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There has been, to date, little discussion about the defining features and measures of wildlife health in the literature or legislation. Much wildlife health work focuses on the detection and response to infectious or parasitic diseases; this perspective has been reinforced by the focus of the One Health initiative on wildlife as sources of emerging infections. The definition of health as “the absence of disease” lags 70 yr behind modern concepts of human health and emerging concepts of wildlife health in terms of vulnerability, resilience, and sustainability. Policies, programs, and research that focus on the integration of wildlife health with natural resource conservation, ecosystem restoration, and public health need a working definition of health that recognizes the major threats to fish and wildlife are the result of many other drivers besides pathogens and parasites, including habitat loss, globalization of trade, land-use pressure, and climate change. A modern definition of wildlife health should emphasize that 1) health is the result of interacting biologic, social, and environmental determinants that interact to affect capacity to cope with change; 2) health cannot be measured solely by what is absent but rather by characteristics of the animals and their ecosystem that affect their vulnerability and resilience; and 3) wildlife health is not a biologic state but rather a dynamic social construct based on human expectations and knowledge.
Helicobacter infection in cetaceans was first reported from the US in 2000 when the isolation of a novel Helicobacter species was described from two Atlantic white-sided dolphins (Lagenorhynchus acutus). Since then, Helicobacter species have been demonstrated in cetaceans and pinnipeds from around the world. Since 1990, the Animal Health and Veterinary Laboratories Agency Polwhele, Truro, has been involved in the UK Cetacean Strandings Investigation Programme to establish the cause of death of cetacean species stranded along the coast of Cornwall, England. We describe the isolation of Helicobacter cetorum in a striped dolphin (Stenella coeruleoalba) and evidence of H. cetorum infection in cetaceans from European waters.
Age-related differences in susceptibility to infectious disease are known from a wide variety of plant and animal taxonomic groups. For example, the immature immune systems of young vertebrates, along with limited prior exposure to pathogens and behavioral factors, can place juveniles at greater risk of acquiring and succumbing to a pathogen. We studied the ontogenetic susceptibility of terrestrial direct-developing frogs (Eleutherodactylus coqui) to the fungal pathogen, Batrachochytrium dendrobatidis (Bd), which is responsible for the decline of amphibian species worldwide. By exposing juvenile and adult frogs to the same dose and strain of Bd, we uncovered ontogenetic differences in susceptibility. Froglets exposed to the pathogen had significantly lower survival rates compared with control froglets, while adult frogs largely cleared infection and had survival rates indistinguishable from control frogs, even when exposed to a much higher dose of Bd. The high disease-induced mortality rate of juveniles may explain ongoing population declines in eastern Puerto Rico, where Bd is endemic and juveniles experience higher prevalence and infection intensity compared to adults. Our results have important implications for understanding and modeling the decline, possibly to extinction, of amphibian populations and species.
During July–August 2010, 28 Christmas Island flying foxes (Pteropus melanotus natalis) were captured and anesthetized for examination, sample collection, and release to determine the potential role of disease in recent population declines. Measurements and samples were taken for morphologic, hematologic, biochemical, and parasitologic analysis. These are the first blood reference ranges reported for this species. These data are being used to inform investigations into conservation status and population management strategies for the Christmas Island flying fox.
We inventoried and assessed historical anthrax outbreak data from 1962–2008 in wild wood bison (Bison bison athabascae) in Wood Buffalo National Park and the Slave River Lowlands (SRL), Northwest Territories, Canada. We compared these results with a 2010 outbreak in the SRL. Anthrax outbreaks have occurred in 12 of the years between 1962 and 2008 in wild wood bison with 1,515 anthrax deaths detected. The average number of carcasses found each outbreak year was 126 (range 1–363), though local averages varied. The numbers of animals found dead per outbreak declined over the past four decades. Outbreaks varied in duration from 16–44 days (average length 25.5 days). The length of an outbreak was not a determinant of the number of dead bison found, but outbreaks starting in July had more deaths than those staring in June. Males were more likely to be detected in an outbreak, outbreaks were likely not random events, and there was no relationship between outbreak size or length and location. Future surveillance activities may benefit from targeting bulls and planning surveillance activities for more than 3 wk after outbreak detection. Coordinating data collecting and recording efforts between jurisdictions may overcome historical challenges in inconsistent record keeping.
Danielle B. Araujo, Luzia A. Martorelli, Ana Paula G. A. Kataoka, Angélica C. A. Campos, Camila S. Rodrigues, Luiz F. Sanfilippo, Elenice S. Cunha, Edison L. Durigon, Silvana R. Favoretto
Rabies causes thousands of human and animal deaths worldwide each year. The emergent importance of rabies in wild animals demonstrates the necessity of epidemiologic studies of infection in these species toward the development of better strategies for prevention and control of rabies. We analyzed the circulation of rabies virus among wildlife species from a native rainforest in São Paulo State, Brazil. We used the rapid fluorescent focus inhibition test (RFFIT) to test for rabies virus-neutralizing antibodies in 139 captured terrestrial mammals and the fluorescent antibody test (FAT), mouse inoculation test (MIT), and reverse-transcriptase (RT)-PCR to test for virus in samples from the central nervous system of 53 animals found dead. The percentage of samples positive by RFFIT was 10.8%. All samples tested by FAT, MIT, and RT-PCR were negative. Research should be continued to obtain a better understanding of the role of wildlife in the circulation and transmission of rabies virus.
