Disease control tools are needed to mitigate the effect of the fungal pathogen Batrachochytrium dendrobatidis (Bd) on amphibian biodiversity loss. In previous experiments, Bd metabolites (i.e., noninfectious chemicals released by Bd) have been shown to induce partial resistance to Bd when administered before live pathogen exposure and therefore have potential as an intervention strategy to curb Bd outbreaks. In the wild, however, amphibians inhabiting Bd-endemic ecosystems may have already been exposed to or infected with Bd before metabolite administration. It is therefore critical to evaluate the efficacy and safety of Bd metabolites applied postexposure to live Bd. We tested whether Bd metabolites administered postexposure would induce resistance, exacerbate infections, or have no effect. The results confirmed that Bd metabolites applied before pathogen exposure significantly reduced infection intensity, but Bd metabolites applied after pathogen exposure neither protected against nor exacerbated infections. These results reveal the importance of timing the application of Bd metabolites early in the transmission season for Bd-endemic ecosystems and emphasize that Bd metabolites prophylaxis may be a useful tool in captive reintroduction campaigns where Bd threatens the success of re-establishing endangered amphibian populations.

Antimicrobial resistance (AMR) is a critical challenge of the 21st century for public and animal health. The role of host biodiversity and the environment in the evolution and transmission of resistant bacteria between populations and species, and specifically at the wildlife-livestock-human interface, needs to be further investigated. We evaluated the AMR of commensal Escherichia coli in three mammalian herbivore species—impala (Aepyceros melampus), greater kudu (Tragelaphus strepsiceros), and plains zebra (Equus quagga)—targeting populations living under two conditions: captivity (French zoos) and free ranging (natural and private parks in Zimbabwe). From 137 fecal samples from these three host species, 328 E. coli isolates were isolated. We measured the AMR of each isolate against eight antibiotics, and we assessed the presence of AMR genes and mobile genetic element class 1 integrons (int1). Isolates obtained from captive hosts had a higher probability of being resistant than those obtained from free-ranging hosts (odds ratio, 293.8; confidence interval, 10–94,000). This statistically higher proportion of AMR bacteria in zoos than in natural parks was especially observed for bacteria resistant to amoxicillin. The percentage of int1 detection was higher when isolates were obtained from captive hosts, particularly captive impalas. Ninety percent of bacterial isolates with genes involved in antibiotic resistance also had the int1 gene. The sul1, sul2, blaTEM, and stra genes were found in 14, 19, 0, and 31%, respectively, of E. coli with respective antibiotic resistance. Finally, plains zebra carried AMR significantly more often than the other species.

A popular species in the pet trade, and therefore in the illegal wildlife trade, the diamond-backed terrapin, Malaclemys terrapin terrapin, population has seen significant declines. Associated with the illegal wildlife trade, occasions arise in which terrapins are confiscated, and no standard operating procedures exist for repatriation into the wild. To develop such procedures, an understanding of the pathogens circulating in the wild diamond-backed terrapin population in New Jersey is needed. We sampled 30 wild female diamond-backed terrapins for herpesvirus, Mycoplasmopsis, ranavirus, and intestinal and blood parasites and performed white blood cell counts and differentials and evaluated biochemistry values. Terrapins had an average age of 10 yr (8–15 yr), and 70% were gravid at the time of sampling. Thirty-three percent of the sampled northern diamond-backed terrapins were positive for Mycoplasmopsis sp., and all were negative for ranavirus and herpesviruses. Occasional blood parasites were found, and few intestinal parasites were noted. There was no significant difference between gravid status and any of the blood parameters (P<0.05). Blood chemistry values appeared to vary according to feeding activity; no differences were noted in the values in relation to gravid status. Four terrapins had heterophil to lymphocyte (H:L) ratios above 4.5, significantly higher than the other terrapins sampled, which may indicate inflammation. Two of the four had Mycoplasmopsis, one sample was contaminated by other bacteria and was discarded, and one was negative. No significant difference was found between Mycoplasmopsis infection status and H:L ratio (P=0.926). Our findings, though conducted on a small number of female terrapins at a specific time point, provide data on the pathogens that may be circulating in this population, adding to the current body of knowledge and helping to guide decision making for the reintroduction of confiscated diamond-backed terrapins into New Jersey's wild population.

