Physiologic traits are promising indicators of population health in the face of rapidly changing environments. We obtained values of diverse physiologic parameters for Two-banded Plovers (Charadrius falklandicus) in coastal sites in Patagonia, Argentina, with the objectives of determining the timeline in which these parameters become affected by the stress of capture and handling and of obtaining reference values for future monitoring of these populations. We analyzed packed cell volume, white blood cell profile, heterophil/lymphocyte ratio, bacterial agglutination titer, and total protein, glucose, triglyceride, and cholesterol levels in apparently healthy birds. Glucose, total white blood cell count, lymphocytes, and eosinophil levels showed changes with handling times >60 min after capture. The remaining parameters did not manifest significant alterations in response to capture and handling of up to 232 min (average=105.2, SD=56.7). Therefore, although researchers should attempt to obtain blood samples as soon as possible after capture, inclusion of physiologic parameters in monitoring studies of species not easily sampled in a few minutes, such as Two-banded Plovers and other shorebird species during migration, should not be discouraged. Here we provide a physiologic report for the species that can be considered as reference values during the nonbreeding season at Patagonian coastal sites.

Seventy-two free-ranging American martens (Martes americana) in Michigan, US were immobilized using isoflurane from 2011 to 2015. In total, 129 anesthetic procedures were performed with no mortalities. Hypothermia and hyperthermia were the most common anesthetic complications, and the mean rectal temperatures were significantly higher during summer than in winter. Dental abnormalities were common; the majority of abnormal findings were broken or discolored teeth attributed to previous dental trauma and were not trap-induced. Blood (n=72) was analyzed from 53 martens for venous blood gas, lactate, hematocrit, and/or selected serum biochemistry analytes. Lactate concentration was measured by two different devices (VetScan i-STAT 1 and Lactate Plus) and compared for clinical agreement for 26 samples. Both methods for lactate measurement provided statistically similar results. Using domestic feline reference ranges, the acid-base status and relative arterial oxygen saturation of anesthetized martens in this study were normal as determined by blood pH and pulse oximetry, respectively. Serum biochemistry parameters, multiple environmental parameters, and marten-specific attributes were evaluated for their influence on lactate in American martens using linear regression and an information-theoretic approach with model averaging. Blood urea nitrogen was in all of the top models and was positively related to lactate (β=0.02, 95% confidence interval: 0.00–0.04). Initial body temperature, ambient temperature, and time from trap discovery until immobilization of martens were informative predictors for lactate level. Recommendations for the live-trapping and isoflurane anesthesia of free-ranging martens include using caution during warmer summer months, minimizing disturbance prior to induction, monitoring lactate in addition to vital rates, and being prepared to prevent or treat both hypothermia and hyperthermia during any time of year.

We tested serum samples from 387 free-ranging wolves (Canis lupus) from 2007 to 2013 for exposure to eight canid pathogens to establish baseline data on disease prevalence and spatial distribution in Minnesota's wolf population. We found high exposure to canine adenoviruses 1 and 2 (88% adults, 45% pups), canine parvovirus (82% adults, 24% pups), and Lyme disease (76% adults, 39% pups). Sixty-six percent of adults and 36% of pups exhibited exposure to the protozoan parasite Neospora caninum. Exposure to arboviruses was confirmed, including West Nile virus (37% adults, 18% pups) and eastern equine encephalitis (3% adults). Exposure rates were lower for canine distemper (19% adults, 5% pups) and heartworm (7% adults, 3% pups). Significant spatial trends were observed in wolves exposed to canine parvovirus and Lyme disease. Serologic data do not confirm clinical disease, but better understanding of disease ecology of wolves can provide valuable insight into wildlife population dynamics and improve management of these species.

Rabbit hemorrhagic disease virus (RHDV) was released in Australia as a biocontrol agent for wild European rabbits (Oryctolagus cuniculus) in 1995–96; however, its effects were variable across Australia with the greatest population reductions seen in lower annual rainfall areas (<400 mm). There is speculation that the reduced effectiveness observed at higher annual rainfall sites is at least partially due to the presence of a nonpathogenic calicivirus (RCV-A1). The RCV-A1 is related to RHDV and confers partial and transient protection against lethal RHDV infection in laboratory tests. What is not well understood is where, how, and to what degree RCV-A1 impedes the effect of RHDV-mediated rabbit control under field conditions. We investigated seven wild rabbit populations across six states and territories representing different seasonal rainfall zones across Australia, four times during 2011–12, to investigate if the presence and prevalence of RCV-A1 coincided with a change in RHDV immunity status within these populations. Besides serology, tissue samples from both trapped and shot rabbits were collected for virus detection by reverse transcription PCR. Overall, 52% (n=258) of the total samples (n=496) tested positive for RHDV antibodies and 42% (n=208) positive for RCV-A1 antibodies; 30% (n=150) of the sera contained antibodies to both viruses. The proportion of rabbits with RHDV antibodies increased significantly at sites where RCV-A1 antibodies were present (χ²₁, α=0.1, P<0.001). Evidence that preinfection of RCV-A1 may lead to a higher proportion of sampled rabbits with antibodies to both viruses was found at only one site.

