Free-living wildlife across Africa is found across expansive rangelands, frequently interacting with pastoral communities, their livestock, and domestic dogs (Canis familiaris). African wildlife populations are threatened by development, poaching and disease. Infectious diseases have caused significant declines, sabotaging conservation efforts. Canine distemper virus (CDV) infections have increased in incidence in wildlife over the past four decades. Sympatric domestic dogs have been presumed to be the reservoirs of the virus for wildlife. A systematic review using Preferred Reporting Items for Systematic Reviews and Meta-Analyses 2020 guidelines was carried out to investigate CDV infections in wildlife in Africa, to 1) analyze the conservation concerns associated with the disease, 2) identify the dynamics of the virus across different ecoregions, and 3) ascertain the source of the virus for free-living wildlife. The database searches identified 65 articles relevant to the study questions and an additional 43 valuable for wider discussion of the topic area. Canine distemper outbreaks were reported to occur in wildlife in six African countries, with mortality rates between 30% and 94% across all species affected. Eleven wildlife species were identified as susceptible, 64% of which (7/11) are classified as threatened by the International Union for Conservation of Nature. Disease dynamics varied between ecoregions because of differences in land use; virus strains; climate change and concurrent infections; ecotones; domestic and wildlife population densities; wildlife monitoring capacity; and wildlife movement. Nine countries reported outbreaks in domestic dog populations in or near wildlife habitats, but none reported confirmed transmission to wildlife. Of 23 reports investigating the role of domestic dogs as CDV reservoirs, 14 (61%) concluded that domestic dogs acted as either reservoirs or sources of CDV for wildlife, four (17%) did not identify dogs as reservoirs, and five (22%) were not sure of the role played by domestic dogs. This review highlights the importance of integrating active CDV surveillance in wildlife conservation programs.

Sensitive and specific antemortem diagnostic tests are a prerequisite for effective management of chronic wasting disease (CWD). Paired with readily accessible samples that accurately reflect CWD status, the real-time quaking-induced conversion (RT-QuIC) assay has the potential to enable more effective CWD surveillance and interventions. We evaluated the feasibility of RT-QuIC as a CWD diagnostic test using 6-mm ear tissue biopsies from elk (Cervus canadensis). First, we evaluated the effect of ear spatial location on seeding activity. We observed an effect of ear punch spatial location on the amyloid formation rate (AFR): Samples collected from the periphery of the ear evidenced a statistically significant increase in AFR relative to ear punches from the ventral midline. Gross microdissection of an ear pinna suggested that there was more small nerve innervation around the periphery of the ear. Second, we evaluated the diagnostic sensitivity, specificity, and predictive value of RT-QuIC using ear punches from elk that had been previously diagnosed via ELISA testing. We evaluated the impact of nonstatistical and statistical approaches on diagnostic accuracy. Specificity and positive predictive value were perfect when statistical analyses were used to evaluate the binomial distribution (CWD positive versus CWD negative) of the data. Conversely, sensitivity and negative predictive value were modest, independent of the application of statistical analysis, indicating that RT-QuIC may be susceptible to false-negative data in this context. Taken together, our data support the idea that RT-QuIC, when paired with US Department of Agriculture-approved diagnostic tests, may provide more time to stakeholders for making major management decisions.

The European hedgehog (Erinaceus europaeus), common in rural and suburban environments, is perceived as declining within its native range, prompting conservation concerns. This study of hedgehogs admitted to a French rehabilitation center aimed to identify causes of death, detect potential emerging diseases, and systematically screen dead hedgehogs for toxicants and hemopathogens. Using clinical information, necropsy examination, and ancillary testing in 159 dead hedgehogs, we identified the primary cause of death of 92% of them. The leading causes of death were impaired general condition (30%); bacterial infections (26%), frequently caused by Salmonella Enteritidis; and trauma (20%). Additional factors contributing to death were identified in 78%, including a high proportion with significant parasite infestations. Toxicologic screening revealed that 42% of hedgehogs had been exposed to anticoagulant rodenticides, including 6.4% hedgehogs with levels compatible with acute intoxication. These hedgehogs were also exposed to trace metals, especially lead, but no pesticides residues were detected. Hemopathogen screening revealed a low diversity and prevalence of blood pathogens with 3.6, 2.9, and 2.9% individuals infected with Anaplasma phagocytophilum, Mycoplasma turicensis, and Mycoplasma wenyonii, respectively. The variety of diagnoses suggests that there is no unique prominent or emerging disease responsible for the mortality of these hedgehogs. This study also highlighted the high exposure of hedgehogs to various toxicants. To further investigate the extent and causes of population declines, health information issued from several centers should be combined with demographic data as well as ecologic assessments on the availability and quality of natural resources.

