Although cause-of-death information on bottlenose dolphins (Tursiops truncatus) can be located in the literature, few citations include mortality data over a long period of time covering a broad geographic region. This study describes major pathologic findings and probable causes of death of bottlenose dolphins over a 14-yr period (1993–2006) for the coastal region of South Carolina. Probable causes of death for 97 cases were determined based on gross pathology and histopathology. In an additional 30 cases, probable cause of death was apparent from gross pathology alone, and carcass condition precluded histopathology. Of the 97 dolphins examined grossly and histologically, 30 (31%) likely died of infectious disease and 46 (47%) of noninfectious disease; the cause of death was unknown in 21 (22%). Bacterial infections accounted for the large majority of fatal infections and emaciation was the leading cause of noninfectious mortality. Twelve dolphins were killed by human interactions. Of the 30 dolphins diagnosed from gross examination alone, 23 likely died from human interaction and seven were killed by stingray-spine inflictions. Although the absence of consistent use of microbiology, biotoxin analysis and contaminant testing decreases the conclusiveness of the findings, this study has broad implications in establishing baseline data on causes of death of bottlenose dolphins for future studies and for the detection of emerging diseases.

Necropsy reports for 28 stranded, cold-stunned Kemp’s ridley sea turtles (Lepidochelys kempii) that died between 2001 and 2006 were reviewed retrospectively. Gross and microscopic lesions were compiled to describe the pathologic and parasitologic findings in turtles that were found freshly dead on the beach or that died within 48 hr of stranding. Anatomic lesions of varying severity were identified in each of the examined turtles and were identified in tissues of the alimentary, respiratory, integumentary, nervous and sensory, and urogenital systems in order of decreasing frequency. Necrotizing enterocolitis and bacterial or fungal pneumonia were the most frequently encountered lesions that were considered clinically significant. Parasites and parasitic lesions were identified primarily in tissues of the alimentary system and included intestinal cestodiasis and parasitic granulomas containing larval cestodes or nematodes. Postlarval cestodes were also found in the coelom of two turtles. In many cases, the extent and severity of lesions were judged to be insufficient to have solely caused mortality, suggesting that additional factors such as metabolic, respiratory, and electrolyte derangements; hypothermia; and drowning may be important proximate causes of death in cold-stunned turtles. Results of this study provide insight into pathologic conditions that may be of clinical relevance to rehabilitation efforts for cold-stunned sea turtles.

Between 1993 and 1999, encephalitis caused by morbillivirus was diagnosed by immunohistochemistry and histology in six lynx (Lynx canadensis) and one bobcat (Lynx rufus) in the eastern Canadian provinces of New Brunswick and Nova Scotia. Five of the six cases in lynx occurred within an 11-mo period in 1996–97. A second bobcat with encephalitis caused by unidentified protozoa and a nematode larva also had immunohistochemical evidence of neurologic infection by morbillivirus. The virus was identified as canine distemper virus (CDV) by reverse transcriptase polymerase chain reaction and nucleotide sequencing in four of five animals from which frozen tissue samples were available, and it was isolated in cell culture from one of them. To our knowledge, this is the first report of disease caused by CDV in free-living felids in North America.

A portion of retropharyngeal lymph nodes from 6,824 wild cervids, comprising 1,458 white-tailed deer (Odocoileus virginianus), 5,345 mule deer (Odocoileus hemionus), 11 unidentified deer (Odocoileus spp.), and 10 elk (Cervus elaphus), were examined histologically for evidence of inflammation. Focal granulomatous lymphadenitis was detected in 0.3% of white-tailed deer, 1.3% of mule deer, and in one of 10 elk. Material consistent with Splendore-Hoeppli phenomenon was present in 70 (93%) of 75 cases of lymphadenitis. Replicate sections stained with Gram, acid fast, and methenamine silver stains were examined for organisms. Organisms morphologically consistent with Actinomyces spp. were found in one white-tailed deer; no organisms were detected in any of the other cases of lymphadenitis. Aerobic and anaerobic culture from frozen tissue from the lymph nodes was negative, as was polymerase chain reaction (PCR) for Mycobacterium spp. and Mycobacterium bovis. The method of sampling likely underestimated the prevalence of lymphadenitis.

Chytridiomycosis is an often fatal fungal disease of amphibians caused by Batrachochytrium dendrobatidis. This disease has been implicated in the worldwide decline of many anuran species, but studies of chytridiomycosis in wild salamanders are limited. Between August 2006 and December 2006, we tested wild amphibians in North Carolina, USA (n=212) by polymerase chain reaction (PCR). We identified three PCR-positive animals: one Rana clamitans and two Plethodontid salamanders. We experimentally infected two species of native North Carolina Plethodontid salamanders, the slimy salamander (Plethodon glutinosus) and the Blue Ridge Mountain dusky salamander (Desmognathus orestes) with 1,000,000 zoospores of B. dendrobatidis per animal. Susceptibility was species dependent; all slimy salamanders developed clinical signs of chytridiomycosis, and one died, whereas dusky salamanders remained unaffected. In a second experiment, we challenged naïve slimy salamanders with either 10,000 or 100,000 motile zoospores per animal. Clinical signs consistent with chytridiomycosis were not observed at either dose or in uninfected controls during the 45 days of this experiment. All animals inoculated with B. dendrobatidis in both experiments, regardless of dose, tested positive by PCR. Our study indicates that slimy salamanders are more susceptible to clinical chytridiomycosis than dusky salamanders, and in a laboratory setting, a dose greater than 100,000 zoospores per animal is required to induce clinical disease. This study also indicates that PCR is a very sensitive tool for detecting B. dendrobatidis infection, even in animals that are clinically unaffected, thus positive results should be interpreted with caution.

