Feline leukemia virus (FeLV) was not detected in Florida pumas (Puma concolor coryi) in almost 20 yr of surveillance; however, the finding of two FeLV antigen-positive pumas during the 2002–2003 capture season led to an investigation of FeLV in the population. Between January 1990 and April 2007, the proportion of pumas testing FeLV antibody positive increased, with antibody-positive pumas concentrated in the northern portion of puma range. Five of 131 (4%) pumas sampled between July 2000 and April 2007 were viremic, with all cases clustered in Okaloacoochee Slough (OKS). Clinical signs and clinical pathology at capture were absent or included lymphadenopathy, moderate-to-severe anemia, and lymphopenia. All viremic pumas died; causes of death were septicemia (n=2), intraspecific aggression (n=2), and anemia/dehydration (n=1). Outcome after FeLV exposure in pumas was similar to that in domestic cats, with evidence of regressive, latent, and persistent infections. Management of the epizootic included vaccination, and as of April 2007, 52 free-ranging pumas had received one or more inoculations. Vaccinations were concentrated in OKS and in a band between OKS and the remainder of the puma population. There have been no new cases since July 2004; however, the potential for reintroduction of the virus remains.

Ohio's oral rabies vaccination (ORV) program was established to prevent the westward spread of the raccoon (Procyon lotor) rabies virus (Lyssavirus, Rhabdoviridae) in Ohio, USA. The program, which targets raccoons, distributes vaccine-bait units (VBU) at a target density of 75 units/km². Few studies have examined the relationship of VBU density and target population density to the prevalence of rabies virus-neutralizing antibodies (RVNA). We conducted experimental VBU distributions in August 2003 and August 2004, 150 km west of the ORV zone where there was no history of raccoon rabies. We measured change in RVNA titers in blood collected from live-trapped raccoons before and after VBU distributions. A closed population mark–recapture estimate of the size of the target population was 91 raccoons/km², compared to the realized VBU distribution density of 70 units/km². Surprisingly, 41% of 37 serum samples were RVNA-positive (≥0.05 IU/ml) before VBU distribution in 2003, but all titers were <0.25 IU/ml. Although viable VBUs were distributed in August 2003, only 21% of 315 samples were RVNA-positive before VBU distribution in 2004, but 9% had titers ≥0.25 IU/ml. Tetracycline (biomarker in bait) prevalence in teeth indicated that 57% of raccoons ingested VBUs after distribution in 2003, and 54% ingested VBUs after distribution in 2004. However, only 8% and 11% of sera were positive for RVNA (≥0.05 IU/ml) after VBU distribution in 2003 and 2004, respectively. Only 4–5% of sera collected after bait distribution had titers ≥0.25 IU/ml each year. The standard distribution density of 75 VBUs/km² was insufficient to produce a population-wide immunoprotective response against rabies infection in our high-density target population. Presence of RVNA in a presumed naïve population before baiting demonstrates that estimating prevalence of RVNA after oral rabies vaccination can be problematic without knowledge of background titers and seasonal changes in prevalence of RVNA before and after baiting.

The Tsaatan (or Dhuka) peoples of northern-western Mongolia are one of the few remaining reindeer-herding cultural groups in the world. Recently a disease condition that involves sudden death of reindeer and cases involving fever, lethargy, and pale mucous membranes has been reported. Examination of blood smears collected in the 2005 field season resulted in the identification of intra-erythrocytic inclusions resembling Anaplasma spp. in smears from clinically sick animals. Using universal polymerase chain reaction (PCR) primers for the amplification of the 60 kDa chaperonin gene (cpn60, also known as hsp60 or groEL), we detected sequences corresponding to Anaplasma ovis in reindeer blood samples. Species-specific PCR primers for A. ovis were designed and validated and used to screen blood samples from Mongolian reindeer. Screening of 66 blood samples collected in the 2006 field season resulted in the detection of A. ovis in 80% of the samples. Our results indicate a high prevalence of A. ovis in the Tsaatan reindeer herds and an association with clinical disease that is likely to be anaplasmosis. To our knowledge this is the first report of natural A. ovis infection in reindeer.

Johne's disease, a fatal and contagious gastrointestinal infection caused by Mycobacterium avium subsp. paratuberculosis (Map), was first diagnosed in an endangered Florida Key deer (Odocoileus virginianus clavium) in 1996 and later in six additional Key deer deaths from 1998 to 2004. We investigated the geographic distribution of Map in the Lower Florida Keys from February 2005 through May 2006 via collection of blood and fecal pellets from 51 live-captured deer, collection of 550 fecal samples from the ground, and by necropsies of 90 carcasses. Tissue and fecal samples also were submitted from 30 raccoons (Procyon lotor), three feral cats (Felis catus), an opossum (Didelphis virginiana), and a Lower Keys marsh rabbit (Sylvilagus palustris hefneri). Mycobacterium avium subsp. paratuberculosis was identified in 23 Key deer fecal samples collected from the ground, tissue samples from two clinically ill Key deer, and from the mesenteric lymph node of a raccoon. The results of this study indicate that Map persists in the Key deer population and environment at a low prevalence, but its distribution currently is limited to a relatively small geographic area within the range of Key deer.

