There are many known species of Bartonella, Gram-negative bacteria that can cause febrile illness and fatality in humans and animals. These pathogens are often transmitted through hematophagous arthropod vectors such as fleas and lice. Despite increasing awareness about Bartonella spp. and their zoonotic potential, as well as existing literature on Bartonella spp. in cervids, little is known about the diversity of Bartonella spp. in white-tailed deer (Odocoileus virginianus) and their associated keds in the southeastern US. We examined the prevalence and diversity of Bartonella spp. in an enclosed herd of white-tailed deer and their ectoparasites, deer keds (Lipoptena mazamae), in Alabama. The overall prevalence of Bartonella infection in this population of deer was 16% (10/63) and 24% (23/96) in keds associated with deer that we sampled. Three species of Bartonella were identified in both deer and their keds: Bartonella bovis, Bartonella schoenbuchensis, and Bartonella sp. 1. Additionally, Bartonella melophagi was detected in white-tailed deer but not in the sampled keds. The detection of four Bartonella species in one population of white-tailed deer, three of which have known zoonotic potential, highlights the importance of Bartonella diversity within host species.

Our aim was to investigate the occurrence and distribution of Chlamydia suis and other Chlamydiaceae in the wild boar (Sus scrofa) population of Switzerland and Northern Italy and the detection of tetracycline resistance genes by PCR. We collected a total of 471 conjunctival swabs (n=292), rectal swabs (n=147), and lung tissue samples (n=32) belonging to 292 wild boars. The prevalence of Chlamydiaceae in the investigated wild boar populations was very low (1.4%, 4/292). We found C. suis in rectal or conjunctival swabs but not in lung samples. The low chlamydial prevalence might be attributed to limited contacts between wild boars and outdoor domestic pigs due to strict biosecurity measures or limited numbers of rural pig herds. The tetA(C) gene fragment was detected in six samples, which were all negative for Chlamydiaceae, and was probably not of chlamydial origin but more likely from other bacteria. The low tetracycline resistance rate in wild boar might be explained by the lack of selective pressure. However, transmission of resistance genes from domestic pigs to wild boar or selective pressure in the environment could lead to the development and spread of tetracycline-resistant C. suis strains in wild boars.

Orphaned eastern cottontail rabbits (ECRs; Sylvilagus floridanus) often present to wildlife clinics within their geographic range and require considerable dedication of time and resources. The objective of this analytical cross-sectional study was to assess initial examination findings to be used as prognostic indicators for orphaned neonatal and juvenile ECRs. The medical records of the University of Illinois Wildlife Medical Clinic were searched for ECRs presenting between 2012 and 2018. This criterion identified 1,256 ECRs that were then classified as survivors (survived and released) or as nonsurvivors (euthanized or natural death) within 72 h of admission. Presenting weight, body system abnormalities, hydration status, intervention prior to presentation, and singleton versus group presentation were categorically recorded for each individual ECR. The data were modeled using a series of logistic regression models fitted using the general linear model. Individuals were significantly more likely to be nonsurvivors if they presented as singletons (P<0.0001), presented with moderate/severe (P<0.001) or mild integumentary signs (P=0.0261), presented with multi-organ disease (P<0.001), presented with neurologic signs (P<0.0003), or had treatment provided prior to presentation (P=0.031). Factors that did not predict survival status in ECRs included body weight (P=0.210), presence of respiratory signs (P=0.674), and presence of dehydration (P=0.356). These findings may be used at wildlife medical clinics to make triage criteria for euthanasia as well as dedicate limited funds and labor to cases with the best prognosis for survival.

The American White Ibis (Eudocimus albus) is a nomadic wading bird common to wetland habitats in the southeastern US. In south Florida, US, habitat depletion has driven many ibis to become highly urbanized. Although they forage in neighborhood parks, artificial wetlands, backyards, and golf courses, the majority continue to nest in natural wetlands, often in dense, mixed species colonies. Adults and juveniles commonly disperse thousands of kilometers to other breeding colonies along the Gulf and southeast Atlantic coasts, presenting the potential for close contact with humans, domestic animals, and other wild bird species. Historically, wading birds were not considered to be significant hosts for influenza A virus (IAV), yet as ibis regularly move among various human, domestic animal, and wildlife interfaces, their potential to be exposed to or infected with IAV deserves attention. We experimentally challenged wild-caught, captive-reared White Ibis (n=20) with IAV, tested wild White Ibis for IAV, and serologically tested wild White Ibis for antibodies to IAV. White Ibis were highly susceptible to experimental challenge with H6N1 and H11N9 IAVs, with cloacal shedding lasting an average of 6 d. All 13 infected birds seroconverted by 14 d postinfection as determined by microneutralization. In contrast, no birds challenged with H3N8 were infected. We tested 118 swabs and 578 serum samples from White Ibis captured in southeastern Florida for IAV infection and antibodies to IAV, respectively. Although no IAVs were isolated, 70.4% serum samples were antibody positive by blocking enzyme-linked immunosorbent assay (bELISA). Neutralizing antibodies to H1–H12 were detected in 96.0% of a subset of bELISA positive birds (n=196) and 81.0% tested antibody positive to two or more hemagglutinin subtypes, indicating that exposure to multiple IAVs is common. These results provide evidence that White Ibis are susceptible and naturally infected with IAV and may represent a component of the IAV natural reservoir system.

