide range of animals, and so-called high-risk PVs contribute to malignant progression. In marine mammals, the incidence and prevalence of PV infection, transmission pathways, and persistence of infection are largely unknown. Using virus-like particles of bottlenose dolphin PV type 1 (TtPV1) as the antigen, enzyme-linked immunosorbent assay (ELISA) studies were conducted to evaluate PV antibody prevalence in bottlenose dolphins. In total, sera obtained from 115 dolphins were examined. Fifty-one percent of captive dolphins (n=18 of 35) and 90% of free-ranging dolphins (n=72 of 80) were antibody positive. Higher ELISA reactivity was observed among males compared with females. Sexually immature dolphins appeared more likely to seroconvert with age. Besides determining their PV antibody prevalence, each animal was also assessed for the presence of orogenital tumors. Interestingly, the mean age of free-ranging dolphins with tumors (n=21) was 11.2 yr compared with 29.9 yr in captive dolphins with tumors (n=9). Results from the current study suggest PV infection in bottlenose dolphins is common, that the main route of PV transmission among them may be horizontal, and that orogenital neoplasia may develop in early life stages of certain free-ranging bottlenose dolphins.

We investigated the responses of endoparasites in red-backed voles (Myodes gapperi) to fire in a boreal forest ecosystem. Because fire affects the environmental conditions and biodiversity of the forest ecosystem, the life cycle of parasites may also be affected because of the absence of intermediate hosts in the environment. We hypothesized that the prevalence of endoparasites would be influenced by the parasites' life cycle and habitat characteristics (forest vs. burned). We found that prevalence of endoparasites was different between forested and burned habitats (χ²=37.49, P<0.001). Cestodes, nematodes, and coccidia showed different responses to habitat alteration (χ²=37.43, P<0.001). There was a higher prevalence of cestodes in forested (53.5%) than burned habitats (35.0%). However, there was higher prevalence of coccidia in burned (55.0%) than forested (42.9%) habitats. Furthermore, although prevalence of cestode infection was lower in burned than forested habitat, individuals in both habitats had similar intensities of cestodes. Our study showed that habitat can significantly affect the parasite communities, depending on specific parasite life cycles.

The last outbreak of foot-and-mouth disease (FMD) in the United States occurred in 1929. Since that time, numbers and distribution of feral swine (Sus scrofa) have increased greatly, especially in the southern states. This creates a potential risk to livestock production because swine are susceptible to, and can be carriers of, several economically harmful diseases of livestock. Most importantly, swine are potent amplifiers of FMD virus. In this study, global positioning system (GPS) collars were placed on rangeland cattle (Bos indicus × taurus) and feral swine to determine shared habitat use by these species on a large ranch in south Texas from 2004 to 2006. The aim was to identify locations and rates of interspecies contact that may result in effective transfer of FMD virus, should an outbreak occur. In shrubland and riparian areas, animals were dispersed, so contacts within and between species were relatively infrequent. Indirect contacts, whereby cattle and feral swine used the same location (within 20 m) within a 360-min period, occurred primarily at water sources, and seasonally in irrigated forage fields and along ranch roads. Direct contacts between species (animals <20 m apart and within 15 min) were rare and occurred primarily at water sources. Changes in ranch management practices are suggested to reduce interspecies contact should an FMD disease outbreak occur. This information can also be used to improve current epidemiologic models to better fit free-ranging animal populations.

Fleas were collected from live-captured small mammals to identify potential flea-borne pathogens, seasonal prevalence of flea species, and host preference as part of the US military rodent-borne diseases surveillance program conducted at one US military installation and 10 military training sites, northern Gyeonggi Province, Republic of Korea. During 2003–04, 948 fleas (563 females and 385 males) were recovered from 2,742 small mammals (seven rodent and one insectivore species). Apodemus agrarius (striped field mouse) accounted for 88.9% (2,439/2,742) of the small mammals, followed by Crocidura lasiura (4.2%), Mus musculus (2.9%), Microtus fortis (2.2%), Myodes regulus (0.6%), Micromys minutus (0.5%), Tscherskia triton (0.5%), and Rattus norvegicus (0.3%). Small mammal infestation rates (number with fleas/number captured) ranged from 7.7% (M. minutus and T. triton) to 31.3% (M. regulus). Flea indices were highest for M. regulus (0.69/captured rodent), followed by C. lasiura (0.54), M. fortis (0.41), A. agrarius (0.34), and R. norvegicus (0.33). Overall, Ctenophthalmus congeneroides (51.3%) was more frequently collected, followed by Stenoponia sidimi (42.6%), Rhadinopsylla insolita (5.5%), Neopsylla bidentatiformis (0.4%), Rhadinopsylla concava (0.1%), and Doratopsylla coreana (0.1%). Ctenophthalmus congeneroides was more frequently collected from small mammals during the spring and summer, while S. sidimi was more frequently collected during the winter season. Rickettsia typhi, the causative agent of murine typhus, was detected in 3.2% of specimens (7/220 pools from 654 fleas; minimum field infection rate [number of positive pools/total number of fleas] was 1.1%).