Nineteen map turtles (Graptemys spp.) maintained under natural conditions were investigated because of chronic shell abnormalities. Animals were evaluated using a novel shell scoring system that divided the 54 scutes into six regions, with each region scored for lesion extent and severity, and summated to produce a total shell disease score (TSDS). Complete blood counts and various biochemistry analytes (total protein, albumin, globulin, urea, uric acid, 25-hydroxycholecalciferol, phosphorus, and ionized and total calcium) were measured. Under ketamine-medetomidine-morphine anesthesia, cytology tape strips and full thickness shell biopsies were collected aseptically for microbiologic, histologic (including scoring of biopsy quality), and ultrastructural evaluations. The TSDSs were low and ranged from 4 to 22 (median=9) out of a possible score of 54. There were no correlations between TSDS and any hematologic or biochemistry parameter. The histologic quality of shell biopsies was good, and normal shell structure, by both light and electron microscopy, is described. Small clefts and pitting lesions were noted in 8/19 sections. There was no evidence of erosion, ulceration, inflammation, or infectious agents, but algae and diatoms were observed. Six biopsies yielded aerobic isolates (Chryseobacterium indologenes, Aeromonas hydrophila, Ralstonia pickettii, and Morganella morganii), whereas 11 shell samples grew various clostridial anerobes. No fungal organisms were cultured. Although the etiology of the lesions described remains unknown, the use of a scoring system in conjunction with full thickness biopsies is suggested to help standardize investigations into chelonian shell disease in the future.

The objective of the current study was to elucidate the within-host dynamics of bovine viral diarrhea virus (BVDV) type-1 infection to better understand how this virus could be maintained in white-tailed deer (Odocoileus virginianus, WTD) populations. The BVDV type-1 used in this study was originally isolated from a free-ranging WTD in Indiana. Four fawns were intranasally inoculated with 2 ml BVDV type-1 strain 544 WTD at a 10⁶ tissue culture infectious dose (TCID₅₀)/ml. Two fawns were inoculated with sham inoculum (negative controls). Animals were bled on days −7, 0, 1, 7, and 14 postinoculation (PID) for a complete blood count, chemistry panel, buffy coat (BC), real-time RT-PCR, and virus neutralization (VN). On days 7 and 14 PID, nasal and rectal swabs were obtained for RT-PCR and two of the virus-inoculated fawns and one of the negative controls fawns were euthanized. At necropsy, multiple samples were obtained for histopathology and in situ hybridization (ISH). Quantitative RT-PCR was performed on serum, BC, nasal, and rectal swabs. All animals tested negative for BVDV type 1 neutralizing antibodies on day 0 and animals in the control group remained seronegative throughout the study. No gross lesions were observed at necropsy. BVDV was isolated from lung and pooled lymph nodes from all BVDV-inoculated fawns on days 7 and 14 PID. Infected deer had lymphoid depletion, apoptosis, and lymphoid necrosis in the Peyer’s patches and mesenteric lymph nodes. BVDV was detected in lymphoid tissues of infected animals by ISH. No lesions or virus were identified in control fawns. On day 7 PID, samples from two virus-inoculated fawns were positive for BVDV by virus isolation and RT-PCR from BC and nasal swab samples. One fawn was also positive on a rectal swab. Nasal and rectal swabs from all animals were negative on day 14. Results indicate that infection of WTD with BVDV is possible, and leads to histologic lesions in variety of tissues. In addition, virus shedding into the environment through feces and other secretions is likely.

Plasma protein electrophoresis is recognized as a reliable diagnostic tool in avian medicine; however, the influence of circannual phenomena such as molt on protein electrophoregrams is poorly documented. The molt is a period of heavy hormonal and metabolic change in birds. The purpose of this study was to investigate the effects of molt on total protein concentration and electrophoresis patterns in birds. Blood samples were taken from 19 Bar-headed Geese (Anser indicus) from mid-May to mid-August, at 15-day intervals. At the same time, molting stage of each bird was recorded. Total protein concentrations were measured and plasma agarose gel electrophoresis was performed on these samples. The Bar-headed Goose was chosen as a model, because they molt over a very short period. The total protein concentration and albumin, alpha-2, beta, and gamma fractions were at their minimum values during molt, whereas the prealbumin and alpha-1 fractions rose to their maximum levels. This study provides baseline information relevant to changes occurring in avian proteinograms throughout the molt. The increase in prealbumin and alpha-1 fractions may be related to an increase in plasma thyroid hormones during molt. The decrease observed in albumin, alpha-2, beta, and gamma fractions may be related to protein and energy shifts toward feather growth, as well as to an expansion of the circulatory system located around the feather follicles with secondary dilutional effects on protein fractions. From a clinical point of view, the observed changes associated with molting were less significant than initially expected, and would not likely results in incorrect diagnoses based on interpretation of the protein electrophoretic patterns.