Freshwater turtles are sensitive to environmental changes and can serve as sentinel species for ecosystem health assessments. The Efroymson Restoration at Kankakee Sands in northwestern Indiana, USA has been restored in the past 25 yr from primarily agricultural land to a mosaic of prairie and wetland habitats. Health assessments of 40 free-ranging painted turtles (Chrysemys picta) at Kankakee Sands were performed in May 2021 to evaluate overall health status, screen for infectious diseases, and obtain baseline clinical pathology values in this population. Assessment of each turtle included a physical examination, complete blood count, plasma biochemistry panel, blood lactate level, venous blood gas analysis, serum trace mineral panel, serum vitamin D₃ level, and plasma protein electrophoresis. Oral and cloacal swabs were tested for adenoviruses, herpesviruses, frog virus 3, and Mycoplasmopsis species by PCR in 39 painted turtles. Four turtles were positive for adenovirus, which shared 100% homology to Sulawesi tortoise adenovirus. Two turtles were herpesvirus-positive with 100% homology to emydid herpesvirus 1. No Mycoplasmopsis spp. or frog virus 3 was detected. Female turtles had significantly higher manganese, prealbumin, uric acid, triglycerides, and ionized calcium levels, while male turtles had significantly higher cholesterol, glutamate dehydrogenase, and CO₂ levels. These baseline data can be used for future research into freshwater turtle health in restored wetland habitats.

Infections can have far-reaching sublethal effects on wildlife, including reduced maintenance of external structures. For many wildlife taxa, daily maintenance of external structures (termed preening in birds) is critical to fitness, but few studies have examined how infections alter such maintenance. Mycoplasma gallisepticum is a common pathogen in free-living House Finches (Haemorhous mexicanus), where it causes mycoplasmal conjunctivitis. Despite documented behavioral changes associated with M. gallisepticum infections in finches, no studies have examined how preening behavior may change with infection and how potential differences in preening may affect feather quality. To test this, we experimentally inoculated captive House Finches with M. gallisepticum or a control treatment, and we collected behavioral and feather quality data to detect potential changes in feather maintenance due to infection. We found that finches infected with M. gallisepticum preened significantly less often, and within the infected treatment, birds with the highest conjunctivitis severity preened the least often. However, there was no difference in the quality scores for secondary flight feathers collected from control versus infected birds. We also assayed feather water retention and found that the degree of water retention correlated with our feather quality scores, such that feathers with poor scores retained more water. However, as with quality scores, feather water retention did not differ with infection; this may be due to the controlled environment that the birds experienced while in captivity. Our data suggest that, in addition to sickness behaviors previously observed in finches, M. gallisepticum infection decreases other behaviors critical to survival, such as preening. While the consequences of reduced preening on feather maintenance were not apparent in captive conditions, further work is needed to determine whether House Finches in the wild that are infected with M. gallisepticum experience a fitness cost, such as increases in ectoparasite loads, due to this reduced feather maintenance.