Infection with Salmonella spp. has long been recognized in avian wildlife, although its significance in causing avian mortality, and its zoonotic risk, is not well understood. This study evaluates the role of Salmonella spp. in wild bird mortality events in the United States from 1985 through 2004. Analyses were performed to calculate the frequency of these events and the proportional mortality by species, year, month, state, and region. Salmonellosis was a significant contributor to mortality in many species of birds; particularly in passerines, for which 21.5% of all mortality events involved salmonellosis. The proportional mortality averaged a 12% annual increase over the 20-yr period, with seasonal peaks in January and April. Increased salmonellosis-related mortality in New England, Southeastern, and Mountain-Prairie states was identified. Based on the results of this study, salmonellosis can be considered an important zoonotic disease of wild birds.

The raccoon (Procyon lotor) is the definitive host of Baylisascaris procyonis, a large intestinal roundworm that is zoonotic and can result in fatal or severe central nervous system disease in young children. Prevalence of infection among raccoon populations often is high, and in the midwestern United States, B. procyonis has been reported in 68–82% of raccoons. Raccoon populations have increased in response to changes in human land use, and often reach higher densities in urban and suburban landscapes than rural landscapes. However, shifts in foraging behavior among urban raccoons could impact the transmission of B. procyonis if small vertebrate intermediate hosts are not a significant part of the raccoon diet. The objective of this study was to compare prevalence of B. procyonis infection between urban and rural raccoon populations on a regional scale. Necropsy was done on 204 raccoons collected from September through February during 2000–2005 from seven states across the Midwest (regional sample). Baylisascaris procyonis was found in 54% of examined raccoons. Prevalence differed between land-use types (χ²=11.56, df=1, P=0.0007), and was higher among animals collected from rural locations (65%) than those collected in urban locations (41%). Intensity of infection also differed (F=5.52, df=1, P=0.02), with rural raccoons having greater worm burdens (x¯=29.63±36.42) than urban raccoons (x¯=13.85±18.47). Despite high densities of raccoons in urban landscapes, fewer urban raccoons were infected with B. procyonis, suggesting decreased dependence on intermediate hosts as a food source. This possible explanation was supported by a similar trend in prevalence among subsamples of raccoons collected from three Chicago-area populations (local samples) with differing levels of urbanization, population densities, and foraging behavior that had been intensively monitored during 1995–2002. Decreased transmission of B. procyonis in urban landscapes may be due to decreased predation of intermediate hosts, and contact of juvenile raccoons with B. procyonis eggs may be an important factor in maintaining infections within such populations.

Two ferrets (Mustela putorius furo) were experimentally infected with phocine distemper virus (PDV), from the 1988 seal epizootic in Europe, in order to determine whether the stable transfected Vero cell line (Vero.DogSLAMtag) expressing canine “signaling lymphocyte activation molecules” (SLAM; CD150) receptors, was more suitable for isolating and characterizing PDV when compared with Vero (American Type Culture Collection # C1008) and primary seal kidney (PSK) cells. Both ferrets displayed characteristic clinical signs of distemper, including fever and rash, 10 days postinoculation (dpi) and, due to increased morbidity, they were euthanized 12 dpi. Histologic lesions, suggestive of infection with morbilliviruses, were observed in tissues from both ferrets, and the tissues stained positive using immunohistochemistry. Isolation of PDV from isolated peripheral blood lymphocytes (PBLs), taken at 5 and 10 dpi, was achieved by cocultivation with Vero and PSK cells, following several passages. Cytopathic effects (CPE) were observed in Vero cell cultures at 29 dpi and in PSK cell cultures at 22 dpi. Phocine distemper virus was isolated from frozen infected ferret lung tissue within 48 hr, when isolation was attempted using the Vero.DogSLAMtag cell line. In addition, a reverse transcriptase polymerase chain reaction (RT-PCR) test was developed to detect a 114 base pair (bp) portion of the nucleocapsid gene found only in PDV. This RT-PCR methodology was used to confirm the identity of the virus subsequently isolated from the ferrets. Viral isolates from the infected ferrets, as well as cultures of virus originally isolated from a dolphin and a porpoise and maintained in Vero cells, also replicated faster and produced higher titers of virus when propagated in Vero.DogSLAMtag cells. These results indicate that Vero.DogSLAMtag cells offer a substantial improvement (including faster viral replication resulting in primary viral isolation in a shorter period of time, and higher yield of virus finally obtained) over traditional cell culture methodologies for isolation and characterization of marine mammal morbilliviruses.