Since 2006, bat populations in North America have suffered devastating mortality from an emerging disease known as white-nose syndrome (WNS). The causal agent of WNS is the fungus Pseudogymnoascus destructans. In April 2014, WNS was discovered in little brown bats (Myotis lucifugus) in Michigan, US, and has since been documented in 12 counties. Because current surveillance for WNS focuses primarily on mine-hibernating species in winter, it is subject to geographic, species, and seasonal bias. To investigate species affected and potential associations of gender, seasonal life cycle, and region with P. destructans prevalence, 1,040 rabies-negative bats were sampled from May 2014 to May 2015 from animals submitted as part of statewide rabies surveillance. The vast majority (96%) of the sample population consisted of big brown bats (Eptesicus fuscus), a noncavernicolous species. Two methods were used to detect P. destructans: fluorescence of the muzzle, wing, and tail membranes under ultraviolet light and PCR targeting genomic DNA on wing samples. Only five bats (0.5%), all M. lucifugus, were confirmed positive after nucleic acid sequencing of PCR amplicons. No other species were infected. All infected bats were collected from April to May, coinciding with their emergence from hibernation. As P. destructans and WNS spread westward, novel surveillance streams may provide a useful tool for wildlife management agencies seeking to detect the fungus where winter hibernacula such as caves and mines are absent or otherwise inaccessible.

The Galapagos sea lion (Zalophus wollebaeki), an endangered species, experiences high pup mortality (up to 100%) in years when El Niño events reduce food supply in the Galapagos Islands. Mortality of pups in non-El Niño years is estimated to be 5% in undisturbed colonies. From 2009 to 2012 we observed high pup mortality (up to 67%) in colonies close to the Galapagos capital, Puerto Baquerizo Moreno, where contact with humans, domestic animals, and rats is frequent. Gross postmortem findings from 54 pups included hemorrhagic lesions in liver and congestion in lungs; histopathology suggested a possible association with infectious diseases. Evidence of Leptospira infection was found in five out of seven samples collected in 2010. Canine distemper viral (CDV) RNA was detected in tissues from six sea lions (in 2011–12), four of which were confirmed by nucleotide sequencing. The absence of CDV antibodies in 109 juvenile animals tested in 2014 at urban and remote colonies could indicate that the CDV infection observed in 2011 was likely confined to a few animals. Our results indicated that Galapagos sea lions have been exposed at least to two pathogens, Leptospira and CDV; however, the impact of these infections on the sea lions is unclear.

Lymphoproliferative disease virus (LPDV) is an oncogenic avian retrovirus that was previously thought to exclusively infect domestic turkeys but was recently shown to be widespread in Wild Turkeys (Meleagris gallopavo) throughout most of the eastern US. In commercial flocks, the virus spreads between birds housed in close quarters, but there is little information about potential risk factors for infection in wild birds. Initial studies focused on distribution of LPDV nationally, but investigation of state-level data is necessary to assess potential predictors of infection and detect patterns in disease prevalence and distribution. We tested wild turkey bone marrow samples (n=2,538) obtained from hunter-harvested birds in New York State from 2012 to 2014 for LPDV infection. Statewide prevalence for those 3 yr was 55% with a 95% confidence interval (CI) of 53–57%. We evaluated a suite of demographic, anthropogenic, and land cover characteristics with logistic regression to identify potential predictors for infection based on odds ratio (OR). Age (OR=0.16, 95% CI=0.13–0.19) and sex (OR=1.3, 95% CI=1.03–1.24) were strong predictors of LPDV infection, with juveniles less likely to test positive than adults, and females more likely to test positive than males. The number of birds released during the state's 40-yr translocation program (OR=0.993, 95% CI=0.990–0.997) and the ratio of agriculture to forest cover (OR=1.13, 95% CI=1.03–1.19) were also predictive of LPDV infection. Prevalence distribution was analyzed using dual kernel density smoothing to produce a risk surface map, combined with Kulldorff's spatial scan statistic and the Anselin Local Moran's I to identify statistically significant geographic clusters of high or low prevalence. These methods revealed the prevalence of LPDV was high (>50%) throughout New York State, with regions of variation and several significant clusters. We revealed new information about the risk factors and distribution of LPDV in New York State, which may be beneficial to game bird managers and producers of organic or pasture-raised poultry.

Pathogens of wildlife can have direct impacts on human and livestock health as well as on biodiversity, as causative factors in population declines and extinctions. The World Organization for Animal Health (OIE) seeks to facilitate rapid sharing of information about animal diseases to enable up-to-date risk assessments of translocations of animals and animal products. The OIE also produces manuals of recommended methods to standardize diagnostic testing. Ranaviruses are important amphibian pathogens that may have spread through international trade, and infections became notifiable to OIE in 2009. We surveyed and reviewed published literature for data on sampling, diagnostic testing, and reporting of ranavirus during 2009–14. We also investigated attitudes and awareness of the OIE and its recommendations for best practice. We found that sampling effort is uneven and concentrated in the northern hemisphere. We also identified citizen science projects that have the potential to improve the quantity and quality of data on the incidence of ranavirus infection and the circumstances surrounding disease outbreaks. We found reporting of infection to be inconsistent: reporting was split between the published literature (where it was subject to a 2-yr lag) and the OIE with little overlap, results of negative diagnostic tests were underreported, and scientific researchers lacked awareness of the role of the OIE. Approaches to diagnostic screening were poorly harmonized and heavily reliant on molecular methods. These flaws in the mechanisms of ranavirus detection and reporting hamper the construction of a comprehensive disease information database.