The Sri Lanka Wildlife Health Centre was created in 2011 as an intersectoral surveillance system intended to address gaps in emerging and zoonotic disease preparedness in the country. This study, which was conducted during August 2015 to November 2015, examined perceived obstacles and opportunities to undertake wildlife disease surveillance in Sri Lanka. Responses to a pretested postal questionnaire by 315 field officers from the Department of Wildlife Conservation (DWC) and Department of Animal Production and Health (livestock development instructors [LDIs] and field veterinary surgeons [FVSs]) were used in the analysis. Responses came from all 25 administrative districts in the country. The reported obstacles (and proportion of respondents) included lack of transport facilities (80%), skilled staff (75%), infrastructure (65%), human resources (62%), training (67%), and a mandate for getting involved in wildlife disease surveillance (28%). These obstacles have resulted in rare or sporadic collection and submission of samples and their testing for surveillance purposes. A focus on rabies surveillance found that 84 (78%) DWC officers whose mandate was to deal with wild animals and 41 (40%) LDIs whose mandate was to deal with agricultural animals had inadequate knowledge to inform rabies surveillance activities and actions. Despite being outside of their mandate, 68% of FVSs and 72% of LDI s were willing to contribute to wild animal disease surveillance. Overcoming legal impediments to handling wildlife was cited as an essential step to establishing a multisectoral surveillance system involving government branches, universities, and citizens.

We evaluated the health of 31 (eight males, 23 females) founder eastern quolls (Dasyurus viverrinus), translocated to a fenced reserve in the Australian Capital Territory between February 2016 and July 2017. Quolls were wild caught in Tasmania (16 animals) or captive bred at Mount Rothwell Biodiversity Interpretation Centre, Victoria (15 animals). Quolls were assessed for the presence of selected potential pathogens (Toxoplasma gondii, herpesviruses, Salmonella serovars, hemoprotozoa, and ectoparasites). We assessed the relationships among sex, provenance (captive or free ranging), T. gondii or herpesvirus infection, weight, and hematologic and biochemical variables. Six of 21 quolls (29%) tested were seropositive for antibodies to T. gondii. Seropositive quolls weighed significantly more and had significantly lower potassium levels, anion gaps, and urea and triglyceride levels than seronegative quolls had. Eighteen of 31 (58%) combined conjunctival-pharyngeal-cloacal swabs collected from quolls were PCR positive for a newly identified gammaherpesvirus, tentatively named dasyurid gammaherpesvirus 3. There were no significant differences among hematologic and biochemical variables or body weights from PCR-positive and PCR-negative quolls. Eighteen of 18 (100%) of rectal-swab samples were culture negative for Salmonella serovars. Three species of tick (Ixodes tasmani, Ixodes fecialis, and Ixodes holocyclus), two species of mite (Andreacus radfordi, one unidentified), and four species of flea (Pygiopsylla hoplia, Acanthopsylla rothschildi rothschildi, Uropsylla tasmanica, and Stephanocircus dasyuri), were detected on wild-caught quolls, whereas a fifth species of flea, Echidnophaga myremecobii, was detected only on captive-bred quolls. Five of 15 blood samples (33%) were positive for hemoprotozoan DNA via PCR, a novel Hepatozoon species, a novel Theileria species, Theileria paparinii, and Trypanosoma copemani were detected. Despite the presence of several potential pathogens known to be associated with disease in other marsupials, the quolls were considered to be in good general health, suitable for translocation, and a viable population was subsequently established.

