Since 1999, large-scale mortalities of fish-eating birds have been observed on the Great Lakes, and more specifically on Lake Erie. Type E botulism has been established as the primary cause of death. The mechanism of type E botulism exposure in fish-eating birds is unclear. Given that these birds are thought to eat live fish exclusively, it seems likely that their prey play a key role in the process, but the role of fish as potential transport vectors of botulinum neurotoxin type E (BoNT/E) to birds has not been adequately investigated. Between June 2003 and April 2004 a methodological model for exposing fish to Clostridium botulinum was developed and used to compare the sensitivity of rainbow trout (Oncorhynchus mykiss), round goby (Neogobius melanostomas), walleye (Stizostedion vitreum), and yellow perch (Perca flavescens) to four doses (0, 800, 1,500, and 4,000 Mouse Lethal Doses) of Clostridium botulinum type E neurotoxin. Each fish species expressed unique changes in both behavior and skin pigmentation prior to death. Yellow perch survived significantly longer (P<0.05) than the three other species at all toxin treatments. Results of this study suggest that live fish can represent a significant vector for transfer of BoNT/E to birds.

In a 4-mo study (June 2004–September 2004), round gobies (Neogobius melanostomas) were dosed orally every 72 hr for up to 21 days with Clostridium botulinum neurotoxin type E (BoNT/E) at one of four doses: 0, 50, 250, and 500 mouse lethal doses (MLD). Fish were observed for changes in pigmentation and behavior for the duration of the experiment. Mortality was observed with all treatments, with the exception of the 0 MLD control. Clinical signs observed were consistent with prior research and appeared to occur in a threshold manner. The mean times to death and percent mortalities were dose dependant. Hazard ratios were determined to have a significant positive (parameter estimate=0.03) linear relationship with dose. The hazard ratio showed that per one unit dose increase, the instantaneous probability of a fish dying increased 1.02%. Postmortem analysis of experimental fish demonstrated that 11% (3/27) of fish contained detectable BoNT/E in their visceral fraction. The other 89% tested negative for BoNT/E, despite the fact that all fish died as a result of BoNT/E exposure. Therefore, botulism should not necessarily be ruled out as the cause of a fish kill, even if the fish test negative for BoNT/E.

Avian vacuolar myelinopathy (AVM) is a neurologic disease of unknown etiology that affects bald eagles (Haliaeetus leucocephalus), American coots (Fulica americana), and several species of waterfowl. An unidentified neurotoxin is suspected as the cause of AVM, which has been documented at several reservoirs in the southeastern United States. We conducted diagnostic and epidemiologic studies annually during October–March from 1998–2004 at Clarks Hill/Strom Thurmond Lake on the Georgia/South Carolina border to better understand the disease. Avian vacuolar myelinopathy was confirmed or suspected as the cause of morbidity and mortality of 28 bald eagles, 16 Canada geese (Branta canadensis), six American coots, two great-horned owls (Bubo virginianus), and one killdeer (Charadrius vociferus). Active surveillance during the outbreaks yielded annual average prevalence of vacuolar lesions in 17–94% of coots, but not in 10 beavers (Castor canadensis), four raccoons (Procyon lotor), and one gray fox (Urocyon cinereoargenteus) collected for the study. Brain lesions were not apparent in 30 Canada geese collected and examined in June 2002. The outbreaks at this location from 1998–2004 represent the most significant AVM-related bald eagle mortality since the Arkansas epornitics of 1994–95 and 1996–97, as well as the first confirmation of the disease in members of Strigiformes and Charadriiformes.

