White-nose syndrome (WNS) has notably affected the abundance of Myotis lucifugus (little brown myotis) in North America. Thus far, substantial mortality has been restricted to the eastern part of the continent where the cause of WNS, the invasive fungus Pseudogymnoascus destructans, has infected bats since 2006. To date, the state of Washington is the only area in the Western US or Canada (the Rocky Mountains and further west in North America) with confirmed cases of WNS in bats, and there the disease has spread more slowly than it did in Eastern North America. Here, we review differences between M. lucifugus in western and eastern parts of the continent that may affect transmission, spread, and severity of WNS in the West and highlight important gaps in knowledge. We explore the hypothesis that western M. lucifugus may respond differently to WNS on the basis of different hibernation strategies, habitat use, and greater genetic structure. To document the effect of WNS on M. lucifugus in the West most effectively, we recommend focusing on maternity roosts for strategic disease surveillance and monitoring abundance. We further recommend continuing the challenging work of identifying hibernation and swarming sites to better understand the microclimates, microbial communities, and role in disease transmission of these sites, as well as the ecology and hibernation physiology of bats in noncavernous hibernacula.

We describe two cases of wound infections of koalas (Phascolarctos cinereus), one wild and one captive, in which Lonepinella-like organisms were involved. The wild adult koala was captured with bite wound injuries, as part of a koala population management program in Queensland, Australia. In both cases, there was evidence of physical trauma causing the initial wound. The captive koala suffered injury from the cage wire, and the wild koala had injuries suggestive of intermale fighting. Gram-negative bacteria isolated from both cases proved to be challenging to identify using routine diagnostic tests. The wound in the captive koala yielded a pure culture of an organism shown by whole genome sequence (WGS) analysis to be a member of the genus Lonepinella, but not a member of the only formally described species, L. koalarum. The wound of the wild koala yielded a mixed culture of Citrobacter koseri, Enterobacter cloacae and an organism shown by WGS analysis to be Lonepinella, but again not Lonepinella koalarum. Both cases were difficult to treat; the captive koala eventually had to have the phalanges amputated, and the wild koala required removal of the affected claw. The two Lonepinella isolates from these cases have a close relationship to an isolate from a human wound caused by a koala bite and may represent a novel species within the genus Lonepinella. Wound infections in koalas linked to Lonepinella have not been reported previously. Wildlife veterinarians need to be aware of the potential presence of Lonepinella-like organisms when dealing with wound infections in koalas, and the inability of commercial kits and systems to correctly identify the isolates.

Increasing reports of canine distemper virus (CDV) in a variety of hosts, and changing CDV dynamics, have led to renewed interest in the ecology of CDV infections in wildlife. Longitudinal serologic studies provide insights into intrapopulation and intraindividual pathogen dynamics, but few studies in wildlife have been conducted. We used data from 235 raccoons (Procyon lotor) captured on more than one occasion between May 2011 and November 2013 to investigate CDV dynamics in Ontario, Canada. Using mixed multivariable logistic regression, we found that juvenile raccoons were more likely to be seronegative from August to November than from May to July. Using paired titers from CDV-exposed individual raccoons, we determined that the winter breeding season, when there is high intraspecific contact and an increase in susceptible juveniles, may be a period of high risk for CDV exposure. Interestingly, CDV seropositive adult raccoons had nondetectable titers ranging from 1 mo to 1 yr later. Based on our preliminary investigation using two different statistical approaches, CDV exposure was associated with a decrease in parvovirus titer. This result raises important questions about whether virus-induced immune amnesia occurs after CDV exposure, which has been described for measles virus, a closely related pathogen. Overall, our results provide significant insights into CDV dynamics. Further research is needed to investigate whether CDV-induced immune amnesia occurs in raccoons and to determine the potential impacts of a reduced population immunity that may occur secondary to CDV exposure, particularly as it relates to rabies control efforts.