Wild carnivores serve as reservoirs of several zoonotic Bartonella species such as Bartonella henselae, Bartonella vinsonii subsp. berkhoffii, and Bartonella rochalimae. The raccoon dog (Nyctereutes procyonoides viverrinus) is the most common native carnivore in Japan, but epidemiologic studies of Bartonella infections have not been performed in this animal species yet. Here, we report a molecular survey of B. rochalimae prevalence in 619 wild raccoon dogs captured from 2009 to 2017 in western Japan. Bartonella rochalimae DNA was detected in 7.1% (44/619) of the raccoon dogs examined by PCR targeting the rpoB and ssrA genes. All of the sequences obtained were identical in each of the genes. The prevalence of B. rochalimae by sex of the animals was 6.1% (21/344) in male and 8.4% (23/275) in female. The prevalence by year varied from 2% (1/45) in 2011 to 14% (4/28) in 2016. The prevalence (7.9%) of B. rochalimae in the raccoon dogs with sarcoptic mange tended to be higher than the prevalence (4.0%) in the animals without the infestation of mites, although the differences were not significant. Sequence analysis indicated that Japanese raccoon dogs in the area examined were infected with B. rochalimae carrying identical sequences in the rpoB and ssrA genes. In addition, the raccoon dog strain had few sequence variations in both genes compared to other known B. rochalimae strains detected in other parts of the world.

Chronic wasting disease (CWD) is an infectious disease, but reported associations suggest several metals—especially copper (Cu) and manganese—potentially play a role in this and other prion diseases. To assess the utility of dietary Cu supplementation in protecting white-tailed deer (Odocoileus virginianus) from CWD, we compared incidence and disease course among individuals naturally exposed to CWD while being maintained on sustained-release Cu boluses or unsupplemented (control). Oral Cu supplementation increased liver tissue Cu concentrations compared to controls but did not affect susceptibility to CWD or survival after natural exposure in the captive white-tailed deer we studied. Over the 27 mo study, 89% (8/9) of the Cu-supplemented deer and 86% (6/7) of control deer became CWD-infected. Survival to 27 mo postexposure did not differ between Cu-supplemented and control deer: model-averaged survival probabilities to 27 mo were 0.45–0.47 for all combinations of Cu treatment and PRNP gene haplotype presence. The PRNP gene haplotype influenced the probability of deer remaining biopsy negative for at least 17 mo but did not affect overall susceptibility.

Trematode infestation of Mallards (Anas platyrhynchos) varies between northern Poland and the Czech Republic. We determined the trematode fauna of juvenile and adult Mallards from Poland in 2010–16 (n=79 Mallards) and from the Czech Republic in 1964–2001 (n=296). Infracommunity diversity of trematodes from juvenile Mallards, defined by the Brillouin index and mean number of species, was significantly greater in Poland than it was in the Czech Republic. There were more species and greater biodiversity in the trematode communities of Mallards from Poland, where the environment is more natural and less altered by human activity than it is in the Czech Republic. In the trematode community of juvenile Mallards, the Simpson's index of biodiversity was higher in the Czech Republic than it was in Poland. The Berger-Parker dominance index was comparable in both countries. In terms of prevalence, the structure of the trematode fauna was hierarchic, and trematode species had different positions in this hierarchy in the two countries. Statistically significant differences in the epidemiologic indices between infection of juvenile Mallards from Poland and the Czech Republic were noted for Bilharziella polonica, Echinostoma miyagawai, Echinostoma revolutum, Prosthogonimus cuneatus, Prosthogonimus ovatus, Notocotylus attenuatus, Echinoparyphium recurvatum, Australapatemon minor, Apatemon gracilis, Cyathocotyle prussica, Hypoderaeum conoideum, Metorchis xanthosomus, Psilochasmus oxyurus, and Cotylurus cornutus.

White-tailed deer (Odocoileus virginianus) serve as a host for cattle fever ticks (Rhipicephalus [Boophilus] microplus and Rhipicephalus [Boophilus] annulatus; CFTs); therefore, deer are a concern for CFT control programs in southern Texas, US. Systemic (oral delivery of ivermectin) and topical (permethrin on pelage) treatment devices have been developed for white-tailed deer; however, the efficacy of these treatment options has not been determined for CFTs in southern Texas. Our objectives were to evaluate the effectiveness of CFT treatment strategies by 1) measuring exposure rates of deer to the acaricides permethrin and ivermectin, 2) determining the relationship between CFTs on deer and exposure to the acaricides, and 3) determining if photos from remote cameras at medicated bait sites can be used as a measure of acaricide treatment. We captured 327 deer at four sites in southern Texas. Deer visitation to medicated bait sites was monitored using remote cameras from March 2010 to February 2012. There was no relationship between the presence of permethrin and the probability of being infested with CFTs (P≥0.336). The probability of infestation with CFTs decreased as serum ivermectin levels increased for male (n=18, P=0.098) and female (n=33, P<0.001) deer. Our results indicate ivermectin may be more effective in treating CFTs than permethrin; thus it would be worthwhile to develop topical acaricides other than permethrin for treating white-tailed deer in southern Texas.