To determine whether Bartonella species may perpetuate in sites where ticks are absent, we analyzed blood and spleen samples from small mammals trapped near Cantwell, Alaska during September 1999. Blood smears from seven of 48 Myodes (Clethrionomys) rutilus suggested a Bartonella species infection; five of the seven yielded cultures consistent with Bartonella species; PCR of rrs, gltA, and rpoB genes confirmed Bartonella infection in four of these culture samples. Phylogenetic analysis of rrs, gltA, and rpoB genes demonstrated that the sequences clustered with the Bartonella vinsonii clade. Based on PCR, two additional blood samples and four of 22 spleen samples also contained DNA of this Bartonella species for an overall prevalence of 14% (10/70) among the 70 M. rutilus sampled in this study. We conclude that Bartonella species may commonly infect M. rutilus in a site where ticks are absent.

To estimate the prevalence of Bartonella in small mammals of different species, during different seasons, and at different study sites, and to provide baseline data for the risk assessment of human Bartonella infection, we captured small mammals using snap traps in Zhejiang Province, China. Bartonella species were detected in small-mammal samples by polymerase chain reaction and positive amplicons were sequenced. Bartonella DNA was detected in 47% (90/192) of Apodemus agrarius, 31% (14/45) of Rattus losea, 16% (7/43) of Rattus norvegicus, 24% (9/37) of Eothenomys melanogaster, 4% (1/28) of Niviventer confucianus, 30% (7/23) of Suncus murinus, 22% (2/9) of Microtus fortis, 27% (2/7) of Rattus tanezumi, and 29% (2/7) of Apodemus peninsulae. No Bartonella DNA was detected in 27 unidentified Soricidae or nine Mus musculus. This is the first report of Bartonella DNA detected in E. melanogaster and N. confucianus. The prevalence of Bartonella DNA varied among small-mammal species, study sites, and seasons; the prevalence of Bartonella DNA between genders did not vary significantly within a species. The sequences we report were most similar to Bartonella grahamii.

Surveys for disease agents were conducted in introduced free-ranging elk (Cervus elaphus nelsoni) in Arkansas and Kentucky. Elk had been captured in Colorado and Nebraska and released in Arkansas during 1981–1985. From 1997 through 2002 elk were captured in Arizona, Kansas, North Dakota, New Mexico, Oregon, and Utah and released in southeastern Kentucky. Specimens were collected from 170 hunter-killed elk in Arkansas during 1998–2006, and 44 elk in Kentucky during 2001–2004. Significant findings included isolation of Mycobacterium avium paratuberculosis from one elk in Kentucky and evidence of previous or current infections by Parelaphostrongylus tenuis in several animals in Arkansas. Serological tests provided evidence of previous infection by epizootic hemorrhagic disease virus, bluetongue virus, bovine viral diarrhea virus, infectious bovine rhinotracheitis virus, parainfluenza-3 virus, and multiple serovars of Leptospira interrogans. Mycobacterium bovis, Brucella abortus, chronic wasting disease (CWD), and hemoparasites such as Anaplasma spp. were not detected. Results from elk obtained through these surveys were consistent with exposure to disease agents endemic in livestock and wildlife in Arkansas and Kentucky.