Water deer (Hydropotes inermis) are among the most common wildlife to approach farmhouses and livestock barns in Korea. We collected 305 water deer from Gangwon (n = 168), South Chungcheong (n = 89), and Gyeongsang (n = 48) provinces in 2010–12 and used PCR and serologic tests to screen the deer for pathogens. In 2010, tests for bovine viral diarrhea virus (BVDV), rotavirus, and Brucella abortus were positive in 8% (5/60), 2% (1/60), and 59% (33/56) of the animals, respectively. In 2010, the water deer were negative for foot-and-mouth disease virus, coronaviruses, and Mycobacterium bovis. All samples collected in 2011 and 2012 were negative for all pathogens analyzed. These results suggest that at least two of the investigated pathogens, BVDV and B. abortus, circulate among water deer in South Korea.
An outbreak of salmonellosis in wild passerines caused mass mortality of Eurasian Tree Sparrows (Passer montanus) in Hokkaido, Japan, 2005–06; however, the etiology was poorly understood. In winter 2008–09, sparrow mortality again occurred in Hokkaido, and 202 deaths in 100 incidents at 94 sites were reported. We conducted a comprehensive investigation to evaluate the cause and impact on sparrow populations. We collected 26 carcasses at 13 sites, including a zoological park. In addition, Salmonella screening of zoo animals was conducted as a biosecurity measure. Salmonella Typhimurium was isolated from multiple organs in all examined sparrows; they were diagnosed with septicemic salmonellosis. Eleven sites (85%) were related to wild bird feeding and six of eight sparrow fecal samples, including from the zoo, were S. Typhimurium-positive. No infection was detected in zoo animals. Isolates belonged to three phage types: DT40 (88%), DT110 (8%), and DT120 (4%). Pulsed-field gel electrophoresis patterns were the same in all isolates, regardless of phage type. Biochemical characteristics and antibiotic-resistance profiles of DT40 were similar in all isolates, indicating a single origin. The mortality was likely associated with that in 2005–06 because the isolates had the same profiles. Tissue levels of sodium, calcium, and magnesium (the main components of chemical deicer suspected to be the major cause of poisoning deaths in 2005–06 mortality) were not higher in the affected sparrows. We conclude that an emerging epidemic infection with S. Typhimurium DT40 related to bird feeding was the cause of sparrow mortality in 2008–09 and suggest that this causative strain is host-adapted to sparrows in Japan. The mortality might have had some impact on the local population, but its influence was limited.
Capybaras (Hydrochoerus hydrochaeris) are the world's largest rodents and play an epidemiologic role in the transmission of zoonotic pathogens, including the causative agents of Brazilian spotted fever, leptospirosis, and others. We surveyed the health of 31 free-ranging capybaras at the Alberto Löfgren State Park, São Paulo, Brazil using a variety of diagnostic methods. Hematology and serum chemistry were consistent with mild malnutrition and parasitism but did not indicate severe physiologic imbalance or disease. All animals were serologically negative for Rickettsia rickettsii, Leishmania spp., and Trypanosoma sp., but antibodies against rabies virus (71%), Leptospira sp. (26%), and Toxoplasma sp. (23%) were detected. Salmonella sp. was not cultured from fecal samples. Frequently cultured enterobacteria included Escherichia coli (61%), Enterococcus casseiflavus (35%), Enterococcus faecalis (35%), Enterobacter aerogenes (32%), Klebisella pneumoniae (32%), and Serratia marcescens (32%). No potentially pathogenic fungi were cultured from hair samples. Fecal parasitology revealed infection by Protozoophaga sp. (58%), Viannella spp. (23%), Strongyloides spp. (10%), and Ancilostomatidae (10%). A total of 218 ticks was retrieved from the animals: Amblyomma sp. larvae and nymphs (43%), A. dubitatum adults (52%), and A. cajennense adults (5%). The capybaras were free from most potentially zoonotic pathogens evaluated; however, the presence of Amblyomma spp. ticks (potential vectors of Rickettsia spp.) and indirect evidence of exposure to the rabies virus, Leptospira sp., and Toxoplasma sp. warrant the maintenance of public health programs and wildlife health monitoring.
Factors that alter the dynamics of ecologic systems can influence transmission of infectious diseases and may lead to decreases in natural populations. Leptospirosis is a cosmopolitan disease of zoonotic importance that affects most mammals. At the southern Gulf of Mexico, Antillean manatees (Trichechus manatus manatus) inhabit highly variable environments, with extended floods during the rainy season and drought conditions during the dry season that affect food availability and the thermal environment for manatees. We tested for changes in prevalence and titers of antibodies to 12 serovars of Leptospira interrogans in manatees between dry and rainy seasons. We determined titers for L. interrogans through microscopic agglutination tests (MAT) from 10 manatees, six during the dry season (DS), and six during the rainy season (RS) in Laguna de las Ilusiones, a landlocked lake hosting a population of about 20 manatees. All individuals were antibody positive (titers ≥100) to at least one serovar. The serovars bataviae, bratislava, canicola, and icterohaemorrhagiae had overall prevalences ≥50%; bataviae, bratislava, and canicola had prevalences ≥50% during both seasons. Serovars icterohaemorrhagiae and pyrogenes had prevalences ≥50% during DS and pomona, tarassovi, wolfii, and autumnalis during RS. Significant differences in prevalence between seasons were found for pomona, tarassovi, and autumnalis. Titers of tarassovi, wolfii, autumnalis, and bataviae were significantly higher during RS. There was a high prevalence of L. interrogans during the RS independent of high availability of plant foods, coinciding with the epizootiology of the bacteria that are endemic to tropical regions. Another factor possibly influencing prevalence is high anthropogenic pressure at the lake, causing an increase in potential sources of infection. Because of possible cross-reaction in MAT, further research is needed on the molecular discrimination of serovars in animals in the lake.
