In the Ngorongoro Conservation Area (NCA), Tanzania, where wildlife and livestock interaction is intense, greater potential for intra- and interspecies disease transmission is expected. We assessed the prevalence of bovine tuberculosis in African buffalo (Syncerus caffer) residing on the valley floor of the crater in the NCA. Apparently healthy animals were randomly selected from herds in nine sites of the Ngorongoro Crater. Syncerus caffer buffalo herds were located using very high-frequency radio-aided rangers positioned in various observation points around the crater in the NCA. A total of 102 African buffalo from 16 herds were immobilized from the ground using a cocktail of 4–10 mg etorphine hydrochloride (M99) and 60–150 mg azaperone tartrate. The M99 was reversed using 10–25 mg diprenorphine hydrochloride depending on age of animals. An interferon gamma assay was performed on harvested plasma samples using sandwich enzyme linked immunosorbent assay. Of the 102 animals sampled, two (2%) African buffalo tested positive for bovine tuberculosis. These results corroborate those of the skin test done recently in cattle in the NCA. The presence of bovine tuberculosis in livestock and wildlife suggested the possibility of cross-species transmission of the disease, indicating the need for appropriate intervention measures.

Our objectives were to evaluate the effects of the 1) season, and 2) postmortem changes on serum biochemistries related with metabolism in Pyrenean chamois (Rupicapra pyrenaica pyrenaica). Serum samples from 98 animals obtained from 2009 to 2012 were included. To investigate seasonal influences on blood parameters, the Pyrenean chamois were captured in drive-nets during the feed abundant (FA; n=32) and food deficient (FD; n=35) seasons. To evaluate the possible differences in biochemistry analytes when sampling live or dead animals, we used serum samples from 32 captured animals and 31 dead animals (obtained during controlled hunting) in the FA season. Significant increases in high-density lipoprotein cholesterol (24%), nonesterified fatty acids (NEFA, 190%), total antioxidant capacity (68%), and haptoglobin (33%) were observed in FD when compared with FA seasons. Albumin and insulin-like growth factor-1 (IGF-1) showed statistically significant decreases of 10% and 11%, respectively, in samples taken in the FD season compared to the FA season. Statistically significant higher concentrations were found in serum low-density lipoprotein cholesterol (22%), triglycerides (28%), acetylcholinesterase (50%), NEFA (383%), albumin (18%), IGF-1 (53%), cortisol (959%), and paraoxonase-1 (20%) in samples collected from live animals compared to samples collected from dead ones. We demonstrated that season should be taken into account when evaluating serum biochemistries in Pyrenean chamois because, in the FD season, these animals present lipid mobilization, decreased albumin and IGF-1, and increased total antioxidant capacity compared with the FA season. In addition, if samples are taken from dead animals, observed decreases in serum low-density lipoprotein cholesterol, triglycerides, albumin, paraoxonase-1, acetylcholinesterase, NEFA, cortisol, IGF-1, and an increase in haptoglobin should be expected.

Bats occupying hibernacula during summer are exposed to Pseudogymnoascus destructans (Pd), the causative agent of white-nose syndrome (WNS), and may contribute to its dispersal. Furthermore, equipment and clothing exposed to cave environments are a potential source for human-assisted spread of Pd. To explore dispersal hazards for Pd during the nonhibernal season, we tested samples that were collected from bats, the environment, and equipment at hibernacula in the eastern US between 18 July–22 August 2012. Study sites included six hibernacula known to harbor bats with Pd with varying winter-count impacts from WNS and two hibernacula (control sites) without prior history of WNS. Nucleic acid from Pd was detected from wing-skin swabs or guano from 40 of 617 bats (7% prevalence), including males and females of five species at five sites where WNS had previously been confirmed as well as from one control site. Analysis of guano collected during summer demonstrated a higher apparent prevalence of Pd among bats (17%, 37/223) than did analysis of wing-skin swabs (1%, 4/617). Viable Pd cultured from wing skin (2%, 1/56) and low recapture rates at all sites suggested bats harboring Pd during summer could contribute to pathogen dispersal. Additionally, Pd DNA was detected on clothing and trapping equipment used inside and near hibernacula, and Pd was detected in sediment more readily than in swabs of hibernaculum walls. Statistically significant differences in environmental abundance of Pd were not detected among sites, but prevalence of Pd differed between sites and among bat species. Overall, bats using hibernacula in summer can harbor Pd on their skin and in their guano, and demonstration of Pd on clothing, traps, and other equipment used at hibernacula during summertime within the WNS-affected region indicates risk for pathogen dispersal during the nonhibernal season.