Sylvatic plague is one of the major impediments to the recovery of the black-footed ferret (Mustela nigripes) because it decimates their primary prey species, prairie dogs (Cynomys spp.), and directly causes mortality in ferrets. Fleas are the primary vector of Yersinia pestis, the causative agent of sylvatic plague. The goal of this research was to better understand the flea fauna of ferrets and the factors that might influence flea abundance on ferrets. Fleas from ferrets were tested for Y. pestis in a post hoc assessment to investigate the plausibility that some ferrets could act as incidental transporter hosts of fleas infected with Y. pestis. Fleas were collected from ferrets captured on the Lower Brule Indian Reservation in central South Dakota, US from 2009 to 2012. A total of 528 fleas collected from 67 individual ferrets were identified and tested for the presence of Y. pestis with a nested PCR assay. The predominant flea recovered from ferrets was Oropsylla hirsuta, a species that comprises 70–100% of the fleas recovered from prairie dogs and their burrows in the study area. Yersinia pestis was detected at low levels in fleas collected from ferrets with prevalence ranging from 0% to 2.9%; male ferrets harbored significantly more fleas than female ferrets. Six of 67 ferrets vaccinated against plague carried fleas that tested positive for Y. pestis, which suggests ferrets vaccinated against plague could inadvertently act as incidental transporter hosts of Y. pestis–positive fleas.

Over the past century, the southern sea otter (SSO; Enhydra lutris nereis) population has been slowly recovering from near extinction due to overharvest. The SSO is a threatened subspecies under federal law and a fully protected species under California law, US. Through a multiagency collaborative program, stranded animals are rehabilitated and released, while deceased animals are necropsied and tissues are cryopreserved to facilitate scientific study. Here, we processed archival tissues to enrich particle-associated viral nucleic acids, which we randomly amplified and deeply sequenced to identify viral genomes through sequence similarities. Anelloviruses and endogenous retroviral sequences made up over 50% of observed viral sequences. Polyomavirus, parvovirus, and adenovirus sequences made up most of the remaining reads. We characterized and phylogenetically analyzed the full genome of sea otter polyomavirus 1 and the complete coding sequence of sea otter parvovirus 1 and found that the closest known viruses infect primates and domestic pigs (Sus scrofa domesticus), respectively. We tested archived tissues from 69 stranded SSO necropsied over 14 yr (2000–13) by PCR. Polyomavirus, parvovirus, and adenovirus infections were detected in 51, 61, and 29% of examined animals, respectively, with no significant increase in frequency over time, suggesting endemic infection. We found that 80% of tested SSO were infected with at least one of the three DNA viruses, whose tissue distribution we determined in 261 tissue samples. Parvovirus DNA was most frequently detected in mesenteric lymph node, polyomavirus DNA in spleen, and adenovirus DNA in multiple tissues (spleen, retropharyngeal and mesenteric lymph node, lung, and liver). This study describes the virome in tissues of a threatened species and shows that stranded SSO are frequently infected with multiple viruses, warranting future research to investigate associations between these infections and observed lesions.

Columbid herpesvirus-1 (CoHV-1) is widespread in feral pigeons in North America and Europe. We used a PCR assay to detect CoHV-1 DNA in oral and cloacal tissues and oral swabs from naturally infected pigeons. Fifty-three feral pigeons from five flocks in Australia (n=3 from south-central Victoria and n=2 from Sydney) were examined for CoHV-1 DNA. We detected CoHV-1 DNA in oral mucosa and cloacal mucosa, with higher concentrations in the oral mucosa. The sensitivity of testing oral swabs was the same as testing the tissue, indicating that testing of oral swabs from live birds is an effective means of screening flocks for CoHV-1 infection. Infection was found in all five of the flocks examined and the prevalence of infection ranged from 70% to100%. Most positive birds could be detected with a single-amplification PCR, but a nested amplification was required to detect others. Oral swabs from Australian native doves and pigeons (n=18) and the introduced Collared Dove (Streptopelia chinensis; n=2) were also tested by the nested PCR and all were negative for CoHV-1 DNA. We describe a fatal infection of CoHV-1 in a wild Powerful Owl (Ninox strenua) that was observed feeding on feral pigeons. This is the first known case of CoHV-1 causing death in a wild bird of prey in Australia. Our data suggest that CoHV-1 is widespread in feral pigeon flocks in Australia but we did not find it in native doves and pigeons. Spillover into native avian predator species may be occurring.

Ljungan virus (LV) is a picornavirus originally isolated from Swedish bank voles (Myodes glareolus) in 1998. The association of LV with human disease has been debated ever since, but fundamental data on the ecology of the virus are still lacking. Here we present results of the first intensive study on the prevalence of LV in bank voles trapped in Fennoscandia (Sweden and Finland) from 2009–12 as determined by PCR. Using an LV-specific real-time reverse transcriptase PCR, LV was detected in the liver of 73 out of 452 (16.2%) individuals and in 13 out of 17 sampling sites across Sweden and Finland (mean per site prevalence 16%, SE 3%, range 0–50%). We found more infected animals in autumn compared to spring, and lighter and heavier individuals had a higher prevalence than those with intermediate body masses. The result that LV prevalence is also lower in heavier (i.e., older) animals suggests for the first time that LV infection is not persistent in rodents.