Eastern and western spotted skunks (Spilogale putorius and Spilogale gracilis) serve as definitive hosts for skunk cranial worm (Skrjabingylus chitwoodorum), a metastrongylid nematode that spends its adult stage inhabiting the frontal sinuses of the host cranium. Skunk cranial worm can cause severe damage to the skull of spotted skunks, and this damage is identifiable in preserved specimens. We visited six mammal collections between 2016 and 2018 and used 578 spotted skunk skull specimens to identify patterns in prevalence and severity of skunk cranial worm damage in spotted skunks. Specimens were collected between 1895 and 1981 from across the US. We assessed effects of host genetic clade, collection year, precipitation, and sex on prevalence and severity rates. We hypothesized that the midwestern genetic clade (plains spotted skunk, Spilogale putorius interrupta), which experienced a range-wide population decline in the mid-1900s, would experience the highest infection and severity rates. We expected precipitation, but not sex to influence infection and severity. Our top models indicated that host genetic clade, precipitation in the year prior to specimen collection, and the year prior to specimen collection best predicted prevalence and severity of skunk cranial worm. We suggest the positive association with precipitation is a result of local availability of gastropods, the intermediate host for skunk cranial worm. There was a negative association between prior year and prevalence, but the severity of damage increased over time for all clades. Given concerns over population declines of spotted skunks as well as observations of increasing damage over the past century, there is a need for further research on the impacts of cranial worm infection on individual-scale behavior, reproduction and survival, and on population-scale demographics.

Carnaby's Cockatoos (Calyptorhynchus latirostris) are in decline in SW Western Australia from several processes, including habitat loss and fragmentation. However, in recent years, a disease syndrome has also emerged as a significant population threat. Emerging diseases in wildlife have the potential for catastrophic effects on population numbers, especially if a species is experiencing existing pressure from other threatening processes. This article describes an investigation into a hindlimb paralysis syndrome that has occurred in the summer and autumn since 2012 in 84 wild Carnaby's Cockatoos. Recovery from the syndrome has been demonstrated in 21 of 33 cases when supportive therapy was applied. Although a definitive diagnosis has not been obtained, the hypothesized etiology is an organophosphate-induced delayed-onset neuropathy. The syndrome may indicate that interaction between the cockatoos and inland agricultural practices are affecting this migratory species in ways that are, so far, poorly understood.

Attenuated strains of rabies virus (RABV) have been used for oral vaccination of wild carnivores in Europe and North America. However, some RABV vaccines caused clinical rabies in target animals. To improve the safety of attenuated RABV as an oral vaccine for field use, strategies using selection of escape mutants under monoclonal antibody neutralization pressure and reverse genetics–defined mutations have been used. We tested the safety, immunogenicity, and efficacy of one RABV construct, ERA-g333, developed with reverse genetics by intramuscular (IM) or oral (PO) routes in big brown bats (Eptesicus fuscus). Twenty-five bats received 5×10⁶ mouse intracerebral median lethal doses (MICLD₅₀) of ERA-g333 by IM route, 10 received 5×10⁶ MICLD₅₀ of ERA-g333 by PO route, and 22 bats served as unvaccinated controls. Twenty-one days after vaccination, 44 bats were infected by IM route with 10^2.9 MICLD₅₀ of E. fuscus RABV. We report both the immunogenicity and efficacy of ERA-g333 delivered by the IM route; no induction of humoral immunity was detected in bats vaccinated by the PO route. Two subsets of bats vaccinated IM (n=5) and PO (n=3) were not challenged, and none developed clinical rabies from ERA-g333. Scarce reports exist on the evaluation of oral rabies vaccines in insectivorous bats, although the strategy evaluated here may be feasible for future application to these important RABV reservoirs.

Reticuloendotheliosis viruses (REVs) are known to cause immunosuppressive and oncogenic disease that affects numerous avian species. Reticuloendotheliosis viruses are present worldwide and recently have been reported in South America with cases of infected commercial flocks in Argentina. We surveyed for the presence of REV in birds from a state in the northern region of Brazil using real-time PCR. We report here the presence of REV in Brazil, detected in Muscovy Ducks (Cairina moschata), Wild Turkeys (Meleagris gallopavo), and chickens (Gallus gallus) at a relatively high prevalence (16.8%). Phylogenetic analysis indicated a close relationship of these strains to variants in the US. This study provides evidence of REV in the Amazon biome and provides a baseline for future surveillance of the virus in the region and throughout Brazil.