Seabirds are prominent components of the North Atlantic marine environment, and their parasites offer an insight into seabird ecologic interactions. Parasites also provide vital information on historic biogeography of host associations and thus may reveal broad changes in the marine ecosystem. Helminths of Common Murres (Uria aalge) and Thick-billed Murres (Uria lomvia) in the northwest Atlantic marine environment were assessed to determine parasite community composition and changes in their parasite fauna since the 1960s. In total, 623 helminths, representing Digenea, Eucestoda, Nematoda, and Acanthocephala, were recorded from 100 Common and Thick-billed Murres collected from breeding colonies along the coasts of Labrador, Newfoundland, and Greenland. Parasite communities differed from those reported from the 1960s, and over 85% of the specimens were tapeworms (mostly in the genus Alcataenia). The high prevalence (26%) and mean intensity (14.6) of A. longicervica, a Pacific species recorded recently from Newfoundland, indicates that this tapeworm was established in the Atlantic by 2006. Significantly higher A. longicervica prevalence (>53%) and mean intensity (27.3) in the murres from Greenland and in wintering murres compared to murres from breeding colonies in Labrador and Newfoundland suggest a mechanism for the introduction of this species to the Atlantic. Periodic mixing of populations of Thysanoessa species, the euphausiid intermediate host of Alcataenia, occurs along the seas adjacent to the North Pacific and those along the Siberian Arctic. The mixing of infected Thysanoessa likely exposed North Atlantic and Arctic murres, which are geographically isolated from Pacific murres, to this tapeworm. The greater prevalence of A. longicervica in Thick-billed Murres was consistent with diet analyses, which revealed a greater proportion of euphausiids.

In order to determine potential definitive hosts of the digenetic trematode, Bolbophorus damnificus, two American White Pelicans (Pelecanus erythrorhynchos), two Double-crested Cormorants (Phalacrocorax auritus), two Great Blue Herons (Ardea herodias), and two Great Egrets (Ardea alba) were captured, treated with praziquantel, and fed channel catfish (Ictalurus punctatus) infected with B. damnificus metacercariae. Patent infections of B. damnificus, which developed in both American White Pelicans at 3 days postinfection, were confirmed by the presence of trematode ova in the feces. Mature B. damnificus trematodes were recovered from the intestines of both pelicans at 21 days postinfection, further confirming the establishment of infection. No evidence of B. damnificus infections was observed in the other bird species studied. This study provides further evidence that Double-crested Cormorants, Great Blue Herons, and Great Egrets do not serve as definitive hosts for B. damnificus.

A female southern right whale (Eubalaena australis) neonate was found stranded on the Western Cape coast of southern Africa. Skin samples were taken the same day from three different locations on the animal’s body and stored at −20 C. Isolation through repetitive culture of these skin sections yielded a single yeast species, Candida zeylanoides. Total genomic DNA also was isolated directly from skin samples. Polymerase chain reaction analysis of the internal transcribed spacer region of the fungal ribosomal gene cluster revealed the presence of Filobasidiella neoformans var. neoformans, the teleomorphic state of Cryptococcus neoformans. Fungal infections in cetaceans seem to be limited when compared to infections caused by bacteria, viruses and parasites. However, Candida species appear to be the most common type of fungal infection associated with cetaceans. To our knowledge this is the first report of a C. zeylanoides infection in a mysticete, as well as the first report of a dual infection involving two opportunistic pathogenic yeast species in a cetacean.

The increasing importance of wildlife diseases in conservation efforts places an additional importance on research study design, data analysis, and interpretation. In this paper, we explore the design and analysis of wildlife disease data with regard to hypothesis testing, statistical power, sample sizes, the relative costs of type I versus type II errors, and effect size. To illustrate these ideas, we conducted a literature review of the Journal of Wildlife Diseases (JWD), ran computer simulations that estimate type II error rates for statistical techniques commonly used in JWD, and reanalyzed previously published data on disease prevalence. Many studies published in JWD used chi-squared analysis on prevalence data, but only 19% reported estimates of the observed effect size. Furthermore, 10% of studies had pooled sample sizes ≤40, and many had potentially high costs of type II relative to type I errors. Our computer simulations suggest that many articles published in JWD lack sufficient statistical power, and this, coupled with our findings that many studies often ignore high costs of type II errors, argues for increased attention to statistical power. Finally, our data reanalysis shows how the presentation of observed effect sizes could allow a better assessment of the biologic significance of findings reported in JWD. We conclude with some general guidelines to assist wildlife disease researchers in the design of future studies and the statistical analysis of their data.

Surveys for evidence of exposure to pseudorabies virus (PRV), Brucella suis, swine influenza virus (SIV; human-like H1N1, reassortant type H1N1, H1N2-like H1N1 and H3N2), porcine circovirus 2 (PCV 2), and porcine respiratory and reproductive syndrome virus (PRRSV) in feral swine (Sus scrofa) were conducted in areas where feral swine were geographically associated with high densities of transitional swine premises in South Carolina and high densities of commercial swine production in North Carolina. In South Carolina, 10/50 (20.0%), 7/50 (14.0%), and 29/49 (59.2%) feral swine tested antibody positive for PRV, B. suis, and PCV-2, respectively. Antibodies to PRRSV (0/49) and SIV (0/49) were not detected. In North Carolina, antibodies to PRV and B. suis were not detected in serum samples from 120 feral swine; however, antibodies to PRRSV (1/120 [0.8%]), PCV-2 (86/120 [71.7%]; these included 80 positives plus six suspects), and SIV (108/119 [90.7%]) were present. The presence of PRV and B. suis in South Carolina may have been due to the introduction of infected feral swine into the area or to a previous association of feral swine with infected transitional swine. Their absence in the North Carolina populations may have been due to the absence of these disease agents in the feral swine originally introduced into the area and the lack of a potential for contact with infected commercial swine. Feral swine associated with commercial swine in North Carolina may have been exposed to SIV subtypes circulating in commercial swine via airborne spread of SIV from commercial swine facilities. Feral swine seropositive for PCV-2 were prevalent in both states, which may indicate efficient transmission from commercial swine and transitional swine, or that PCV-2 is widespread in feral swine. The low prevalence of animals with antibodies against PRRSV may indicate a less-than-efficient means of transmission from commercial to feral swine. Additional epidemiologic studies are needed to understand the risks and mechanisms of transmission of disease agents among commercial, transitional, and feral swine, and the role of feral swine as reservoirs of these disease agents.