A captive colony of adult Big Brown Bats (Eptesicus fuscus) was experimentally infected with a rabies virus (RABV) variant isolated from the salivary glands of a naturally infected Big Brown Bat and passaged once through murine neuroblastoma cell culture. Bats were divided into 11 groups, which were composed of one to three noninfected and one to three infected individuals each. Twenty of 38 animals were infected intramuscularly into both left and right masseter muscles; they received a total of 10^3.2 median mouse intracerebral lethal dose (MICLD₅₀) of Big Brown Bat RABV variant. Experimental outcome after viral exposure was followed in the bats for 140 days postinoculation (PI). Of 20 infected bats, 16 developed clinical rabies, and the mean incubation period was 24 days (range: 13–52 days). Three infected bats never seroconverted and succumbed early to infection (13 days). Four infected bats that survived until the end of the experiment without any signs of disease maintained detectable antibody titers until the third month PI, peaking between days 13 and 43, and consequent drop-off below the threshold for detection occurred by day 140. Limited excretion of virus in saliva of infected bats during the clinical course of disease was observed in two individuals on days 13 and 15 PI (<24 hr prior to onset of clinical illness). No bat-to-bat transmission of RABV to noninfected bats was detected.

We report two separate naturally occurring cases of fatal herpesviral disease in Cooper's Hawks (Accipiter cooperii). Gross lesions included splenomegaly and hepatomegaly, with diffuse pale mottling or scattered small white foci. Histologic lesions included splenic and hepatic necrosis associated with eosinophilic intranuclear inclusion bodies characteristic of herpesvirus. In one case, necrosis and inclusions were also noted in bone marrow, thymus, bursa of Fabricius, thyroid gland, parathyroid gland, ceca, and the enteric system. Transmission electron microscopy demonstrated viral particles typical of herpesvirus within hepatocyte nuclei and budding from the nuclear membrane. Herpesviral DNA was amplified via polymerase chain reaction (PCR) of paraffin-embedded liver and spleen, and sequence data were consistent with columbid herpesvirus-1, an alphaherpesvirus of Rock Pigeons (Columba livia). PCR results provide evidence that this disease is transmitted to raptors via Rock Pigeons, most likely through ingestion of Rock Pigeons as prey.

Tissues and serum from 59 raccoons (Procyon lotor), 42 coyotes (Canis latrans), and seven Striped Skunks (Mephitis mephitis) collected in Dane and Iowa Counties, Wisconsin, USA, between October 2005 and March 2006 were microscopically and serologically examined for the presence of Trichinella spp. Encapsulated larvae were found on compression slides prepared from tongue tissues from a few animals. Complete tissue digestion of tongues revealed that 19% of the raccoons, 26% of the coyotes, and none of the seven skunks tested were infected with Trichinella spp. Cats were subsequently experimentally infected by feeding them the raccoon tissues containing muscle larvae, and muscle larvae isolated from the collected tongues were experimentally transmitted to mice. Multiplex polymerase chain reaction analysis of the isolated muscle larvae demonstrated two distinct bands migrating at 127 base pairs (bp) and 316 bp in all samples, which together are diagnostic for Trichinella murrelli; the isolates were assigned Istituto Superiore di Sanita (ISS) codes ISS1656 through ISS1667, and ISS1708 through ISS1710 by the International Trichinella Reference Centre. These findings extend the geographic range of T. murrelli into Wisconsin, USA.

The huemul (Hippocamelus bisulcus), an endemic Patagonian deer, has been endangered for decades. Although conservation in Argentina has been directed at the 350–600 remaining huemul, the population has not recovered. In initial studies on the potential effects of diseases on huemul population dynamics, skeletal remains collected between 1993 and 2007 in the Andes (41–45°S, 71.5–72°'W) were examined macroscopically for osteopathologic changes. Bones from six huemul were free of lesions, findings were inconclusive in 13 huemul with less than three bones, and osteopathologic processes were detected in 13 adults. Considering the limited remains/case, the prevalence of osteopathy (52%) among adults probably is conservative; 63% showed mandibular, 100% maxillary, and 78% appendicular lesions. Although predation represented the actual cause of death, the observed skeletal lesions would affect predator avoidance, possibly explaining the low average adult age (3.1 yr) and lack of population recovery. Compared with other studies in ungulates, huemul were affected at a younger age, and they had more severe pathologic changes. Due to the chronic nature of disease, low huemul population density, physiognomy, and spatiotemporal pattern of lesions, we discard senescence; gender; fulminating infections; congenital anomalies; metabolic, endocrine, genetic, or neurologic disorders; parasitism or marasmus; and fluorosis as primary etiologic factors. We hypothesize that generalized secondary chronic alveolar osteomyelitis and osteoarthritis in huemul is related to the nutritional ecology of these animals. Selenium deficiency, which impairs bone metabolism and causes periodontitis in ruminants, occurs in the region and it is more prevalent at high altitudes. Traditional winter grounds at low elevations, sometimes far from high mountains, have been converted to livestock production, which has eliminated migratory behavior and keeps huemul in remote high-elevation refuges. Although this descriptive study contributes to huemul conservation, additional approaches are needed to investigate the etiology of this osteopathy and to close other gaps in knowledge on biology and ecology of huemul.