Soon after the appearance of West Nile virus (WNV) in North America, a number of public health authorities designated the American Crow (Corvus brachyrhynchos) a sentinel for WNV detection. Although preliminary studies have suggested a positive association between American Crow mortality and increased risk of WNV infection in humans, we still know little about dynamic variation in American Crow mortality, both baseline levels and mortality associated with WNV. We hypothesized that the complex social behavior of American Crows, which is shaped by age and seasonal factors, influences both baseline mortality and WNV mortality in American Crow populations. We examined American Crow mortality data from Quebec for the 2005 WNV surveillance year, which lasted from 5 June to 17 September 2005. The variables of interest were age, gender, body condition index, time of year, and land cover. We used a log-linear model to examine baseline mortality. Logistic regression and general linear regression models were constructed to examine variables associated with mortality due to WNV. We found that both age and time of year were key variables in explaining baseline mortality. These two variables were also risk factors for WNV mortality. The probability that a carcass tested positive for WNV increased with the age of the dead bird and as summer progressed. WNV-positive carcasses also had a lower body condition index than WNV-negative carcasses. We believe that the first major wave of American Crow mortality observed in the early summer of 2005 was the result of natural mortality among young American Crows. Because this mortality was not linked to WNV, it appears that American Crow may not be a good species for early detection of WNV activity. Our data also suggest that second-year American Crows play a major role in propagating WNV during their movements to urban land covers during midsummer.

Although vesicular stomatitis has been present for many years in the Americas, many aspects of its natural history remain undefined. In this study, we challenged five adult Virginia opossums (Didelphis marsupialis) with vesicular stomatitis New Jersey serotype virus (VSNJV). Opossums had no detectable antibodies against VSNJV prior to being inoculated with 10^6.5 median tissue culture infective doses (TCID₅₀) of VSNJV by two routes; intraepithelial/subepithelial (IE/SE) inoculation and scarification in the muzzle (SM). Clinical response was monitored daily and animals were tested for viral shedding. All infected animals developed vesicles and ulcers on the tongue and inflammation of the nasal alar folds. Virus was isolated from esophagus-pharynx, nasal, and from ocular swabs and lesions samples. The failure to detect viremia in these animals indicates that a source other than blood may be required for transmission to insect vectors. Our results suggest that D. marsupialis could play a role in the maintenance of VSNJV outside of domestic animal populations and could provide a model to study vesicular stomatitis virus pathogenesis.

The brush-tailed rock-wallaby (Petrogale penicillata) is listed as vulnerable on the International Union for Conservation of Nature (IUCN) Red List of Threatened Species. Parasitic diseases have been proposed as possible contributing factors to the decline of the species, but very little is known about the effects of parasites on this host. This study determined the antibody prevalence of the protist Toxoplasma gondii in a wild brush-tailed rock-wallaby population from three neighboring colonies in southeast Queensland, Australia. Fecal egg and oocyst count, tick count, severity of skin rash, and presence of lice and microfilariae were also monitored during four or five trapping periods over 1 yr. Antibodies against T. gondii were detected in 5% of animals (3/64). Fecal egg and oocyst counts were highly variable, but fecal egg counts were lower in subadult animals relative to adults. Neither fecal egg count nor oocyst count was associated with variation in blood variables or condition index, but a negative association between fecal egg count and oocyst count was observed. Microfilariae (Breinlia spelaea), lice (Heterodoxus octoseriatus), and skin lesions were seen more frequently during the November trapping period. A mite, Thadeua sp., was more likely to be detected in these skin lesions than in skin of unaffected wallabies. Tick (Ixodes holocyclus and Haemaphysalis bancrofti) counts also varied between trapping periods and were lowest in the April/May trapping period. This study provides the most detailed account to date of parasite burdens in a vulnerable macropodid, but no clear evidence emerged linking parasites to adverse impact on the host.

There is growing evidence that populations of the northern flying squirrel (Glaucomys sabrinus) are declining in the eastern United States, perhaps due to competition with the southern flying squirrel (Glaucomys volans). Potential causes include parasite-mediated or apparent competition from the shared intestinal nematode, Strongyloides robustus, which has been shown to detrimentally affect the northern flying squirrel but not the southern flying squirrel. To investigate this hypothesis, we conducted a preliminary study on the parasite community of both flying squirrel species from sites in Pennsylvania where the two species occur sympatrically and where G. sabrinus is now considered endangered at the state level. We compared these parasite communities with those from northern flying squirrels from northern New York where the southern flying squirrel is absent. We found eight species of gastrointestinal parasites (Pterygodermatites peromysci, Lemuricola sciuri, Syphacia thompsoni, Syphacia spp., Capillaria spp., Citellinema bifurcatum, Strogyloides robustus, and an unidentifiable cestode species) in both species of flying squirrels examined for our study. The parasite-mediated competition hypothesis was partially supported. For example, in Pennsylvania, S. robustus was overdispersed in southern flying squirrels, such that a small proportion of the hosts carried a large proportion of the worm population. In addition, we found S. robustus to be present in northern flying squirrels when the species were sympatric, but not where southern flying squirrels were absent in New York. However, there was no association between S. robustus and the body condition of flying squirrels. We detected a potential parasite community interaction, as S. robustus abundance was positively associated with P. peromysci.