Comprehensive disease surveillance has not been conducted in elk (Cervus canadensis) in Tennessee, US, since their reintroduction to the state 20 yr ago. We identified causes of death, estimated annual survival, and identified pathogens of concern in elk at the North Cumberland Wildlife Management Area (NCWMA), Tennessee, US. In 2019 and 2020, we captured 29 elk (21 females, eight males) using chemical immobilization and fitted individuals with GPS collars with mortality sensors. Elk that died between February 2019 and February 2022 were necropsied to identify causes of death; these included disease associated with meningeal worm (Parelaphostrongylus tenuis; n=3), poaching (n=1), vehicular collision (n=1), legal hunter harvest (n=1), and unknown due to carcass degradation (n=3). Using data from GPS collars and known-fate survival models, we estimated an average yearly survival rate of 80.2%, indicating that survival had not significantly increased from soon after elk reintroduction (79.9%). We collected blood, tissue, feces, and ectoparasites opportunistically from anesthetized elk for health surveillance. We identified lone star ticks (Amblyomma americanum; n=53, 85.5%; 95% confidence interval [CI], 73.72–92.75), American dog ticks (Dermacentor variabilis; n=8, 12.9%; 95% CI, 6.13–24.40), and black-legged ticks (Ixodes scapularis; n=1, 1.6%; 95% CI, 0.08–9.83). We detected evidence of exposure to Anaplasma marginale (100%; 95% CI, 84.50–100.00), Leptospira interrogans (70.4%; 95% CI, 49.66–85.50), Toxoplasma gondii (55.6%; 95% CI, 35.64–73.96), epizootic hemorrhagic disease virus (51.9%; 95% CI, 32.35–70.84), and Theileria cervi (25.9%; 95% CI, 11.78–46.59). Johne's disease (Mycobacterium avium subsp. paratuberculosis) is potentially established within the population, but has not been previously documented in eastern elk populations. Disease associated with P. tenuis was a primary cause of death, and more research is needed to understand its ecology and epidemiology. Research to determine population implications of other detected pathogens at the NCWMA is warranted.

Cytauxzoonosis is a fatal tick-borne disease in domestic cats caused by infection with the apicomplexan Cytauxzoon felis. Bobcats are the natural wild-vertebrate reservoirs for C. felis, and infections are typically subclinical and chronic in this species. The present study was done to determine the prevalence and geographic distribution of C. felis infection in wild bobcats from Oklahoma and the occurrence in northwestern Texas. Tongue samples from 360 bobcats were collected from 53 counties in Oklahoma and 13 samples from three counties in Texas. For DNA extracted from each tongue sample, a probe-based droplet digital PCR assay was performed targeting the C. felis mitochondrial gene cytochrome c oxidase subunit III (cox3). Prevalence of C. felis infection was calculated for each county sampled, and data from individual counties were combined according to geographic regions and compared using chi-square tests. Overall prevalence of C. felis in bobcats from Oklahoma was 80.0% (95% confidence interval [CI], 75.6–83.8). The prevalence of infection was >90% for bobcats from central, northeastern, south-central, and southeastern regions of Oklahoma, but <68% for bobcats from northwestern and southwestern regions. Bobcats from central counties in Oklahoma were 25.693 times more likely to be infected with C. felis compared to all other bobcats sampled from the state. Higher prevalence estimates of C. felis in bobcats appeared to be in counties where known tick vectors are most common. Occurrence of C. felis in bobcats from northwestern Texas was 30.8% (95% CI, 12.4%–58.0%) based on 13 samples. Results of this study support the utilization of bobcats as sentinel animals to identify geographic areas with risk of C. felis infection to domestic cats.