Despite the devastating collapse of three vulture populations on the Asian subcontinent as a result of their exposure to diclofenac, there is little available information on the normal physiology of many vulture species, including the African White-backed Vulture (Gyps africanus). Such information is needed to fully understand mechanisms for toxicity and to identify and prevent future health problems. The aim of this study was to establish baseline parameters for hematologic and selected serum chemistry parameters for this model species for further studies into the toxicity of diclofenac. Captive nonreleasable and wild African White-backed Vultures were used to determine reference values. For hematology, erythrocyte counts, hemoglobin concentration, hematocrit, packed cell volume, mean corpuscular volume, mean corpuscular hemoglobin concentration, and total and differential leukocyte counts were measured. Chemical analytes measured included sodium, potassium, calcium, albumin, and globulin concentrations, aspartate aminotransferase, creatine kinase, and alanine aminotransferase activities. Uric acid and urea concentrations and the urea:uric acid ratio also were evaluated. Values are presented as means, standard deviations, and reference intervals. The serum chemistry parameters selected may provide a starting point for the evaluation of changes in renal and hepatic function; these organ systems are most severely affected by diclofenac. Results were also compared with values reported for G. africanus nestlings, and from these results it is evident that the clinical pathologic parameters are age related. This indicates that the use of nestling values for the evaluation of clinical pathologic findings in adults may be unreliable and could lead to incorrect assumptions.

Development of p-aminopropiophenone (PAPP) as a toxicant for pest predator management in New Zealand and Australia prompted investigation of its toxicity to potential nontarget species. Acute oral toxicity of PAPP in brushtail possums (Trichosurus vulpecula), dama wallabies (Macropus eugenii), and Mallards (Anas platyrhynchos) was estimated in pen trials, carried out between February 2000 and September 2001. The susceptibility of possums (LD₅₀≥500 mg kg⁻¹) and wallabies (LD₅₀ 89 mg kg⁻¹) to PAPP was low in comparison to noncarnivorous placental mammal species, but ducks (LD₅₀ 38 mg kg⁻¹) were more susceptible than other bird species. These results suggest that the nontarget hazard to possums and wallabies from PAPP bait applied for pest predator control would be low. However, future development of PAPP as a vertebrate pest control agent should include rigorous assessments of the hazard posed by bait formulations to bird species and provision for delivery techniques that could mitigate exposure of nontarget birds.

Previously, we reported that wild eastern box turtles (Terrapene carolina carolina) with aural abscesses contained higher body burdens of organochlorine (OC) compounds than those without the lesion. This lesion in captive chelonians is associated with turtles that are fed diets deficient in vitamin A. To examine the pathophysiology of this lesion and evaluate the relationship between OC burdens and vitamin A metabolism, we maintained red-eared sliders (Trachemys scripta elegans) under different conditions of OC exposure and dietary vitamin A concentrations from August 2005 to February 2006. Dietary vitamin A concentration (0 or 5 international units/g in the diet) and OC exposure (no OC compound or the mixture of 2 mg/kg chlordane, 0.25 mg/kg aroclor, and 1 mg/kg lindane) did not affect histologic score based on degree of squamous metaplasia of the tympanic epithelium or levels of plasma or liver vitamin A among the study groups. The results of this study suggest that 6 mo of exposure to the selected OC compounds, or similar duration of reduced dietary vitamin A concentrations do not influence the formation of squamous metaplasia and aural abscesses in red-eared sliders. Further studies are required to determine whether the duration of the experiment was insufficient, the OC compounds selected were inappropriate, the dosing was incorrect, and whether there are other unknown mechanisms causing the reported association between OC exposure and aural abscesses seen in eastern box turtles.

The Alligator Snapping Turtle (Macrochelys temminckii) is a large freshwater turtle endemic to river systems that drain into the Gulf of Mexico. Turtle populations were sharply reduced by commercial harvest in the 1970s and 1980s; however, the species has yet to be protected under the Endangered Species Act. While anthropogenic stressors such as habitat fragmentation and degradation and illegal capture continue to threaten populations, the degree to which disease may be contributing to any decline of the Alligator Snapping Turtle is unknown. Data were collected from 97 free-ranging Alligator Snapping Turtles in nine waterways in Florida and Georgia from 2001 to 2006. Eleven turtles were captured more than once, resulting in a total sample pool of 123. Reference ranges were established for complete blood count, plasma biochemistry values, trace metals (mercury, zinc, copper, lead, and arsenic), and nutrient parameters (vitamins A, E, D, and selenium). Variations by capture location, sex, and season were detected and likely resulted from external factors such as habitat and diet. Turtles sampled in one location were positive for tortoise herpesviral antibodies. Blood mercury values also differed among populations. This study provides justification for the use of these long-lived aquatic turtles as biologic monitors of the health of local freshwater ecosystems.

