A Global Perspective on Oral Vaccination of Wildlife against Rabies.

The long-term mitigation of human-domestic animal-wildlife conflicts is complex and difficult. Over the last 50 yr, the primary biomedical concepts and actualized collaborative global field applications of oral rabies vaccination to wildlife serve as one dramatic example that revolutionized the field of infectious disease management of free-ranging animals. Oral vaccination of wildlife occurred in diverse locales within Africa, Eurasia, the Middle East, and North America. Although rabies is not a candidate for eradication, over a billion doses of vaccine-laden baits distributed strategically by hand, at baiting stations, or via aircraft, resulted in widespread disease prevention, control, or local disease elimination among mesocarnivores. Pure, potent, safe, and efficacious vaccines consisted of either modified-live, highly attenuated, or recombinant viruses contained within attractive, edible baits. Since the late 1970s, major free-ranging target species have included coyotes (Canis latrans), foxes (Urocyon cinereoargenteus; Vulpes vulpes), jackals (Canis aureus; Lupulella mesomelas), raccoons (Procyon lotor), raccoon dogs (Nyctereutes procyonoides), and skunks (Mephitis mephitis). Operational progress has occurred in all but the latter species. Programmatic evaluations of oral rabies vaccination success have included: demonstration of biomarkers incorporated within vaccine-laden baits in target species as representative of bait contact; serological measurement of the induction of specific rabies virus neutralizing antibodies, indicative of an immune response to vaccine; and most importantly, the decreasing detection of rabies virus antigens in the brains of collected animals via enhanced laboratory-based surveillance, as evidence of management impact. Although often conceived mistakenly as a panacea, such cost-effective technology applied to free-ranging wildlife represents a real-world, One Health application benefiting agriculture, conservation biology, and public health. Based upon lessons learned with oral rabies vaccination of mesocarnivores, opportunities for future extension to other taxa and additional diseases will have far-reaching, transdisciplinary benefits.

Highly Pathogenic Avian Influenza A(H5N1) Virus Clade 2.3.4.4b Infections in Wild Terrestrial Mammals, United States, 2022.

We describe the pathology of natural infection with highly pathogenic avian influenza A(H5N1) virus of Eurasian lineage Goose/Guangdong clade 2.3.4.4b in 67 wild terrestrial mammals throughout the United States during April 1‒July 21, 2022. Affected mammals include 50 red foxes (Vulpes vulpes), 6 striped skunks (Mephitis mephitis), 4 raccoons (Procyon lotor), 2 bobcats (Lynx rufus), 2 Virginia opossums (Didelphis virginiana), 1 coyote (Canis latrans), 1 fisher (Pekania pennanti), and 1 gray fox (Urocyon cinereoargenteus). Infected mammals showed primarily neurologic signs. Necrotizing meningoencephalitis, interstitial pneumonia, and myocardial necrosis were the most common lesions; however, species variations in lesion distribution were observed. Genotype analysis of sequences from 48 animals indicates that these cases represent spillover infections from wild birds.

Predation risk drives aposematic signal conformity.

Contrary to expectations regarding efficient predator education mediated by lack of ambiguity and enhanced prey recognition, aposematic signals often show considerable intraspecific variability. For example, some striped skunks (Mephitis mephitis) are almost entirely white, others have black-and-white stripes of equivalent thicknesses, yet others are mostly black. We tested the ecological correlates of this variation in patterning using 749 museum skins collected across North America. Skunks had longer white-black borders and more bilaterally symmetrical stripes in areas with a greater number of potential predator species, and this effect was more marked for mammalian than avian predators, the latter of which may be less deterred by noxious defenses. Skunks from locations with greater predator diversity were less variable in the extent of whiteness on their dorsa and less variable in the length of their white-black borders, suggesting strong selection from predators leads to greater conformity in stripe patterns, even at the same location, but weak selection from predators leads to relaxed selection on pattern conformity. Skunks exhibited greater areas of black pelage in areas of greater humidity conforming to Gloger's rule. Our results indicate that relaxed predation pressure is key to warning signal variation in this iconic species, whereas stronger pressure leads to signal conformity and stronger signals.

Pathogenic Leptospira are widespread in the urban wildlife of southern California.