In April 2020, nearly 5,000 free-ranging common frogs (Rana temporaria) were found dead on the surface of the water in a retention pond in the Swiss Alps. Macroscopic and microscopic lesions revealed multisystem emphysema, affecting multiple organs. The most severe lesions were seen in the skin, eyes, and blood vessels of internal organs and were secondary to the sudden massive distension of the skin and other affected organs. All frogs had similar lesions consistent with those described associated with gas bubble disease. No obvious pre-existing conditions potentially priming the occurrence of the observed lesions could be detected. All the examined frogs were negative by PCR for Batrachochytrium dendrobatidis, Ranavirus and Ranid Herpesvirus 3 (now Batravirus ranidallo 3). The proposed etiology is considered to be an undetermined physical event, leading to an abrupt change in the molecular or physical characteristics of the water (namely pressure and oxygen or other gas supersaturation), resulting in the occurrence of the observed lesions in the frogs. No obvious pumping system malfunction was recorded in the Mägisalp ponds before the mass mortality, but a sudden and temporary undetected change in the water flow, which then quickly rebalanced, cannot be excluded. Other hypotheses include weather conditions, such as lightning strikes in the water, or a device detonating in the water.

Herpesviruses have been detected in bat species from several countries, with a limited number of studies examining herpesviruses in Pteropus spp. (flying foxes) and no investigation of herpesviruses in Australian flying foxes. We examined the presence and prevalence of herpesviruses in the four mainland Australian flying fox species. A nested PCR targeting highly conserved amino acid motifs in the DNA polymerase (DPOL) gene of herpesviruses was used to analyze 564 samples collected from 514 individual Pteropus scapulatus, Pteropus poliocephalus, Pteropus alecto, and Pteropus conspicillatus. The prevalence of herpesvirus DNA in blood, urine, oral, and fecal swabs from the four species was 17% in P. scapulatus, 11% in P. poliocephalus, 10% in P. alecto, and 9% in P. conspicillatus (31% in P. conspicillatus spleen tissue). Five putative novel herpesviruses were detected. Following PCR amplicon sequence analysis, four of the herpesviruses grouped phylogenetically with the gammaherpesviruses, with nucleotide identities between 79% and 90% to gammaherpesviruses from Asian megabats. A betaherpesvirus was detected in P. scapulatus with 99% nucleotide identity to the partial DPOL gene sequence of an Indonesian fruit bat betaherpesvirus. This study lays the foundation for future epidemiology research of herpesviruses in Australian Pteropus spp. and adds to the discussion of hypotheses surrounding the evolutionary epidemiology of bat-borne viruses on a global scale.

Paslahepevirus balayani (hepatitis E virus [HEV]) is the causative agent of hepatitis E, a worldwide zoonosis involving a wide range of hosts among domestic and wild animals. This species is characterized by a great genomic heterogeneity and includes eight genotypes, HEV-1 to HEV-8. The HEV-3 genotype is one of the most common types circulating in Italy in humans and Suidae. Although domestic and wild Sus scrofa and deer (Cervidae) are recognized as the main reservoirs of HEV, several other wild species are potential carriers. A total of 228 liver samples were collected from nonungulate wild animals, found dead, in the framework of the regional passive surveillance program in Umbria and Marche regions (central Italy) during 2018–20. These were tested using real-time reverse-transcriptase PCR (RT-PCR) for detection of RNA of HEV-1 to HEV-4 and confirmed by nested RT-PCR assay. One of the 11 samples collected from crested porcupines (Hystrix cristata) tested positive for the presence of HEV RNA; all other samples were negative. Sequence analysis based on the full-length genome revealed that this isolate, 49434/UM/2018 (accession no. OL658617), belongs to the HEV-3e subtype. These findings suggest a potential role of crested porcupines as a carrier of HEV infection.

African swine fever is a viral hemorrhagic disease of pigs (Sus scrofa) caused by a double-stranded DNA virus, African swine fever virus (ASFV). With up to 100% mortality in pigs and no vaccine, the socioeconomic impacts of this disease are immense. In sub-Saharan Africa, warthogs (Phacochoerus africanus) are the original vertebrate host for the virus, which is transmitted among wild suids via the soft tick Ornithodoros moubata. In Malawi, O. moubata is thought to be widely distributed, with the potential to spread ASFV beyond its historical enzootic zone in the Central Region. We surveyed 76 domestic pig farms, 18 active warthog burrows, and three bushpigs (Potamochoerus larvatus) resting sites for O. moubata in the Central Region of Malawi, and tested the ticks for ASFV using PCR. We found a limited distribution for O. moubata: 75 ticks were discovered at a single farm in the Mchinji District. Eleven percent of these ticks were ASFV positive. This suggests that wildlife and O. moubata play a limited role in the epidemiology of ASF in Malawi; thus, other factors for disease spread, such as fomites and pig-to-pig infection, need to be considered.