Group A rotaviruses infect and cause diarrhea in the young of a broad range of terrestrial mammals, but it is unknown, to our knowledge, whether they infect marine mammals. During February and March of 2002 and 2003, we collected 125 serum samples and 18 rectal swab samples from Galapagos sea lion pups (GSL, Zalophus wollebaeki), and 22 serum samples from Galapagos fur seal pups (GFS, Arctocephalus galapagoensis) from nine islands of the Galapagos archipelago, Ecuador. Sera were tested for antibodies (immunoglobulin G [IgG]) to rotavirus by an enzyme immunoassay using rhesus rotavirus as the capture antigen. In addition, rectal swabs were analyzed for the presence of rotavirus genomic double-stranded RNA by silver-stained polyacrylamide gel electrophoresis. Antibodies to rotavirus were detected in 27 GSL pups (22%) and five GFS pups (23%), and rotavirus RNA was detected in the fecal sample from one GSL pup (6%). These results provide the first evidence that rotavirus infections are prevalent at an early age in Galapagos sea lions and Galapagos fur seals.

Our objective in this prospective study was to determine the natural course of Brucella abortus infection in cohorts of seropositive and seronegative, female bison (Bison bison) and their offspring in Yellowstone National Park (YNP) for 5 yr. We collected specimens from 53 adult females and 25 calves at least once and from 45 adults and 22 calves more than once. Annual seroconversion rates (negative to positive) were relatively high (23% for calves and juvenile bison, 6% in the total sample of adult female bison in our study, and 11% in the adult females that began the study as seronegatives). Antibody was not protective against infection, even for calves that passively received antibody from an infected mother’s colostrum. Antibody levels stayed remarkably constant, with only a slow decline over time. We found only two seroconversions from a weak positive status to negative. Infected bison aborted and shed viable bacteria. Risk of shedding infective Brucella was highest for bison in the 2 yr following seroconversion from negative to positive. In one bison, we detected shedding for 3 yr following seroconversion. Regardless of serostatus of dams and neonates, most calves were seronegative by 5 mo of age. There was no relationship between the antibody status of the dam and the tendency of a calf to seroconvert to positive during the duration of the study.

Wild bird surveillance for highly pathogenic avian influenza (HPAI) H5N1 virus from 2004 to 2007 in Thailand indicated that the prevalence of infection with avian influenza H5N1 virus in wild birds was low (1.0%, 95% confidence interval [CI]: 0.7–1.2, 60/6,263 pooled samples). However, the annual prevalence varied considerably over this period, with a peak of 2.7% (95% CI: 1.4, 4.1) in 2004. Prevalence dropped to 0.5% (95% CI: 0.3, 0.8]) and 0.6% (95% CI: 0.3, 1.0) in 2005 and 2006, respectively, and then increased to 1.8% (95% CI: 1.0, 2.6) in 2007. During this period, 16 species from 12 families of wild birds tested positive for H5N1 virus infection. All samples from juvenile birds were negative for H5N1 virus, whereas 0.6% (95% CI: 0.4, 0.9) of pooled samples from adult birds were positive. Most positive samples originated from peridomestic resident species. Infected wild bird samples were only found in provinces where poultry outbreaks had occurred. Detection of H5N1 virus infection in wild birds was reported up to 3 yr after eradication of the poultry outbreaks in those provinces. As observed with outbreaks in poultry, the frequencies of H5N1 outbreaks in wild birds were significantly higher in winter. Further understanding of the mechanisms of persistence and ongoing HPAI H5N1 transmission between wild birds and domestic poultry is needed.

Five cases of fatal babesiosis in free-ranging chamois (Rupicapra r. rupicapra) attributed to infections with Babesia capreoli were recently recorded in two regions of the Swiss Alps. To investigate the ecologic factors that possibly lead to those fatal B. capreoli infections in chamois, blood, ticks, and demographic data of 46 roe deer (Capreolus c. capreolus), 48 chamois, and nine red deer (Cervus elaphus) were collected in 2006 and 2007 in both affected regions. Whereas no parasitic inclusions were found by microscopical examination of blood smears, B. capreoli was identified by polymerase chain reaction/sequencing in blood of 12 roe deer (26%, 95% confidence interval [CI]: 14.3–41.1), one chamois (2%, CI: 0–6.1), and one red deer (11%, CI: 0.3–48.2). Prevalence of B. capreoli was significantly higher in roe deer compared with chamois (P<0.001). All 214 ticks were identified as Ixodes ricinus, and significantly more roe deer (63%, CI: 47.5–76.8) were infested compared with chamois (21%, CI: 10.5–35.0, P<0.001). Overall, prevalences of both tick infestation and Babesia infection increased significantly (P<0.001) with decreasing altitude, and Babesia-positive samples were detected significantly more often from animals with tick infestation compared with animals without ticks (P=0.040). Our results indicate that roe deer may play an important reservoir role for B. capreoli. It is hypothesized that the expansion of the presumed vector I. ricinus to higher elevations and its increased abundance in overlapping habitats of roe deer and chamois may favor the spillover of B. capreoli from roe deer to chamois.