Twenty-eight hog-nosed skunks (Conepatus leuconotus), 23 striped skunks (Mephitis mephitis), and nine spotted skunks (Spilogale gracilis) from an area of sympatry in west-central Texas were examined for helminth parasites. Shared helminth species among all three host species were one nematode (Physaloptera maxillaris), two acanthocephalans (Pachysentis canicola, Macracanthorhynchus ingens), and one cestode (Mathevotaenia mephitis). Two nematodes (Gongylonema sp. and Filaria taxidaea) occurred in both the striped and hog-nosed skunks. One nematode (Filaroides milksi) and one acanthocephalan (Oncicola canis) were collected only from C. leuconotus. The most common helminth infections for striped and hog-nosed skunks were P. maxillaris and P. canicola. Helminth species richness was highest in hog-nosed skunks, but striped skunks had the highest prevalences and intensities of all the common helminth species. The helminth fauna of spotted skunks was markedly depauperate in terms of species richness and helminth abundance compared to the other two host species. Differences in helminth communities across these three sympatric skunks may be related to differences in their relative abundance, behavior, food habits, and geographic range.

Piroplasmosis has been identified as a possible cause of mortality in reintroduced Przewalski's horses (Equus ferus przewalskii) in the Dsungarian Gobi (Mongolia). A cross-sectional and a longitudinal study were conducted in a representative sample (n=141) of the resident domestic horse population and in 23 Przewalski's horses to assess the prevalence of Theileria equi and Babesia caballi. Piroplasms were detected in blood by light microscopy in 6.7% (95% confidence interval [CI]: 3.6–12.2%) of the domestic horse samples. Antibody prevalence was 88.6% (95% CI: 82.4–92.9%) for T. equi and 75.2% (95% CI: 67.4–81.6%) for B. caballi. Antibody prevalence did not change over time, but antibody prevalence for both piroplasms were significantly lower in animals less than 1 yr of age. For both piroplasms, the prevalence of presumably maternal antibodies (falling titers) in foals was 100%. Only one of 16 foals seroconverted against T. equi during the study period, despite that piroplasms were found in two other individuals. The incidence density (ID) of T.equi in foals was therefore 0.0012 seroconversions per horse day (95% CI: 0.00029–0.0057). In contrast, yearlings had an ID of 0.0080 (95% CI: 0.0049–0.010) for T. equi and 0.0064 (95% CI: 0.0036–0.0093) for B. caballi, and in seven individuals piroplasms were detected. The seroprevalence of both piroplasms rose from 20% in spring to 100% in autumn. Comparison of domestic and Przewalski's horses resulted in a standardized prevalence ratio (SPR) of 0.98 (95% CI: 0.80–1.24, not significant) for B. caballi; in contrast, the prevalence of T. equi in Przewalski's horses was significantly lower than expected (SPR=0.51, 95% CI: 0.50–0.64).

Documenting the occurrence of Parelaphostrongylus odocoilei has historically relied on the morphological examination of adult worms collected from the skeletal muscle of definitive hosts, including deer. Recent advances in the knowledge of protostrongylid genetic sequences now permit larvae to be identified. Dorsal-spined larvae (DSLs) collected in 2003–2004 from the lung and feces of six Columbian black-tailed deer (Odocoileus hemionus columbianus) from Oregon were characterized genetically. The sequences from unknown DSLs were compared to those from morphologically validated adults and larvae of P. odocoilei at both the second internal transcribed spacer (ITS-2) of ribosomal DNA and the mitochondrial cytochrome oxidase II gene. We provide the first unequivocal identification of P. odocoilei in Columbian black-tailed deer from Oregon. The broader geographic distribution, prevalence, and pathology of P. odocoilei are not known in populations of Oregon deer.

Our abilities to assess health risks to free-ranging dolphin populations, to treat live-stranded or captive dolphins, and to evaluate the risks of disease transmission between humans and dolphins have suffered from a lack of basic information on microorganisms associated with normal, presumably healthy free-ranging individuals. In order to provide these data, we sampled free-ranging bottlenose dolphins (Tursiops truncatus) off Florida, Texas, and North Carolina during 1990–2002. Blowhole and anal/fecal samples yielded 1,871 bacteria and yeast isolates and included 85 different species or groups of organisms. Vibrios, unidentified pseudomonads, Escherichia coli, Staphylococcus spp., and a large group of nonfermenting Gram-negative bacteria represented >50% of isolates. Vibrio alginolyticus and Vibrio damsela were the most commonly recovered bacteria from both anal/fecal and blowhole samples. Many organisms occurred sporadically in dolphins that were sampled repeatedly, but some were consistently isolated from individual animals and may indicate the carrier state. Vibrios were common, but some geographic variability in the presence of these and other organisms was noted. Potential pathogens of significance to humans and other animals were recovered.