We developed a stochastic susceptible–exposed–infectious–removed (SEIR) model to simulate a range of plausible morbillivirus outbreak scenarios in a randomly mixing population of 170 endangered Hawaiian monk seals (Neomonachus schauinslandi). We then modeled realistic vaccination and quarantine measures to determine the potential efficacy of such mitigation efforts. Morbillivirus outbreaks represent substantial risk to monk seals—91% of simulated baseline outbreaks grew (R₀>1), and in one-third of the scenarios all, or nearly all, individuals were infected. Simulated vaccination efforts in response to an outbreak were not effective in substantially reducing infections, largely because of the prolonged interval between vaccination and immunity. Prophylactic vaccination, in contrast, could be an effective tool for preventing outbreaks. Herd immunity is practically achievable because of the small sizes of monk seal populations and the animals' accessibility on shore. Adding realistic spatial structure to the model, as informed by movement of seals tracked in the main Hawaiian Islands with the use of telemetry, greatly reduced the simulated impact of outbreaks (≤10 seals were infected in 62% of spatially structured simulations). Although response vaccination remained relatively ineffective, spatial segregation allowed herd immunity to be achieved through prophylactic vaccination with less effort. In a randomly mixing population of 170 seals, 86% would need to be vaccinated to achieve herd immunity in 95% of simulated outbreaks, compared to only approximately 60% in three spatially segregated subgroups with the same combined abundance. Simulations indicate that quarantining a modest number (up to 20) of ill seals has the potential to extinguish even fast-growing outbreaks rapidly. The efficacy of quarantine, however, is highly dependent upon rapid detection and response. We conclude that prophylactic vaccination combined with a quarantine program supported by vigilant surveillance and rapid, reliable diagnosis could greatly mitigate the threat of a morbillivirus outbreak in Hawaiian monk seals.

A total of 600 wild birds were analyzed for the causes of mortality in the Republic of Korea (ROK) from 2011 to 2013. Avian poxvirus (APV) infections were identified as the primary cause of mortality in 39% (29/74) Oriental Turtle Doves (Streptopelia orientalis). At necropsy, all 29 S. orientalis birds, of which, 76% (22/29) were juveniles, had severe diphtheritic lesions in their oral and nasal cavities and on their eyelids, which were the lesions of APV that resulted in mortality. We detected APV infection by chorioallantoic membrane inoculation and molecular study of the partial region of the P4b gene. All isolates belonged to the same APV strain and were identical to strains isolated from several different pigeon species in South Africa. Phylogenetically, the APV strain identified in S. orientalis belonged to subclade A2, which includes isolates from several species of pigeons from different parts of the world, including the United Kingdom, Germany, India, Egypt, Hawaii, Georgia, Hungary, South Africa, Tanzania, and the ROK. This identity indicated that this diphtheritic APV strain may be a potential pathogen of other pigeon species in the ROK and neighboring countries throughout the range of S. orientalis. However, reticuloendotheliosis virus insertion into the APV genome was not detected by PCR in any of the 29 APV infections. An identical strain of APV observed in S. orientalis was also detected in Culicoides arakawae (biting midge), with annual peak populations corresponding to the presence of APV in S. orientalis. Culicoides arakawae may be a primary vector of APV in S. orientalis. Active surveillance of APVs in wild birds and C. arakawae is needed to better understand the epidemiology of APVs, host-vector relationships, and its ecological effects on S. orientalis in the ROK.

Raccoons (Procyon lotor) are found worldwide. They are frequently seen in crowded inner cities as well as in forests or wooded areas, often living in proximity to humans and their pets. We examined sera from 100 wild raccoons in Japan for antibodies to six canine viruses with veterinary significance to assess their potential as reservoirs. We also aimed to understand the distribution of potentially infected wildlife. We found that 7% of samples were seropositive for canine distemper virus (CDV), 10% for canine parvovirus type 2, 2% for canine adenovirus type 1, 6% for canine adenovirus type 2, and 7% for canine coronavirus. No samples were found to be seropositive for canine parainfluenza virus. Seropositivity rates for canine distemper virus and canine parvovirus type 2 were significantly different between areas, and younger raccoons (<1 yr old) were more frequently seropositive than older raccoons. Because raccoons belong to the suborder Caniformia, similar to dogs (Canis lupus familiaris), our results suggest that they can act as reservoirs for some of these important canine viruses and might be involved in viral transmission. Further study should include isolation and analysis of canine viruses in wild raccoons from a wider area.