Influenza A viruses (IAVs) of the subtypes H13 and H16 are primarily found in gulls (Larus spp., order Charadriiformes). Although the gull-adapted subtypes replicate efficiently during infection, gulls usually remain apparently healthy during infection. Avian influenza virus isolates are generally separated into two distinct populations, North American and Eurasian, because of the limited gene flow between the continents. Reassortment between these lineages does occur occasionally; however, direct intercontinental transmission of all eight gene segments is rare. Extensive research has been done to understand the ecology of IAV subtypes that naturally circulate in ducks (order Anseriformes), but the ecology of H13 and H16 IAVs in gulls remains far less studied. In Finland, gulls were screened for IAVs for passive (dead and diseased gulls) and active (clinically healthy gulls) surveillance purposes during the years 2005–10. During that period, 11 H13, two H16 viruses, and one H3N8 IAV were detected. We sequenced partial and full-length hemagglutinin genes of these gull-origin IAVs for phylogenetic assessments. All but one of the H13 genes clustered together with northern European and northeastern Asian viruses, whereas one virus clustered with North American viruses. Interestingly, a high rate (10/14) of these low-pathogenic IAVs was detected in dead or diseased gulls. The atypical clinical status of the IAV-positive gulls and previous observations of circovirus-like inclusion bodies in diseased gulls during autopsies, led us to screen for concurrent circovirus infections in our samples. The DNA of circovirus, an immunosuppressive pathogen of both birds and mammals, was detected in 54% (7/13) of the tested IAV-positive gulls, whereas only 25% (14/56) of our panel of IAV-negative gulls tested positive by circovirus PCR.

There are three major lineages of marine mammal strains of Brucella spp.: Brucella ceti ST23, found predominantly in porpoises; B. ceti ST26, in pelagic delphinids and ziphiids; and Brucella pinnipedialis ST24/25, predominantly in seals. The isolation of Brucella spp. in mysticetes has been described only in common minke whales (Balaenoptera acutorostrata) in Norway and Scotland. We report a third case of Brucella infection and isolation in a minke whale associated with a large abscess. In contrast to the two previous reports that involved isolates of B. pinnipedialis ST24 or the porpoise-associated B. ceti complex ST23, this case was associated with the dolphin-associated B. ceti ST26. Thus, minke whales can be infected naturally with members of all the distinct major lineages of Brucella associated with marine mammals. This report is unique in that the B. ceti ST26 did not originate from a pelagic delphinid or a beaked whale.

Species that scavenge on dead animals are exposed to enhanced disease risks. Eight hypotheses have been suggested to explain how scavengers avoid becoming sick from their diet. We conducted a systematic review of the literature and found correlative support for four of the eight hypotheses but limited evidence of systematic studies of the hypotheses. We found no support that using urine to sterilize carcasses, having bald heads, eating rapidly, or food-washing behavior reduced disease risk in carrion eaters. With the exception of food washing, none of these hypotheses have been properly evaluated as an adaptation to avoid sickness from carrion. There is some support for having a specialized microbiome, having enhanced immunologic defenses, avoiding rotten food, and maintaining a low gastric pH to eliminate pathogens. Specialized immunologic defenses and having a low pH have the most support, but the diversity of mechanisms suggests that there is a great opportunity for even more detailed study. Increased knowledge in these mechanisms may provide biomimetic insights to help combat foodborne illnesses and enhance health.

West Nile virus (WNV) was first detected in New York in 1999 and, during its expansion across the continental US, southern Canada, and Mexico, members of the Corvidae (ravens, crows, magpies, and jays) were frequently infected and highly susceptible to the virus. As part of a behavioral study of Steller's Jays (Cyanocitta stelleri) conducted from 2011–14 in the coastal California counties of San Mateo and Santa Cruz, 380 Steller's Jays were captured and tested for antibodies to WNV. Using the wild bird immunoglobulin G enzyme linked immunoassay, we failed to detect antibodies to WNV, indicating either that there was no previous exposure to the virus or that exposed birds had died.

Reticuloendotheliosis virus (REV) infects a wide range of avian species. Since 1998, when it was first reported in a captive flock of the endangered Attwater's Prairie-chicken (Tympanuchus cupido attwateri; APC), REV has plagued APC recovery efforts. While REV frequently occurs in captive bird flocks throughout the world, including commercial poultry, the reservoir for initial infection of flocks is poorly understood. From 2008–16, 412 blood samples and 216 liver samples collected from 32 species of birds on or near Attwater Prairie Chicken National Wildlife Refuge in Colorado County, Texas, US, and 89 blood samples obtained from a Texas game farm that provides thousands of Northern Bobwhites (Colinus virginianus) and Ring-necked Pheasants (Phasianus colchicus) for hunting throughout Texas, were tested for REV by real-time PCR. Of the 717 samples, one liver sample from a Savannah Sparrow (Passerculus sandwichensis) and three blood samples from game farm Ring-necked Pheasants tested positive for REV. These data, although limited, indicate a low prevalence of REV in birds sharing or in close proximity to APC habitat. More-extensive surveillance testing is warranted to determine the spatial and temporal dynamics of REV in wild bird populations and the relative role these birds may play as potential reservoirs for maintaining REV infections in both the wild and captive setting.