Heads of 64 white-tailed deer (Odocoileus virginianus) fawns, harvested in the vicinity of Magrath, Alberta, Canada, (49°24′782″N, 112°52′113″W) were examined for the presence of nasal bots and lice. The deer were collected between 8–30 January 2004 as part of a government-approved herd reduction protocol. The entire surface of each head was scanned visually for the presence of lice. Each head was split longitudinally, and the nasal passages, sinuses, and ethmoid region were washed for recovery of nasal bots. First instar Cephenemyia spp. were recovered from 17 heads (27%). Intensity of infestation ranged from 1–18 larvae (mean intensity 4.8). Among fawns, there were no significant differences in prevalence or mean intensity between the sexes. Two species of nasal bots were identified. Smaller larvae, tentatively identified as C. jellisoni, were present in 16 of 17 infested deer while larger specimens, tentatively identified as C. phobifera, were found in four deer; and in three of the four it co-occurred with C. jellisoni. The presence of C. phobifera in Alberta would represent a range extension for this species, which has not been known to occur west of North Dakota. Thirty-one fawns (48%) were infested with the sucking louse Solenopotes ferrisi. One infested fawn also had one specimen of the chewing louse, Tricholiopeurus lipeuroides.

The ectoparasite fauna of reintroduced roe deer (Capreolus capreolus) was surveyed in a Mediterranean forest in Israel. Ectoparasites were collected from four female hand-reared deer during 2004 and 2005. Seasonality, predilection sites of infestation, and the apparent effect of the parasites are presented. This is the first study of roe deer parasites in the East Mediterranean. The ectoparasite fauna included three hippoboscid fly (Lipoptena capreoli, Hippobosca equina, and Hippobosca longipennis), four tick (Rhipicephalus sanguineus, Rhipicephalus turanicus, Rhipicephalus kohlsi, and Hyalomma marginatum), and one unidentified trombiculid mite species. For most of these ectoparasites, this is the first record on roe deer. All ectoparasite species were documented in Israel prior to the reintroduction program; exotic ectoparasites were not detected.

Specimens of gastric mucosa and liver of 25 free-ranging Eurasian lynx (Lynx lynx), and four red foxes (Vulpes vulpes) shot in Sweden during 1999–2000, were investigated for the presence of Helicobacter species. Histopathology, bacteriologic culture and urease test, Helicobacter genus-specific 16S rDNA PCR analysis, and DNA sequence analysis were applied. Numerous Helicobacter-like organisms were observed histologically in the gastric mucosa of one fox. Helicobacter spp. were detected in the stomach by PCR analysis in 17 (68%) of the lynx and in three (75%) of the foxes. Seven of the positive lynx were also positive in the urease test. PCR fragments, amplified from lynx and foxes, were sequenced and compared with those of known Helicobacter species. PCR products from lynx were closely related (≥98% homology) to H. heilmannii, and PCR fragments from foxes demonstrated close homology to H. heilmannii and H. salomonis. No Helicobacter spp. or Helicobacter-like organisms could be cultured. The PCR analysis of the liver was negative for all animals. The pathologic significance of the presence of Helicobacter spp. in the stomach of free-ranging lynx and foxes remains uncertain.

A wild-caught, adult red fox (Vulpes vulpes) from Doñana National Park (southern Spain), in very poor condition, died during anesthesia. At necropsy, the submandibular, retropharyngeal, mediastinal, axillary, mesenteric, and popliteal lymph nodes were enlarged, and the right submandibular and mesenteric lymph nodes had hard, white-colored concretions (calcifications). Multiple white pinpoint foci were observed in the lungs, and abscesses were present in the left dorsal lung lobes. No lesions were seen in tonsils, liver, or spleen. On histopathology multiple tuberculous granulomas, with Ziehl-Neelsen– positive bacilli, were observed in the lung, and multifocal necrotic areas with calcification were present in the submandibular and mesenteric lymph nodes. Nucleic acid amplification from lymph node samples demonstrated the presence of mycobacteria belonging to the Mycobacterium tuberculosis complex. One strain was isolated by culture and identified as Mycobacterium bovis. The suspected route of infection was oral, probably after repeated scavenging of infected wild ungulate carcasses. This is the first report of generalized bovine tuberculosis (bTB) in a wild canid. This finding raises concerns about bTB as a disease risk for protected species, livestock, and humans in Mediterranean Spain.