Leptospirosis, the most widespread zoonotic disease in the world, is broadly understudied in multi-host wildlife systems. Knowledge gaps regarding Leptospira circulation in wildlife, particularly in densely populated areas, contribute to frequent misdiagnoses in humans and domestic animals. We assessed Leptospira prevalence levels and risk factors in five target wildlife species across the greater Los Angeles region: striped skunks (Mephitis mephitis), raccoons (Procyon lotor), coyotes (Canis latrans), Virginia opossums (Didelphis virginiana), and fox squirrels (Sciurus niger). We sampled more than 960 individual animals, including over 700 from target species in the greater Los Angeles region, and an additional 266 sampled opportunistically from other California regions and species. In the five target species seroprevalences ranged from 5 to 60%, and infection prevalences ranged from 0.8 to 15.2% in all except fox squirrels (0%). Leptospira phylogenomics and patterns of serologic reactivity suggest that mainland terrestrial wildlife, particularly mesocarnivores, could be the source of repeated observed introductions of Leptospira into local marine and island ecosystems. Overall, we found evidence of widespread Leptospira exposure in wildlife across Los Angeles and surrounding regions. This indicates exposure risk for humans and domestic animals and highlights that this pathogen can circulate endemically in many wildlife species even in densely populated urban areas.

Biodiversity indices and Random Forests reveal the potential for striped skunk (Mephitis mephitis) fecal microbial communities to function as a biomarker for oral rabies vaccination.

Wildlife disease surveillance and monitoring poses unique challenges when assessing rates of population vaccination, immunity, or infection prevalence. Non-invasively detected biomarkers can help reduce risk to both animal and field personnel during wildlife disease management activities. In this study, we investigated the utility of fecal microbiome data collected from captive striped skunks (Mephitis mephitis) in predicting rabies virus vaccination and infection status. We sequenced the hypervariable region 4 (V4) of the bacterial 16S gene and estimated alpha and beta diversity across timepoints in three groups of skunks: vaccination then rabies virus infection, sham vaccination then rabies virus infection, and rabies virus infected without vaccination. Alpha diversity did not differ among treatment groups but beta diversity between treatments was statistically significant. The phyla Firmicutes and Proteobacteria were dominant among all samples. Using Random Forests, we identified operational taxonomic units (OTUs) that greatly influenced classification of fecal samples into treatment groups. Each of these OTUs was correlated with fecal volatile organic compounds detected from the samples for companion treatment groups in another study. This research is the first to highlight striped skunk microbiome biodiversity as a vaccination biomarker which pushes the frontier on alternative methods for surveillance and monitoring of vaccination and disease in wildlife populations.

Amdoparvovirus-associated disease in striped skunks (Mephitis mephitis).

Disease caused by the archetypical amdoparvovirus (APV), Aleutian mink disease virus (AMDV), has been well studied, but APV infections in other carnivores are poorly understood. Skunk amdoparvovirus (SKAV), one of a handful of newly discovered APVs, is apparently species-specific in striped skunks (Mephitis mephitis) and has a high prevalence across North America. We have evaluated the infection status and viral tissue distribution in a cohort of 26 free-ranging California skunks from a single rehabilitation facility who were euthanized due to poor prognosis for recovery from neurologic disease. SKAV was detected in the majority of this cohort, and virus was associated with a spectrum of lesions including tubulointerstitial nephritis, meningoencephalitis, myocarditis, and arteritis. Affected tissue and patterns of inflammation were partially overlapping with those of AMDV infection but were notably distinct in the kidney.

First Report of Skunk Amdoparvovirus (Species Carnivore amdoparvovirus 4) in Europe in a Captive Striped Skunk (Mephitis mephitis).

Skunk amdoparvovirus (Carnivore amdoparvovirus 4, SKAV) is closely related to Aleutian mink disease virus (AMDV) and circulates primarily in striped skunks (Mephitis mephitis) in North America. SKAV poses a threat to mustelid species due to reported isolated infections of captive American mink (Neovison vison) in British Columbia, Canada. We detected SKAV in a captive striped skunk in a German zoo by metagenomic sequencing. The pathological findings are dominated by lymphoplasmacellular inflammation and reveal similarities to its relative Carnivore amdoparvovirus 1, the causative agent of Aleutian mink disease. Phylogenetic analysis of the whole genome demonstrated 94.80% nucleotide sequence identity to a sequence from Ontario, Canada. This study is the first case description of a SKAV infection outside of North America.

Defining County-Level Terrestrial Rabies Freedom Using the US National Rabies Surveillance System: Surveillance Data Analysis.