A pedunculated cauliflower-like mass was detected on the left posterior limb of a subadult male red deer (Cervus elaphus) after a hunt in Portugal. Histologically the lesion was classified as cutaneous fibropapilloma. The identification of Cervus elaphus papillomavirus (CePV-1 variant) was based on sequencing of the L1 gene. The L2 sequence revealed a nine-nucleotide deletion, as already reported in the Italian and Hungarian CePV-1, further supporting the theory that this is a distinctive genomic characteristic of this viral variant, as this feature has been found in distinct cases from geographically distant countries. In addition, a coinfection with bovine papillomavirus was evidenced by amplification and sequencing of the E5 gene, confirming the ability of Delta papillomaviruses to cross-infect different animal species and providing more evidence that wildlife may act as reservoir for papillomaviruses affecting domestic species. Papillomavirus infection in red deer has been sporadically described in different European countries; in this work, we describe the identification of a CePV-1 variant infection associated with a red deer fibropapilloma in Portugal.

Rodent-borne hantaviruses have been reported in many of the countries surrounding Ukraine; however, to date we have no knowledge of the viral strains circulating in Ukraine within reservoirs such as the striped field mouse (Apodemus agrarius), the yellow-necked field mouse (Apodemus flavicollis), and the bank vole (Myodes glareolus). To determine the prevalence of hantaviruses in Ukraine, we captured 1,261 mammals, of which 1,109 were rodents, in 58 field sites within the province of Volyn in western Ukraine. Foci of the striped field mouse tended to occur in the eastern and southern parts of the province, whereas the bank vole were clustered in western and northern regions. The striped field mouse and bank vole had detectable serum antibodies to Puumala virus (PUUV) or Dobrava virus (DOBV) antigens at 7% or 2%, respectively, using an indirect immunofluorescence assay. Antibody prevalence among the bank vole males and females was equivalent, whereas for the striped field mouse, the prevalence among males was 5% versus 1% for females. In two bank vole specimens, we were able to detect partial nucleotide sequences that showed identity to PUUV. In summary, this study suggests that two human pathogens, PUUV and DOBV, cocirculate in the bank vole and the striped field mouse, respectively, in Ukraine. Future studies will focus on new rodent collections that will enable obtaining the complete genome sequences of the PUUV and DOBV strains circulating in Ukraine to provide guidance on the design of optimal molecular diagnostics that can enable insight into the potential contribution of hantaviruses to human disease in Ukraine.

A lethargic juvenile male harp seal (Pagophilus groenlandicus) in poor nutritional condition was found on the beach on the north shore of Prince Edward Island, Canada, in June 2017. Microscopic examination revealed a severe nonsuppurative encephalitis positive for morbillivirus antigen on immunohistochemistry. Virus isolation attempts were negative. However, phocine distemper virus (PDV) was detected in brain tissue RNA extracts by a seminested reverse transcription PCR that targeted the paramyxovirus RNA–dependent RNA polymerase (pol) gene. Comparison of the resulting partial PDV pol nucleotide sequence revealed it was nearly identical to PDV strains isolated from eastern Atlantic harbor seals (Phoca vitulina vitulina) during a 1988 epizootic in the Wadden and Irish seas, and a western Atlantic harbor seal (Phoca vitulina concolor) that stranded in Maine, US, in 2006. Our study confirmed that closely related PDV strains are circulating in multiple seal species along the coastlines of North America and Europe.

There is limited information about virus epidemiology of shorebirds (family Charadriidae and Scolopacidae) in the East Asia-Australasia flyway. We investigated the prevalence of avian influenza viruses (AIVs) in shorebirds in Hokkaido, Japan, the stopover site of the flyway, to understand the ecology of AIV translocation in the flyway from 2006 to 2010. In total, 1,698 shorebirds belonging to 26 species were captured and released into two different sites using mist nets. Tracheal and cloacal swabs were collected from each bird using cotton swabs. The RNA of influenza A viruses was detected using reverse transcription loop-mediated isothermal amplification. One AIV-positive sample was obtained from a Lesser Sand Plover (Charadrius mongolus) captured in September 2010 at Lake Komuke. Full lengths of hemagglutinin (HA), neuraminidase (NA), polymerase acidic protein, nucleoprotein, matrix protein 1, and nuclear export protein genes were successfully amplified from the AIV-positive sample. All sequences showed the highest identity with sequences obtained from virus strains from Anseriformes species. Shorebirds migrated to Japan 1 mo earlier than did Anseriformes species. Therefore, the Lesser Sand Plover could have been infected by the virus from Anseriformes species on the breeding grounds. The HA sequence showed the highest identity with the H10 sequence whereas the NA sequence exhibited the highest identity with the N7 sequence. Phylogenic analysis showed that the detected subtype H10N7 belongs to the Eurasia lineage and the related strain might have widely spread in Asia in 2009.