Posture, ventilation, and acid-base balance using auricular venous blood values (pH, lactate, base excess [BE], HCO₃⁻, PO₂, SO₂, and PCO₂), oxygen saturation of hemoglobin (SpO₂), and end-tidal carbon dioxide (P_ETCO₂) were compared between sternal (STE) and lateral (LAT) recumbency in free-ranging black rhinoceros (Diceros bicornis bicornis) receiving oxygen insufflation. Data are reported as median, minimum, and maximum (median [minimum, maximum]). Thirty-six desert-adapted black rhinoceros (20 male, 16 female; age 8 [1.5, 33] yr) were immobilized in Namibia in March and April of 2008, from a helicopter, by remote intramuscular injection with etorphine HCl, azaperone, and hyaluronidase. Time from darting to recumbency was 6.0 (3, 15.5) min. Data were organized into two sampling periods: sample period 1 (P1, collected within 0–20 min postdarting; 13 [6.5, 19] min) and sample period 2 (P2, collected between 20–40 min postdarting; 32 [22.3, 39] min). All animals were acidemic (pH 7.24 [7.07, 7.32]) and hypoxemic (PO₂ 51 [38, 95.2]; SO₂ 78 [64, 96] mmHg) after capture. Lactate at P1 was 7.2 (3.2, 16.8) mmol/l and decreased (P=0.01) to 4.6 (1.2, 10.9) mmol/l at P2. At P2, lactate was less (P=0.06) in LAT 3.5 (1.2, 8.6) mmol/l than in STE posture 7.4 (3.1, 10.9) mmol/l. In P2, PO₂, SO₂, and SpO₂ were higher (P=0.02, 0.10, and 0.01, respectively) in STE than in LAT. End-tidal carbon dioxide in LAT was 38 (26, 47) mmHg and increased (P<0.001) rapidly to 48 (37, 55) mmHg when animals were moved into STE; no corresponding change in PCO₂ was observed. These preliminary findings suggest that STE posture in recumbent black rhinoceros reduces dead-space ventilation and improves oxygenation. Lateral posture was associated with lower blood lactate, quicker lactate recovery, or both. It is possible that the posture of recumbent rhinoceros after capture affects lactate accumulation and clearance, or both, and procedures should consider positioning in order to enhance perfusion.

We assessed the efficacy and safety of hyaluronidase to improve chemical immobilization of free-ranging polar bears (Ursus maritimus) captured from helicopter by remote drug delivery along the Ontario coast line of northwestern James Bay and southern Hudson Bay during September 2005 and October 2007. We used a single blind study design in which one person prepared and loaded all darts without the shooter knowing whether hyaluronidase (150 IU per dart) or sterile water was added to the immobilizing drug mixture of xylazine and zolazepam-tiletamine (XZT). We found that we often required more than one dart to immobilize bears in the control group (XZT sterile water; >1 dart for 15 of 28 captures) versus the treatment group (XZT hyaluronidase; >1 dart for seven of 26 captures). As a consequence, treatment bears were generally immobilized with smaller XZT dosages (7.9 vs. 9.4 mg/kg; P=0.08) and shorter induction (10 vs. 15 min; P=0.004) than control bears. We found no differences in vital rates and serum biochemistry results between control and treatment bears. We did find, however, that induction times correlated directly with rectal temperature at ≤15 min after immobilization (r=0.39, P=0.004), which suggests that use of hyaluronidase also helped prevent development of high body temperature (hyperthermia) in polar bears. Overall we found hyaluronidase to be effective and safe for capture of polar bears. We recommend further study to determine whether effects of hyaluronidase are dose dependent and recommend that others involved with capture of seasonally fat species such as polar bears consider use of hyaluronidase to improve chemical immobilization.