Bovine tuberculosis (Tb), due to infection with virulent Mycobacterium bovis, represents a threat to New Zealand agriculture due to vectorial transmission from wildlife reservoir species, principally the introduced Australian brushtail possum (Trichosurus vulpecula). An oral-delivery wildlife vaccine has been developed to immunize possums against Tb, based on formulation of the human Tb vaccine (M. bovis BCG) in edible lipid matrices. Here BCG bacilli were shown to be stable in lipid matrix formulation for over 8 mo in freezer storage, for 7 wk under room temperature conditions, and for 3–5 wk under field conditions in a forest/pasture margin habitat (when maintained in weatherproof bait-delivery sachets). Samples of the lipid matrix were flavored and offered to captive possums in a bait-preference study: a combination of 10% chocolate powder with anise oil was identified as the most effective attractant/palatability combination. In a replicated field study, 85–100% of wild possums were shown to access chocolate-flavored lipid pellets, when baits were applied to areas holding approximately 600–800 possums/km². Finally, in a controlled vaccination/challenge study, chocolate-flavored lipid vaccine samples containing 10⁸ BCG bacilli were fed to captive possums, which were subsequently challenged via aerosol exposure to virulent M. bovis: vaccine immunogenicity was confirmed, and protection was identified by significantly reduced postchallenge weight loss in vaccinated animals compared to nonvaccinated controls. These studies indicate that, appropriately flavored, lipid delivery matrices may form effective bait vaccines for the control of Tb in wildlife.

A 22.4-ha impoundment experienced an outbreak of Largemouth bass (Micropterus salmoides) virus (LMBV) disease in the summer of 2006. All dead or dying largemouth bass observed throughout the entire event were recorded and removed. In this study, we estimated mortality and examined size distribution, condition, and biomass following the outbreak. Boat-mounted electrofishing was used to collect largemouth bass for a mark-recapture population estimate and other population metrics. Fish samples were examined for evidence of LMBV, other infectious diseases, and physical abnormalities. Cell cultures inoculated with samples from moribund fish developed cytopathic effects typical of LMBV, and polymerase chain reaction (PCR) confirmed the presence of LMBV. The total number (N±95% confidence interval) of stock-size largemouth bass remaining was estimated to be 2,301±528 fish (103 bass/ha). The total observed mortality, including dead and dying individuals, during the LMBV outbreak was 176 largemouth bass (7% of the initial population). The total biomass remaining was estimated at 1,592 kg of stock-size bass and a relative biomass of 71.5 kg of stock-size largemouth bass per hectare. Largemouth bass size structure was dominated by quality and preferred (300–510 mm) size classes, with very few memorable-size or larger (>510 mm) fish, and the relative weight of largemouth bass was unusually variable. These results demonstrate that largemouth bass abundance and biomass in the reservoir remained very high despite mortalities attributed to a LMBV outbreak.

Proactive and reactive tactics have been utilized in Ontario, Canada, to prevent raccoon rabies from becoming established. A total of 96,621 raccoons (Procyon lotor) and 7,967 striped skunks (Mephitis mephitis) were live captured using 1,221,044 trap nights, vaccinated against rabies by injection, and released, during proactive Trap-Vaccinate-Release (TVR) programs in southern Ontario during 1994–2007. During those years, on average, 43% to 83% of the raccoon populations were vaccinated against rabies. In addition, 20,129 raccoons and 2,735 skunks were vaccinated against rabies, and 8,311 raccoons and 1,449 skunks were euthanized, using 576,359 trap nights, during reactive Point Infection Control (PIC) operations in eastern Ontario during 1999–2005. A significant correlation was detected between trapping effort and the percentage of the raccoon population that was vaccinated. Between 1999 and 2007, 132 cases of raccoon variant rabies (130 raccoons, two striped skunks) were reported in eastern Ontario. The last case occurred on 23 September 2005 with Ontario being free of reported raccoon rabies to 10 November 2008, proving that TVR and PIC are effective tactics for the control of this disease.

Toll-like receptor (TLR) 3 recognizes double-stranded RNA (dsRNA) and activates a signal transduction pathway that results in the release of a variety of chemokines and cytokines and apoptotic activity. Variability in TLR3 expression may play an important role in disease susceptibility of white-tailed deer (WTD; Odocoileus virginianus) to bluetongue and epizootic hemorrhagic disease viruses, which are dsRNA viruses. Because little is known about TLR3 in WTD, our objective was to sequence WTD TLR3 mRNA and to determine baseline levels of tissue expression. A 209-base pair sequence of TLR3 mRNA was obtained from WTD peripheral blood mononuclear cells. Dot blots confirmed that the sequence obtained was part of total WTD mRNA. Variable expression or ligand binding of TLR3 may contribute to observed susceptibility differences between populations of WTD, so the level of TLR3 in small intestine, skin, spleen, heart, cecum, rumen, lymph node, lung, kidney, and liver from WTD fawns (n=2) was analyzed using real-time reverse transcriptase-polymerase chain reaction. Tissue expression of TLR3 mRNA relative to the housekeeping gene β-actin was highest in spleen, heart, skin, and lung.