We surveyed free-ranging Canada Geese (Branta canadensis), Trumpeter Swans (Cygnus buccinator), Mute Swans (Cygnus olor), and Mallards (Anas platyrhynchos) to estimate the prevalence of antibodies to avian bornavirus (ABV) and of cloacal shedding of ABV RNA in southern Ontario, Canada. Blood samples and cloacal swabs were collected from 206 free-ranging Canada Geese, 135 Trumpeter Swans, 75 Mute Swans, and 208 Mallards at 10 main capture sites between October 2010 and May 2012. Sera were assessed for antibodies against ABV by enzyme-linked immunosorbent assay and swabs were evaluated for ABV RNA using real-time reverse-transcription PCR. Serum antibodies were detected in birds from all four species and at each sampling site. Thirteen percent of the geese caught on the Toronto Zoo site shed ABV RNA in feces compared with 0% in geese sampled at three other locations. The proportions of shedders among Mute Swans, Trumpeter Swans, and Mallards were 9%, 0%, and 0%, respectively. Birds that were shedding viral RNA were more likely to have antibodies against ABV and to have higher antibody levels than those that were not, although many birds with antibodies were not shedding. We confirmed that exposure to, or infection with, ABV is widespread in asymptomatic free-ranging waterfowl in Canada; however, the correlation between cloacal shedding, presence of antibodies, and presence of disease is not fully understood.
Monkeypox (MPX) is a re-emerging zoonotic disease that is endemic in Central and West Africa, where it can cause a smallpox-like disease in humans. Despite many epidemiologic and field investigations of MPX, no definitive reservoir species has been identified. Using recombinant viruses expressing the firefly luciferase (luc) gene, we previously demonstrated the suitability of in vivo bioluminescent imaging (BLI) to study the pathogenesis of MPX in animal models. Here, we evaluated BLI as a novel approach for tracking MPX virus infection in black-tailed prairie dogs (Cynomys ludovicianus). Prairie dogs were affected during a multistate outbreak of MPX in the US in 2003 and have since been used as an animal model of this disease. Our BLI results were compared with PCR and virus isolation from tissues collected postmortem. Virus was easily detected and quantified in skin and superficial tissues by BLI before and during clinical phases, as well as in subclinical secondary cases, but was not reliably detected in deep tissues such as the lung. Although there are limitations to viral detection in larger wild rodent species, BLI can enhance the use of prairie dogs as an animal model of MPX and can be used for the study of infection, disease progression, and transmission in potential wild rodent reservoirs.
Grey seals (Halichoerus gryphus), the main final host of the gastric parasitic nematode Contracaecum osculatum in the Baltic, have recently recolonized the southwestern Baltic Sea. This colonization could lead to an increase in prevalence and intensity of third-stage larvae of C. osculatum in livers of Baltic cod (Gadus morhua), which serve as transport host for this helminth. We performed a parasitologic study of cod in spring 2012 and compared the results with previously unpublished data from 1982/1983. Additionally, grey seals were counted annually from 2000 to 2011 at three haul-out sites in the southwestern Baltic. Of 97 cod livers examined in the 1982/1983 survey, 22% harbored C. osculatum larvae, whereas 55.1% of the examined cod livers (n = 185) were infected in 2012; the mean intensity and mean abundance increased from 4.3 and 0.9 to 20.2 and 11.1, respectively. Molecular identification (PCR) confirmed the identity of the larvae. The grey seal population increased markedly during the 12-yr period. We suggest that the elevated parasitism of cod livers is associated with the successful re-establishment of grey seals in the southwestern Baltic.
Winter ticks (Dermacentor albipictus) on elk (Cervus elaphus canadensis) have recently increased in numbers in the Yukon, Canada, potentially posing risks to other indigenous host species in the region. To evaluate the regional source of winter ticks in the Yukon, we sequenced one nuclear (ITS-2) and two mitochondrial (16SrRNA and COI) genes, and genotyped 14 microsatellite loci from 483 winter tick specimens collected across North America. We analyzed genetic variation across the geographic and host ranges of this tick species with the use of variance partitioning, Bayesian clustering, and standard population genetic analyses. Based on our results, winter ticks on elk in the Yukon could have originated either by translocation from central Alberta or by northward range expansion of more geographically proximate populations in northern Alberta and British Columbia. Although there was some genetic structuring of winter ticks on different hosts in the same region, we found little evidence of host specificity in winter ticks from five ungulate host species, suggesting that the winter ticks on elk in the Yukon could potentially become established on other locally available host species such as moose (Alces alces).
We captured 36 Northern Bobwhites (Colinus virginianus) in Mitchell County, Texas in June–September 2013, and examined them for the eyeworm Oxyspirura petrowi. We recovered 334 eyeworms from 28 of 29 adult bobwhites (97%); infections ranged from 1–40 worms and mean (±SD) abundance of 11.9±13.0. Three of seven juveniles were infected, and those infected had one eyeworm each. Prevalence of eyeworms was similar among months. However, mean abundance of eyeworms peaked in July and August (3.3±2.1, 13.5±15.0, and 16.9±15.5), and decreased in September (6.3±3.0). We suggest that several previous studies may underreport prevalence and abundance because in those studies only the eye surface and nictitating membrane were examined, and not eye-associated tissue, ducts, glands, or sinuses.