Sarcoptic mange epidemics erupted in two of the remaining populations of endangered San Joaquin kit foxes (Vulpes macrotis mutica). Both populations are in urban habitats in the cities of Bakersfield and Taft, California, USA. The risk of disease spread from the two urban populations to nearby nonurban populations, and then throughout the species range, is of considerable conservation concern. To date, mange has not been detected in any nonurban populations despite considerable surveillance effort. The reasons for the lack of detections of mange among nonurban foxes are unknown. We monitored urban kit fox movements using geographic positioning system (GPS) collars to test the hypothesis that urban foxes were not venturing into nonurban habitats. Of 24 foxes monitored December 2018 to November 2019, 19 (79%) made excursions from urban into nonurban habitats from 1–124 times. The mean number of excursions per 30 d was 5.5 (range 0.1–13.9 d). The mean proportion of locations in nonurban habitats was 29.0% (range 0.6–99.7%). The mean maximum distance that foxes traveled into nonurban areas from the urban-nonurban interface was 1.1 km (range 0.1–2.9 km). Mean number of excursions, proportion of nonurban locations, and maximum distance into nonurban habitats were similar between Bakersfield and Taft, females and males, and adults and juveniles. At least eight foxes apparently used dens in nonurban habitats; shared use of dens may be an important mode of mange mite transmission between conspecifics. Two of the collared foxes died of mange during the study and two others had mange when captured at the end of the study. Three of these four foxes had made excursions into nonurban habitats. These results confirm a significant potential for mange to spread from urban to nonurban kit fox populations. We recommend continued surveillance in nonurban populations and continued treatment efforts in the affected urban populations.

A combination of tiletamine–zolazepam, medetomidine, and azaperone was used to immobilize captive Chacoan peccaries (Catagonus wagneri) for health assessments and biological sample collection at the Centro Chaqueño para la Conservación e Investigación (CCCI) in the Paraguayan Chaco during July in 2017 and 2018. In total, 83 peccaries kept in 0.25–1.50 hectare enclosures were immobilized via dart-administered anesthetic. Mean animal weight was 33.89±3.74 kg (standard deviation; n=77). The mean intramuscular (IM) anesthetic drug and dosages were 0.03±0.00 mg/kg of medetomidine, 0.91±0.10 mg/kg of Zoletil 50 (tiletamine–zolazepam), and 0.30±0.03 mg/kg azaperone. The mean time to recumbency after darting was 6.07±2.65 min. The mean time to reach the anesthetic plane postdarting was 10.00±2.00 min. Muscle relaxation was adequate to allow minor veterinary procedures. A mean dosage of 0.15±0.02 mg/kg of atipamezole was given IM to reverse the medetomidine. Recoveries were smooth and animals were standing by 59.17±30.18 min postreversal. Full recovery and release back to enclosures occurred 90±30 min postreversal. A single dose of this drug combination provided adequate anesthesia for 88% of adult Chacoan peccaries; 12% needed a supplemental dose of tiletamine–zolazepam because of failure to receive the full dose from the anesthetic dart. Sex and age did not impact the dosage required to achieve immobilization. Confinement during recovery from anesthesia is required with this protocol. Aside from mild hypoxemia, no adverse effects from anesthesia were observed. However, oxygen supplementation as a part of this protocol is recommended to support circulatory and respiratory capacity.

Wild animals are commonly captured for conservation, research, and wildlife management purposes. However, capture is associated with a high risk of morbidity or mortality. Capture-induced hyperthermia is a commonly encountered complication believed to contribute significantly to morbidity and mortality. Active cooling of hyperthermic animals by dousing with water is believed to treat capture-induced pathophysiological effects, but remains untested. This study aimed to determine the pathophysiological effects of capture, and whether cooling by dousing with cold water effectively reduces these effects in blesbok (Damaliscus pygargus phillipsi). Thirty-eight blesbok were randomly allocated into three groups: a control group that was not chased (Ct, n=12), chased not cooled (CNC, n=14), and chased plus cooled group (C+C, n=12). The CNC and C+C groups were chased for 15 min prior to chemical immobilization on day 0. Animals in the C+C group were cooled with 10 L of cold water (4 C) for 10 min during immobilization. All animals were immobilized on days 0, 3, 16, and 30. During each immobilization, rectal and muscle temperatures were recorded, and arterial and venous blood samples collected. Blesbok in the CNC and C+C groups presented with capture-induced pathophysiological changes characterized by hyperthermia, hyperlactatemia, increased markers of liver, skeletal, and cardiac muscle damage, hypoxemia, and hypocapnia. Cooling effectively returned body temperatures to normothermic levels, but neither the magnitude nor the duration of the pathophysiological changes differed between the CNC and C+C groups. Therefore, at least in blesbok, capture-induced hyperthermia appears not to be the primary cause of the pathophysiological changes, but is more likely a clinical sign of the hypermetabolism resulting from capture-induced physical and psychological stress. Although cooling is still recommended to prevent the compounding cytotoxic effects of persistent hyperthermia, it is unlikely to prevent stress- and hypoxia-induced damage caused by the capture procedure.