We investigated the effectiveness of culture-independent molecular methods for determining host-associated microbial diversity in bighorn sheep (Ovis canadensis). Results from bacterial culture attempts have been the primary source of information on host-associated bacteria, but studies have shown that culture-based results significantly underestimate bacterial diversity in biological samples. To test the effectiveness of culture-independent methods, we extracted DNA from nasal and oropharyngeal swab samples collected from bighorn sheep in four different populations. From these samples, we amplified, cloned, and sequenced small subunit (16S) ribosomal DNA (rDNA) to identify the scope of microbial diversity in bighorn respiratory tracts. Phylogenetic analysis of these rDNA gene sequences revealed organismal diversity an order of magnitude higher than was determined by culture methods. Pasteurellaceae bacteria were the most diverse phylogenetic group in live bighorn sheep, and members of bacterial genera often associated with respiratory disease were found in all the samples. Culture-independent methods were also able to directly detect leukotoxin (lktA) gene sequences in swab and lung tissue samples. Overall, our results show the power of culture-independent molecular methods for identifying microbial diversity in bighorn sheep and the potential for these methods to detect the presence of virulence genes in biological samples.

Bacterial kidney disease (BKD), caused by Renibacterium salmoninarum, is a widespread problem with major implications for salmonid fish species. The mechanisms by which the bacterium has reached high levels of infection previously unrecorded in the Laurentian Great Lakes are presently unknown. Research involving reservoirs and mechanisms of R. salmoninarum transmission in fish is lacking because of the ecologic complexity of heterogeneous habitats and the lack of adequate funding. Herein, we report on the isolation of R. salmoninarum from the kidneys of the sea lamprey (Petromyzon marinus). The bacterium was cultured from kidneys of 16% and 4% of lampreys collected from two locations within the Lake Ontario watershed in 2003 and 2004, respectively. The identity of bacterial colonies was verified with the nested polymerase chain reaction and quantitative enzyme-linked immunosorbent assay.

The isolation and characterization of two bacterial species, Streptococcus agalactiae and Lactococcus garvieae, previously unreported in wild marine mammals are described from a freshly dead bottlenose dolphin, Tursiops truncatus, from Kuwait Bay, Kuwait, in September 2001. Conventional and rapid identification systems were used to determine that isolates from muscle and kidney were S. agalactiae and L. garvieae, respectively. The isolates were gram-positive, catalase-negative, oxidase-negative, nonhemolytic cocci. The S. agalactiae was serotyped to group antigen B, whereas the L. garvieae could not be assigned to any serogroup. These Kuwait isolates displayed considerable homogeneity with corresponding American Type Culture Collection (ATCC) type isolates. Although the dolphin S. agalactiae isolate was nonhemolytic, it was biochemically similar to S. agalactiae isolated from mullet sampled in the concurrent Kuwait Bay fish kill. Some biochemical heterogeneity was observed between the dolphin isolates and corresponding mammalian ATCC type isolates, especially with Voges Proskauer, alaninephenylanaline-proline arylamidase, and alpha-galactosidase tests. Nile tilapia, Oreochromis niloticus, experimentally infected with the dolphin S. agalactiae and L. garvieae isolates experienced 90% and 0% mortalities, respectively. This is the first isolation of S. agalactiae and L. garvieae from a wild marine mammal, and the microbial characteristics established here provide pertinent information for the future isolation of these bacteria.

Blood samples were collected from 2,635 caribou (Rangifer tarandus), 1,238 grizzly bears (Ursus arctos), and 930 wolves (Canis lupus) from throughout mainland Alaska during 1975–98. Sera were tested for evidence of exposure to Brucella spp. Serum antibody prevalences were highest in the northwestern region of the state. In any specific area, antibody prevalences for caribou and wolves were of a similar magnitude, whereas antibody prevalence for bears in these same areas were two to three times higher.