Radiotelemetry transmitters have become critical to studies of wildlife ecology. However, little is known about how transmitter implantation surgery affects the mobility of some species, including the timber rattlesnake, Crotalus horridus. Tracking snake movement can provide insights into the effects of transmitter implantation. During 2007–11, 71 radio transmitters were surgically implanted intracoelomically in 47 timber rattlesnakes. Over 20 of these snakes underwent surgery at least twice in 5 yr to replace old transmitters. Surgeries were performed under general anesthesia with a local nerve block at the site of implantation, 20 cm cranial to the cloaca. Snakes were also administered postsurgical meloxicam and enrofloxacin every 24 h for three doses. Two to five days after surgery, snakes were released at their original locations and radiotracked regularly during the active seasons (April–October 2007–11). Average daily movement data (distance traveled) were compiled for each snake. Snakes undergoing transmitter surgery in a given year did not differ significantly in distance traveled compared to snakes that had transmitters but did not have surgery in that year. Distance traveled for each snake did not differ before or after surgery or between weeks 1 and 2 postsurgery, indicating that the transmitter implantation did not alter snake movement.

We evaluated the use of vaginal implant transmitters (VITs) as a means of detecting, capturing, and radio collaring Rocky Mountain bighorn sheep (Ovis canadensis canadensis) lambs to estimate survival and to facilitate carcass recovery to assess causes of mortality. We focused on one of several bighorn herds in Colorado, US, suffering from depressed recruitment that was not preceded by a classic all-age die-off. We captured, radio-collared, diagnosed pregnancy by ultrasound examination, and inserted VITs into 15 pregnant ewes from a herd residing near Granite, Colorado. We were subsequently able to collar a lamb from each of 13 VITs, and two additional lambs opportunistically from ewes without transmitters. As lambs died, we recovered and submitted carcasses for necropsy and laboratory assessment. All lambs captured and one additional lamb (carcass found opportunistically) were dead by about 130 d of age: 11 died of apparent pneumonia (all within 8−10 wk of age), one died from trauma after being kicked or trampled, one was killed by a mountain lion (Puma concolor), and three died of starvation likely caused by abandonment after capture. Pneumonic lambs had involvement of Mycoplasma ovipneumoniae and leukotoxigenic Bibersteinia trehalosi. The use of VITs and lamb collars enabled us to efficiently identify pneumonia as the predominant cause of depressed lamb recruitment in this herd; however, we urge care in neonatal lamb handling to minimize abandonment.

African swine fever (ASF) is one of the most important and complex infectious diseases affecting pigs (Sus scrofa). The disease has been present in Sardinia, Italy, since 1978. Factors influencing the presence of the disease on the island are the presence of illegally bred pigs, uncontrolled movements of animals, and local traditions. Implementation of public health programs is essential for controlling ASF. The use of new diagnostic techniques on both wild boar (WB) and illegally bred pigs would provide tools for faster and more inexpensive control of the disease. We evaluated a commercial serological test kit (Pen-side [PS]) for use in the field. We sampled 113 hunter-harvested WB during the 2014–15 season, collecting blood and lung samples to conduct serological analyses and to screen for the ASF virus. Although the sensitivity (81.8%) and specificity (95.9%) of tests performed in the field were reduced compared to the same test in laboratory, they nevertheless allowed for rapid diagnosis and reduced unnecessary carcass destruction. The test, conducted in the field, was less expensive than in the laboratory and required less manpower. Therefore, we conclude that the combined use of antibody PS test and antigen PS test may be a valuable emergency management method during an outbreak as well as a useful tool for conducting regular monitoring activities as a preventive policy.

During the winter of 2013–14, 22 Canada geese (Branta canadensis) were admitted to the Wildlife Clinic at the Cummings School of Veterinary Medicine at Tufts University with nonspecific neurologic abnormalities and emaciation. Five of these geese, along with three geese that were submitted dead, were evaluated via histopathology, immunohistochemistry, and reverse transcription PCR (RT-PCR) for bornaviruses. Histopathologically, six of the eight birds had lymphoplasmacytic encephalitis. One bird, which also had encephalitis, had a dilated esophagus. Lead poisoning, West Nile virus, avian influenza, and avian paramyxovirus infection were excluded from the diagnosis. Brain tissue from all eight geese was positive for bornaviral N-antigen on immunohistochemistry. Frozen brain tissue from five birds was available for bornavirus RT-PCR. Three of the five birds were positive for the bornavirus M gene. Formalin-fixed paraffin-embedded brain tissue was evaluated on the remaining three geese via RT-PCR, with one of these geese testing positive. A bornavirus was subsequently cultured in duck embryo fibroblasts from the brain of one Canada Goose. This virus genome was sequenced, and the virus was identified as aquatic bird bornavirus 1. We were unable to identify any unusual features of this genome that would account for its apparent pathogenicity, given that subclinical infection with bornavirus in waterfowl is common in North America.