Hepatitis E virus (HEV) is a globally distributed pathogen that causes acute hepatitis in people. Recent human cases of HEV arising after contact with urban rats (Rattus spp.) have raised concerns regarding whether rats may be a source of HEV infection. We investigated whether urban Norway rats (Rattus norvegicus) could be a source of HEV in an underserved urban neighborhood of Vancouver, Canada. We found that 15% of rats tested positive for rat HEV, and that HEV status was associated with increasing rat body length and family relationships. Rat HEV isolates were clustered according to their location on either the east or west side of a busy roadway bisecting this neighborhood, suggesting that this street is a barrier to HEV spread. Widespread distribution of HEV among rats in this neighborhood poses potential human health risks, emphasizing the need to reduce close contact of people with rats and their excreta.

Emydomyces testavorans is an onygenalean fungus associated with shell and skin lesions in chelonians and is increasingly linked to morbidity and mortality. Effective disinfection methods are a critical component of biosecurity protocols to prevent and manage emydomycosis; however, disinfectant efficacy against E. testavorans is unknown. This study determined the in vitro efficacy of 13 disinfectants across a range of concentrations and exposure times used in veterinary, wildlife, and environmental settings against E. testavorans. Pure E. testavorans culture material was exposed to each disinfectant at manufacturer-recommended concentration and exposure times in triplicate. Disinfectant was removed via centrifugation and cultures were inoculated onto inhibitory media plates, maintained at room temperature (20-22 C), and examined weekly for fungal growth. After 6 wk of incubation, ethanol, Virkon S (active ingredient potassium peroxymonosulfate 21.41%), benzalkonium chloride, and desiccation proved ineffective at completely inhibiting fungal growth. All other agents, including bleach, chlorhexidine, and accelerated hydrogen peroxides, effectively inhibited fungal growth. These data suggest that several effective disinfection options are available for use against E. testavorans. These findings can be used in managed care and field settings to reduce transmission of E. testavorans between turtles through contaminated surfaces and equipment.

Transmissible spongiform encephalopathy, or prion disease, poses a serious threat to wildlife; however, the susceptibility of apex predators is still being assessed. We investigated variation in the prion protein gene in Florida panthers (Puma concolor coryi) and found that admixture from Central American pumas probably introduced a novel, albeit benign, prion allele.

Wildlife may be infected with a high diversity of piroplasms and may serve as hosts or reservoirs for piroplasms of importance for domestic animals and public health. In the US, some piroplasms (e.g., Babesia spp. and Theileira cervi) have been associated with disease in native and exotic ungulates. Blood from 447 ruminants in Nevada were PCR tested for piroplasms, with 115 individuals from five species or subspecies found positive: 28/46 California bighorn sheep (Ovis canadensis californiana), 1/16 desert bighorn sheep (Ovis canadensis nelson), 70/255 mule deer (Odocoileus hemionus), 14/53 elk (Cervis canadensis nelsoni), and 2/55 pronghorn (Antilocapra americana). Six mountain goats (Oreamnos americanus) and four moose (Alces alces) were negative. Four piroplasm species were detected-Babesia duncani, Babesia sp. RD63, Babesia odocoilei, and Theileria cervi-like spp.-but three B. duncani strains were detected, giving seven unique species or strains. Babesia duncani, the most common piroplasm detected (83%), occurred in California and desert bighorn sheep, elk, and mule deer. Bighorn sheep were only infected with B. duncani, whereas mule deer and elk were each infected with three piroplasm species or strains. The single Theileria sp. and B. odocoilei detections were in pronghorn. Free-ranging ruminants in Nevada are infected with a high diversity of piroplasms, several of which are zoonotic or reported to cause disease in both wild and domestic ruminants.