Two West Indian manatees (Trichechus manatus spp.) were reported with severe emaciation. One animal was a Florida manatee from the Everglades; the other was an Antillean manatee from Cuba. On necropsy, both animals had nephrolithiasis, pyelonephritis, and moderate to severe renomegaly. Histopathology revealed multifocal to diffuse pyelonephritis, interstitial nephritis, and nephrocalcinosis. The stones were analyzed and consisted primarily of calcium carbonate. Serum chemistry values for the Florida animal revealed no renal abnormalities. The mechanism of calculus formation remains unclear in manatees. In horses, another hindgut fermenter, the most common urolith is also calcium carbonate. Urinalyses performed on manatees are very similar to those of horses (i.e., alkaline urine, low specific gravity, and calcium carbonate crystals). Formation of uroliths in manatees may have a pathogenesis similar to equine urolithiasis.

The chytridiomycete fungus Batrachochytrium dendrobatidis is known to be focally distributed across Europe, but has only been linked to chytridiomycosis at a few locations in Spain. Here we report the second occurrence of chytridiomycosis in European amphibians. We found a population of endangered Sardinian newts (Euproctus platycephalus) exhibiting clinical signs of disease including loss of digits and patchy, discolored skin. Molecular examination of skin samples tested positive for B. dendrobatidis. The population of E. platycephalus has been in decline on a timescale consistent with the global emergence of chytridiomycosis, and the ecology of this salamander suggests that the disease in this species warrants concern.

The pathogenic chytrid fungus Batrachochytrium dendrobatidis, which causes the disease chytridiomycosis, has been implicated in declines of amphibian populations throughout the world, including declines and extinctions of local populations of mountain yellow-legged frogs, Rana muscosa, in the California Sierra Nevada. Previous studies have shown B. dendrobatidis achieves its maximum growth rate in culture in the temperature range of 17–25 C, and exposure to very high temperatures can clear frogs of B. dendrobatidis infection. Here we present the results of a laboratory experiment in which experimentally infected R. muscosa tadpoles were followed through metamorphosis at temperatures of 17 and 22 C. All infected animals developed clinical disease within a similar time frame. However, frogs housed at 22 C exhibited a significantly lower mortality than those housed at 17 C. Within 35 days after metamorphosis, 50% of the frogs housed at 22 C died, while 95% of the frogs housed at 17 C died. Clinical signs subsided in the surviving frogs at 22 C, despite persistent infection. Because both temperatures are within the optimal thermal range for growth of B. dendrobatidis, we propose that the difference in outcome indicates the effect of temperature on the host's resistance to chytridiomycosis, rather than an effect on the fungus alone.

An outbreak of anthrax in the Jwana Game Reserve in Jwaneng, Botswana, was first observed when three cheetahs (Acinonyx jubatus) died of the disease in November 2004. In the aftermath of this event, banked serum samples collected from 23 wild-caught cheetahs were examined, by the inhibition enzyme-linked immunoassay (ELISA), for antibodies to the protective antigen (PA) of Bacillus anthracis. Of the 23 cheetahs, 16 regularly accessed the reserve. Antibodies to PA were detected in one cheetah collected in May 2004, indicating the disease was occurring well before it was first noticed. This appears to be the first demonstration of naturally acquired anthrax antibodies in cheetahs. The finding of one antibody-positive animal amongst at least 16 potentially exposed individuals is consistent with existing reports that it is uncommon for cheetahs to develop natural immunity to anthrax.

Plague, caused by the bacterium Yersinia pestis, causes die-offs of colonies of prairie dogs (Cynomys ludovicianus). It has been argued that other small rodents are reservoirs for plague, spreading disease during epizootics and maintaining the pathogen in the absence of prairie dogs; yet there is little empirical support for distinct enzootic and epizootic cycles. Between 2004 and 2006, we collected blood from small rodents captured in colonies in northern Colorado before, during, and for up to 2 yr after prairie dog epizootics. We screened 1,603 blood samples for antibodies to Y. pestis, using passive hemagglutination and inhibition tests, and for a subset of samples we cultured blood for the bacterium itself. Of the four species of rodents that were common in colonies, the northern grasshopper mouse (Onychomys leucogaster) was the only species with consistent evidence of plague infection during epizootics, with 11.1–23.1% of mice seropositive for antibody to Y. pestis during these events. Seropositive grasshopper mice, thirteen-lined ground squirrels (Spermophilus tridecemlineatus), and deer mice (Peromyscus maniculatus) were captured the year following epizootics. The appearance of antibodies to Y. pestis in grasshopper mice coincided with periods of high prairie dog mortality; subsequently, antibody prevalence rates declined, with no seropositive individuals captured 2 yr after epizootics. We did not detect plague in any rodents off of colonies, or on colonies prior to epizootics, and found no evidence of persistent Y. pestis infection in blood cultures. Our results suggest that grasshopper mice could be involved in epizootic spread of Y. pestis, and possibly, serve as a short-term reservoir for plague, but provide no evidence that the grasshopper mouse or any small rodent acts as a long-term, enzootic host for Y. pestis in prairie dog colonies.