Brucellosis (Brucella bovis) in sika deer (Cervus nippon) can cause enormous losses to stag breeding, especially in areas in which stag breeding has become an important industry. It also poses a threat to humans because it is a zoonotic disease. Use of the loop-mediated isothermal amplification (LAMP) assay has been poorly described in the diagnosis of brucellosis in deer. We developed a LAMP assay targeting the omp25 gene sequence to detect brucellosis in sika deer. The reaction can be completed in 60 min at 63 C and, with a detection limit of 17 pg, it was more sensitive than conventional PCR, with its detection limit of 1.7 ng. No cross-reactivity was observed with four bacteria: Escherichia coli, Salmonella enterica subsp. enterica, Clostridium pasteurianum, and Pseudomonas aeruginosa. We used 263 samples of blood to evaluate the reaction. The percentage of agreement between LAMP and PCR reached 91%; relative specificity reached 87%, and relative sensitivity reached 100%. The results indicate LAMP can be a simple and rapid diagnostic tool for detecting brucellosis in sika deer, particularly in the field, where it is essential to control brucellosis in deer with a rapid and accurate diagnosis for removal of positive animals.

An outbreak of a novel reassortant of highly pathogenic avian influenza A (H5N2) virus (HPAIV) decimated domestic turkeys (Meleagris gallopavo) from March through mid-June, 2015 in the state of Minnesota, US. In response, as part of broader surveillance efforts in wild birds, we designed a pilot effort to sample and test hunter-harvested Wild Turkeys (Meleagris gallopavo) for HPAIV in Minnesota counties with known infected poultry facilities. We also collected opportunistic samples from dead Wild Turkeys or live Wild Turkeys showing neurologic signs (morbidity and mortality samples) reported by the public or state agency personnel. Cloacal and tracheal samples were collected from each bird and screened for avian influenza virus (AIV) RNA by real-time reverse transcription PCR. From 15 April to 28 May 2015, we sampled 84 hunter-harvested male Wild Turkeys in 11 Minnesota counties. From 7 April 2015 through 11 April 2016, we sampled an additional 23 Wild Turkeys in 17 Minnesota counties. We did not detect type A influenza or HPAIV from any samples, and concluded, at the 95% confidence level, that apparent shedding prevalence in male Wild Turkeys in central Minnesota was between 0% and 2.9% over the sampling period. The susceptibility of wild turkeys to HPAIV is unclear, but regular harvest seasons make this wild gallinaceous bird readily available for future AIV testing.

Corynebacterium pseudotuberculosis is an intracellular bacteria and the etiologic agent of caseous lymphadenitis in domestic and wildlife species. We report C. pseudotuberculosis infection in Patagonian huemul (Hippocamelus bisulcus) from the Cerro Castillo National Reserve, Region of Aysen, Chile. Subcutaneous abscesses in the abdominal and pectoral regions from two animals were sampled and bacteriologic isolation was performed. In both cases, we isolated a C. pseudotuberculosis strain belonging to the ovine genotype. In addition, one isolate was resistant to ciprofloxacin and streptomycin. We report that H. bisulcus is a susceptible species to this bacterium, which is transmitted by direct or indirect contact with domestic sheep (Ovis aries) and which represents a potential conservation threat to populations of H. bisulcus. Additional research and prevention efforts should be addressed.

Bighorn sheep (Ovis canadensis) are more susceptible to pneumonia caused by Mannheimia haemolytica than are domestic sheep (Ovis aries). Leukotoxin produced by M. haemolytica is the principal virulence factor involved in pneumonia pathogenesis. Although leukotoxin is cytolytic to all subsets of ruminant leukocytes, neutrophils are the most susceptible subset. Bighorn sheep neutrophils are four- to eightfold more susceptible to leukotoxin-induced cytolysis than are domestic sheep neutrophils. We hypothesized that the higher susceptibility of bighorn sheep neutrophils, in comparison to domestic sheep neutrophils, is due to higher expression of CD18, the receptor for leukotoxin on leukocytes. Our objective was to quantify CD18 expression on neutrophils of bighorn sheep and domestic sheep. Cell-surface CD18 expression on bighorn sheep and domestic sheep neutrophils was measured as antibody binding capacity of cells by flow cytometric analysis with two fluorochrome-conjugated anti-CD18 monoclonal antibodies (BAQ30A and HUH82A) and microspheres. Contrary to our expectations, CD18 expression was higher (P<0.0001) with monoclonal antibody BAQ30A and was higher (P<0.0002) as well with monoclonal antibody HUH80A on domestic sheep neutrophils in comparison to bighorn sheep neutrophils. These findings suggest that the higher in vitro susceptibility to leukotoxin of bighorn sheep neutrophils compared to domestic sheep neutrophils is not due to higher expression of the leukotoxin receptor CD18 on bighorn sheep neutrophils.