Serum samples from 52 free-ranging brown bears (Ursus arctos) collected in Croatia over a period of 10 yr (1998–2007) were tested by microscopic agglutination test for specific antibodies (Ab) to 12 Leptospira spp. pathogenic serovars. At titers ranging from 1:100 to 1:2,000, 19 samples (36.5%) were Ab-positive to at least one serovar. Antibodies for 10 Leptospira spp. serovars were detected: Icterohaemorrhagiae, Australis, Sejroe, Canicola, Poi, Hardjo, Ballum, Saxkoebing, Pomona, and Grippotyphosa. In comparison to previous reports, the prevalence of Ab to serovar Icterohaemorrhagiae (52.6%) was significantly higher. Other common serovars were Australis (47.4%) and Sejroe (42.1%). High Ab titers for serovars Canicola (1:500) and Grippotyphosa (1:1,000) were detected for the first time in free-ranging bears from Croatia. A significant correlation between the age of the bears and detection of Ab to Leptospira spp. serovars suggested the presence of pathogenic agents in the natural habitats, whereas increasing trends of Ab prevalence for specific serovars (Icterohaemorrhagiae, Australis, and Sejroe) confirmed cohabitation of bears with rats and other small terrestrial mammals on garbage dumps and at bear feeding stations. To prevent cohabitation of bears and rodents, improvements in Croatian waste treatment, big game management, and rodent control programs are strongly recommended, especially in Lika and Gorski Kotar, regions that have high-quality natural habitats for brown bears in Croatia.

Papillomatous skin lesions from a green lizard (Lacerta viridis) were examined histologically, using electron microscopy and DNA was isolated from the lesions for herpesviral DNA detection. Histology confirmed the lesions to be squamous epithelial papillomas. Using electron microscopy, no virus particles were detected. The specific sequence of herpesviral DNA-directed DNA polymerase (EC 2.7.7.7) was amplified using degenerate primers in a nested format. The 235–base-pair (bp) sequence was sequenced and compared with previously published DNA-directed DNA polymerase sequences from various reptile herpesviruses. The sequence from the green lizard showed significant similarity with sequence of fibropapilloma-associated turtle herpesviruses from sea turtles.

Identified in 1998, Ljungan virus (LV; Picornaviridae) causes type 1 diabetes–like symptoms and myocarditis in bank voles (Myodes glareolus) from Sweden and Denmark, and may be a zoonotic agent of several important diseases (e.g., intrauterine fetal death, type 1 diabetes, Guillain-Barré syndrome, and myocarditis). Using a real-time reverse transcriptase–polymerase chain reaction assay and sequence analysis, we detected LV in bank voles, and for the first time, in yellow-necked mice (Apodemus flavicollis) collected during 2006 from a site in northern Italy. The global distribution of LV and its role as a mammalian pathogen deserve further attention.

During lyssavirus surveillance, 350 brains from four species of fruit bats and one species of insectivorous bat were collected from seven locations in Northern Nigeria during May to October, 2006. Lyssavirus antigen was not detected in the brains, and isolation attempts in mice were unsuccessful. However, serologic tests demonstrated the presence of lyssavirus-neutralizing antibodies in bat sera. Of 140 sera tested, 27 (19%) neutralized Lagos bat virus, and two of these additionally neutralized Mokola virus. The positive samples originated from the straw-colored fruit bat (Eidolon helvum) and the Gambian epaulet bat (Epomophorus gambianus). No neutralizing activity was detected against other lyssaviruses including rabies, Duvenhage, and West Caucasian bat viruses.

A fowl adenovirus was isolated from wild Black Kites (Milvus migrans) that died around Kashipur, Uttarakhand, India. This virus isolate produced cytopathic effects in chicken embryo liver (CEL) cells and reacted with fowl adenovirus 4 (FadV-4) antiserum in agar gel immunodiffusion and immunofluorescence tests. The virus isolate was neutralized by FadV-4 antiserum with a neutralization titer of 800. Electron microscopy of infected CEL cells showed the presence of hexagonal virion particles measuring in size about 80–100 nm. An amplicon of 1,223 base pairs was detected using polymerase chain reaction with primers designed to target the hexon gene of FadV-4.