To investigate pseudorabies-virus (PrV) –antibody and viral-DNA prevalence, we collected blood, nasal and genital swabs, and tonsillar and lymph-node tissue samples from 139 wild boars (Sus scrofa; 39 piglets, 30 juveniles, and 70 adults), during the hunting season of 2010–2011 in Tuscany, Central Italy. We performed immunohistochemistry with anti-PrV monoclonal antibodies on selected tissue samples. Forty-three of 139 (30.9%) boars were PrV-antibody positive and a 1,954–base-pair PrV-specific product was amplified from nine nasal (6.5%) and 26 genital (18.7%) swabs. Sequence analysis of PrV-positive PCR products revealed identity scores of 99–100% with Suid herpesvirus 1 strain Becker (JF797219) and confirmed the identification of PrV DNA in tested swabs. There was significantly higher antibody prevalence in adults than in juveniles and in piglets than in juveniles. The prevalence of viral DNA was significantly higher in genital swabs than in nasal specimens. The percentage of positive nasal swabs did not differ among age classes. Piglets had a higher percentage of PCR-positive genital swabs than juvenile and adult subjects (30.8% vs. 13.3% and 14.3%, respectively). Results confirmed that PrV infection is widespread in the wild boar population in the study area. The presence of anti-PrV antibodies and of the PrV virus in piglets could be related to vertical transmission of the virus. This hypothesis was also supported by a higher presence of viral genome in genital swabs than in nasal swabs. This field study supports the importance of vertical transmission of PrV, and the high prevalence of virus in genital swabs supports venereal transmission in adult feral boars.
Gregory G. Turner, Carol Uphoff Meteyer, Hazel Barton, John F. Gumbs, DeeAnn M. Reeder, Barrie Overton, Hana Bandouchova, Tomáš Bartonička, Natália Martínková, Jiri Pikula, Jan Zukal, David S. Blehert
Definitive diagnosis of the bat disease white-nose syndrome (WNS) requires histologic analysis to identify the cutaneous erosions caused by the fungal pathogen Pseudogymnoascus [formerly Geomyces] destructans (Pd). Gross visual inspection does not distinguish bats with or without WNS, and no nonlethal, on-site, preliminary screening methods are available for WNS in bats. We demonstrate that long-wave ultraviolet (UV) light (wavelength 366–385 nm) elicits a distinct orange–yellow fluorescence in bat-wing membranes (skin) that corresponds directly with the fungal cupping erosions in histologic sections of skin that are the current gold standard for diagnosis of WNS. Between March 2009 and April 2012, wing membranes from 168 North American bat carcasses submitted to the US Geological Survey National Wildlife Health Center were examined with the use of both UV light and histology. Comparison of these techniques showed that 98.8% of the bats with foci of orange–yellow wing fluorescence (n = 80) were WNS-positive based on histologic diagnosis; bat wings that did not fluoresce under UV light (n = 88) were all histologically negative for WNS lesions. Punch biopsy samples as small as 3 mm taken from areas of wing with UV fluorescence were effective for identifying lesions diagnostic for WNS by histopathology. In a nonlethal biopsy-based study of 62 bats sampled (4-mm diameter) in hibernacula of the Czech Republic during 2012, 95.5% of fluorescent (n = 22) and 100% of nonfluorescent (n = 40) wing samples were confirmed by histopathology to be WNS positive and negative, respectively. This evidence supports use of long-wave UV light as a nonlethal and field-applicable method to screen bats for lesions indicative of WNS. Further, UV fluorescence can be used to guide targeted, nonlethal biopsy sampling for follow-up molecular testing, fungal culture analysis, and histologic confirmation of WNS.
Hypoxemia is anticipated during wildlife anesthesia and thus should be prevented. We evaluated the efficacy of low flow rates of supplemental oxygen for improvement of arterial oxygenation in anesthetized brown bears (Ursus arctos). The study included 32 free-ranging brown bears (yearlings, subadults, and adults; body mass 12–250 kg) that were darted with medetomidine-zolazepam-tiletamine (MZT) from a helicopter in Sweden. During anesthesia, oxygen was administered intranasally from portable oxygen cylinders at different flow rates (0.5–3 L/min). Arterial blood samples were collected before (pre-O2), during, and after oxygen therapy and immediately processed with a portable analyzer. Rectal temperature, respiratory rate, heart rate, and pulse oximetry-derived hemoglobin oxygen saturation were recorded. Intranasal oxygen supplementation at the evaluated flow rates significantly increased the partial pressure of arterial oxygen (PaO2) from pre-O2 values of 9.1±1.3 (6.3–10.9) kPa to 20.4±6.8 (11.1–38.7) kPa during oxygen therapy. When oxygen therapy was discontinued, the PaO2 decreased to values not significantly different from the pre-O2 values. In relation to the body mass of the bears, the following oxygen flow rates are recommended: 0.5 L/min to bears <51 kg, 1 L/min to bears 51–100 kg, 2 L/min to bears 101–200 kg, and 3 L/min to bears 201–250 kg. In conclusion, low flow rates of intranasal oxygen were sufficient to improve arterial oxygenation in brown bears anesthetized with MZT. Because hypoxemia quickly recurred when oxygen was discontinued, oxygen supplementation should be provided continuously throughout anesthesia.