Congenital deformities and neoplasia are poorly documented in wildlife, owing to the difficulty of detection in wild populations. Congenital deformities may lead to premature mortality, thus reducing the chances of thorough documentation. Importantly, neoplasia diagnoses depend on either sampling suspicious lesions from living individuals or access to fresh, undisturbed carcasses, which can prove challenging. We describe five cases of suspected congenital cranial deformities (midfacial cleft, wry nose, and brachygnathia inferior) and two possible cases of cranial neoplasia (orbital bone mass and a soft tissue mass) opportunistically observed in wild giraffe (Giraffa spp.) across their range in Africa. Although cases are largely limited to subjective description because physical examination is often not possible, it is critical to document such observations to help identify and track potential health concerns in wild giraffe populations.

Wildlife diseases are a major threat for species conservation, and there is a growing need to implement more comprehensive disease response programs to better identify these diseases of concern. During March 2017, we observed moribund and dead eastern newts, Notophthalmus viridescens, in a single pond in middle Tennessee. All moribund individuals were emaciated. We euthanized and processed all individuals immediately on-site and later performed histopathology and quantitative PCR for ranavirus, the protist Perkinsea, and chytrid fungi Batrachochytrium dendrobatidis and Batrachochytrium salamandrivorans. One newt was positive for ranavirus. Histopathology showed no evidence of ranavirosis but did reveal overwhelming coccidiosis. Overlapping partial sequences of coccidian 18S subunit DNA showed a 96.4% match with Eimeria steinhausi, suggesting that lesions were due to a previously undescribed Eimeria sp. In 2019, two more moribund newts were encountered at the same pond. Histopathology revealed the same suspicious parasitic organisms, and one individual was positive for B. dendrobatidis. Further research on how seasonal and other environmental parameters may influence coccidia-associated morbidity and mortality is warranted. These events highlight the importance of histopathologic evaluation of mortality events and provide guidance for investigation of future outbreaks.

The Galapagos sea lion (Zalophus wollebaeki), an endemic and endangered pinniped, faces an increasing threat due to infectious diseases related to domestic animals. Dirofilaria immitis, the parasite responsible for canine heartworm disease, is one such threat, as canine infections on the archipelago have been documented. We used a canine heartworm antigen test kit to analyze the blood from 25 juvenile Galapagos sea lions for D. immitis. Two (8%) sea lions tested positive for D. immitis antigen. Using morphologic and genetic assessments, we evaluated 20 filarial-like worms collected from within the heart of an adult male Galapagos sea lion during a previous routine postmortem examination. The intracardiac worms were morphologically consistent with adult D. immitis, and sequence analysis of targeted PCR amplicons confirmed their identity. This is the first report of D. immitis infection in Galapagos sea lions, which could become a major health problem for these pinnipeds. Further studies are necessary to confirm the level of threat from this parasite; however, widespread adoption of routine heartworm testing, prevention, and treatment in the canine population, and the control of mosquitos, could potentially reduce the disease impact on this endangered pinniped species.

During a survey in wetlands from southern Lima, Peru, two non-O1/non-O139 Vibrio cholerae isolates were obtained from samples collected from an American Oystercatcher (Haematopus palliatus) and a Wren-like Rushbird (Phleocryptes melanops). Vibrio cholerae was identified by amplification and sequencing of 16S rRNA, differentially grown on CHROMagar Vibrio media, and confirmed by ompW amplification. Isolates were confirmed to be non-O1/non-O139 serotypes and to lack the ctxA gene as inferred using PCR. Susceptibility to eight antimicrobial agents was assessed, with one isolate being resistant to azithromycin, doxycycline, tetracycline, and furazolidone. Our results indicate the utility of surveillance for V. cholerae in wetlands in the metropolitan Lima area.