Chronic wasting disease (CWD) is a fatal, emerging disease of cervids associated with transmissible protease-resistant prion proteins. The potential for CWD to cause dramatic declines in deer and elk populations and perceived human health risks associated with consuming CWD-contaminated venison have led wildlife agencies to embark on extensive CWD control programs, typically involving culling to reduce deer populations. We characterized the spatial distribution of CWD in white-tailed deer (Odocoileus virginianus) in Wisconsin to facilitate CWD management. We found that CWD prevalence declined with distance from a central location, was locally correlated at a scale of 3.6 km, and was correlated with deer habitat abundance. The latter result is consistent with patterns expected for a positive relationship between density and prevalence of CWD. We recommend management activities focused on culling in geographic areas with high prevalence to have the greatest probability of removing infected individuals. Further research is needed to elucidate the factors involved in CWD spread and infection rates, especially the role of density-dependent transmission.

During 1999–2003, 127 cases of raccoon variant rabies were reported in raccoons (Procyon lotor) and striped skunks (Mephitis mephitis) in Ontario, Canada. Raccoons accounted for 98% (125/127) of the reported cases with behaviors/conditions including aggression, fighting with dogs, ataxia, vocalizations, appearance of being sick, and the presence of porcupine (Erethizon dorsatum) quills. Seventy-eight percent of the rabid raccoons were adults. Juveniles were underrepresented (22%) compared with the adult/juvenile ratios found in nonrabid Ontario raccoon populations. Of the known aged raccoons, 83% were ≤3 yr of age, and 22% of the rabid adult female raccoons had evidence of having had a litter during the year in which they were found to be rabid. The majority of rabid raccoons were reported during the fall, winter, and spring, suggesting a relationship between raccoon behavioral activities such as denning and breeding and the timing of rabies outbreaks. Multiple cases of raccoon rabies occurred at several barns, suggesting that those structures serve as focal points of rabies transmission as a result of denning activities. Movements of five rabid raccoons (range 1,564–4,143 m) were not different from movements of nonrabid raccoons in Ontario. Sixty-six percent of the rabid animals were submitted by government staff, stressing the importance of those agencies in rabies control and surveillance operations. Increased knowledge of the behaviors of rabid raccoons should assist in the development of management strategies for rabies.

Serological and genetic material collected over 15 years (1990–2004) from 207 cougars (Puma concolor) in four populations in the Rocky Mountains were examined for evidence of current or prior exposure to feline immunodeficiency virus (FIV), feline parvovirus (FPV), feline coronavirus (FCoV), feline calicivirus (FCV), canine distemper virus (CDV), feline herpesvirus (FHV), and Yersinia pestis. Serologic data were analyzed for annual variation in seroconversions to assess whether these pathogens are epidemic or endemic in cougars, and to determine whether family membership, age, sex, or location influence risk of exposure. FIV and FPV were clearly endemic in the studied populations, whereas exposure to FCoV, FCV, CDV, and Y. pestis was more sporadic. No evidence was found for FHV. Age was the most consistent predictor of increased exposure risk, often with no other important factors emerging. Evidence for transmission within family groups was limited to FIV and FCoV, whereas some indication for host sex affecting exposure probability was found for FIV and Y. pestis. Overall, cougar populations exhibited few differences in terms of pathogen presence and prevalence, suggesting the presence of similar risk factors throughout the study region.