BACKGROUND: Rabies is a deadly zoonotic disease with nearly 100% fatality rate. In the United States, rabies virus persists in wildlife reservoirs, with occasional spillover into humans and domestic animals. The distribution of reservoir hosts in US counties plays an important role in public health decision-making, including the recommendation of lifesaving postexposure prophylaxis upon suspected rabies exposures. Furthermore, in surveillance data, it is difficult to discern whether counties have no cases reported because rabies was not present or because counties have an unreported rabies presence. These epizootics are monitored by the National Rabies Surveillance System (NRSS), to which approximately 130 state public health, agriculture, and academic laboratories report animal rabies testing statistics. Historically, the NRSS classifies US counties as free from terrestrial rabies if, over the previous 5 years, they and any adjacent counties did not report any rabies cases and they tested ≥15 reservoir animals or 30 domestic animals. OBJECTIVE: This study aimed to describe and evaluate the historical NRSS rabies-free county definition, review possibilities for improving this definition, and develop a model to achieve more precise estimates of the probability of terrestrial rabies freedom and the number of reported county-level terrestrial rabies cases. METHODS: Data submitted to the NRSS by state and territorial public health departments and the US Department of Agriculture Wildlife Services were analyzed to evaluate the historical rabies-free definition. A zero-inflated negative binomial model created county-level predictions of the probability of rabies freedom and the expected number of rabies cases reported. Data analyzed were from all animals submitted for laboratory diagnosis of rabies in the United States from 1995 to 2020 in skunk and raccoon reservoir territories, excluding bats and bat variants. RESULTS: We analyzed data from 14,642 and 30,120 county-years in the raccoon and skunk reservoir territories, respectively. Only 0.85% (9/1065) raccoon county-years and 0.79% (27/3411) skunk county-years that met the historical rabies-free criteria reported a case in the following year (99.2% negative predictive value for each), of which 2 were attributed to unreported bat variants. County-level model predictions displayed excellent discrimination for detecting zero cases and good estimates of reported cases in the following year. Counties classified as rabies free rarely (36/4476, 0.8%) detected cases in the following year. CONCLUSIONS: This study concludes that the historical rabies freedom definition is a reasonable approach for identifying counties that are truly free from terrestrial raccoon and skunk rabies virus transmission. Gradations of risk can be measured using the rabies prediction model presented in this study. However, even counties with a high probability of rabies freedom should maintain rabies testing capacity, as there are numerous examples of translocations of rabies-infected animals that can cause major changes in the epidemiology of rabies.

Coinfection of Canine Distemper Virus and Rabies Virus in Wildlife Samples Submitted for Routine Rabies Testing.

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.

Rabies surveillance in the United States during 2021.

OBJECTIVE: To provide epidemiological information on the occurrence of animal and human rabies in the US during 2021 and summaries of 2021 rabies surveillance for Canada and Mexico. PROCEDURES: State and territorial public health departments and USDA Wildlife Services provided data on animals submitted for rabies testing in 2021. Data were analyzed temporally and geographically to assess trends in domestic animal and wildlife rabies cases. RESULTS: During 2021, 54 US jurisdictions reported 3,663 rabid animals, representing an 18.2% decrease from the 4,479 cases reported in 2020. Texas (n = 456 [12.4%]), Virginia (297 [8.1%]), Pennsylvania (287 [7.8%]), North Carolina (248 [6.8%]), New York (237 [6.5%]), California (220 [6.0%]), and New Jersey (201 [5.5%]) together accounted for > 50% of all animal rabies cases reported in 2021. Of the total reported rabid animals, 3,352 (91.5%) involved wildlife, with bats (n = 1,241 [33.9%]), raccoons (1,030 [28.1%]), skunks (691 [18.9%]), and foxes (314 [8.6%]) representing the primary hosts confirmed with rabies. Rabid cats (216 [5.9%]), cattle (40 [1.1%]), and dogs (36 [1.0%]) accounted for 94% of rabies cases involving domestic animals in 2021. Five human rabies deaths were reported in 2021. CLINICAL RELEVANCE: The number of animal rabies cases reported in the US decreased significantly during 2021; this is thought to be due to factors related to the COVID-19 pandemic.

Prevalence of Trypanosoma cruzi, the Etiologic Agent of Chagas Disease, Infection in Texas Skunks (Mammalia: Mephitidae).