Dennis Slate, Richard B. Chipman, Timothy P. Algeo, Samuel A. Mills, Kathleen M. Nelson, Christopher K. Croson, Edward J. Dubovi, Kurt Vercauteren, Randall W. Renshaw, Todd Atwood, Shylo Johnson, Charles E. Rupprecht
In 2011, we conducted a field trial in rural West Virginia, USA to evaluate the safety and immunogenicity of a live, recombinant human adenovirus (AdRG1.3) rabies virus glycoprotein vaccine (Ontario Rabies Vaccine Bait; ONRAB) in wild raccoons (Procyon lotor) and striped skunks (Mephitis mephitis). We selected ONRAB for evaluation because of its effectiveness in raccoon rabies management in Ontario and Quebec, Canada, and significantly higher antibody prevalence rates in raccoons compared with a recombinant vaccinia-rabies glycoprotein (V-RG) vaccine, Raboral V-RG®, in US–Canada border studies. Raccoon rabies was enzootic and oral rabies vaccination (ORV) had never been used in the study area. We distributed 79,027 ONRAB baits at 75 baits/km2 mostly by fixed-wing aircraft along parallel flight lines at 750-m intervals. Antibody prevalence was significantly higher at 49.2% (n = 262) in raccoons after ONRAB was distributed than the 9.6% (n = 395) before ORV. This was the highest antibody prevalence observed in raccoons by US Department of Agriculture Wildlife Services for areas with similar management histories evaluated before and after an initial ORV campaign at 75 baits/km2 with Raboral V-RG. Tetracycline biomarker (TTCC) was significantly higher among antibody-positive raccoons after ONRAB baiting and was similar among raccoons before ORV had been conducted, an indication of vaccine-induced rabies virus–neutralizing antibody production following consumption of bait containing TTCC. Skunk sample size was inadequate to assess ONRAB effects. Safety and immunogenicity results supported replication of this field trial and led to a recommendation for expanded field trials in 2012 to evaluate safety and immunogenicity of ground-distributed ONRAB at 150 baits/km2 in residential and commercial habitats in Ohio, USA and aerially distributed ONRAB at 75 baits/km2 in rural habitats along US–Quebec border.
The signaling lymphocyte activation molecule (SLAM) is a receptor for morbilliviruses. To understand the recent host range expansion of canine distemper virus (CDV) in carnivores, we determined the nucleotide sequences of SLAMs of various carnivores and generated three-dimensional homology SLAM models. Thirty-four amino acid residues were found for the candidates binding to CDV on the interface of the carnivore SLAMs. SLAM of the domestic dog (Canis lupus familiaris) were similar to those of other members of the suborder Caniformia, indicating that the animals in this group have similar sensitivity to dog CDV. However, they were different at nine positions from those of felids. Among the nine residues, four of domestic cat (Felis catus) SLAM (72, 76, 82, and 129) and three of lion (Panthera leo persica) SLAM (72, 82, and 129) were associated with charge alterations, suggesting that the felid interfaces have lower affinities to dog CDV. Only the residue at 76 was different between domestic cat and lion SLAM interfaces. The domestic cat SLAM had threonine at 76, whereas the lion SLAM had arginine, a positively charged residue like that of the dog SLAM. The cat SLAM with threonine is likely to have lower affinity to CDV-H and to confer higher resistance against dog CDV. Thus, the four residues (72, 76, 82, and 129) on carnivore SLAMs are important for the determination of affinity and sensitivity with CDV. Additionally, the CDV-H protein of felid strains had a substitution of histidine for tyrosine at 549 of dog CDV-H and may have higher affinity to lion SLAM. Three-dimensional model construction is a new risk assessment method of morbillivirus infectivity. Because the method is applicable to animals that have no information about virus infection, it is especially useful for morbillivirus risk assessment and wildlife conservation.
Pamela A. Tuomi, Michael J. Murray, Michael M. Garner, Caroline E. C. Goertz, Robert W. Nordhausen, Kathleen A. Burek-Huntington, David M. Getzy, Ole Nielsen, Linda L. Archer, Heather T. D. Maness, James F. X. Wellehan, Thomas B. Waltzek
Small superficially ulcerated skin lesions were observed between October 2009 and September 2011 during captive care of two orphaned sea otter pups: one northern (Enhydra lutris kenyoni) in Alaska and one southern (Enhydra lutris nereis) in California. Inclusions consistent with poxviral infection were diagnosed by histopathology in both cases. Virions consistent with poxvirus virions were seen on electron microscopy in the northern sea otter, and the virus was successfully propagated in cell culture. DNA extraction, pan-chordopoxviral PCR amplification, and sequencing of the DNA-dependent DNA polymerase gene revealed that both cases were caused by a novel AT-rich poxvirus. Bayesian and maximum likelihood phylogenetic analyses found that the virus is divergent from other known poxviruses at a level consistent with a novel genus. These cases were self-limiting and did not appear to be associated with systemic illness. To our knowledge, this is the first report of a poxvirus in a mustelid species. The source of this virus, mode of transmission, zoonotic potential, and biological significance are undetermined.
Monitoring circulating pathogens in wildlife populations is important in evaluating causes and sources of disease as well as understanding transmission between wild and domestic animals. In spring 2010, a sudden die-off in a chamois (Rupicapra rupicapra) population sharing habitat with livestock occurred in northeastern Austria. Nineteen animals were submitted for examination. Necropsy and pathohistologic and bacteriologic results yielded lesions associated with Pasteurellaceae species. Additional testing included enterobacterial repetitive intergenic consensus and random amplification of polymorphic DNA PCR analysis to evaluate the circulating strains. The isolated strains were most closely related to Mannheimia glucosida and Bibersteinia trehalosi. Reports of mass mortalities in chamois due to pneumonia have been reported previously in the northern Alpine area of Italy. To the authors' knowledge, this is the first report of acute mortality due to strains of Mannheimia and Bibersteinia in Austrian chamois.