During the autumn of 1999 (mid-August–late September), an outbreak of hemorrhagic disease in white-tailed deer (Odocoileus virginianus) caused by epizootic hemorrhagic disease virus serotype 1 (EHDV-1) occurred along the east coast of the United States from Georgia to New Jersey. An EHDV-1 epizootic of such magnitude had not been described in this region since 1975. To determine the genetic relatedness among the 1999 viruses, as well as among additional EHDV-1 isolates from the eastern and western United States, portions of the S10 and L2 gene segments were sequenced and compared utilizing phylogenetic analyses. Nearly all of the 1999 eastern isolates were identical in nucleotide sequence at one or both loci. Additionally, confirmed cases of EHDV-1 in white-tailed deer occurred in a south (Georgia)-to-north (New Jersey/Virginia) progression over a short period of approximately six weeks. Taken together, these results indicate that this outbreak resulted from the spread of a single viral strain. The phylograms derived from analysis of the entire sample set displayed eastern and western region–specific clusterings (topotypes), as well as an eastern versus western difference in branch lengths, which may reflect the influence of epizootic versus enzootic transmission patterns on viral genetic diversity.

The Galápagos penguin (Spheniscus mendiculus) is an endangered species endemic to the Galápagos Islands, Ecuador. In 2003 and 2004, 195 penguins from 13 colonies on the islands of Isabela and Fernandina in the Galápagos archipelago were examined. Genetic sexing of 157 penguins revealed 62 females and 95 males. Hematology consisted of packed cell volume (n=134), white blood cell differentials (n=83), and hemoparasite blood smear evaluation (n=114). Microfilariae were detected in 22% (25/114) of the blood smears. Female penguins had significantly higher eosinophil counts than males. Serum chemistry on 83 penguins revealed no significant differences between males and females. Birds were seronegative to avian paramyxovirus type 1–3, avian influenza virus, infectious bursal disease virus, Marek's disease virus (herpes), reovirus, avian encephalomyelitis virus, and avian adenovirus type 1 and 2 (n=75), as well as to West Nile virus (n=87), and Venezuelan, western and eastern equine encephalitis viruses (n=26). Seventy-five of 84 (89%) penguins had antibodies to Chlamydophila psittaci but chlamydial DNA was not detected via polymerase chain reaction in samples from 30 birds.

Twenty-nine free-ranging Himalayan tahr (Hemitragus jemlahicus) were darted in the Sagarmatha National Park (Nepal) using different combinations of xylazine and ketamine. Animals in Group 1 (n=4) received a mean xylazine–ketamine dose of 2.77±0.99 mg/kg xylazine plus 3.32±0.19 mg/kg ketamine in males and 2.39±0.10 mg/kg xylazine plus 4.29±0.17 mg/kg ketamine in females. Animals in Group 2 (n=25) received a mean xylazine–ketamine dose of 1.70±0.41 mg/kg xylazine plus 3.06±0.74 mg/kg ketamine in males and 1.82±0.29 mg/kg xylazine plus 3.29±0.52 mg/kg ketamine in females. No anesthetic-related mortality was recorded. Anesthesia was reversed by a standard dose of 11 mg/animal of atipamezole administered by intramuscular injection. Although all anesthetic dosages immobilized free-ranging tahr successfully, a quick and smooth recovery was obtained (11.1±5.6 min) only with the dosages of Group 2.

Three captive Shira's moose (Alces alces shirasi) were orally inoculated with a single dose (5 g) of whole-brain homogenate prepared from chronic wasting disease (CWD)–affected mule deer (Odocoileus hemionus). All moose died of causes thought to be other than CWD. Histologic examination of one female moose dying 465 days postinoculation revealed spongiform change in the neuropil, typical of transmissible spongiform encephalopathy. Immunohistochemistry staining for the proteinase-resistant isoform of the prion protein was observed in multiple lymphoid and nervous tissues. Western blot and enzyme-linked immunosorbent assays provided additional confirmation of CWD. These results represent the first report of experimental CWD in moose.