Over the last four decades, Barred Owls (Strix varia) have expanded their range to include much of western North America, including California. This expansion is suspected to have contributed to declining populations of a closely related species, the federally threatened Northern Spotted Owl (Strix occidentalis caurina). As a result, understanding potential health threats to Barred Owls has implications for Spotted Owl health and recovery. From 2016 to 2020, 69 Barred Owls were collected to determine the apparent prevalence of periorbital nematode infection, to identify the parasite species present, and to investigate the potential pathologic effects on their hosts. The nematodes were morphologically identified as Oxyspirura and Aprocta spp. On the basis of phylogenetic analyses, they were clearly divergent from published sequences of other species within these genera. Overall, 34 (49%) Barred Owls were infected with periorbital nematodes, with Oxyspirura sp. infections being much more common (94%) than Aprocta sp. (18%). Histopathology revealed varying severity of conjunctivitis in infected owls. Despite the frequency of infection and subsequent inflammation, parasite burden was not associated with reduced body weight in these owls. As a result, the potential health effect of these nematodes is unclear. Further taxonomic characterization is needed to determine potential novelty of these nematodes.

Canine distemper virus (also known as Canine morbillivirus), the etiologic agent of canine distemper, is a highly contagious pathogen causing a multisystemic infection in carnivores globally. Canine distemper may be clinically indistinguishable from rabies, and outbreaks of either disease are major concerns. In the US, both diseases are endemic and managed by parenteral vaccination in domestic animals. In wildlife, oral vaccination and trap-vaccinate-release programs are available for rabies prevention, but no such strategies exist for canine distemper. We evaluated the prevalence at which canine distemper virus occurred concurrently in animals infected with rabies virus. Real-time quantitative reverse transcriptase PCR (qRT-PCR) was performed on specimens previously diagnosed with rabies during 2017–19 by the New York State Rabies Laboratory. Real-time qRT-PCR detected concurrent canine distemper virus infection in 73 of 1,302 animals with rabies virus. Coinfection rates were approximately 9% in Procyon lotor, 2% in Vulpes vulpes, and 0.4% in Mephitis mephitis, with an overall prevalence of 5.6%. As comorbidities in wildlife occur, laboratory-based surveillance and confirmatory testing are critical to rapid decision making for disease prevention. Rabies virus incursions are expensive and difficult to manage, and spillover events create health risks to humans and domestic animals as well as to free-roaming wildlife.

South American sea lions (Otaria byronia) and Peruvian fur seals (Arctocephalus australis) are sympatric species inhabiting the coastal Peruvian marine ecosystem. Declining abundance has prompted population health monitoring programs, including temporal monitoring of blood parameters. Several methods exist to determine total leukocyte count, yet no studies have evaluated agreement between methods in pinnipeds. We assessed agreement between total leukocyte counts determined by blood film estimate, Leuko-TIC, HemoCue, and UNOPETTE methods by using archival results from pinnipeds at Punta San Juan, Peru. Blood film estimates were prospectively performed, and resulting data were compared with retrospective leukocyte counts obtained from both species between 2009 and 2019 by using the other methodologies. Agreement in hematologic counts between methods was evaluated using Passing-Bablok regression and Bland-Altman plots (α=0.05). In total, 295 individuals (201 A. australis and 94 O. byronia) were included in the analysis. The blood film estimate method resulted in the highest leukocyte values (P<0.0001). Leuko-TIC counts were significantly higher than HemoCue counts (P<0.0001). Constant and proportional error was present in the agreement between the blood film estimate method and the other methods. Given the variation demonstrated between the different methodologies, additional research is needed to further evaluate agreement between these methodologies. The results underscore the importance of maintaining consistency in leukocyte count methodology for monitoring trends in population health over time. Method consistency may be the more important clinical consideration for assessing changes in leucocyte count over time and avoiding apparent changes depending on the methodology used.