Three hundred and fourteen red foxes (Vulpes vulpes) in the province of Soria, Spain, were examined for hantavirus and lymphocytic choriomeningitis virus (LCMV) infection (and were likely to have been infected by feeding on infected rodents). Immunofluorescence and western blot assays confirmed 3.5% (11/314) to have antibodies to hantaviruses, and the immune fluorescence assay showed 2.2% (7/314) to have antibodies to LCMV. The serologic status of the animals showed no statistically significant association with sex or age. Although studies on the prevalence of hantaviruses and LCMV normally focus on rodents, our results showed that foxes can provide complementary information in determined areas.

Blastocystis is a genus of parasitic protozoans that live in humans, mammals, and birds and which has been widely studied due to its low host specificity. Limited data are available, however, regarding its presence in wildlife, particularly in South Korea. Contact between wild boars (Sus scrofa) and livestock or humans has steadily increased as wild boars venture down from the mountains to farms and residential areas. In this study, we examined the status and subtypes (STs) of Blastocystis in wild boars in South Korea and confirmed its zoonotic potential. From March 2016 to November 2018, we collected 433 fecal samples throughout the country from trapped or road-killed wild boars. The 18S rRNA gene was used for molecular identification and subtyping and the proportion of PCR-positive samples was 10.4%. We then assessed positive samples for associations with sex, region, and seasonal infection; however, no statistical significance was observed for any variable other than season. Phylogenetic analyses revealed that all sequences belonged to subtype 5 and had 99.5–99.9% identity with sequences obtained from Japanese cattle (Bos taurus) and 97.1% identity with sequences obtained from Chinese. Subtype 5 has been implicated in zoonoses, indicating that Korean wild boars could transmit Blastocystis to humans and other livestock. Our results, in accordance with the One Health concept, strongly support continued interest and efforts by public health and disease control organizations toward transmission prevention.

Rhinella alata is a small terrestrial bufonid that occurs in Ecuador, Colombia, and Panama. Between January 2014 and October 2015, we inspected 339 R. alata from Panama and report myiasis in eight of these toads. All infested toads were male and presented with mobile dark fly larvae visible beneath the ventral skin. At necropsy, we identified the larvae as belonging to the family Sarcophagidae (flesh flies). Flesh flies have been variously considered as predators, parasitoids, and parasites of anurans. There are at least four species of flesh flies that infect adult amphibians in the Neotropics, with the most common and widespread being Lepidodexia bufonivora. Myiasis has only rarely been reported in Panamanian anurans. Anuran cases of sarcophagid myiasis are usually fatal and we suspect myiasis as the cause of death for the R. alata that died in the current study.

We reviewed necropsy records of 124 Whooping Cranes (Grus americana) recovered following reintroduction of 268 individuals from 2001 to 2016 in the eastern US. Causes of death were determined in 62% (77/124) of cases facilitated by active monitoring that limited decomposition and scavenging artifact. The greatest proportions of mortality were caused by predation (0.468; 95% confidence interval 0.356–0.580; 36/77), collision with power lines or vehicles (0.260; 0.162–0.358; 20/77), and gunshot (0.169; 0.085–0.253; 13/77). Six deaths were attributed to infection (0.078; 0.018–0.138; 6/77), including bacterial and fungal etiologies. Lead analysis of 50 liver samples yielded two results with elevated concentrations (3.65 and 10.97 ppm wet weight), and 10 bone samples from partial carcasses lacking suitable liver tissue resulted in one elevated result (48.82 ppm dry weight). These data indicate that underlying subclinical or clinical lead toxicosis may be a factor in up to 5% of deaths attributed to predation or impact trauma. Brain cholinesterase activity testing indicated no exposure to organophosphate or carbamate pesticides (mean±SD=17.32±2.90 µmol/min/g, 31/71). The causes of death and potential underlying factors summarized in this study constitute the first definitive mortality survey of migratory Whooping Cranes based on a high carcass recovery rate. Causes of death by infectious etiologies remained comparatively rare in this study, and occurred as single cases with no evidence of sustained transmission among reintroduced Whooping Cranes.