Through evaluation of serum and plasma buterylcholinesterase (BChE) and brain acetylcholinesterase (AChE) activity, we investigated the possibility of the involvement of an acute organophosphate toxicosis in the pathogenesis of ongoing annual outbreaks of paresis and paralysis that in some cases progress to death, in endangered Western Australian Carnaby's cockatoos (Zanda latirostris). The condition, named Carnaby's hindlimb paralysis syndrome (CHiPS), was first described in 2012. Following initial investigations involving clinical, epidemiologic, toxicologic, gross necropsy, and histologic evaluation, a toxic etiology, specifically an organophosphate toxicosis, was considered most likely. The study aimed to validate the BChE assay for use in serum and plasma in Carnaby's cockatoos. This study found no evidence of changes in serum or plasma BChE or brain AChE that indicate an acute organophosphate toxicosis as the cause of CHiPS. Although these results render an acute organophosphate toxicosis unlikely, an organophosphate-induced delayed neuropathy has not been ruled out. Based on the results from the BChE validation study, the authors can recommend this assay for the evaluation of BChE measurement in plasma and serum from Carnaby's cockatoos with results showing excellent accuracy and precision.

Infectious diseases can have detrimental effects on wildlife populations, particularly those that persist at small sizes, have low genetic diversity, and are affected by fragmented habitat. One such example is the endangered Florida panther (Puma concolor coryi), which has been intensively managed since the early 1980s, with the current population ranging between 120 and 230 individuals. For more than three decades, panthers have been captured, demographics recorded, and blood samples collected to monitor for multiple infectious diseases; however, an updated comprehensive study of many of these pathogens has not occurred since 1991. Our goal was to identify temporal patterns and spatial clustering in seroprevalence; determine if the pathogens of interest tend to co-occur; and describe relationships between an individual's genetic assignment (admixed or canonical) and seropositivity. We analyzed serology data for eight pathogens representing different modes of transmission (direct, indirect, vector borne) and infection duration (acute, chronic) from 232 panthers collected between 1992 and 2017. Panthers held consistently high seropositivity for feline calicivirus (62.3%) and panleukopenia virus (79.7%) throughout the study, whereas feline herpesvirus and feline leukemia virus were at lower prevalence (3.1% and 2.4%, respectively), although neither had been noted prior to 1992. Panthers were frequently seropositive for canine distemper virus and feline immunodeficiency virus, and seroprevalence fluctuated through time. West Nile virus seropositivity increased over the study period following its introduction in North America in 1999. Panthers were consistently negative for feline coronavirus, which causes feline infectious peritonitis. Genetics and demographics (sex and age) had little influence on serostatus, and coexposure among pathogens did not tend to occur. Both feline immunodeficiency virus and feline leukemia virus appeared to have spatial clusters of seropositive individuals. Our findings enhance the understanding of pathogen exposure in panthers, informing and supporting ongoing surveillance efforts for timely detection and management of potential disease threats in vulnerable populations.