As part of a species recovery program, 129 Canada lynx (Lynx canadensis) originating from British Columbia, the Yukon, Manitoba, and Quebec, Canada, and Alaska, USA, were reintroduced to southwestern Colorado, USA, from 1999 to 2003. Of 52 lynx mortalities documented by October 2003, six lynx, including a female and her 5-mo-old kitten, had evidence of Yersinia pestis infection as determined by fluorescent antibody test and/or culture. Postmortem findings in these lynx were characterized by pneumonia, ranging from acute suppurative pneumonia, to multifocal necrotizing pneumonia, to fibrinous bronchopneumonia. Histopathologic examination of lung revealed multiple areas of inflammation and consolidation, areas of edema and hemorrhage, and bacteria surrounded by extensive inflammation. Spleens had severe lymphoid depletion and hypocellular red pulp. Lymphadenomegaly was observed in only one plagueaffected lynx. We hypothesize that these Canada lynx were exposed to Y. pestis by infected prey, and these are the first reports of plague in this species.

An adult female harbor seal (Phoca vitulina richardsi) stranded in northern California on 25 June 2004, exhibited progressive weakness, disorientation, and seizures, and despite therapy, died within 4 days. On pathologic examination, a lead fishing sinker was in the stomach, and changes in the brain, heart, kidney, liver, lymph nodes, and spleen were supportive of acute lead toxicosis. The diagnosis was made on the basis of concentrations of lead in the sinker (90–98% lead), antemortem whole blood (0.66 ppm), and postmortem tissues (84 ppm, wet weight liver). This first documented case of lead toxicosis in a wild marine mammal demonstrates an additional way in which human fishing activities can harm marine mammals.

Koi herpesvirus (KHV), a highly contagious and lethal virus that affects both koi (Cyprinus carpio koi) and common carp (Cyprinus carpio), was isolated in 1998 from two outbreaks of koi suffering mass mortality in New York State, USA, and in Israel. The disease had been described as early as 1996 in Europe. In July 2004, this virus was found associated with a mass mortality event in wild common carp in the Chadakoin River, New York, USA (42°07′N, 79°W). Affected fish typically showed marked hyperplasia of gill tissues, abdominal adhesions, and severe multifocal to diffuse external hemorrhages. The virus isolated in this outbreak was somewhat unusual in that it initially replicated well in fathead minnow cell cultures, which is typical of spring viremia of carp virus. Testing at the National Veterinary Services Laboratories, Ames, Iowa, USA, confirmed the virus's identity to be KHV. Koi herpesvirus is not currently on the OIE (World Organisation for Animal Health) list of notifiable diseases; however, it is capable of causing mass mortality in susceptible fish at permissive temperatures.

Oral vaccination of free-ranging wildlife is a promising technique in rabies control. The small Asian mongoose (Herpestes javanicus) is an important reservoir of rabies on several Caribbean islands, but no vaccines have been evaluated for this species. Captive mongooses were used to test the safety and efficacy of the commercially licensed vaccinia-rabies glycoprotein (V-RG) recombinant vaccine and a newly developed genetically engineered oral rabies virus vaccine (SPBNGA-S). In one study using V-RG, no vaccinated animals developed detectable rabies virus–neutralizing antibodies, and all but one died after experimental challenge with rabies virus. In contrast, all animals given SPBNGA-S demonstrated seroconversion within 7 to 14 days after vaccination and survived rabies virus challenge. On the basis of these preliminary results indicating the greater efficacy of SPBNGA-S vs. V-RG vaccine, additional investigations will be necessary to determine the optimal dose and duration of vaccination, as well as incorporation of the SPBNGA-S vaccine into edible bait.

Serum samples from 14 lions (Panthera leo) from Queen Elizabeth National Park, Uganda, were collected during 1998 and 1999 to determine infectious disease exposure in this threatened population. Sera were analyzed for antibodies against feline immunodeficiency virus (FIV), feline calicivirus (FCV), feline herpesvirus 1 (feline rhinotracheitis: FHV1), feline/canine parvovirus (FPV/CPV), feline infectious peritonitis virus (feline coronavirus: FIPV), and canine distemper virus (CDV) or for the presence of feline leukemia virus (FeLV) antigens. Ten lions (71%) had antibodies against FIV, 11 (79%) had antibodies against CDV, 11 (79%) had antibodies against FCV, nine (64%) had antibodies against FHV1, and five (36%) had antibodies against FPV. Two of the 11 CDV-seropositive lions were subadults, indicating recent exposure of this population to CDV or a CDV-like virus. No lions had evidence of exposure to FeLV or FIPV. These results indicate that this endangered population has extensive exposure to common feline and canine viruses.