An adult female white-tailed deer (Odocoileus virginianus) with a history of shaking, ataxia, and severe debilitation was submitted for examination. Macroscopic lesions included severe emaciation, severe abdominal and mesenteric lymphadenopathy, and several rumen-associated masses. Microscopically, the ruminal masses and lymph nodes were infiltrated by pleomorphic neoplastic lymphocytes. Similar lymphoblasts were associated with the leptomeninges, choroid plexus, and the intestinal mucosa; these cells were intensely positive for CD3 antigen, indicating their T-cell origin. Lymphoproliferative viruses (bovine leukemia virus and malignant catarrhal fever virus) or epizootic hemorrhagic disease virus were not detected by polymerase chain reaction. To our knowledge, this case represents the first report of the immunophenotype of a multicentric lymphosarcoma, metastasis involving the brain, and epitheliotropic lymphoblasts in a white-tailed deer.

Periodic pneumonia outbreaks cause large-scale die-offs that threaten the viability of bighorn sheep (Ovis canadensis) populations. Bighorns are highly susceptible to pneumonia, and in some cases this susceptibility may be exacerbated by trace mineral deficiencies. To evaluate responses to injectable selenium supplementation, eight captive bighorn sheep were treated with either an injectable sodium selenite supplement or a saline control. We collected 6-ml blood aliquots before and at 1, 6, and 12 wk posttreatment. We submitted one set of aliquots immediately to measure selenium (Se) and zinc (Zn) concentrations and glutathione-peroxidase (GSH-Px) activity; additional aliquots were held at about 22 C and then submitted at 1, 3, and 7 days postcollection to assess effects of storage on these measures. Neither Se nor GSH-Px were affected by selenite injections. Both Se and GSH-Px demonstrated small linear decays over the 7-day storage period (0.011 ppm/day [SE=0.0027] and 15.78 mmole/l/sec/day [SE=6.88], respectively); in contrast, Zn concentrations in stored samples increased logarithmically (0.35 ppm/day on the natural log scale). Blood Se and GSH-Px were not correlated in sampled bighorns; however, because all values for both measures were within normal limits, lack of correlation did not affect interpretation of these data in our study.

The Ile du Grand Connétable nature reserve is a rocky island off the Northern Atlantic coast of South America that hosts a unique population of Magnificent Frigatebirds (Fregata magnificens, Pelecaniformes). A high chick mortality, associated with nodular proliferative lesions, involving featherless areas, such as legs, neck, eyelids, and beak, was recorded during a consecutive 2 yr and affected almost half of the generation. Investigations were, therefore, conducted to determine the cause of these epidemics. Although histopathologic investigations suggested that malnutrition, because of fewer resources in the Frigates’ fishing area, could be the cause of the epidemic, a novel alphaherpesvirus, tentatively called Fregata magnificens herpesvirus, was detected in cutaneous crusts on the diseased birds. Although in this study, we do not prove the causal link of this new virus to the symptoms observed, it can nevertheless be suggested that in debilitated hosts, a productive herpesvirus infection might accelerate, and/or be accelerated by, population declines. These results emphasize the need to take into consideration the possible role of herpesviruses in weakened populations of wild birds in conservation management plans.

On 10 October 2007, a Black Vulture (Coragyps atratus) was presented to the Wildlife Center of Virginia, Waynesboro, Virginia, USA, because of an inability to fly. Examination revealed multiple swollen, fluctuant joints. The bird suffered from lead toxicosis and had a prominent leukocytosis. Histopathologic evaluation revealed an acute fibrinoheterophilic polyarthritis, and results of routine aerobic and anaerobic culture of joint fluid were negative, although Mycoplasma sp. sequence-specific polymerase chain reaction was positive. Amplification of a portion of the 16S rRNA and subsequent phylogenetic analysis of the amplicon identified Mycoplasma corogypsi. This is the first report of polyarthritis being diagnosed in association with a Mycoplasma sp. in a vulture species. However, fulfilling Koch’s postulates through experimental infections is required to draw conclusions concerning an etiologic diagnosis.

We documented the antibody prevalence to three arboviruses, St. Louis encephalitis virus (SLEV), eastern equine encephalitis virus (EEEV), and West Nile virus (WNV), in Crested Caracaras (Caracara cheriway; n=80) in Florida from 2007 to 2008. Antibody prevalence to WNV was higher (9%) than for the other viruses. Most seropositive birds were adults (≥3 yr of age), with 55% of adults testing positive for antibodies to at least one virus. Adults were significantly more likely to have antibodies to WNV than nonadults (P<0.001). Prevalence of SLEV and EEEV antibodies among Crested Caracaras was 3% for each virus, and three adult caracaras had indistinguishable anti-flavivirus antibodies. The susceptibility of Crested Caracaras to adverse effects of WNV, SLEV, or EEEV infection remains unknown; however, we observed that some free-ranging individuals survived infection and successfully fledged young. Knowledge of arboviral infection among Florida’s Crested Caracara, which is both state and federally threatened, is valuable considering increasing pressure on this population from rapid and extensive habitat alterations.