Dredging is commonly used to maintain navigational channels in freshwater lakes such as Lake Erie, and the dredged material may be discarded in confined disposal facilities (CDFs). The effects of these CDFs on wildlife health are largely unknown. We compared health assessments of painted turtles (Chrysemys picta) at two sites in southwestern Lake Erie, Ohio, USA in May 2022: CDF for dredged material (n=27), and a protected coastal marsh (PCM; n=24). Each turtle underwent a physical examination, blood collection for hematology and plasma biochemistry profiles, and testing for Chlamydia spp., herpesviruses, ranavirus (frog virus 3), and Mycoplasmopsis spp. via oral and cloacal swabs. Six turtles were positive for Chlamydia spp. (11.8%; 95% confidence interval [CI]: 4.4%-23.9%), two from the CDF and four from the PCM. One Chlamydia-positive turtle was co-infected with herpesvirus (2.0%; 95% CI: 0%-10.5%) with 96.3% homology with emydid herpesvirus. No ranavirus or Mycoplasmopsis species were detected (0%, 95% CI: 0%-6.7%). Turtles captured from the CDF had significantly higher heterophil percentages, heterophil/lymphocyte ratios, and glucose levels, but significantly lower total protein and calcium/phosphorus ratios than turtles from PCM. Turtles living in the CDF were hyperglycemic, potentially indicating increased stress levels compared to the normoglycemic turtles from PCM. As humans continue to alter wetland habitats, the impact of anthropogenic sites such as CDFs on freshwater chelonian health and welfare should be monitored.

Molecular approaches are becoming more prevalent for the diagnosis of neurodegenerative diseases in human medicine and can be extended to diagnosis of wildlife diseases such as chronic wasting disease and other prion diseases. These diseases have been associated with exosome-bound molecular biomarkers of disease progression, such as proteins and micro RNA molecules (miRNA). We tested and optimized a method for exosomal miRNA isolation from minimally invasive, small-volume serum samples obtained from white-tailed deer (Odocoileus virginianus). We confirmed the isolation of exosomes and optimized a commercially available benchtop kit to obtain sufficient and pure RNA for miRNA sequencing. The selected method for RNA extraction combines two 500-μL serum aliquots into one elution column and re-eluting the final product of the column. We identified 137 miRNA present in healthy white-tailed deer that can be used as a baseline to identify putative miRNA biomarkers of disease progression and mechanisms of infection in future comparative disease studies. This approach to biomarker discovery may help to inform biological processes in wildlife populations and provide alternatives to invasive or postmortem samples.

Effects on physiology were evaluated in 49 free-ranging moose (Alces alces), 11 adult males, 36 females, two of unknown sex) in Minnesota, USA, immobilized by helicopter darting with thiafentanil (10 mg) and xylazine (30 mg) in February 2020 and March 2021. Pursuit time (PT), induction time (IT), recumbency time (RT), recovery time (RC), temperature (T), and body position were recorded. For 14 females, respiratory rate (RR), pulse rate (PR), and pulse oximetry (SpO2) were obtained before and at 1, 5, and 10 min after 4 L/min nasal oxygen supplementation was started; pH, lactate (Lac), arterial oxygen (PaO2), and carbon dioxide (PaCO2) tensions were obtained pre-treatment and 5 min post-treatment. Immobilizations were reversed with naltrexone (200 mg) and tolazoline (800 mg) intramuscularly. Median PT and IT were 4 min; a longer dart needle significantly reduced IT (P=0.0274). Most (98%) remained sternal; 77% held their head upright. Median RC was 3 min. Temperature remained ≤41.2 C. Median RR, PR, and SpO2 were RR=20 breaths/min, PR=70 beats/min, SpO2=91% pre-treatment and RR=30 breaths/min, PR=72 beats/min, SpO2=97% 5-min post-treatment. Median blood gas values were pH=7.45, Lac=5.62 mmol/L, PaCO2=44 mmHg, and PaO2=65 mmHg pre-treatment and pH=7.48, Lac=4.99 mmol/L, PaCO2=41.1 mmHg, and PaO2=78 mmHg 5-min post-treatment. Physiologic improvement from oxygen supplementation was clinically relevant but not statistically significant. All moose survived immobilization. Thiafentanil and xylazine combination provided safe and effective immobilization in free-ranging moose.