Wild rodents (n=330) were trapped around the villages of Altındere and Coşandere (Maçka, Trabzon Province), Ayder, Ortan, and Yolkiyi (Çamlıhemşin, Rize Province), and Bozdağ (Ödemiş, İzmir Province) in northeastern and western Turkey during April 2004. Samples were tested for arenavirus, hantavirus, and cowpox virus (family Poxviridae, genus Orthopoxvirus, CPXV) antibodies by using immunofluorescence assays (IFAs). Antibodies against arenaviruses were found in eight of 330 (2.4%) rodents. Arenavirus-sero-positive animals were found from all study sites. Antibodies to Puumala virus (family Bunyaviridae, genus Hantavirus, PUUV) were detected in four of 65 Microtus voles tested. Of the PUUV-IFA-positive voles, one Microtus guentheri lydius was caught from Izmir, and one Microtus roberti and two Microtus rossiaemeridionalis were captured near Trabzon. All 264 Apodemus spp. mice tested negative for antibodies to Saaremaa virus (family Bunyaviridae, genus Hantavirus, SAAV); the single Dryomys nitedula tested negative for both PUUV and SAAV antibodies. Only one (0.3%) of the rodents, an Apodemus sylvaticus from Trabzon area, tested seropositive to CPXV. This is the first serologic survey for rodent-borne viruses in their natural hosts in Turkey. Although these preliminary results support presence of several virus groups with zoonotic potential, additional studies are needed to identify the specific viruses that are present in these populations.

A free-ranging adult female eastern box turtle (Terrapene carolina carolina) was presented to the University of Tennessee in October 2003 because of suspected trauma and blindness. Physical examination revealed lethargy, clear ocular and nasal discharges, and white oral and laryngeal plaques. Intracytoplasmic inclusions within heterophils and large mononuclear leukocytes were observed on routine blood smear examination. Postmortem findings included necrosis of epithelial and parenchymal cells with intracytoplasmic inclusions. Ultrastructurally, the leukocyte inclusions consisted of variably electron-dense granular material and viral particles consistent with the Iridoviridae family of viruses. The virus shared 100% sequence identity to a 420-base pair sequence of frog virus 3 (family Iridoviridae, genus Ranavirus) as determined by polymerase chain reaction and gene sequencing targeting a portion of the Ranavirus major capsid protein gene.

Roe deer (Capreolus capreolus), chamois (Rupicapra rupricapra rupicapra), and domestic sheep in the Orobie Alps, Italy, were serologically tested for antibodies to selected pathogens that may be transmitted across species. Antibodies against Brucella spp. and bovine herpesvirus 1 (roe deer and chamois only) were not detected in any species. In roe deer, antibodies were detected against Toxoplasma gondii (13%) and Neospora caninum (3%). Chamois tested positive for antibodies to T. gondii (5%), N. caninum (21%), bovine respiratory syncytial virus (BRSV) (41%), bovine parainfluenza type-3 virus (17%), pestiviruses (18%), and Mycoplasma conjunctivae (17%). In the sheep, particularly high antibody prevalence rates were found for T. gondii (78%), Chlamydophila spp. (20%), pestiviruses (90%), BRSV (82%), and M. conjunctivae (81%).