Rodents (and their fleas) that are associated with prairie dogs are considered important for the maintenance and transmission of the bacterium (Yersinia pestis) that causes plague. Our goal was to identify rodent and flea species that were potentially involved in a plague epizootic in black-tailed prairie dogs at Thunder Basin National Grassland. We collected blood samples and ectoparasites from rodents trapped at off- and on-colony grids at Thunder Basin National Grassland between 2002 and 2004. Blood samples were tested for antibodies to Y. pestis F-1 antigen by a passive hemagglutination assay, and fleas were tested by a multiplex polymerase chain reaction, for the presence of the plague bacterium. Only one of 1,421 fleas, an Oropsylla hirsuta collected in 2002 from a deer mouse, Peromyscus maniculatus, tested positive for Y. pestis. Blood samples collected in summer 2004 from two northern grasshopper mice, Onychomys leucogaster, tested positive for Y. pestis antibodies. All three positive samples were collected from on-colony grids shortly after a plague epizootic occurred. This study confirms that plague is difficult to detect in rodents and fleas associated with prairie dog colonies, unless samples are collected immediately after a prairie dog die-off.

This study aimed to describe the occurrence of Yersinia pestis, Rickettsia rickettsii, Anaplasma phagocytophilum, and ectoparasites in a wild rodent community in the eastern Sierra Nevada. From May to September 2006, rodents were live-trapped, examined for ectoparasites, and blood was collected. All rodents were serologically tested for antibodies to Y. pestis, R. rickettsii, and A. phagocytophilum; in addition, blood samples and ectoparasites were tested by PCR to detect the presence of these zoonotic agents. Overall, 89 rodents, 46 fleas, and four ticks were collected. Antibody prevalence rates observed for rodents were 14% for R. rickettsii or antigenically related spotted-fever group rickettsiae, and 8% for A. phagocytophilum. No samples were positive for antibodies to Y. pestis. Positive PCR results included one yellow-pine chipmunk for Y. pestis (C_T=32.8), one golden-mantled ground squirrel for R. rickettsii (C_T=33), and one flea found to be co-infected with both R. rickettsii (C_T=17) and A. phagocytophilum (C_T=36). The results of this study provide evidence of multiple zoonoses overlapping within a single, located rodent community.

Mannheimia haemolytica is an important cause of pneumonia in bighorn sheep (BHS; Ovis canadensis). Leukotoxin (Lkt), the primary virulence determinant of M. haemolytica, induces cytolysis of all subsets of leukocytes. Previously, we have shown that CD18, the β subunit of β₂-integrins, mediates Lkt-induced cytolysis. However, it is not clear whether CD18 of all three β₂-integrins, LFA-1, Mac-1, and CR4, mediates Lkt-induced cytol-ysis. The objective of this study was to determine whether BHS LFA-1 (CD11a/CD18) serves as a receptor for Lkt. Plasmids encoding cDNA for BHS CD11a and CD18 were cotransfected into Lkt-resistant HEK-293 cells. Flow cytometric analysis of transfectants confirmed cell surface expression of BHS LFA-1, Lkt-LFA-1 binding and Lkt-induced intracellular calcium elevation. More importantly, the transfectants were efficiently lysed by Lkt in a concentration-dependent manner. Collectively, these results indicate that BHS LFA-1 serves as a functional receptor for M. haemolytica Lkt.

Sixteen captive and five free-ranging black bears (Ursus americanus) were immobilized with a combination of butorphanol, azaperone, and medetomidine (BAM). The BAM drug combination was premixed using 0.5 ml butorphanol (30 mg/ml), 0.25 ml aza-perone (50 mg/ml), and 0.25 ml medetomidine (20 mg/ml) per milliliter to yield a final mix of (15 mg butorphanol 12.5 mg azaperone 5 mg medetomidine)/ml. This combination, dosed at 0.4 ml BAM/∼23 kg estimated body weight, provided a mean induction time of 10 min (95% confidence interval [CI] = 2 min), consistent anesthesia without apparent adverse effects, and smooth recovery (mean = 15 min, 95% CI = 4 min) after antagonism with atipamezole (5 mg/mg medetomidine) alone or in combination with naltrexone (5 mg/mg butorphanol). Based on our initial observations, BAM appears to be a reversible and accessible drug combination for immobilizing black bears that merits further evaluation for field use.

Atipamezole, azaperone, black bear, butorphanol, chemical immobilization, medetomidine, naltrexone, Ursus americanus.