During rehabilitation, acute renal failure due to leptospirosis occurred in eight male northern elephant seals (Mirounga angustirostris) that stranded along the central California coast in 2011. Characteristic histologic lesions including renal tubular degeneration, necrosis, and mineralization, and mild lymphoplasmacytic interstitial nephritis were noted in the six animals examined. Immunohistochemistry, bacterial culture, and PCR were positive in 2/3, 2/3, and 3/4 seals, respectively, and 6/8 had high serum antibody titers to Leptospira interrogans serovar pomona. Pulsed-field gel electrophoresis confirmed one isolate as serovar pomona. Variable number tandem repeat (VNTR) analysis showed both elephant seal isolates were identical to each other but distinct from those isolated from California sea lions (Zalophus californianus). The time from stranding to onset of azotemia was 1 to 38 (median = 24) days, suggesting some seals were infected at the rehabilitation facility. Based on temporal and spatial incidence of infection, transmission among elephant seals likely occurred during rehabilitation. Molecular (VNTR) analysis of the two isolates indicates there is a unique L. interrogans serovar pomona genotype in elephant seals, and sea lions were not the source of infection prior to or during rehabilitation. This study confirms the susceptibility of northern elephant seals to leptospirosis, indicates intraspecies transmission during rehabilitation, and reports the first isolation and preliminary characterization of leptospires from elephant seals.
Capillaria hepatica is a parasitic nematode that infects the liver of rats (Rattus spp.), and occasionally other mammalian species, including humans. Despite its broad geographic distribution and host range, the ecology of this parasite remains poorly understood. We characterized the ecology of C. hepatica in urban Norway rats (Rattus norvegicus) in Vancouver, Canada. The overall prevalence of C. hepatica among Norway rats was 36% (241/671); however, there was significant variation in prevalence among city blocks. Using a generalized linear mixed model to control for clustering by block (where OR is odds ratio and CI is confidence interval), we found C. hepatica infection was negatively associated with season (spring [OR = 0.14, 95% CI = 0.05–0.39]; summer [OR = 0.14, 95% CI = 0.03–0.61]; winter [OR = 0.34, 95% CI = 0.13–0.84], compared to fall) and positively associated with sexual maturity (OR: 7.29, 95% CI = 3.98–13.36) and presence of cutaneous bite wounds (OR = 1.87, 95% CI = 1.11–3.16). Our understanding of the ecology of C. hepatica in rats is hindered by a paucity of data regarding the main mechanisms of transmission (e.g., environmental exposure vs. active cannibalism). However, associations among infection, season, maturity, and bite wounds could suggest that social interactions, possibly including cannibalism, may be important in transmission.
In May 2012, an adult, male bottlenose dolphin (Tursiops truncatus) was found stranded and dead on the Spanish Mediterranean coast. At necropsy, several areas of malacia were macroscopically observed in the periventricular parenchyma of the cerebrum. Microscopically a severe, diffuse, pyogranulomatous, and necrotizing meningoencephalomyelitis was associated with numerous intralesional highly pleomorphic fungal structures. After culture, the fungus, Cunninghamella bertholletiae, was identified by culture and PCR. To our knowledge, this is the first reported case of central nervous system mucormycosis due to Cunninghamella bertholletiae in a cetacean.
Epizootic hemorrhagic disease virus (EHDV) causes a highly infectious noncontagious hemorrhagic disease in wild and captive deer (Cervidae) populations in the US. Although rapid and accurate identification of the disease is important, identification of the serotype is equally important for understanding the epidemiology of the disease in white-tailed deer (Odocoileus virginianus) populations. We developed a one-step multiplex reverse transcriptase PCR assay for rapid differentiation and identification of EHDV serotypes 1, 2, and 6 in cell culture and clinical samples by targeting the viral gene segment 2 (L2) that encodes for the structural protein VP2. From 2009 to 2012, 427 clinical samples including tissue and blood (in ethylenediaminetetraacetic acid) from white-tailed deer, found EHDV positive by real-time PCR, were used to evaluate this subtyping assay. Eighteen percent of the positive samples tested were EHDV-1, 59% were EHDV-2, and 21% were EHDV-6; 2% of the samples were positive for more than one subtype, indicating mixed infection. This assay provides a rapid, sensitive, specific diagnostic tool for differentiation and identification of EHDV serotypes in field samples and virus isolates.
To assess implications for public health we compared the resistance of Enterococcus spp. strains to antibacterial drugs in wild and exotic animals with strains originating in domesticated animals and characterized correlations between Enterococcus species, the source of the isolate, and the degree of resistance to selected antibiotics. All strains, regardless of source, were susceptible to β-lactams, gentamicin, linezolid, and teicoplanin; the highest resistance was to kanamycin, quinupristin, and rifampicin. Thirteen strains from undomesticated animals were resistant to vancomycin, and one strain, from a fox, was resistant to streptomycin (high-dose). Multidrug-resistant strains accounted for 46% of the strains from wild animals and 59% of the strains from an exotic animal (the Russian tortoise; Testudo horsfieldii). Despite the relatively low level of resistance in the strains isolated from wild and exotic animals, the large number of intermediately susceptible strains in these groups is an indication of the evolutionary character of the development of resistance, suggesting that these animals may be potential reservoirs of Enterococcus strains resistant to a wide panel of currently used antibiotics.