Poisoning of waterfowl due to ingestion of lead pellets is a worldwide problem in areas that are subject to hunting. No studies have assessed exposure of waterbirds to this heavy metal in Argentina, in spite of intense hunting activity, and the fact that only lead ammunition is commercially available. The objective of this study was to evaluate duck exposure to lead by examining gizzard and bone samples collected from 30 wild ducks, 16 Rosy-billed Pochard (Netta peposaca), and 14 Fulvous Whistling-Duck (Dendrocygna bicolor), provided by hunters in northern Santa Fe Province, Argentina, in July 2007. Radiographs, followed by dissection of the gizzards, showed that 31% of the Rosy-billed Pochards and 29% of the Fulvous Whistling-Ducks had ingested lead pellets (between one and four per animal). Lead in bone was found at concentrations associated with detrimental health effects. In spite of the small number of samples in this project, these results indicate high levels of lead exposure (both recent and chronic) in these species. This is the first report of a problem in Argentina that could represent a threat to the health and conservation of native aquatic species, their predators, and the wetlands they inhabit.

In avian medicine, hematologic and biochemical laboratory investigations are still in their infancy, because of the difficulty involved in collecting data. This has led to a lack of reference values and a nonstandardized approach to specimens obtained in critical conditions. The Eurasian Buzzard (Buteo buteo) is one of the most common raptors in Italy, yet little is known about the physiologic blood parameters of this species. Serum biochemistry and electrophoretic investigations were performed in 40 healthy Eurasian Buzzards in different Italian wildlife rescue centers waiting to be released after recovering from trauma injuries. Mean values for biochemistry parameters were pancreatic amylase 626.9 IU/l, uric acid 7.5 mg/dl, aspartate aminotransferase 330.9 IU/l, glucose 375.1 mg/dl, lipase 26.3 IU/l, total protein 38.4 g/l, total bilirubin 0.04 mg/ dl, lactate dehydrogenase 2,008.4 IU/l, creatinine kinase 1,604.1 IU/l, alanine aminotransferase 40.4 IU/l, alkaline phosphatase 89.8 IU/l, magnesium 2.3 mg/dl, calcium 10.2 mg/dl, phosphorus 2.02 mg/dl, cholesterol 192.2 mg/ dl, triglyceride 116.4 mg/dl, albumin 14.5 g/l, creatinine 0.1 mg/dl. Mean electrophoretic values were prealbumin 1.4 g/l, albumin 14.2 g/l, alpha 1 globulin 5.9 g/l, alpha 2 globulin 4.7 g/l, beta globulin 7.5 g/l, gamma globulin 3.6 g/l, albumin/globulins ratio 0.8 g/l.

We report an unusual snaring of the larynx in an adult, female common bottlenose dolphin (Tursiops truncatus). The dolphin was observed swimming and diving in Haifa Port, Israel, but was found dead the next day, 60 km south, on the coast. Postmortem examination revealed stranded-cordage, nylon filaments wrapped around the larynx, cutting through the soft tissue, and extending down into the forestomach, where a large mass of netting was found. The cachectic state of the dolphin and the subacute to chronic, hyper-plastic response of soft tissue surrounding the filaments lodged around the larynx, suggest a prolonged period of starvation, which led to the final weakness and wasting of the dolphin.

Necropsy of an Atlantic bottlenose dolphin (Tursiops truncatus) neonate that stranded dead on Folly Beach, Charleston County, South Carolina, USA, on 17 November 2007, revealed multiple congenital heart malformations. Cardiac anomalies included a hypertrophic right ventricle, ventricular septal defect (VSD), aortic dilation, atrial septal defect (ASD) between a functionally common atrium and a left atrial remnant, subvalvular pulmonic stenosis, and a hypoplastic pulmonary artery and mitral valve. Few incidences of abnormal cardiac development in cetaceans have been published. The case study serves to document a novel congenital heart malformation not previously reported, to our knowledge, in free-ranging bottlenose dolphins.

Reference range clinical serum chemistry values were established for free-ranging lactating northern fur seals (Callorhinus ursinus). Fur seals sampled for this study were part of a healthy and growing population in the Kuril Islands of far-east Russia. Blood was collected from 45 females between June and August 2005 to 2007. Fresh serum was assayed for 16 components. Packed-cell volume was determined from fresh whole blood. Results are made available for future comparisons with the declining population of northern fur seals on the Pribilof Islands and are compared with published values for other otariid species.

The objective of this study was to document the presence of the fungal granulomatous skin disease lacaziosis in stranded Indian River Lagoon (IRL) bottlenose dolphins (Tursiops truncatus). From 1 January 2007 through 31 December 2007, stranded dolphins from the northern part of the IRL were thoroughly examined, and appropriate tissue samples were collected. The intralesional fungal agent (Lacazia loboi) was identified histologically in three bottlenose dolphins. Histologically, lacaziosis has been previously documented in IRL dolphins inhabiting the southern portion of the lagoon. Our findings suggest that the disease occurs throughout the lagoon. Enhanced monitoring of the prevalence of lacaziosis in dolphins throughout the IRL is needed to assess changes in population health.