The European Bison (Bison bonasus) is the largest mammal in Europe and is classified as an endangered species. Leishmaniosis is a vector-borne disease caused by the protozoan Leishmania infantum. In general, this infection has been associated with dogs, cats, and humans. However, epidemiologic studies and reports confirm that the parasite is able to infect many other mammalian species. Recent evidence has demonstrated that ruminants in endemic areas are exposed to L. infantum infection. Moreover, climate change has allowed the northward spread of vector species, causing the expansion of L. infantum infection in regions traditionally classified as nonendemic in Europe. The aim of this study was to determine the presence or absence of anti-L. infantum antibodies in serum samples from 343 European bison in Poland, collected from 2017 to 2023. For this purpose, the presence of anti-Leishmania antibodies was analyzed using an in-house multispecies ELISA. Anti-Leishmania antibodies were detected in four animals, an overall seroprevalence of 1.17%. The results provide scientific evidence of serologic exposure to the parasite in Poland, a country previously considered nonendemic for L. infantum infection.

We investigated the prevalence of arthropod-borne viral diseases in a population of free-ranging mantled howler monkeys (Alouatta palliata) in Costa Rica in 1998. Blood samples were opportunistically collected from monkeys anesthetized for another study. Serology was performed on 64 individuals to assess exposure of this population to vesicular stomatitis virus, equine encephalitis viruses, Mayaro virus, St. Louis encephalitis virus, yellow fever virus, and dengue virus. The New Jersey serotype of vesicular stomatitis (VSV-NJ) was the only pathogen for which the population tested positive (44% [28/64]). This is the first report of antibodies against VSV-NJ in nonhuman primates in Costa Rica.

Butorphanol-azaperone-medetomidine (BAM) is commonly used for white-tailed deer (Odocoileus virginianus) immobilization in captive and free-ranging populations. It is a federally regulated controlled substance requiring stringent regulatory compliance, complicating field application. A prescription-only drug combination, nalbuphine-medetomidine-azaperone® (NalMed-A) provides a less-regulated alternative for use by wildlife professionals. Efficacy and safety of these drug combinations for immobilization of deer have not been compared in a controlled trial, and reports of dose-specific effects of NalMed-A on white-tailed deer physiology are lacking. Additionally, residual effects of these drugs on deer behavior, food consumption, and stress response have not been reported. In February through April 2021, we immobilized 30 captive female, adult white-tailed deer in three treatment groups (n=10 each). Hand-injected doses were 1.5 mL BAM intramuscularly (IM; 41.0 mg butorphanol, 13.6 mg azaperone, 16.4 mg medetomidine), 1.5 mL NalMed-A IM (60.0 mg nalbuphine, 15.0 mg medetomidine, 15.0 mg azaperone), and 2.0 mL NalMed-A IM (80.0 mg nalbuphine, 20.0 mg medetomidine, 20.0 mg azaperone). We compared quality of immobilizations and reversals and times to induction and reversal among treatments, collected biological samples to measure stress hormones and blood gases, and conducted observations to determine treatment-related variations in behaviors. When an effective dose was administered, both BAM and NalMed-A produced rapid and smooth immobilization and recovery after reversal. All treatments in combination with manual restraint caused some degree of hyperthermia, hypoxemia, hypercarbia, bradycardia, respiratory and metabolic acidosis, and elevated lactate and serum cortisol. At 60 d, all deer were still alive, with no apparent residual effects. Vital signs of deer exposed to manual restraint and these drug combinations should be monitored closely, with supportive therapy provided when needed. We suggest BAM and NalMed-A are safe for immobilizing deer in situations similar to our trials, although doses may perform differently in deer remotely injected without manual restraint.

Leptospirosis caused by the pathogenic serovars of Leptospira spp. is a zoonotic disease of global importance. Brown rats (Rattus norvegicus), due to their worldwide distribution and reservoir host status, are considered the most important reservoir for environmental leptospiral persistence and infections in animals. We aimed to isolate, identify, and characterize Leptospira spp. circulating among brown rat populations in Zaria, Kaduna state, Nigeria. Kidney samples from 305 brown rats from different locations in Zaria were collected April-June 2022 and cultured using bacteriologic methods. We also extracted DNA from 24 kidney tissue samples for molecular detection of pathogenic Leptospira spp. and for phylogenetic analysis. Leptospira spp. was detected in 57.7% of the samples by using culture methods and in 91.7% of the samples by using PCR. The phylogenetic tree revealed two distinct clusters among the reference sequences, aligning with the known P1 and P2 subclades. The sequences from this study all clustered together on the phylogenetic tree and were most similar to Leptospira interrogans serovar Pomona strain RZ 11. The high prevalence of Leptospira spp. detected in Zaria suggests that further surveillance and assessment of the public risk are warranted.