Asian-lineage H5 highly pathogenic avian influenza viruses (HPAIV) have caused recurrent outbreaks in poultry and wild birds. In January 2014, H5N8 HPAIV caused outbreaks in South Korea and subsequently spread to East Asia, Europe, and North America. We report the isolation of an H5N8 HPAIV strain from wild birds in Seoul, the most-developed city in South Korea. We analyzed the complete genome sequence of this isolate and estimated its origin using a phylogenetic analysis. The Seoul H5N8 isolate clustered phylogenetically with strains isolated from migratory wild birds but was distinct from Korean poultry isolates. This H5N8 virus was likely introduced into the urbanized city by migratory wild birds. Therefore, wild bird habitats in urbanized areas should be carefully monitored for HPAIV.

The Asiatic black bear (Ursus thibetanus; ABB) is a globally endangered species for which a restoration program has been ongoing in South Korea since 2001. However, there is little information on immobilization protocols for ABBs. We evaluated the use of medetomidine-zolazepam-tiletamine for their immobilization. During 2005–13, we anesthetized 60 ABBs (32 males, 28 females; 7 mo to 12 yr old) with medetomidine 0.03–0.045 mg/kg and zolazepam-tiletamine 1.54–2.3 mg/kg; reversal of anesthesia was done with atipamezole 0.15–0.225 mg/kg administered intravenously alone or intravenously and intramuscularly (50:50). Mean (and SD) for physiologic collected for 373 immobilizations of at least 60 min were: time to sedation, 7.8 (5.4) min; anesthesia induction time, 13.7 (8.1) min; complete recovery time, 14.8 (12.4) min; respiratory rate, 14 (7) breaths/min; heart rate, 51 (16) beats/min; rectal temperature, 37.3 (1.3) C; and hemoglobin oxygen saturation, 88% (6%). Few cardiopulmonary side effects occurred during immobilization and adequate depth of anesthesia was maintained for >60 min without need for supplementation. The dosage and drug combination used was effective for immobilization of ABBs with minimal adverse effects on vital signs and can be recommended in most clinical applications.

Clostridium perfringens is a Gram-positive, anaerobic, spore-forming bacterium that can induces gas gangrene or enteritis in poultry and humans and many other mammalian species. Here, we report an outbreak of C. perfringens type A and type C coinfection in wild boars (Sus scrofa). In February 2016, 10 dead wild boars, including two fresh carcasses, were found in Zhaosu County, Xinjiang Province, People's Republic of China. The two fresh carcasses were included in this study. Two strains of C. perfringens were isolated, identified, genotyped, and phylogenetically analyzed. Based on postmortem examination, bacterium isolation and identification, enterotoxin detection, and auxiliary tests, we made a diagnosis that the wild boar were killed by C. perfringens. Our findings provide the evidence that wild boar can be killed by C. perfringens intoxication. Wild boars are important reservoirs for many zoonotic agents. Therefore, more actions should be taken on the surveillance, prevention, and control of wild pig-borne diseases.

Giardia duodenalis is an intestinal protozoan capable of causing gastrointestinal disease in a range of vertebrate hosts. It is transmitted via the fecal-oral route. Understanding the epidemiology of G. duodenalis in animals is important, both for public health and for the health of the animals it infects. We investigated the occurrence of G. duodenalis in wild Swedish red foxes (Vulpes vulpes), with the aim of providing preliminary information on how this abundant predator might be involved in the transmission and epidemiology of G. duodenalis. Fecal samples (n=104) were analysed for G. duodenalis using a commercially available direct immunofluorescent antibody test. Giardia duodenalis cysts were found in 44% (46/104) of samples, with foxes excreting 100 to 140,500 cysts per gram of feces (mean, 4,930; median, 600). Molecular analysis, using PCR with sequencing of PCR amplicons, was performed on 14 samples, all containing over 2,000 cysts per gram feces. Amplification only occurred in four samples at the tpi gene, sequencing of which revealed assemblage B in all four samples. This study provides baseline information on the role of red foxes in the transmission dynamics of G. duodenalis in Sweden.

Green sea turtles (Chelonia mydas) with cutaneous fibropapillomatosis (FP) occur in some populations worldwide, and the survivability of these individuals may be compromised depending on the disease severity score. Furthermore, populations may be negatively affected in areas with a high prevalence of the disease. The stress responses (corticosterone, glucose, lactate, and hematocrit) to capture and handling were assessed in animals with different FP severity scores. From 18 January 2013 to 31 July 2014, 33 juvenile (sex was not determined) C. mydas were collected from the effluent of a steel mill in the state of Espírito Santo, Brazil, by casting a net from the bank of a canal in the morning. The body conditions of animals with FP scores of FP2 and FP3 were poor, which suggests that these turtles were unable to adequately feed themselves, and animals with an FP score of FP3 exhibited an impaired corticosterone response. This may compromise the fitness of animals in populations with a high incidence of advanced-stage FP.