Background: Chagas disease is one of the world's most neglected tropical diseases, infecting over six million people across the Americas. The hemoparasite Trypanosoma cruzi is the etiological agent for the disease, circulating in domestic, peridomestic, and sylvatic transmission cycles that are maintained by triatomine vectors and a diversity of wild and synanthropic hosts. Public health and wildlife management interventions targeting the interruption of T. cruzi transmission rely on an understanding of the dynamics driving the ecology of this zoonotic pathogen. One wildlife host that purportedly plays a role in the transmission of Chagas disease within the southern United States is the striped skunk (Mephitis mephitis), although infection prevalence in this species is poorly understood. Materials and Methods: To this end, we conducted a PCR-based surveillance of T. cruzi in 235 wild skunks, representing 4 species, across 76 counties and 10 ecoregions in Texas, United States, along with an evaluation of risk factors associated with the infection. Results: We recovered an overall T. cruzi prevalence of 17.9% for all mephitid taxa aggregated, ranging between 6.7% for plains spotted skunks (Spilogale putorius interrupta) and 42.9% for western spotted skunks (Spilogale gracilis). We report the first cases of T. cruzi infection in plains spotted and American hog-nosed skunks (Conepatus leuconotus), of important note for conservation medicine since populations of both species are declining within Texas. Although not statistically significant, we also detected trends for juveniles to exhibit greater infection risk than adults and for differential sex biases in T. cruzi prevalence between taxa, which align with variations in species-specific seasonal activity patterns. No geographic or taxonomic risk factors were identified. Conclusion: Our study contributed key data for population viability analyses and epidemiologic models in addition to providing a baseline for future T. cruzi surveillance among skunks and other wildlife species.

Arthrostoma supriatnai sp. nov. (Nematoda: Ancylostomatidae) parasitic in Mydaus javanensis (Carnivora: Mephitidae) from Mount Ciremai, Java, Indonesia.

Arthrostoma supriatnai sp. nov. was described from Mydaus javanensis obtained from Mount Ciremai, Java, Indonesia. It is characterized by having a buccal capsule with ten articulated plates including a pair of additional lateral plates. To date, the genus Arthrostoma consists of eleven species that are native to Asia. Of them, only A. miyazankiense and A. tunkanati have ten articulated plates including the lateral plates. However, the present species has a much stouter body, being readily distinguishable from these two species. Moreover, its male has much shorter spicules than A. miyazakiense, and an arrow-shaped gubernaculum, differing from A. tunkanati, in which gubernaculum is distally bifid. In the female, the present species is readily distinguished from these two species in the shape and number of vulval swellings.

Field Evaluation of Tiletamine-Zolazepam-Medetomidine for Immobilization of Raccoons (Procyon lotor) and Striped Skunks (Mephitis mephitis).

The effectiveness of tiletamine-zolazepam (1.7 mg/kg) plus medetomidine (0.07 mg/kg; TZM) as an immobilizing combination in raccoons (Procyon lotor; n=43) and striped skunks (Mephitis mephitis; n=7) was evaluated during October 2019. Mean (±SD) induction time for raccoons was 15.0±6.6 min. First signs of recovery (head up) occurred 12.9±6.0 min after receiving atipamezole reversal (0.35 mg/kg) and animals were standing in 30.3±16.1 min. Mean induction time for skunks was 11.7±5.8 min. Following reversal, skunks first raised their heads in 6.7±4.3 min and stood in 17.1±12.9 min. Recovery in skunks and female raccoons was not related to length of time immobilized, but male raccoons that were immobilized for longer periods of time stood faster after reversal. Raccoon heart rate (HR) remained steady during immobilization, but respiration rate (RR) and rectal temperature (RT) declined. The HR and RR were similar among males and females, but RT of male raccoons were, on average, 0.5 C higher than those of females, and rate of temperature decline was slower for males. The HR, RR, and RT of skunks declined during immobilization. Although induction times for both raccoons and skunks were longer than expected, induction and recovery were smooth, side effects were few, analgesia was adequate for nonsurgical procedures, and reversal reduced time in captivity.

Interspecific Asymmetries in Behavioral Plasticity Drive Seasonal Patterns of Temporal Niche Partitioning in an Island Carnivore Community.

Animals vary considerably in the amount of behavioral plasticity they exhibit in daily activity timing and temporal niche switching. It is not well understood how environmental factors drive changes in temporal activity or how interspecific differences in the plasticity of activity timing ultimately manifest in free-living animals. Here, we investigated the temporal structure and organization of activity patterns of two insular mammalian carnivores living in sympatry, the island fox (Urocyon littoralis) and island spotted skunk (Spilogale gracilis amphiala). Using collar-mounted accelerometers, we assessed the plasticity of behavioral activity rhythms in foxes and skunks by investigating how environmental factors drive the distribution of locomotor activity across the day and year, and subsequently examined the dynamics of temporal niche overlap between the two species. We documented that foxes express phenotypic plasticity in daily activity timing across the year, ranging from nocturnal to diurnal to crepuscular rhythms depending on the individual and time of year. Most notably, foxes increased the proportion of daytime activity as seasonal temperatures decreased. Overall, activity patterns of foxes were consistent with the circadian thermoenergetics hypothesis, which posits that animals that switch their patterns of activity do so to coincide with the most energetically favorable time of day. In contrast to foxes, skunks exhibited little behavioral plasticity, appearing strictly nocturnal across the year. While the duration of skunk activity bouts increased with the duration of night, timing of activity onset and offset extended into daytime hours during summer when the duration of darkness was shortest. Analysis of temporal niche overlap between foxes and skunks suggested that niche overlap was highest during summer and lowest during winter and was dictated primarily by temporal niche switching in foxes, rather than skunks. Collectively, our results highlight how interspecific asymmetries in behavioral plasticity drive dynamic patterns of temporal niche overlap within an island carnivore community.