Anthrax, caused by the spore-forming bacterium Bacillus anthracis, poses a threat to wood bison (Bison bison athabascae) conservation. We used descriptive epidemiology to characterize a large outbreak of anthrax in the Mackenzie bison population in the Northwest Territories, Canada, in 2012 and investigated historical serologic exposure of the bison to the bacterium in nonoutbreak years. Between late June and early August 2012, 451 bison carcasses were detected; mortality peaked from 13–19 July. A substantial number of calves, yearlings, and adult females died in the 2012 outbreak, unlike in two previous anthrax outbreaks in this population that killed mostly mature males. On the basis of the difference in estimates of population size prior to the outbreak (2012) and after the outbreak (2013), it is possible that not all dead bison were found during the outbreak. We assessed serologic history of exposure to B. anthracis by using samples from the Mackenzie wood bison population collected between 1986 and 2009. Overall, 87 of 278 samples were positive (31%). Seroprevalence was lower in females (18%, 10/55) than males (36%, 72/203). The highest proportion of positive submissions (90%) was from 1994, the year following the only anthrax outbreak within the historical data set. Both adult males and females had a higher likelihood of being seropositive than the younger age categories. There was a trend toward declining antibody levels between the 1993 and 2012 outbreak years.

American black bears (Ursus americanus) are commonly infested with ticks throughout their range, but there are few surveys for tick-borne disease agents in bears. To characterize tick infestations and determine the prevalence of current infection with Babesia spp. and past or current infection with Ehrlichia spp. in newly re-established populations of black bears in east central and southeastern Oklahoma, US, we identified adult (n=1,048) and immature (n=107) ticks recovered from bears (n=62). We evaluated serum and whole blood samples from a subset (n=49) for antibodies reactive to, and characteristic DNA fragments of, Ehrlichia spp., as well as characteristic DNA fragments of Babesia spp. Amblyomma americanum, the most common tick identified, was found on a majority (56/62; 90%) of bears and accounted for 697/1,048 (66.5%) of all ticks recovered. Other ticks included Dermacentor variabilis (338/1,048; 32.3%) from 36 bears, Amblyomma maculatum (9/1,048; 0.9%) from three bears, and Ixodes scapularis (4/1,048; 0.4%) from three bears. Antibodies reactive to Ehrlichia spp. were detected in every bear tested (49/49; 100%); maximum inverse titers to Ehrlichia chaffeensis ranged from 64–4,096 (geometric mean titer 1,525). However, PCR failed to identify active infection with E. chaffeensis, Ehrlichia ewingii, or an Ehrlichia ruminantium-like agent. Infection with Babesia spp. was detected by PCR in 3/49 (6%) bears. Together these data confirm that tick infestations and infection with tick-borne disease agents are common in bears in the southern US. The significance of these infestations and infections to the health of bears, if any, and the identity of the Ehrlichia spp. responsible for the antibody reactivity seen, warrant further evaluation.

Chagas disease, also known as American trypanosomiasis, is caused by the flagellate protozoan Trypanosoma cruzi. It is a significant health concern in South and Central America, where millions of people are infected or at risk of infection, and is an emerging health concern in the US. The occurrence of Chagas disease in natural environments is supported by mammal host species, but those primary species may vary based on geographic location. In South Texas, the primary host species for the disease is poorly understood, and required a field study to determine the spatial distribution of T. cruzi prevalence in free-ranging mammals. Our study objectives were to determine the spatial distribution and prevalence of T. cruzi parasites in free-ranging mammals. We compared T. cruzi prevalence among species, among vegetative communities, and among different topographies (i.e., floodplain versus upland). From December 2011 through December 2013, 450 blood and tissue samples from geolocated free-ranging wildlife mammal species were analyzed with the use of polymerase chain reaction to detect protozoan T. cruzi DNA. We also calculated mammal abundance with the use of mark–recapture methodology and recorded capture-site characteristics such as vegetation structure. We found that animals in grasslands had a significantly lower infection rate when summed across all species compared with animals in dense hardwoods and semi-improved woodlands (P=0.001). A higher percentage of infections were found in the lower-elevation floodplain—65% (28/43) of animals sampled, compared to upland areas—25% (9/36) of animals sampled. Our study suggested that common free-ranging meso-mammals supported T. cruzi in natural environments and are of public health concern in South Texas. Mitigation strategies should consider a range of management activities to include vegetation management, selective application of insecticides, and changes in human behavior in high-risk areas.

Between 2007 and 2014, 83 cetaceans were found stranded along the Ligurian coast of Italy, in the Pelagos Sanctuary, the largest marine protected area in the Mediterranean basin. Forty-nine (59%) were submitted to complete or partial necropsy, depending on the conservation status of the carcass. Based on gross and histological pathology and ancillary testing, the cause of death was determined and categorized as anthropogenic or natural (i.e., nonanthropogenic) in origin for 33 animals (67%) and of undetermined origin in the remaining 16 (33%). Natural causes of death, accompanied by either poor or good nutritional status, were attributed to 29 animals (59%), whereas four (8%) were diagnosed with an anthropogenic cause of death, consisting of interaction with fishing activities. Infectious and noninfectious disease was the most common cause of death, involving 29 cetaceans (59%). These data are valuable for understanding health and mortality trends in cetacean populations and can provide information for establishing policies for cetacean conservation and management in such an important protected area of the Mediterranean basin.