Pathogens from domestic canines represent a significant and constant threat to wildlife. This study looked for four common canine pathogens, Babesia vogeli, Ehrlichia canis, Leishmania infantum, and canine parvovirus 2 (CPV-2) in mammals from the Pampa Biome, southern Brazil. Animals killed by vehicular trauma on a road traversing this biome were evaluated over a 1-yr period. Tissues collected from 31 wild mammals and six dogs were further analyzed by specific real-time PCR assays for each pathogen. Babesia vogeli and L. infantum were not detected in any investigated animal. Ehrlichia canis was detected in one dog and CPV-2 in nine animals: four dogs, three white-eared opossums (Didelphis albiventris), one pampas fox (Lycalopex gymnocercus), and one brown rat (Rattus norvegicus). These results demonstrate the occurrence of important carnivore pathogens (E. canis and CPV-2) in domestic dogs and wild mammals from the Pampa Biome in southern Brazil.

We compared the effects of animal gender, species, and intestinal helminth burden on mercury concentrations in rodents. Total mercury concentrations were determined in the liver and kidney tissues of 80 small rodents (44 yellow-necked mice, Apodemus flavicollis, and 36 bank voles, Myodes glareolus) captured in the Ore Mountains (northwest Bohemia, Czech Republic). Overall, 25/80 (32%) of animals were infected by intestinal helminths. The differences in mercury concentration between rodents infected and not infected with intestinal helminths were not statistically significant. Statistically significant differences in mercury concentrations were found only between voles and mice (that were not infected with intestinal helminths). This suggests the differences may be associated with host genetics. Apodemus flavicollis body tissues had significantly lower (P=0.01) mean Hg concentrations (0.032 mg/kg) than Myodes glareolus (0.279 mg/kg), provided that animals were not infected by intestinal helminths; if the animals were infected by intestinal helminths, the difference between both groups was insignificant. The effect of gender in this study was significant only for voles (without helminth infection); for mice (either with or without helminth infection) the differences between genders were not significant. Myodes glareolus males had significantly lower (P=0.03) Hg concentrations in liver and kidney tissues (0.050 mg/kg) than Myodes glareolus females (0.122 mg/kg). These results reveal the importance of considering species and gender when evaluating mercury concentrations.

Free-ranging wild pig (Sus scrofa) populations may cause widespread environmental damage and transmit diseases at the wildlife-livestock interface. For example, African swine fever (ASF) is a highly contagious viral disease of pigs capable of causing catastrophic economic losses. Prevention and preparedness for ASF require understanding wild and domestic pig movements and distribution. We characterized a “grey” swine market and described the risks it poses, contributing to the threats associated with wild pig populations. We monitored www.kijiji.ca, a Canadian internet classified advertising service, for sales of domestic wild boar, pot-bellied pigs, other breeds, and their hybrids across Canada from 28 April to 30 June 2021. Data collected included seller-defined breed, age, sex, number for sale, sexual intactness, presence of identifying tags or tattoos, and the date and location of listings. Advertisement locations were mapped and compared with existing wild pig distributions, identifying areas new populations might be established, and existing populations supplemented or genetically diversified. We identified 151 advertisements on Kijiji: 34% (n=52/151) from Ontario, 29% (n=44/151) from Alberta, 41% (n=62/151) from existing wild pig populations, and 59% (n=89/151) from areas where wild pigs have not yet been identified. We propose requiring the use of individual animal identifiers (tags/tattoos), genetic analysis, and mandatory reporting for all pig sales in Canada to aid in ASF preparedness and to increase regulation and enforcement of the online swine market.