The southern flounder Paralichthys lethostigma is host to two species of philometrid nematodes in the estuarine systems of South Carolina, USA. Histologic studies showed that all worms were hematophagous and that the host-parasite interface varied according to the worm's location in the fish. Individuals of Philometra overstreeti associated with the teeth induced a degradation of the enameloid epithelium but elicited minimal host reaction, whereas those located in the branchial arches induced an intense inflammatory response. Individuals of Philometroides paralichthydis elicited no host reaction. Those associated with the bones of the buccal cavity were contained in a thick collagenous capsule, whereas those located between the depressor and erector muscles of each fin element displaced or thinned the fin inclinator muscles to the extent of eventually causing their complete atrophy. Often the bulk of gravid worms imposed upon uninfected adjacent fin element muscle groups. Damage induced by these worms are likely to affect feeding and swimming behavior of the infected flounder and thus, impact the population structure of this important fish species.

This experimental study was conducted to evaluate the susceptibility of American bison (Bison bison) to liver flukes, Fascioloides magna and Fasciola hepatica. Six bison were each experimentally inoculated with 600 metacercariae of Fascioloides magna, and three were later treated with triclabendazole suspension at 40 mg/kg of body weight. Four additional bison were each experimentally inoculated with 600 metacercariae of Fasciola hepatica. Five control bison were placebo controls. Two controls and all inoculated bison were euthanized 10 mo (Fascioloides magna) and 7 mo (Fasciola hepatica) after inoculation. None of the control bison or the bison inoculated with Fascioloides magna had flukes or lesions characteristic of fluke infection at necropsy. All four bison inoculated with Fasciola hepatica had characteristic liver fluke lesions at necropsy, and three of four bison contained four, 103, and 111 adult flukes, respectively. Fluke eggs were detected in feces of all Fasciola hepatica–inoculated bison during the experiment, but not from the Fascioloides magna–infected bison or control bison. Clinical signs of infection were not observed during the experiment, but hemoglobin and packed cell volumes were lower in the Fasciola hepatica bison when compared to controls, and eosinophil levels were increased. Triclabendazole at 40 mg/kg of body weight appeared to be safe in bison because no toxic reactions were observed. Results from this study indicated bison are susceptible to infection with Fasciola hepatica and are efficient definitive hosts. Because no Fascioloides magna were recovered, bison may have a decreased susceptibility or innate resistance to Fascioloides magna infection, which may account for a lack of reported infections in this host.

An adult, female, free-ranging, sika deer (Cervus nippon yakushimae) from Wicomico County, Maryland, USA, was found circling and having no fear of humans. The animal was euthanized and submitted for a postmortem exam. There were no gross lesions and the deer was negative for rabies. Microscopic examination revealed lymphoplasmacytic, neutrophilic, and eosinophilic meningoencephalitis with intralesional adult nematodes, larvae, and eggs consistent with nematodes in the family Protostrongylidae. Parelaphostrongylus tenuis was identified by polymerase chain reaction and DNA sequencing. To our knowledge, this is the first report of P. tenuis–associated encephalitis in a sika deer.

Overlapping geographic distributions of tick-borne disease agents utilizing the same tick vectors are common, and coinfection of humans, domestic animals, wildlife, and ticks with both Borrelia burgdorferi and Anaplasma phagocytophilum has been frequently reported. This study was undertaken in order to evaluate the prevalence of both B. burgdorferi sensu stricto (hereinafter referred to as B. burgdorferi) and A. phagocytophilum in several species of sciurid rodents from northern California, USA. Rodents were either collected dead as road-kills or live-trapped in four state parks from 13 counties. Thirty-seven western gray squirrels (Sciurus griseus), nine nonnative eastern gray squirrels (S. carolinensis) and an eastern fox squirrel (S. niger), four Douglas squirrels (Tamiasciurus douglasii), and two northern flying squirrels (Glaucomys sabrinus) were tested by polymerase chain reaction (PCR) and serology for evidence of coinfection. Of the 14 individual S. griseus that were PCR-positive for B. burgdorferi, two (14%) also were PCR-positive for A. phagocytophilum and 11 (79%) had serologic evidence of A. phagocytophilum exposure. Two of the four Douglas squirrels were PCR positive for B. burgdorferi and seropositive to A. phagocytophilum. Evidence of coinfection with these zoonotic pathogens in western gray squirrels suggests that both bacteria may be maintained in a similar transmission cycle involving this sciurid and the western black-legged tick Ixodes pacificus, the primary bridging vector to humans in the far-western US.