Congenital malformations have been reported in all classes of vertebrates and may be a determinant of life span and survival. In reptiles, the incidence of congenital malformations can be associated with genetic and environmental causes, including pollution. The characterization of pathological processes involved in the development of congenital malformations of bone in snakes is rare in the literature, but is of great relevance in the field of reptile conservation and environmental health. We describe congenital bone lesions in 50 newborn jararaca (Bothrops jararaca) and 26 South American rattlesnakes (Crotalus durissus terrificus) born from wild-caught pregnant females in Southeastern Brazil. Lesions were evaluated by morphometric quantitative analysis, x-ray microtomography, and histopathologic descriptive analysis. Morphometric analysis showed that jararaca presented more severe axial lesions (kyphosis, scoliosis, and kyphoscoliosis) than rattlesnakes. Female rattlesnakes presented more severe axial lesions than did males. In rattlesnakes, spinal deformities were more frequently diagnosed in the caudal segment of the body. We present x-ray microtomographic assessments and images of malformed snakes (n=9) and characterized novel malformations, such as the agenesis of frontal, parietal, and supraoccipital bones in a jararaca specimen. Histopathologic findings included vertebral body fusion, myositis, coagulation necrosis, and disorganization of periaxial muscle fibers. The new methods and results presented in this study will be useful and informative for future research in pathology, teratology, embryology, and ecotoxicology in snakes.

Lead poisoning of scavenging raptors occurs primarily via consumption of game animal carcasses containing lead, which peaks during fall firearm hunting seasons. We hypothesized that snowfall would mitigate exposure by concealing carcasses. We categorized blood lead level (BLL) for a subsample of Bald Eagles (Haliaeetus leucocephalus) from the Upper Mississippi River Valley and described BLL with respect to age, sex, and snowfall. We captured Bald Eagles overwintering in the Upper Mississippi River Valley (n=55) between December 1999 and January 2002. Individual BLL ranged from nondetectable to 335 μg/dL, with 73% of the samples testing positive for acute exposure to lead. Eagle BLL did not significantly differ between age or sex, but levels were higher immediately following the hunting season, and they were lower when the previous month's snowfall was greater than 11 cm. This study suggests a window of time between the white-tailed deer (Odocoileus virginianus) hunting season and the onset of snow when the population experienced peak exposure to lead. Combining these findings with existing research, we offer a narrative of the annual lead exposure cycle of Upper Mississippi River Valley Bald Eagles. These temporal associations are necessary considerations for accurate collection and interpretation of BLL.

Since its emergence in the 1970s, canine parvovirus (CPV) has been reported in domestic and nondomestic carnivores worldwide with severe implications on their health and survival. Here, we aim to better understand CPV circulation in multihost-pathogens systems by characterizing CPV DNA or viruses in 227 free-ranging wild carnivores of 12 species from Portugal. Collected samples during 1995–2011 were analyzed by PCR and sequence analysis. The canine parvovirus DNA was detected in 4 (2%) animals of two species, namely in wolves (Canis lupus; 3/63, 5%, 95% confidence interval=1.6–3.15) and in a stone marten (Martes foina; 1/36, 3%, 95% confidence interval=0.5–14.2). Viruses in two wolves had VP2 residue 426 as aspartic acid (so-called CPV-2b) and the third had VP2 residue 426 as asparagine (CPV-2a), while the virus in the stone marten uniquely had VP2 residue 426 as glutamic acid (CPV-2c). The comparative analysis of the full-length VP2 gene of our isolates showed other nonsynonymous mutations. The phylogenetic analysis demonstrated that the sequences from wolves clustered together, showing a close relationship with European domestic dogs (Canis lupus familiaris) and wolf strains while the viral sequence from the stone marten grouped with other viruses contained the glutamic acid VP2 426 along with raccoon (Procyon lotor), bobcat (Lynx rufus), and domestic dog strains. This study confirmed that wild carnivores in Portugal are infected by CPV variants, strongly suggesting viral transmission between the wild and domestic populations and suggesting a need for a better understanding of the epidemiology of the disease and its management in wild populations.