Emergent fungal pathogens in herpetofauna are a concern in both wild and captive populations. We diagnosed dermatomycosis by Paranannizziopsis australasiensis in two panther chameleons (Furcifer pardalis) and suspected it in eight others captured from an established free-living nonnative population in Florida, USA. Chameleons developed skin lesions following recent exposure to cold weather conditions while housed in captivity, approximately 10 mo after capture and 12 wk after being placed in outdoor enclosures. Affected animals were treated with oral voriconazole and terbinafine until most cases resolved; however, medications were ultimately discontinued. Paranannizziopsis australasiensis has not previously been described in chameleons, nor in animals originating from a free-ranging population in the USA. Although the source of P. australasiensis infection is uncertain, we discuss several scenarios related to the pet trade and unique situation of chameleon “ranching” present in the USA.

Echinococcus species are zoonotic tapeworms that can impact the health of wildlife, domestic animals, livestock, and humans. Two species of interest in North America are Echinococcus multilocularis and Echinococcus canadensis (Echinococcus granulosus sensu lato). The primary wildlife definitive hosts for E. multilocularis and E. canadensis are similar, including red foxes (Vulpes vulpes), gray foxes (Urocyon cinereoargenteus), coyotes (Canis latrans), and wolves (Canis lupus). These two Echinococcus spp. use different intermediate hosts, including small mammals for E. multilocularis and artiodactylids for E. canadensis. Although historically absent from much of the eastern US, recent reports in new US states (e.g., Virginia, Vermont, Maine, Missouri) highlight the need for Echinococcus spp. surveillance in this region. During 2019–2020, 308 gastrointestinal tracts were collected from wild canids in Pennsylvania and microscopically screened for adult Echinococcus species. Two coyotes (2/155) were co-infected with both E. multilocularis and E. canadensis as determined by molecular confirmation. No red foxes (n=137) or gray foxes (n=16) were positive. These data indicate both Echinococcus species are present in Pennsylvanian coyotes, highlighting the need to better understand the ecological and epidemiological consequences for human and animal health.

Beginning in July 2019, numerous free-ranging brown anoles (Anolis sagrei), an invasive lizard species in Florida, USA, were reported with large, soft, subcutaneous masses and disfiguring facial swellings. Postmortem evaluations of six affected animals, including cytology, histology, and electron microscopy, identified the presence of myriad chain-forming coccoid bacteria surrounded by a prominent clear capsule and abundant lightly basophilic matrix material with minimal associated granulomatous inflammation and effacement of normal tissue. Standard PCR and sequencing of the lesions revealed 100% nucleotide identity to Enterococcus lacertideformus. This bacterium was first observed in 2014 as the cause of a severe, multisystemic infection in several species of lizards (geckos and skinks) on Christmas Island, an Australian external territory in the Indian Ocean. Previously, analysis of E. lacertideformus had been hindered by an inability to grow the bacterium in standard culture conditions. We successfully cultured the organism on primary anole kidney cells. Given the growing recognition of host species diversity and geographic distribution noted for this organism, there is potential concern for spread to native North American lizards, especially the green anole (Anolis carolinensis), whose population numbers have apparently decreased due to introduced brown anoles.

Rabbit hemorrhagic disease virus 2 (RHDV2 or Lagovirus GI.2) began circulating in wild lagomorph populations in the US in March 2020. To date, RHDV2 has been confirmed in several species of cottontail rabbits (Sylvilagus spp.) and hares (Lepus spp.) throughout the US. In February 2022, RHDV2 was detected in a pygmy rabbit (Brachylagus idahoensis). Pygmy rabbits are sagebrush obligates that only occur in the US Intermountain West and are a species of special concern due to the continual degradation and fragmentation of sagebrush-steppe landscapes. The spread of RHDV2 into occupied pygmy rabbit sites may pose a significant threat to their populations because of already declining numbers associated with habitat loss and high mortality rates.