First-stage nematode larvae with a dorsal-spine (DSL) were detected in five of 1,565 fecal samples from gray wolves (Canis lupus) collected in British Columbia, Canada, between 2005 and 2008. Molecular techniques were used to identify the DSL because it was not possible to determine their species identity using morphologic characters. The DSL were identified as Parelaphostrongylus odocoilei based on the results of single-strand conformation polymorphism (SSCP) analyses and DNA sequencing of the ribosomal DNA first and second internal transcribed spacers. Finding DSL of P. odocoilei in the feces of gray wolves was unexpected because P. odocoilei adults are parasites of cervids and bovids. The most likely explanation for the presence of DSL in wolf feces is that they were ingested along with the viscera of recently consumed prey. This was probably black-tailed deer (Odocoileus hemionus columbianus), which are known in the sampling area to be hosts of P. odocoilei. The present study demonstrates the use of SSCP and DNA sequencing for the identification, to the species level, of parasitic nematode larvae in feces.

Canine distemper virus (CDV) has been reported in several wild animal species, but there have been no reports of CDV in hoary fox (Lycalopex vetulus). This paper characterizes the first case of natural CDV infection in hoary fox, including the clinical and pathologic aspects of the disease as well as the viral strain phylogeny.

Chromobacterium violaceum is a gram-negative saprobe bacterium that is a rare opportunistic pathogen in mammals. There are numerous reports in humans including fatalities, but no record exists in free-ranging nonhuman primates. Here we report an infection by C. violaceum in a wild adult male howler monkey (Alouatta palliata) captured at Ballena Marine National Park, in southwestern Costa Rica. The individual had severe skin lesions over its extremities; gross findings included multiple skin ulcers, white foci in liver, and lymphoid hyperplasia. Histologic results included deep dermatitis with presence of necrotic epithelial cells where clusters of coccoid-shaped bacteria were detected. In the liver, numerous neutrophils forming microabscesses, telangiectasia, and focal necrotic areas were observed. Necrotic liver tissue sampled for bacteriologic culture resulted in the isolation of C. violaceum. We could not ascertain the source or mechanism of infection in this case, although infection through skin microabrasions is suspected. To the best of our knowledge, this is the first report for this pathogen in a wild, nonhuman primate. This report also draws attention to this infectious agent as a potential emerging wildlife disease and consideration should be paid by regional veterinary and epidemiologic vigilance services.

In March 2006, a dead, male bottlenose dolphin (Tursiops truncatus) was found in the salt marsh in Charleston, South Carolina, United States. During necropsy, an enterolith was found completely obstructing the intestinal lumen. Further examination of the enterolith revealed a stingray spine nidus. Most terrestrial enteroliths are composed primarily of struvite (magnesium ammonium phosphate); however, the majority of the enterolith discovered in the stranded dolphin was composed of calcium phosphate carbonate. This case provides an interesting comparison of the variation in the mineral composition between terrestrial and marine enteroliths.

Serum samples were collected from wild boars (Sus scrofa) harvested during the 2005–2006 hunting season in Campania, southern Italy. Samples were tested for antibodies to Leptospira interrogan, Brucella spp., Salmonella spp., Aujeszky disease virus (ADV), porcine reproductive and respiratory stress syndrome virus (PRRSV), porcine parvovirus (PPV), classical swine fever virus (CSFV), and swine vesicular disease virus (SVDV). Of the 342 serum samples tested, 15 (4.4%) were seropositive to Brucella spp., nine (2.6%) were seropositive to L. interrogans, 66 (19.3%) were seropositive for Salmonella spp., 105 (30.7%) were seropositive for ADV, 27 (7.9%) were seropositive for PPV, and 129 (37.7%) were seropositive for PRRSV. All sera tested seronegative for SVDV and CSFV antibodies. These results, recorded for the first time in Campania, support the hypothesis that wild boar are reservoirs of certain infectious agents, but some infections in wild boars originate from their domestic counterparts.

Two striped dolphins (Stenella coeruleoalba) were found stranded on the Catalonian Spanish coast. The main pathologic finding in both animals was the existence of multiple granulomatous lesions in the blubber, microscopically composed of macrophages and multinucleated cells containing vacuolar material. This material was identified as ceroid pigment due to its ultrastructural morphology, autofluorescence, and positive staining with periodic acid-Schiff and Ziehl-Neelsen techniques. The special stains and electron microscopy did not reveal any microorganisms associated with the lesions. These findings are very suggestive of ‘‘nutritional panniculitis,'' a well-defined entity associated with vitamin E deficiency that has been rarely described in free-living species.