Waterfowl are the natural hosts of avian influenza virus (AIV) and disseminate the virus worldwide through migration. Historically, surveillance and research efforts for AIV in waterfowl have focused on dabbling ducks. The role of diving ducks in AIV ecology has not been well characterized. In this study, we examined the relative susceptibility and pathogenicity of clade 2.3.4.4 H5 highly pathogenic AIV (HPAIV) in two species of diving ducks. Juvenile and adult Ruddy Duck (Oxyura jamaicensis) and juvenile Lesser Scaup (Aythya affinis) were intranasally inoculated with A/Northern Pintail/WA/40964/2014 H5N2 HPAIV. Additional groups of juvenile Lesser Scaups were inoculated with A/Gyrfalcon/WA/41088/2014 H5N8 HPAIV. The approximate 50% bird infectious doses (BID₅₀) of the H5N2 isolate for adult Ruddy Ducks was <10² 50% egg infectious doses (EID₅₀) and for the juvenile Lesser Scaups it was <10⁴ EID₅₀. There were insufficient juvenile Ruddy Ducks to calculate the BID₅₀. The BID₅₀ for the juvenile Lesser Scaups inoculated with the H5N8 isolate was 10³ EID₅₀. Clinical disease was not observed in any group; however, mortality occurred in the juvenile Ruddy Ducks inoculated with the H5N2 virus (three of five ducks), and staining for AIV antigen was observed in numerous tissues from these ducks. One adult Ruddy Duck also died and although it was infected with AIV (the duck was positive for virus shedding and AIV antigen was detected in tissues), it was also infected with coccidiosis. The proportion of ducks shedding virus was related to the dose administered, but the titers were similar among dose groups. The group with the fewest ducks shedding virus was the adult Ruddy Ducks. There was a trend for the Lesser Scaups to shed higher titers of virus than the Ruddy Ducks. No virus shedding was detected after 7 d postinoculation in any group. Similar to dabbling ducks, Lesser Scaups and Ruddy Ducks are susceptible to infection with this H5 HPAIV lineage, although they excrete lower titers of virus.

Hemorrhagic disease in North America is caused by multiple serotypes of epizootic hemorrhagic disease virus (EHDV) and bluetongue virus (BTV). Diagnostic tests for detection of EHDV and BTV include virus isolation (VI), reverse transcriptase (RT)-PCR, and real-time RT-PCR (rRT-PCR). Our objective was to compare the diagnostic capabilities of three rRT-PCR protocols for detection of EHDV and BTV from naturally infected white-tailed deer (Odocoileus virginianus). We compared the effectiveness of these assays to traditional viral detection methods (e.g., VI) for historic and current clinical cases. Because of the variable nature of tissue collection and storage before diagnostic testing, an evaluation of viral persistence on multiple freeze-thaw events was also conducted. Two of the rRT-PCR assays provided for reliable detection of EHDV and BTV from 100% of clinically affected and VI-confirmed infected animals. Additionally, no significant change in viral titer was observed on multiple freeze-thaw events.

Brucella infection in seals was reported for the first time in 1994 around the coast of Scotland. Since then, marine mammal Brucella infections were found to be widely distributed in the northern hemisphere. Two Brucella species affect marine mammals: Brucella pinnipedialis in pinnipeds and Brucella ceti in cetaceans. We examined the livers of Baltic grey seals (Halichoerus grypus) from the Finnish coast (n=122) hunted, found dead, or killed as by-catch in fishing gear in 2013–15 as part of population health monitoring. We detected B. pinnipedialis in the livers of three grey seals. The bacterium was isolated from livers displaying parasitic cholangitis. We also detected Brucella DNA in liver flukes (Pseudamphistomum truncatum) obtained from a Brucella-infected grey seal, suggesting that flukes might be possible vectors of this pathogen in the marine environment.

The giant panda (Ailuropoda melanoleuca) is the most iconic endangered species in the world, but there is little information about the spatial and temporal distribution of parasites in the wild giant panda population. In total, 193 fecal samples from giant pandas in the Foping National Nature Reserve, People's Republic of China, were analyzed for parasite eggs using a modification of the McMaster technique. The morphology and size of Baylisascaris schroederi eggs were observed under an optical microscope. The prevalence and intensity of B. schroederi infection during the sampling year 2012 were 52.3% (101/193) and 89 eggs/g of feces, respectively, among giant pandas in this population. The prevalence of B. schroederi in the pandas varied during different months of the year, from 7% to 100%, and the prevalences in spring, summer, autumn, and winter were 71, 77, 23, and 18%, respectively. The prevalence was not significantly different between giant pandas that ate two different types of bamboo, but the intensity of infection was higher in the group eating Arundinaria fargesii (P=0.043). Altitude, temperature, and dew point were correlated with the infection intensity (r=−0.224, P<0.001; r=0.328, P<0.001; r=0.328, P=0.028, respectively). There was no correlation between infection intensity and distance to rivers. This study provides a better understanding of B. schroederi prevalence among the wild giant pandas in Foping National Nature, China.