Feral swine (Sus scrofa) are an introduced species to the Great Smoky Mountains National Park (GSMNP), US, and serve as carriers of several diseases that are considered a threat to other wildlife, domestic animals, and humans. During 2013 and 2015, fecal samples from 67 feral swine from the GSMNP within both Tennessee and North Carolina, US, were opportunistically collected as part of a feral swine removal program and submitted to the University of Tennessee College of Veterinary Medicine, Knoxville, Tennessee, for parasite screening by centrifugal sugar flotation. Ten taxa from the phyla Acanthocephala, Apicomplexa, and Nematoda were identified: Ascaris spp., Strongylid-type spp., Capillaria spp., Trichuris suis, Metastrongylus spp., Macracanthorhynchus spp., Coccidia, Sarcocystis spp., and Cryptosporidium spp. In 98.5% of samples, at least one parasite was found. No differences in parasite prevalence or species diversity were noted based on state of collection (Tennessee or North Carolina), sex, or age. The high prevalence of gastrointestinal parasites in these feral swine, some of which are zoonotic, represents a potential public health risk as well as a concern for free-range swine farmers.

Chemical immobilization is widely used by wildlife and veterinary professionals for the safe handling of animals. A combination of nalbuphine (40 mg/mL), azaperone (10 mg/mL), and medetomidine (10 mg/mL), known as NAM, is a low-volume combination with field immobilization practicality and fewer regulations restricting its use in the US than some other drug combinations. We evaluated the safety and effectiveness of NAM as an immobilizing agent for raccoons (Procyon lotor). From May 2021 to February 2022, 16 adult raccoons were captured in cage traps and immobilized with 0.3 mL NAM intramuscularly (12 mg nalbuphine, 3 mg medetomidine, and 3 mg azaperone, regardless of body weight). After administration, time to sedation was measured; body temperature, heart rate, respiratory rate, and oxygen saturation were monitored and recorded every 5 min for 20 min. Each raccoon was weighed; the dose administered was calculated (range 2.2–4.1 mg/kg, mean 3 mg/kg). Mean induction time was 6 min (4–17 min); time to recovery following administration of 15 mg atipamezole, 7.5 mg naltrexone for reversal, was 10 min (6–18 min). Heart rate, oxygen saturation, and respiration rate remained steady during immobilization. Rectal temperature steadily declined. Overall, NAM appeared to be a practical option for raccoon immobilization, providing rapid induction and reversal as well as adequate sedation for short-term handling and minimally invasive sampling.

We examined 82 fresh fecal samples from American White Pelican (Pelecanus erythrorhynchos) at San Pablo Autopan, Toluca Valley, central Mexico, December 2021 to April 2022. Eggs of Contracaecum spp. were observed in 100%, Capillaria spp. in two (2.5%), Strongyloides spp. in eight (9.7%), and coccidial oocysts in 12 (15%).

Two parasites were collected from the epithelial layer of the tongue mucosa of a brown-nosed coati (Nasua nasua) in an area of Atlantic Forest in Minas Gerais State, Brazil. These were identified as female Gongylonema sp. nematodes, not previously reported in Brazilian wild carnivores.

We captured a <1-d-old male elk calf (Cervus canadensis) with a shortened neck. Postmortem examination revealed trauma, meconium aspiration syndrome, and cervical vertebral malformation (partial fusion and narrowed disc spaces). This observation is novel in a wild elk calf, although the gross lesions resembled complex vertebral malformation in neonatal cattle.

Raccoon roundworm (Baylisascaris procyonis) negatively affects woodrat (Neotoma spp.) populations but is not known to occur in the endemic range of endangered Key Largo woodrats (Neotoma floridana smalli). Rectal swabs from 23 raccoons (Procyon lotor) in Key Largo were screened for raccoon roundworm by PCR. All tests were negative, suggesting continued absence.

Mycobacterium tuberculosis complex (MTBC) has rarely been detected in bears (Ursidae). We describe detection of MTBC genetic material using a single-tube, high-multiplex PCR and fluorescence-based detection system in a throat swab collected from a free-living, problem individual during immobilization and telemetry collar deployment. Mycobacterial culture was negative in all samples.

Book reviews express the opinions of the individual authors regarding the value of the book's content for Journal of Wildlife Diseases readers. The reviews are subjective assessments and do not necessarily reflect the opinions of the editors, nor do they establish any official policy of the Wildlife Disease Association.