Baylisascaris procyonis, or raccoon roundworm, is an intestinal nematode parasite of raccoons (Procyon lotor) that is important to public and wildlife health. Historically, the parasite was uncommon in the southeastern US; however, the range of B. procyonis has expanded to include Florida, US. From 2010 to 2016, we opportunistically sampled 1,030 raccoons statewide. The overall prevalence was 3.7% (95% confidence interval=2.5–4.8%) of sampled individuals, and infection intensity ranged from 1 to 48 (mean±standard deviation 9.9±4.0). We found raccoon roundworm in 9/56 (16%) counties sampled, and the percent positive ranged from 1.1% to 13.3% of specimens collected per county. Including previously published data, B. procyonis was detected in 11 Florida counties. We used logistic regression to estimate the contribution of raccoon demographic variables and the presence of the endoparasite Macracanthorhynchus ingens to B. procyonis detection in Florida. Following the model selection process we found housing density, M. ingens presence, and urbanicity to be predictive of raccoon roundworm presence. We also found substantial among-county variation. Raccoon sex and age were not useful predictors. Public health officials, wildlife rehabilitators, wildlife managers, and others should consider any Florida raccoon to be potentially infected with B. procyonis, particularly in areas where housing density is high.

We surveyed the presence of herpesvirus, flavivirus, and coronavirus in 20 Magnificent Frigatebirds (Fregata magnificens) from the protected Alcatrazes Island, Alcatrazes archipelago, Brazil. One adult female was positive for herpesvirus (5% occurrence; 95% confidence interval –5.5 to 15.5), whereas none of the samples were PCR-positive for flavivirus or coronavirus. The obtained herpesvirus was highly similar to the one responsible for annual mortality of Magnificent Frigatebird chicks on Grand Connétable Island, French Guiana; however, no episodes of mass mortality have been recorded in the birds from Alcatrazes. Our findings indicate that this virus may be widespread in Magnificent Frigatebirds of the southwestern Atlantic. The observed differences in morbidity and mortality may be the result of basal immunosuppression of the birds from French Guiana related to environmental or nutritional conditions. The Alcatrazes archipelago sustains the largest frigatebird breeding colony of the southern Atlantic; future monitoring studies with larger sampling sizes are needed to further determine the epidemiologic relevance of the detected herpesviruses, as well as other viruses (e.g., flaviviruses, coronaviruses, avian influenza virus), in seabirds of Alcatrazes Island.

We chemically immobilized a free-ranging moose subspecies, Shiras moose (Alces alces shirasi), using a combination of butorphanol (27.3 mg/mL), azaperone (9.1 mg/mL), and medetomidine (10.9 mg/mL). Ground and helicopter darting with fixed doses of 2 mL and 3 mL, respectively, safely immobilized 13 individuals in Wyoming, USA.

Fibropapillomatosis is a debilitating neoplastic disease associated with Chelonid alphaherpesvirus 5 (ChHV5) infection. We detected the Atlantic variant of ChHV5 associated with a fibropapilloma in a green turtle (Chelonia mydas) found stranded on the western coast of Rio de la Plata, Argentina. This is the southernmost registered case for the southwestern Atlantic.

Chuzan virus is teratogenic in cattle and causes congenital abnormalities, such as hydranencephaly and cerebellar hypoplasia, in calves. Chuzan virus seroprevalence among free-ranging and farmed cervids in South Korea was found to be 4.4% (38/873), confirming exposure to this virus in cervids in South Korea.

A wild lace monitor, Varanus varius, was euthanized due to an inoperable malignant anaplastic sarcoma that resembled hemangiosarcoma. Histopathologic examination identified metastatic spread to the lung, heart, and liver.

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