We report malaria parasite infection (Plasmodium spp.) and associated mortality in a Common Loon (Gavia immer) found dead on Lake Umbagog in New Hampshire, US. Necropsy findings showed the bird to be in good body condition but with pericardial edema and splenomegaly. Histopathological examination of brain and heart revealed intraendothelial and intrahistiocytic proliferation of Plasmodium merozoites with myocarditis. By PCR, the presence of Plasmodium parasites was confirmed from all tissues screened including spleen, muscle, and kidney. Sequencing of nested-PCR products revealed two different Plasmodium lineages, CATUST05 and PADOM11, indicating a mixed malaria parasite infection. Clinical findings strongly support malaria-induced mortality in a Common Loon.

Infection with Brucella spp., long known as a cause of abortion, infertility, and reproductive loss in domestic livestock, has increasingly been documented in marine mammals over the past two decades. We report molecular evidence of Brucella infection in Asian sea otters (Enhydra lutris lutris). Brucella DNA was detected in 3 of 78 (4%) rectal swab samples collected between 2004 and 2006 on Bering Island, Russia. These 78 animals had previously been documented to have a Brucella seroprevalence of 28%, markedly higher than the prevalence documented in sea otters (Enhydra lutris) in North America. All of the DNA sequences amplified were identical to one or more previously isolated Brucella spp. including strains from both terrestrial and marine hosts. Phylogenetic analysis of this sequence suggested that one animal was shedding Brucella spp. DNA with a sequence matching a Brucella abortus strain, whereas two animals yielded a sequence matching a group of strains including isolates classified as Brucella pinnipedialis and Brucella melitensis. Our results highlight the diversity of Brucella spp. within a single sea otter population.

Despite only emerging in the past decade, white-nose syndrome has become among the most devastating wildlife diseases known. The pathogenic fungus Pseudogymnoascus destructans infects hibernating bats and typically leads to high rates of mortality at hibernacula during winter in North America. We developed a set of genetic markers to better differentiate P. destructans isolates. We designed and successfully characterized these 23 microsatellite markers of P. destructans for use in disease ecology and epidemiology research. We validated these loci with DNA extracted from a collection of P. destructans isolates from the US and Canada, as well as from Europe (the likely introduction source based on currently available data). Genetic diversity calculated for each locus and for the multilocus panel as a whole indicates sufficient allelic diversity to differentiate among and between samples from both Europe and North America. Indices of genetic diversity indicate a loss of allelic diversity that is consistent with the recent introduction and rapid spread of an emerging pathogen.

Wellfleet Bay virus (WFBV) is a recently described orthomyxovirus isolated from the tissues of Common Eiders (Somateria mollissima) collected during recurrent mortality events on Cape Cod, Massachusetts, US. Coastal Massachusetts is the only location where disease or mortality associated with this virus has been detected in wild birds, and a previous seroprevalence study found a significantly higher frequency of viral exposure in eiders from this location than from other areas sampled in North America. This suggests that coastal Massachusetts is an epicenter of WFBV exposure, but the reason for this is unknown. Opportunistic sampling of sympatric species and testing of banked serum was used to investigate potential host range and spatiotemporal patterns of WFBV exposure. Antibodies were detected in Herring Gulls (Larus argentatus), Ring-billed Gulls (Larus delawarensis), a White-winged Scoter (Melanitta fusca), and a Black Scoter (Melanitta nigra). These findings demonstrate the likely occurrence of fall/winter transmission, expand our understanding of the host range of the virus, and provide further insight into the epidemiology of WFBV in the northeastern US.

Leptospires are widespread spirochete bacteria that infect mammals, including rodents and humans. We investigated the presence of Leptospira antibodies in two species of rodents from San Pedro River Basin (SPRB) in northwestern Mexico as part of the black-tailed prairie dog (Cynomys ludovicianus) monitoring plan and the North American beaver (Castor canadensis) reintroduction program. We sampled a total of 26 black-tailed prairie dogs and three beavers during October–November 2015. We detected antibodies against Leptospira spp. by microagglutination test in 12 (46%) prairie dogs and in two (67%) beavers. The antibody titers for seropositive rodents varied from 1:100 to 1:200, but none of the animals showed clinical signs of disease. We found seven Leptospira spp. serogroups (Autumnalis, Australis, Bataviae, Canicola, Celledoni, Grippotyphosa, and Sejroe) circulating in rodent species in SPRB. We did not find any differences between sex and age concerning Leptospira-positive rodents. Our findings suggest the presence of endemic cycles and potential risks of Leptospira infection in both species from SPRB. Although the impact of this infection on threatened species remains unclear, human activities and environmental stress might facilitate the emergence or reemergence of leptospirosis disease as has been reported elsewhere.