Arboviruses are important zoonotic agents with complex transmission cycles and are not well understood because they may involve many vectors and hosts. We studied sympatric wild mammals and hematophagous mosquitoes having the potential to act as hosts and vectors in two areas of southern Mexico. Mosquitoes, bats, and rodents were captured in Calakmul (Campeche) and Montes Azules (Chiapas), between November 2010 and August 2011. Spleen samples from 146 bats and 14 rodents were tested for molecular evidence of Venezuelan equine encephalitis virus (VEEV), eastern equine encephalitis virus (EEEV), western equine encephalitis virus (WEEV), and West Nile virus (WNV) using PCR protocols. Bat (Artibeus lituratus, Carollia sowelli, Glossophaga soricina, and Sturnira parvidens) and rodent (Sigmodon hispidus and Oryzomys alfaroi) species were positive for VEEV. No individuals were positive for WNV, EEEV, or WEEV. A total of 1,298 mosquitoes were collected at the same sites, and five of the mosquito species collected were known VEEV vectors (Aedes fulvus, Mansonia indubitans, Psorophora ferox, Psorophora cilipes, and Psorophora confinnis). This survey simultaneously presents the first molecular evidence, to our knowledge, of VEEV in bats and rodents from southern Mexico and the identification of potential sympatric vectors. Studies investigating sympatric nonhuman hosts, vectors, and arboviruses must be expanded to determine arboviral dynamics in complex systems in which outbreaks of emerging and reemerging zoonoses are continuously occurring.

Rabies is a zoonosis that causes thousands of animal and human deaths worldwide. Serological studies provide information concerning rabies virus circulation among animals and humans. We evaluated the circulation of the rabies virus in wildlife in nine municipalities of São Paulo State, Brazil. We took blood samples from 27 terrestrial animals of nine different mammalian species in locations without cases of rabies in human and wild terrestrial mammals. Sera were tested with the use of the rapid fluorescent focus inhibition test (RFFIT) for the detection of rabies virus–neutralizing antibodies (RVNA). The RFFIT was positive in 100% of the samples, with many (81.48%) showing protective titer levels (>0.5 IU/mL) with other samples (18.52%) showing titers representing exposure (<0.5 IU/mL). We report RVNA in novel species (e.g., Alouatta caraya and Tapyrus terrestris). Wild animals were exposed to rabies virus in municipalities without a history of human rabies cases, which demonstrated a need for research to understand the role of these animals in the circulation and transmission of the disease.

Rodents and other small wild mammals are often considered to be pests and vectors for disease in zoos that house small populations of valuable threatened and endangered animals. In 2005, three nonhuman primates at a drive-through zoo in Oregon, US, acquired tularemia from an unknown source. Due to an abundance of California ground squirrels (Spermophilus beecheyi) on zoo grounds, we instituted serosurveillance of this species from July through September 2008 to determine the prevalence of antibodies against pathogens considered to be potentially transmissible to collection animals. Serologic testing was performed for Francisella tularensis; Leptospira interrogans serovars Canicola, Grippotyphosa, Hardjo, Icterohemorrhagiae, and Pomona; Toxoplasma gondii; and Yersinia pestis. All squirrels were seronegative for Yersinia pestis (0%; 0/45) and Toxoplasma gondii (0%; 0/20); there was a prevalence of 2% (1/45) for Francisella tularensis antibodies and 57% (24/42) were positive for various Leptospira serovars. Although it remains unclear whether ground squirrels present a significant risk for transmission of disease to zoo animals, vaccination of high-risk zoo animals against leptospirosis warrants consideration. Beyond this, continued vigilance and persistence with various forms of pest control may reduce the likelihood of disease transmission from wildlife hosts to animals in human care.

We assessed Toxoplasma gondii seroprevalence in 53 free-ranging American black bears (Ursus americanus) in the Central Appalachian Mountains, US. Seroprevalence was 62% with no difference between males and females or between juvenile and adult bears. Wildlife agencies should consider warnings in hunter education programs to reduce the chances for human infection from this source.

This report describes clinical, necropsy, and ancillary diagnostic findings for a mortality event in Rocky Mountain elk (Cervus elaphus nelsoni) calves attributed to malnutrition, pasteurellosis, and an alimentary presentation of adenovirus hemorrhagic disease.

We report natural infections by pathogenic Leptospira of two rodent species endemic to Chile: the degu (Octodon degus) and Darwin's pericote (Phyllotis darwini). We detected Leptospira DNA in kidney and urine samples taken in different years and sites, reaching 33% infection. The effects of infection in these species requires further evaluation.

Technology and infrastructure costs can limit access to oral vaccination tools for wildlife disease control. We describe vaccine bait mass manufacturing employing off-the-shelf technologies. Our approach has helped advance scaling-up of plague vaccination campaigns, but components of this production system could be translated into other wildlife vaccination applications.

We investigated presence of Batrachochytrium dendrobatidis in the Republic of Serbia. Seven out of 88 samples (8%) tested positive, all belonging to the frog genus Pelophylax. Two positive sites were located directly on the Danube River. The Danube River could be an important disease corridor, and distribution of Batrachochytrium dendrobatidis along this river should be further explored.

We tested 772 cloacal swabs from wild ducks to estimate the prevalence of enteric bacteria resistant to extended-spectrum cephalosporins (ESC). We found a low prevalence of the important ESC resistance genotypes, bla_CMY (5.7%) and bla_CTX-M (0.3%). This suggests a minor role for wild ducks in the movement of resistant bacteria in the environment.

We describe a case of myocardial emphysema and necrosis in a bighorn sheep (Ovis canadensis), associated with infection by Clostridium novyi, diagnosed through necropsy, histopathology, and fluorescent antibody testing. We documented rapid onset of disease in an apparently healthy wild sheep and discuss our findings in the context of reported clostridial infections in humans, domestic animals, and wildlife.