Bats have been implicated as potential carriers of Leptospira as a result of surveys, mostly in Australia and South America. We measured the prevalence of pathogenic leptospires in kidneys of bats from Kansas and Nebraska. From 7 August 2012 to 21 August 2012, we extracted DNA from kidneys of 98 big brown bats (Eptesicus fuscus) submitted and found negative for rabies. The DNA was processed in a two-step, seminested PCR assay with a dual-labeled Taqman probe specific for pathogenic leptospires. As a negative control, we used a saprophytic leptospire (Leptospira biflexa Patoc) and, as a pathogenic control, Leptospira interrogans Canicola. All bat kidneys were negative for pathogenic leptospires, suggesting that it is unlikely that the big brown bat, one of the most prevalent bat species in North America, is a reservoir for transmission of leptospires to dogs or humans.
A wide variety of Salmonella serotypes occurs within reptilian hosts, but their ecology is poorly understood. We collected cloacal swabs from tuatara (Sphenodon punctatus), fairy prions (Pachyptila turtur), and skinks (Oligosoma spp.) on Stephens Island, New Zealand, to screen for Salmonella. Soil samples were also collected from inside burrows of tuatara and fairy prions and tested for Salmonella. We sampled repeatedly from October 2009 to October 2011. Cloacal swabs were collected from 620 tuatara, and no intestinal shedding of Salmonella was detected. Similarly, no Salmonella was detected in fairy prions. In contrast, we isolated Salmonella from 6.5% of skinks and 8.4% of soil samples. We identified two serovars of Salmonella from 52 isolates, Salmonella Saintpaul and Salmonella Mississippi. Salmonella Mississippi was isolated from skinks only and S. Saintpaul was found in skinks and soil samples. Salmonella persists in this ecosystem with skinks as the main wildlife reservoir, and an environmental reservoir exists in the soil from burrows used by skinks, tuatara, and fairy prions. Salmonella was absent from skinks and the soil in winter, raising the question of bacteria persisting through winter.
We compared mule deer (Odocoileus hemionus) of two different PRNP genotypes (225SS, 225FF) for susceptibility to chronic wasting disease (CWD) in the face of environmental exposure to infectivity. All three 225SS deer had immunohistochemistry (IHC)-positive tonsil biopsies by 710 days postexposure (dpe), developed classic clinical signs by 723–1,200 dpe, and showed gross and microscopic pathology, enzyme-linked immunosorbent assay (ELISA) results, and IHC staining typical of prion disease in mule deer. In contrast, although all three 225FF deer also became infected, the two individuals surviving >720 dpe had consistently negative biopsies, developed more-subtle clinical signs of CWD, and died 924 or 1,783 dpe. The 225FF deer were “suspect” by ELISA postmortem but showed negative or equivocal IHC staining of lymphoid tissues; both clinically affected 225FF deer had spongiform encephalopathy in the absence of IHC staining in the brain tissue. The experimental cases resembled three cases encountered among five additional captive 225FF deer that were not part of our experiment but also died from CWD. Aside from differences in clinical disease presentation and detection, 225FF mule deer also showed other, more-subtle, atypical traits that may help to explain the rarity of this genotype in natural populations, even in the presence of enzootic CWD.
To determine the prevalence and diversity of Leptospira serogroups circulating in wildlife on farms in Ontario, we tested samples from 51 raccoons (Procyon lotor), seven skunks (Mephitis mephitis), four rats (Rattus norvegicus), and three opossums (Didelphis virginiana) that were trapped on 27 livestock (swine [Sus scrofa], cattle [Bos taurus]) farms in 2010. Seventeen of 51 raccoons (33%; 95% confidence interval [CI], 21–48%) sampled were positive for at least one Leptospira serogroup using the microscopic agglutination test. None of the other 14 animals had detectable Leptospira antibodies. On swine farms, 13 of 30 raccoons (43%; 95% CI, 27–61%) were antibody positive, and on cattle farms, four of 21 raccoons (19%; 95% CI, 8–40%) were positive. Leptospira antibody prevalence in raccoons did not differ between swine and cattle farms. Raccoons were positive to serovars representative of serogroups Grippotyphosa, Australis, Icterohaemorrhagiae, and Pomona and were negative to serovars of serogroups Autumnalis, Canicola, and Sejroe. The prevalence of Leptospira antibodies in raccoons in this study is similar to what has been reported previously; however, the diversity of serogroups was higher in this study than what has been reported in raccoons from an urban area of Ontario, Canada. Understanding the prevalence and distribution of Leptospira serogroups in wildlife in Ontario, Canada, is important for the development and maintenance of appropriate disease management strategies in humans, livestock, and companion animals.
Prevalence of avian influenza virus (AIV) antibodies in the western Atlantic subspecies of Red Knot (Calidris canutus rufa) is among the highest for any shorebird. To assess whether the frequency of detection of AIV antibodies is high for the species in general or restricted only to C. c. rufa, we sampled the northeastern Pacific Coast subspecies of Red Knot (Calidris canutus roselaari) breeding in northwestern Alaska. Antibodies were detected in 90% of adults and none of the chicks sampled. Viral shedding was not detected in adults or chicks. These results suggest a predisposition of Red Knots to AIV infection. High antibody titers to subtypes H3 and H4 were detected, whereas low to intermediate antibody levels were found for subtypes H10 and H11. These four subtypes have previously been detected in shorebirds at Delaware Bay (at the border of New Jersey and Delaware) and in waterfowl along the Pacific Coast of North America.