The susceptibility of wild ruminants, especially cervids, to bovine viral diarrhea virus (BVDV) has remained an enigma. Two white-tailed deer (Odocoileus virginianus) were submitted to the Animal Disease Research and Diagnostic Laboratory (ADRDL) in the fall of 2003 by the South Dakota Game Fish and Parks for chronic wasting disease (CWD) testing. Both animals were CWD negative. The animals were necropsied and histopathology, viral antigen detection, and virus isolation were performed. A noncytopathic (NCP) BVDV was isolated from the lungs and several other tissues of both animals. Formalin-fixed ear notches from both animals were positive for BVDV antigen by immunohistochemistry. The BVDV isolates were typed with the use of polymerase chain reaction in 5′ untranslated region (UTR) and one isolate was typed a Type 2a and the other a Type 1b. Future field surveys to determine the incidence of BVDV along with experimental studies to determine if white-tailed deer fawns can be persistently infected with BVDV are needed.

Transmission of pathogens from domestic animals to wildlife populations (spillover) has precipitated local wildlife extinctions in multiple geographic locations. Identifying such events before they cause population declines requires differentiating spillover from endemic disease, a challenge complicated by a lack of baseline data from wildlife populations that are isolated from domestic animals. We tested sera collected from 12 ocelots (Leopardus pardalis) native to Barro Colorado Island, Panama, which is free of domestic animals, for antibodies to feline herpes virus, feline calicivirus, feline corona virus, feline panleukopenia virus, canine distemper virus, and feline immunodeficiency virus (FIV), typically a species-specific infection. Samples also were tested for feline leukemia virus antigens. Positive tests results were only observed for FIV; 50% of the ocelots were positive. We hypothesize that isolation of this population has prevented introduction of pathogens typically attributed to contact with domestic animals. The high density of ocelots on Barro Colorado Island may contribute to a high prevalence of FIV infection, as would be expected with increased contact rates among conspecifics in a geographically restricted population.

The Camargue area of southern France experienced the re-emergence of West Nile Virus (WNV) in the late summer of 2000 and 2004. Immediately preceding the 2004 outbreak, samples were collected from 432 birds of 32 different species captured in mist nets and from 201 Cattle Egret (Bubulcus ibis) nestlings sampled in their nests between 1 April and 12 June 2004. West Nile virus neutralizing titers of ≥40 were detected in 4.8% (95% confidence limit, 2.9–7.5%) of the adult birds and in 1.6% (0.3–4.6%) of the egret nestlings. Migratory passerines had a higher prevalence of WNV neutralizing antibodies (7.0%) than did resident and short-distance migratory passerines (0.8%), suggesting exposure to WNV or a related flavivirus during overwintering in Africa.

Wild birds are considered to be the natural reservoir of the Newcastle disease virus (NDV; avian paramyxovirus-1) causing Newcastle disease, and are often suspected to be involved in outbreaks in domesticated birds. To assess the epidemiologic status of wild birds living, or overwintering, in Switzerland, 3,049 cloacal swabs covering the period 2003–2006 were screened for NDV, using real time RT-PCR. All samples were negative. This result seems in contrast with previously performed serologic screenings of wild birds.

Blood samples were collected from wild boar (Sus scrofa) shot during the hunting season from 1999 to 2005 in the Czech Republic. Sera were tested by enzyme-linked immunosorbent assay for the presence of antibodies against classical swine fever virus (CSFV), swine vesicular disease virus (SVDV), Aujeszky's disease virus (ADV), and bovine viral diarrhea virus (BVDV). Indirect fluorescence antibody test was used for detection of antibodies against porcine circovirus type 2 (PCV-2) and transmissible gastroenteritis virus (TGEV). Antibodies against ADV, BVDV, PCV-2, and TGEV were detected in 30% (101 of 338), 1% (2 of 352), 43% (57 of 134), and 1% (1 of 134) of wild boars, respectively. Sera of 6,471 and 362 tested wild boars were negative for the presence of antibodies against CSFV and SVDV, respectively. This is the first survey of TGEV antibodies in wild boars and the first serologic survey of viral diseases in wild boars in the Czech Republic. Wild boars in the Czech Republic may act as a potential reservoir of ADV and thus have a role in the epidemiology of this disease.

Type C1 botulism outbreaks in Black-faced Spoonbills (Platalea minor) occurred in Taiwan from 2002 to 2003, and hematologic and biochemistry parameters from botulism-paralyzed birds and recovered birds were compared. Values for creatinine and uric acid were higher (P<0.0025) in birds with botulism than in recovered birds. Lower white blood cell counts (P<0.005) and values for alkaline phosphatase, aspartate aminotransferase, alanine aminotransferase, and triglycerides (P<0.025) were observed in recovered birds. Based on these observations, we suggest that hematologic and biochemistry analyses should be performed to assess the health condition of birds recovering from botulism.