From 12 May 2013 to 29 May 2013, the Gull-billed Tern (Gelochelidon nilotica) colony at the San Diego Bay National Wildlife Refuge, California, US, experienced a mass die-off of at least 92 adults, representing 71–92% of the breeding population on the US west coast. Cause of death was determined to be peritonitis due to perforations of the intestine by a large quantity of acanthocephala (Profilicollis [=Polymorphus] altmani). This is a unique report of P. altmani infecting G. nilotica, and a report of a great impact to a tern population in southern California. Mole crabs (Emerita analoga), the intermediate host for P. altmani and a major component of the Gull-billed Tern diet in San Diego, were found in the stomachs of necropsied terns along with cystacanths, and are the presumed source of the parasite infection. The tern's dietary reliance upon mole crabs likely amplified parasite transmission and infection. We suggest additional research to determine factors that influence parasite infection of intermediate and definitive hosts, particularly mole crabs, given that they are a vital resource for migrating birds within the coastal zone.

The Waved Albatross (Phoebastria irrorata) is a critically endangered seabird in a rapidly shrinking population in the Galápagos Islands, Ecuador. The introduction of novel pathogens and parasites poses a threat to population persistence. Monitoring disease prevalence and guarding against the spread of such agents in endemic taxa are conservation priorities for the Galápagos, where recent increases in the prevalence of avian pox may have contributed to population declines and range contractions in other bird species. During November 2013–January 2014, we identified 14 Waved Albatross nestlings at our study site on Española Island with avian pox-like lesions and clinical signs. Other seabirds, landbirds, and adult Waved Albatrosses were apparently unaffected. Histopathology of tissue samples from five infected nestlings revealed inclusion bodies in all samples, consistent with avipoxvirus infection. We documented higher mortality (6 of 14 nestlings) in affected nestlings than in unaffected young in this small outbreak of avian pox, the first report of its kind in the world's only tropical albatross.

Free-ranging yellow armadillos (Euphractus sexcinctus) from the Brazilian Pantanal were captured for surgical implantation of intra-abdominal radio transmitters. During March 2014, 11 animals were chemically immobilized with a combination of intramuscular ketamine hydrochloride, xylazine hydrochloride, midazolam maleate, and atropine sulfate. The mean induction time and initial heart rate in males was higher than in females. The sex of armadillos did not affect the respiratory rate, which increased from the beginning to end of the monitoring time. The rectal temperatures of females remained almost unchanged, whereas the rectal temperatures of males suffered a slight drop through the procedure. The mean (±SD) total surgical time was 50±6.5 min, and the total anesthetic recovery time was 2.5±0.5 h. The armadillos were monitored for 24 h after surgery and released at their capture site. The anesthetic association was effective, promoting muscle relaxation and analgesia that allowed the performance of surgical procedures.

Epizootic hemorrhagic disease virus (EHDV) is a Culicoides biting midge–transmitted orbivirus (family Reoviridae) of wild and domestic ruminants and is an important pathogen of white-tailed deer (Odocoileus virginianus). Historically, only two serotypes, EHDV-1 and EHDV-2, have been known to be endemic in the US. However, in 2006, an exotic serotype (EHDV-6) was first detected in the US by a long-term passive surveillance system for EHDV and bluetongue viruses. Here we report EHDV-6 detections made through these passive surveillance efforts by the Southeastern Cooperative Wildlife Disease Study (University of Georgia, Athens, Georgia, USA) and the National Veterinary Services Laboratories (US Department of Agriculture, Ames, Iowa, USA) over a 10-yr period (2006–15). The results demonstrated that EHDV-6 was detected from ruminants every year since 2006 and was widespread in the central and eastern US, providing evidence that EHDV-6 is likely now established in the US.

There are approximately 3,000 southern sea otters (Enhydra lutris nereis) in the nearshore environment along the California coast, US, and the species is classified as Threatened under the Endangered Species Act. We tested sera from 661 necropsied southern sea otters sampled from 1997 to 2015 to determine overall exposure to influenza A viruses (IAVs) and to identify subtype-specific antibody responses. Using an enzyme-linked immunosorbent assay (ELISA), antibodies to IAV nucleoproteins were detected in 160 (24.2%) otters, with seropositive animals found in every year except 2008. When the ELISA-positive samples were tested by virus microneutralization, antibody responses were detected to avian-origin hemagglutinin subtypes H1, H3, H4, H5, H6, H7, H9, and H11. Strong antibody responses to pandemic H1N1 (pdmH1N1) were also detected, indicating that epizootic transmission of pdmH1N1 occurred among the southern sea otter population after the emergence of this human-origin virus in 2009. We conclude that southern sea otters are susceptible to infection with avian and human-origin IAV and that exposure to a wide array of subtypes likely occurs during a given otter's 10- to 15-yr life span. Important unanswered questions include what effect, if any, IAV infection has on sea otter health, and how these animals become infected in their nearshore environment.