To examine whether Ostertagia mossi and Ostertagia dikmansi from white-tailed deer (Odocoileus virginianus) are separate species, we obtained DNA sequences for the cytochrome C oxidase subunit I and internal transcribed spacer 2 gene targets for phylogenetic analyses. Neither target revealed separation between morphotypes, confirming they are the same species and providing new data on Ostertagia morphotypes.

Rehabilitation and release of wild birds hinge upon a comprehensive understanding of both biologic and nonbiologic factors, including the medical and physical fitness of the animal. The objectives of this study were to determine reference intervals of packed cell volume and plasma chemistry values for healthy Eurasian Eagle Owls (Bubo bubo) and to compare blood values between age and sex classes. There were significant differences between males and females in total protein and globulin mean values, with males having higher values than females. Adult and juvenile Eagle Owls differed significantly in the mean values of calcium, alkaline phosphatase, and uric acid, with juveniles having higher values than adults. The calculated reference intervals for Eagle Owls can be used in Eurasian Eagle Owls admitted to rescue centers to assist veterinarians and wildlife managers to clinically evaluate and monitor the health of Eagle Owls and make decisions about when to release them after treatment.

Baylisascaris procyonis is a roundworm that does not affect its definitive host, raccoons (Procyon lotor), but may have a devastating effect when ingested by paratenic hosts such as Allegheny woodrats (Neotoma magister). Despite being considered a major factor in woodrat population extirpation and decline in states northward, there have been no large-scale surveys for raccoon roundworm in Virginia, US. From 2022 to 2024, we visited 60 karst or nonkarst rocky habitats known (current or historical) or suspected to be Allegheny woodrat locations in western Virginia. We opportunistically collected raccoon feces as individual piles or subsampled from visible latrines. We used fecal flotation to determine the presence and prevalence of raccoon roundworm eggs in samples collected from 44 sites across 15 counties and 1 independent city. We found evidence of this roundworm at eight (18.2%) sites in six counties. Because woodrats are especially vulnerable to fatal larval infections with B. procyonis, the presence of the parasite at multiple sites suggests conservation measures should be implemented to protect these populations.

Blanding's turtles (Emydoidea blandingii) are a species of conservation concern throughout their natural range. Headstarting is a common chelonian conservation technique in which neonates are reared in managed-care settings before release, but health assessments are rarely incorporated. From 2020 to 2021 we assessed headstarted turtle health pre-release and 1 mo, 1 yr, and 2 yr after release using physical examination, hematology, plasma biochemistry, protein electrophoresis, and pathogen detection in three Illinois counties. Results were compared to wild-reared juveniles in the same habitats. Overall, 767 assessments from 561 turtles were included. Wild-reared and 2 yr post-release headstarts had higher incidence of hemoparasites, asymmetrical nares, and increased creatine kinase and aspartate aminotransferase activities (P<0.05) compared to all other groups. Erythrocyte sedimentation rate and heterophil:lymphocyte ratio were greater, while total leukocyte and lymphocyte counts were lower (P<0.05) in pre-release headstarts compared to wild-reared juveniles. Total solids, albumin, and beta globulins were higher, while the calcium:phosphorous ratio was lower (P<0.05) in pre-release headstarts and wild-reared juveniles vs. other groups. Bile acid levels were highest in pre-release headstarts (P<0.05). Body condition and gamma globulins increased following release, while alpha globulins and the albumin:globulin ratio decreased following release (P<0.05). Two pre-release and one post-release headstart tested positive for Emydomyces testavorans, one post-release headstart was positive for Mycoplasmopsis sp., and nine post-release turtles were positive for adenoviruses. Overall, rearing conditions have a profound and temporally dynamic impact on Blanding's health assessment parameters. Future studies should evaluate long-term impacts on morbidity and mortality to support positive health status and conservation outcomes.