Emerging pathogens may pose additional threats to already vulnerable populations of chelonians, such as gopher tortoises (Gopherus polyphemus). In response to a mortality event on a translocation site in northwest Florida, US during 2013–15, 13 gopher tortoises were necropsied and their tissues were screened for 12 pathogens, including Mycoplasma agassizii, Mycoplasma testudineum, and Frog virus 3–like ranavirus (FV3). The DNA of FV3 was detected via quantitative PCR in the gastrointestinal tract of three tortoises. Subsequently, pathogen surveillance was performed on whole blood and oral–cloacal swab samples of live translocated tortoises from two different enclosures within the site (n=68), rehabilitated tortoises from the site (n=18), and tortoises prior to release on site (n=35) during 2015–17. Mycoplasma spp. were present in all groups and years of live tortoises tested. The DNA of FV3 was detected in 15 individuals both with and without clinical signs of disease in 2016. We recaptured 20 tortoises and captured an additional 20 tortoises in 2017 for surveillance, yet FV3 DNA was no longer detected, even in those that had previously tested positive (n=7). The results of this study contribute to the epidemiology of ranavirus in chelonians and suggests that gopher tortoises could be reservoirs for FV3. We recommend that the status of Ranavirus infection should be included for health screens for gopher tortoises in translocation programs.

Forest musk deer (FMD; Moschus berezovskii) immunoglobulin G efficiently bound to streptococcal G protein (SPG) and weakly bound to staphylococcal A protein. The results suggested that horseradish peroxidase–conjugated SPG could be chosen as an enzyme-labeled antibody substitute and laid a foundation for immunologic research in FMD disease.

The deer ked (Lipoptena cervi), a hematophagous ectoparasite of cervids, is currently spreading in Scandinavia, and the moose (Alces alces) is its main host. However, little is known about the impact of deer keds on moose. We analyzed the hair cortisol concentration (HCC) from 262 moose harvested in the fall in relation to age class, sex, body mass (BM), and deer ked infestation intensity, and BM in relation to age class, sex, and infestation intensity. We found that HCC decreased with increasing deer ked intensity at low ked intensities, but for the higher levels of ked intensities, there was a positive relationship between HCC and ked intensity. The HCC was higher in males than in females and lower in yearlings than in calves and adults. Our failure to find any association between BM and deer ked intensity suggested a negligible impact of deer ked infestation on moose foraging and metabolism at the level of infestation observed early in the infestation, but did not exclude an effect later in winter. Our findings suggested that moose generally tolerated moderate parasitism by keds. However, the increase in HCC at higher ked intensities suggested that the tolerance strategy could be disrupted with further increases in intensities and consequently may negatively affect animal health and welfare.

Tick-borne protozoans of the genus Hepatozoon are obligate hemoparasites that can infect domestic and wild terrestrial vertebrates. Main hepatozoonosis affects canids and involves mainly Hepatozoon canis and Hepatozoon americanum. However, molecular studies revealed the capacity of H. canis to infect a wide range of wild mammals. In July 2018, we conducted an epidemiological survey for tick-borne pathogens in wild hosts, assaying Hepatozoon sp. occurrence in 34 bats captured in different habitats within a conservation unit in the state of Espírito Santo, southeastern Brazil. Blood and spleen tissue DNA samples were submitted to PCR amplifications of Babesia/Theileria and Hepatozoon 18S rRNA gene and 21% (7/34) were positive for Hepatozoon sp. Phylogenetic inferences grouped the obtained sequences from Seba's short-tailed bat (Carollia perspicillata) with the H. canis cluster, and from the great fruit-eating bat (Artibeus lituratus) with rodent-associated Hepatozoon cluster. Further studies are needed to characterize the epidemiological role of Seba's short-tailed bat and the great fruit-eating bat in the wild transmission cycle of these hemoparasites in Brazil.

Brucella ceti has been detected in several species of free-ranging odontocetes, in several geographic areas but it has not been reported in Indo-Pacific finless porpoises (Neophocaena phocaenoides) nor in any odontocetes in waters in the South China Sea. Sampling of odontocetes stranded in Hong Kong Special Administrative Region, People's Republic of China, was carried out as part of a stranding monitoring program to evaluate the pathogens harbored by these threatened species. A real-time PCR method targeting the Brucella genus-specific 31kDa Brucella cell surface salt extractable (bcsp31) gene, gene sequencing, and phylogenetic characterisation produced three PCR products of the expected size and sequence, from two stranded Indo-Pacific finless porpoises. The PCR products were obtained from brain tissue from of a neonate and from mammary fluid from a sexually mature female. Further testing for this pathogen should be performed to determine whether Brucella ceti might have a detrimental effect on reproduction and calf survival in Indo-Pacific finless porpoises and pose a threat to the conservation of this species. The importance of biosafety and biosecurity measures when handling cetaceans or their tissues and products in the South China Sea is also highlighted.