We compared dosages of a combination of sedatives, which included butorphanol tartrate, azaperone tartrate, and medetomidine HCl (BAM) in captive adult Rocky Mountain elk (Cervus elaphus nelsoni). All three BAM dosages (low, medium, and high) effectively immobilized elk and produced an adequate level of sedation in all subjects. Induction times were similar among the three groups (mean±SD: low = 6.9±1.1 min; medium = 6.3±0.9 min; high = 4.7±1.3 min). Most elk became hypoxemic regardless of BAM dosage, but hypoxemia tended to be most severe in the high-BAM group; regardless of BAM dosage, oxygen supplementation improved the percentage of oxygen saturation and stabilized the vital rates. Recovery after administration of antagonists (3 mg atipamezole/mg medetomidine and 2 mg/kg tolazoline) was comparable among groups (range of means = 9±1.5–11.7±1 min). Based on the findings from clinical trials and field data from free-ranging elk immobilizations, we recommend low-dose BAM (2 mL dose; equivalent to 46 mg butorphanol, 30 mg azaperone, and 18 mg medetomidine) and supplemental oxygen for adult elk; immobilization should be antagonized using 3–5 mg atipamezole/mg medetomidine and 2 mg/kg tolazoline, with tolazoline injected about 5–10 min before atipamezole to smooth out recovery.
Recently, 11 cases of animal rabies were reported in the southern region (Suwon and Hwaseong cities) of Gyeonggi Province, South Korea. The cases were temporally separated into two cases in dogs (Canis lupus familiaris) in spring 2012 and nine cases in domestic animals and wildlife in winter 2012–13. All carcasses were submitted for histopathologic examination and viral antigen identification. Sequences of the glycoprotein, nucleoprotein, and glycoprotein–large polymerase protein intergenic noncoding loci of the 11 strains were determined and compared with published reference sequences. All rabies strains were closely related to the Gangwon strains isolated in 2008–09, suggesting that the rabies virus strains isolated in Gyeonggi were introduced from Gangwon Province.
Aleutian mink disease virus (AMDV) occurs in the American mink (Neovison vison) in wild populations and on mink farms and can cause illness and death. The North American river otter (Lontra canadensis) may be exposed to AMDV because of shared space and habitat with mink. Using serology and real-time PCR, we tested river otters across Ontario for AMDV infection. We found no evidence of infection in otters, a surprising finding given the sympatric distribution, niche overlap, and close phylogenetic relationship of the river otter and the American mink. Our results are consistent with the hypothesis that the major point of spillover of AMDV between mink farms and wildlife is manure and composting carcasses on mink farms. Mink farms in Ontario are generally in agricultural landscapes; it is unlikely that river otter use these habitats and thus are likely not exposed to AMDV. We found no evidence that AMD is an important disease for the river otters in Ontario.
On 16 March 2012 a rabid eastern red bat (Lasiurus borealis) was found attached to an evening bat (Nycticeius humeralis) in Randolph County, Arkansas, USA. This appears to be the first confirmed case of a rabid bat attacking a bat of another species.
Serum from Mexican grey squirrels (Sciurus aureogaster) from Mexico City reacted to Orthopoxvirus by enzyme-linked immunosorbent assay. Real-time PCR based on oral swabs and scabs did not detect viral DNA. Antibody prevalence was 30% (n = 366), providing the first evidence of Orthopoxvirus antibodies in Mexican wild rodents.
In 2010, a black-tailed prairie dog (Cynomys ludovicianus) was found dead in Grasslands National Park, Saskatchewan, Canada. Postmortem gross and histologic findings indicated bacterial septicemia, likely due to Yersinia pestis, which was confirmed by molecular analysis. This is the first report of Y. pestis in the prairie dog population within Canada.
We describe mycobacteriosis caused by Mycobacterium peregrinum in Red-crowned Cranes (Grus japonensis) in China. Isolates were identified by bacteriology, molecular identification methods, and phylogenetic analysis. This study shows that M. peregrinum is an important pathogen for mycobacteriosis and could represent a threat to conservation efforts of endangered species.
We describe Aujeszky's disease in a female of red fox (Vulpes vulpes). Although wild boar (Sus scrofa) would be the expected source of infection, phylogenetic analysis suggested a domestic rather than a wild source of virus, underscoring the importance of biosecurity measures in pig farms to prevent contact with wild animals.
Three Northern Mockingbirds (Mimus polyglottos) and one Curve-billed Thrasher (Toxostoma curvirostre) from the Rolling Plains of Texas, USA were sampled for eyeworms in September 2013. All four birds were infected with the eyeworm Oxyspirura petrowi.
Brain samples from 182 red foxes (Vulpes vulpes) from Romania were examined using a standard PCR technique. Results provide evidence of Toxoplasma gondii (11 foxes = 6.0%) and Neospora caninum (1 fox = 0.5%) DNA in red foxes from Romania. No coinfections were found.
Sera from 659 Black-headed Gulls (Chroicocephalus ridibundus) in Dianchi Lake, China were assayed for Toxoplasma gondii antibodies using the modified agglutination test (MAT). Specific T. gondii antibodies were detected in 131 (19.9%) Black-headed Gulls (MAT titer≥1∶5). These results indicate that T. gondii infection is common in Black-headed Gulls.
Epizootic hemorrhagic disease (EHD) virus serotype 2 was identified by reverse-transcription (RT)-PCR in a white-tailed deer (Odocoileus virginianus) found dead in southern Alberta in September 2013. Field observations indicate at least 50 deer, primarily white-tailed deer, and three pronghorn antelope (Antilocapra americana) died during a suspected localized EHD outbreak.
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