The objectives of our research on equine piroplasmosis were to clarify the prevalence of the piroplasms Babesia caballi and Theileria equi among reintroduced Przewalski's horses (takhi, Equus ferus przewalskii) in comparison with horses (Equus caballus) of nomads from around Hustai National Park in Mongolia and to determine the mortality among takhi during 2012 to 2015. The prevalence of T. equi was high in 2014 at 80% among nomadic horses and 84% among takhi. We found no evidence of B. caballi infection among takhi. A homology comparison of the 18S rRNA base sequence of T. equi showed the same sequence in both domestic horses and takhi. According to the national park records, piroplasmosis is a cause of high mortality in young takhi (19%). The results suggested that takhi were infected with T. equi in this park, and the infection is an important issue when reintroducing the takhi into the wild and increasing the numbers of the animals in nature. Continuous monitoring and vector control were noted as important issues in future conservation programs.

We confirmed safety and immunogenicity of mass-produced vaccine baits carrying an experimental, commercial-source plague vaccine (RCN-F1/V307) expressing Yersinia pestis V and F1 antigens. Forty-five juvenile black-tailed prairie dogs (Cynomys ludovicianus) were randomly divided into three treatment groups (n=15 animals/group). Animals in the first group received one standard-dose vaccine bait (5×10⁷ plaque-forming units [pfu]; STD). The second group received a lower-dose bait (1×10⁷ pfu; LOW). In the third group, five animals received two standard-dose baits and 10 were left untreated but in contact. Two vaccine-treated and one untreated prairie dogs died during the study, but laboratory analyses ruled out vaccine involvement. Overall, 17 of 33 (52%; 95% confidence interval for binomial proportion [bCI] 34−69%) prairie dogs receiving vaccine-laden bait showed a positive anti-V antibody response on at least one sampling occasion after bait consumption, and eight (24%; bCI 11–42%) showed sustained antibody responses. The STD and LOW groups did not differ (P≥0.78) in their proportions of overall or sustained antibody responses after vaccine bait consumption. Serum from one of the nine (11%; bCI 0.3–48%) surviving untreated, in-contact prairie dogs also had detectable antibody on one sampling occasion. We did not observe any adverse effects related to oral vaccination.

We studied the etiology of encephalitis in a fin whale (Balaenoptera physalus) that stranded in 2016 on the coast of Denmark. Dolphin morbillivirus (DMV) was detected in the brain and other organs. Phylogenetics showed close relation to DMV isolated from a striped dolphin (Stenella coeruleoalba) from Spain in 2012.

Oral papillomatosis was diagnosed in a gray wolf (Canis lupus) with sarcoptic mange from Minnesota, US found dead in February 2015. Intranuclear inclusion bodies were evident histologically, and papillomaviral antigens were confirmed using immunohistochemistry. Sequencing of the L1 papillomavirus gene showed closest similarity to Lambdapapillomavirus 2.

A subadult male Florida manatee (Trichechus manatus latirostris) stranded dead on Florida's Atlantic coast in January 2015. Necropsy and histopathologic findings confirmed chronic systemic bacterial infection caused by Salmonella enterica serotype IV 50:z4,z23,:- involving renal, respiratory, lymphatic, and skeletal systems. This was a unique case of systemic salmonellosis in a Florida manatee.

We present a novel case of an intestinal cestode infection in a southern sea otter (Enhydra lutris nereis). The cestode species Diphyllobothrium tetrapterum (syn. Diplogonoporus tetrapterus) was confirmed genetically. Stable isotope analysis of whiskers collected from the sea otter did not confirm the consumption of fish as the route of exposure.

Tyzzer's disease was diagnosed as the cause of mortality in free-ranging hatch-year passerine birds in Southern California, US. Birds presented with ataxia, tremors, and opisthotonos and died or were euthanized. Postmortem examination of four birds identified encephalitis on microscopy with characteristic stacked, slender bacterial rods visible with special stains. Infection with Clostridium piliforme was confirmed by real-time PCR.

We describe bilateral true anophthalmia in a juvenile female eastern gray squirrel (Sciurus carolinensis) with histologic confirmation that orbital contents lacked ocular tissues. Additionally, the optic chiasm of the brain was absent and axon density in the optic tract adjacent to the lateral geniculate nucleus was reduced.