Environmental DNA (eDNA) methods provide novel options for the detection of pathogens. The amphibian pathogens Batrachochytrium dendrobatidis (Bd) and Ranavirus have been relatively understudied in Texas, US, so we applied eDNA assays for the surveillance of these pathogens in the upper Brazos River basin near the Texas panhandle. We collected water samples from five urban playa lakes and one reservoir in and around Lubbock, Texas. Quantitative PCR detected both Bd and Ranavirus at one playa lake, representing novel detection of both pathogens in the region. Based on these results, we recommend increased monitoring for the pathogens and symptoms of amphibian disease throughout the region.

Salmonella enterica is one of the main causes of gastrointestinal disease worldwide. Wild birds are capable of harboring a variety of Salmonella serovars, which could have an important role in the epidemiology of salmonellosis in humans and production animals. We tested 519 fecal samples from raptors and aquatic birds from different regions of central (three rehabilitation centers for wildlife and the coastal area) and southern areas of Chile for Salmonella. All samples were obtained in 2015 and 2017, covering all four seasons. Salmonella was isolated from 12 of the 519 samples (2%) analyzed, from two carnivorous birds, four birds with generalist habits, and six waterfowl. Among the isolates obtained, one showed resistance to gentamicin, and one showed a multidrug-resistance phenotype, with resistance to ampicillin, ceftriaxone, ciprofloxacin, chloramphenicol, streptomycin, gentamicin, kanamycin, trimethoprim-sulfamethoxazole, and tetracycline. These results demonstrated the importance of characterizing Salmonella in wild birds because previous studies have shown genetic and phenotypic evidence suggesting interspecies transmission of Salmonella enterica that is resistant to antimicrobials between humans and wild and domestic birds.

Two free-ranging female Sunda colugos (Sunda flying lemur; Galeopterus variegates) were found dead in Singapore in November 2018 and February 2019. Septicemia and mastitis were diagnosed in both animals on postmortem examination. Infectious diseases have not previously been established as a cause of death in wild Sunda colugos.

We generated reference ranges for seasonal leukocyte differential counts of the free-ranging deermouse (Peromyscus maniculatus) from Montana, US. Blood was collected from the retro-orbital capillary sinus of deermice after topical anesthesia with proparacaine. Although season influenced lymphocyte, neutrophil, and monocyte absolute counts, sex and reproductive status did not.

The modified agglutination test was used to study Toxoplasma gondii exposure in 70 eastern Tennessee, US, black bears (Ursus americanus) from 2015 to 2017. Overall, 74% (52/70) of bears were seropositive, and T. gondii was more prevalent in adults than subadults.

Gastrointestinal tracts were obtained from 120 raccoons (Procyon lotor) on properties in the state of Mississippi, US, with and without wildlife baiting to observe the effects of baiting on parasite prevalence and intensity. Raccoons from baited properties had higher prevalence of Gnathostoma procyonis and higher intensities of Physaloptera rara and Macracanthorhynchus ingens, which can cross species barriers.

The pathogenic Leptospira species are very widespread in nature, persisting in the renal tubules of many domestic and wild animal reservoirs. We report the isolation of Leptospira interrogans serovar Pomona in a bottlenose dolphin (Tursiops truncatus) stranded along the coast of Sardinia, Italy, in 2016.

Brucella abortus biovar 1 and atypical rough Brucella were isolated from Himalayan marmot (Marmota himalayana). Multiple-locus variable-number tandem-repeat analysis-16 typing indicated that the isolates both for smooth and atypical rough phenotypes had a common novel multiple-locus variable-number tandem-repeat analysis-16 type (Panel 1: 3-2-7-5-3-4-12-3; Panel 2A: 6-40-8; Panel 2B: 5-2-3-3-3).

We screened raptors (n=188) and columbids (n=2) presenting to a Tampa, Florida, US rehabilitation center from June 2016 to April 2017 for Trichomonas gallinae. One juvenile Cooper's Hawk (Accipiter cooperii) was culture- and PCR-positive and sequencing identified as genotype E1. A subset of culture-negative (10%, n=19) samples were negative via PCR.

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.