The incidence of mycobacterial infections was monitored in brown bears (Ursus arctos) in the National Park Low Tatras in the central European Carpathians in Slovakia. Tissue samples of 20 brown bears were examined microscopically and by culture for the presence of mycobacteria. Acid-fast rods were detected by Ziehl-Neelsen staining in a smear from the kidney of one brown bear, although the culture was negative for mycobacteria. Mycobacterium avium subsp. paratuberculosis, the causative agent of paratuberculosis in ruminants, was isolated from the intestinal mucosa of another two brown bears. The isolates were identified by polymerase chain reaction for the specific insertion sequence IS900. Using standardized IS900 restriction fragment length polymorphism (RFLP) analysis, the M. a. paratuberculosis isolates were classified as RFLP type B-C1, which also were detected in the infected cattle in surrounding area. This study describes the first isolation of M. a. paratuberculosis from a brown bear. Our results confirm that animal species other than ruminants can become infected with M. a. paratuberculosis and can act as potential vectors and/or reservoirs of the infection.

A case of fatal salmonellosis in a Eurasian eagle-owl (Bubo bubo) from Bursa Province (northwestern Turkey) is described. The organs of the bird were examined histo-pathologically and microbiologically. Macroscopic and microscopic findings were consistent with a Salmonella infection. Salmonella enterica subspecies enterica serovar Enteritidis (S. Enteritidis) phage type (PT) 21b was isolated from the liver and spleen in pure culture and from the intestine. The isolate was susceptible to amoxycillin/clavulanic acid, ampicillin, chloramphenicol, gentamicin, kanamycin, tetracycline, and trimethoprim/sulpha-methoxazole. This is the first report of an isolation of salmonellae from a wild bird species from Turkey and the first time S. Enteritidis PT21b has been reported from Turkey.

To determine the prevalence and serovars of Salmonella in free-ranging deer, we cultured feces from white-tailed deer (Odocoileus virginianus) harvested by hunters during a regular firearm season in southeastern Nebraska (USA). We recovered Salmonella from 5 (1%; 95% confidence interval: 0.37–2.20%) of 500 samples and identified four different Salmonella enterica serovars [Litch-field (1), Dessau (1), Infantis (2), and Enteritidis (1)]. Although the prevalence of Salmonella in free-ranging deer appears to be low, the serovars recovered are known to be pathogenic to humans and animals.

We conducted a retrospective se-rologic survey for antibodies against the MPB70 protein of Mycobacterium bovis in wild carnivores from Doñana National Park (southwestern Spain). Serum samples from 118 red foxes (Vulpes vulpes), 39 Iberian lynx (Lynx pardinus), 31 Eurasian badgers (Meles meles), five Egyptian mongoose (Herpestes ichneumon), four European genet (Genetta genetta), and one Eurasian otter (Lutra lutra) were analyzed using an indirect competitive enzyme-linked immunoassay. Antibodies against the MPB70 protein of M. bovis were detected in seven badgers, five foxes, and one lynx. The frequency of positive animals was significantly higher in badger (23%) than in lynx (3%) and fox (4%). Antibodies were not detected in other species. Annual antibody frequency peaked at 38% in badgers and 11% for red fox. These species may contribute to persistence of bovine tuberculosis in Doñana.

Bacteriologic examination of an abscess found between blubber and musculature of a striped dolphin (Stenella coeruleoalba) found dead on 30 March 2002 in the Adriatic Sea, Croatia, was performed and an aerotoler-ant, nontoxogenic Clostridium tertium was isolated in pure culture. National Collections of Industrial Food and Marine Bacteria (NCIMB Ltd., Aberdeen, Scotland, UK) confirmed the results. Sequencing results showed it to be C. tertium with 100% similarity. The strain was named Clostridium tertium strain Zagreb, acceded to the culture collection and assigned the accession number NCIMB 13970. This is the first record of C. tertium in marine mammals.

Disseminated visceral coccidiosis (DVC) was unexpectedly recognized in a wild white-naped crane (Grus vipio) killed by phosphamidon insecticide. On gross pathologic examination, widely disseminated white nodules were found on the serosa of the proventriculus, gizzard, and intestine, as well as on the surface and in the parenchyma of liver, spleen, and cardiac muscle. Microscopically, asexual stages of a coccidia were observed in some nodules. However, the species of coccidia could not be determined because no oocysts were found on fecal examination. This is believed to be the first reported case of DVC in a wild white-naped crane infected with Eimeria spp.