Hematologic and serum chemistry values were determined for 25 free-ranging Florida panther (Puma concolor couguar) neonates from southern Florida sampled from January 2001 to April 2007. The kittens were ≤25 days old, belonging to 12 litters, from 11 different dams. Forty-one blood samples also were collected from 32 free-ranging adult panthers (3–10 yr-old) from southern Florida from November 2000 to February 2007. Male kittens had higher hemoglobin (Hb), hematocrit (HCT), and red blood cell (RBC) counts than did female kittens. Neonates had higher mean serum values of total bilirubin, alkaline phosphatase, cholesterol, calcium, phosphorus, triglycerides, and magnesium than adult panthers but lower mean values of urea nitrogen, creatinine, total protein, albumin, alanine aminotransferase, aspartate aminotransferase, sodium, chloride, creatine kinase, amylase, and total globulin. Neonates also had higher mean values of mean cell volume, mean cell hemoglobin, mean cell hemoglobin concentration, platelets, and the number of lymphocytes and monocytes but lower Hb, HCT, white blood cell count, RBC count, and neutrophils than adult panthers. No other significant differences in serum chemistry or hematology were noted between neonates and adults.

Blood and feces were collected from 34 adult (19 males, 15 females) and seven juvenile (three males, one female, three not reported) free-ranging coyotes (Canis latrans) on the US Department of Energy’s Savannah River Site (South Carolina, USA). Significant (P<0.05) hematologic differences by sex were noted for red blood cell counts, hemoglobin, and hematocrit. Biochemical differences by sex occurred only for albumen (P<0.05). Twenty-one adults were antibody positive for at least one of four viruses: canine adenovirus type 1 (CAV-1; 68%), West Nile virus (WNV; 60%), Eastern equine encephalitis virus (EEEV; 38%), and Canine distemper virus (CDV; 15%). Of the seven Leptospira serovars tested for, seven (25%) of 28 adults were positive for one or more of five serovars: Pomona, Grippotyphosa, Icterohaemorrhagiae, Bratislava, and Autumnalis. Three (43%) of seven juveniles had seropositivity for a virus, one each for CDV, CAV-1, and WNV. No juveniles were seropositive for EEEV or any of the seven Leptospira serovars. Blood smears of 12 adults were positive for Dirofilaria immitis microfilaria, but blood smears from all juveniles were negative. Parvovirus was identified by electron microscopy from the feces of one adult. Ancylostoma spp., Trichuris spp., and Isospora spp. were observed in fecal samples. These data may aid in understanding the role of coyotes in disease ecology.

Pteropus seychellensis comorensis is the only Pteropodidae bat species on the island of Mayotte (Comoros Archipelago), and most aspects of its biology are unknown. In order to catch this large bat, we used a simple and low-cost method, consisting of raised mist nets that were set close to foraging sites. Major factors driving catch success were high food availability, good positioning of mist nets, and careful observation of movement patterns to identify foraging sites where the chances of capture are high. Blood was collected from the alar and humeral veins, which appear to be more practical for this purpose than other parts of the venous system.

In 2007, two new populations of feral swine were discovered in Nance and Valley counties, Nebraska, USA. Necropsies and serologic testing was done on two individuals from the Nance County herd. Results indicated that a lactating sow had positive antibodies for pseudorabies virus (PRV). Investigations conducted by Nebraska Game and Parks Commission Law Enforcement division confirmed that the infected individual was transported illegally to Nebraska, USA, from Texas, USA. All domestic swine herds located within an 8 km radius of the infected individual tested negative for antibodies to PRV. Our results provide a clear example of how diseases can spread because of anthropogenic activities and highlight the need for disease surveillance and monitoring in the import of invasive species.

Twenty adult, free-ranging, female Svalbard reindeer (Rangifer tarandus platyrhynchus) were immobilized with medetomidine-ketamine from 30 September through 9 October 1999 at Svalbard, Norway (78°55'N, 11°56'E). The animals were approached on foot, and the drugs were administered into the heavy muscles of the shoulder or the thigh by dart syringe injection from 15–25 m. The mean (SD) induction time in 10 animals immobilized with 0.113 (0.009) mg/kg of medetomidine and 2.26 (0.19) mg/kg of ketamine (group 2) was significantly shorter (P<0.05) than in 10 animals immobilized with 0.215 (0.043) mg/kg of medetomidine and 1.08 (0.21) mg/kg of ketamine (group 1): 6.5 (3.2) versus 14.3 (10.6) min, respectively. Inductions were calm, major clinical side effects were not detected, and there were no significant differences between groups regarding rectal temperature, pulse rate, respiratory rate, or relative arterial oxygen saturation. The 5 mg of atipamezole/1 mg of medetomidine were given half intramuscularly and half subcutaneously for reversal, and the animals were standing within 9.5 (4.5, group 1) and 13.0 (6.4, group 2) min, respectively, after administration of the antagonist.

A parasitologic study on 129 red foxes (Vulpes vulpes) from Tuscany (central Italy) was carried out in 2004–2006. Five intestinal species were found at necropsy: Dipylidium caninum (prevalence 57.3%), Mesocestoides lineatus (45.4%), Uncinaria stenocephala (39.1%), Toxocara canis (9.1%), and Toxascaris leonina (5.4%). Other parasites not associated with the intestine included Crenosoma vulpis (14.7%), Capillaria aerophila (7.0%), Angiostrongylus vasorum (7.0%), and filarial parasites (17.8%). Coprologic tests were less sensitive and less specific in identifying parasites than direct examinations at necropsy. Trichinella larvae were not found in muscles submitted to artificial digestion. By immunologic assay, antigens of Echinococcus spp. were detected in fecal samples of 20 foxes, but results could not be confirmed by fecal examination or molecular tests.