GEOGRAPHIC SPREAD OF CANINE DISTEMPER IN WILD CARNIVORES IN MICHIGAN, USA: PATHOLOGY AND EPIDEMIOLOGY, 2008-18.

Canine distemper is a widespread disease affecting both domestic and wild carnivores. This investigation of the geographic distribution, wildlife species infected, and relative prevalence rates was conducted over an 11-yr period and helps to document the disease spread, most highly infected wildlife species, and histologic lesions. Animals were collected as found dead, hunter and trapper harvested, and euthanized for displaying signs of abnormal behavior or neurologic disease. This disease appeared to spread from the Lower Peninsula of Michigan into the Upper Peninsula, was most frequently documented in raccoons (Procyon lotor), striped skunks (Mephitis mephitis), and gray fox (Urocyon cinereoargenteus), but also involved additional wildlife species. Three unique wildlife virus strains were identified. Two of these grouped within a separate subclade of the America 2 lineage. A third strain appeared to be a unique sequence type that is not associated with any existing subclade of America 2. We recommend the combined use of routine histology and immunohistochemical staining to confirm the diagnosis, and further recommend that both the lungs and spleen be collected as the optimal tissues to utilize for surveillance purposes.

Rabies surveillance in the United States during 2020.

OBJECTIVE: To provide epidemiological information on animal and human cases of rabies in the US during 2020 and summaries of 2020 rabies surveillance for Canada and Mexico. ANIMALS: All animals submitted for laboratory diagnosis of rabies in the US during 2020. PROCEDURES: State and territorial public health departments and USDA Wildlife Services provided 2020 rabies surveillance data. Data were analyzed temporally and geographically to assess trends in domestic and wildlife rabies cases. RESULTS: During 2020, 54 jurisdictions submitted 87,895 animal samples for rabies testing, of which 85,483 (97.3%) had a conclusive (positive or negative) test result. Of these, 4,479 (5.2%) tested positive for rabies, representing a 4.5% decrease from the 4,690 cases reported in 2019. Texas (n = 580 [12.9%]), Pennsylvania (371 [8.3%]), Virginia (351 [7.8%]), New York (346 [7.7%]), North Carolina (301 [6.7%]), New Jersey (257 [5.7%]), Maryland (256 [5.7%]), and California (248 [5.5%]) together accounted for > 60% of all animal rabies cases reported in 2020. Of the total reported rabid animals, 4,090 (91.3%) involved wildlife, with raccoons (n = 1,403 [31.3%]), bats (1,400 [31.3%]), skunks (846 [18.9%]), and foxes (338 [7.5%]) representing the primary hosts confirmed with rabies. Rabid cats (288 [6.4%]), cattle (43 [1.0%]), and dogs (37 [0.8%]) accounted for 95% of rabies cases involving domestic animals in 2020. No human rabies cases were reported in 2020. CONCLUSIONS AND CLINICAL RELEVANCE: For the first time since 2006, the number of samples submitted for rabies testing in the US was < 90,000; this is thought to be due to factors related to the COVID-19 pandemic, as similar decreases in sample submission were also reported by Canada and Mexico.

Natural disease and evolution of an Amdoparvovirus endemic in striped skunks (Mephitis mephitis).

Striped skunks (Mephitis mephitis) densely populate the human-animal interface of suburbia throughout North America. Skunks share that habitat with numerous related mesocarnivores, where increased contact, competition for shared food and water sources and other stressors contribute to increased exposure and susceptibility to viral infection. The recently identified skunk amdoparvovirus (SKAV) has been detected at high prevalence in skunks and occasionally in mink, but its distribution in North America is unknown. To understand the impact of SKAV in striped skunks and the risk posed to related species, we investigated the geographic distribution of SKAV, analysed its genetic diversity and evolutionary dynamics and evaluated viral distribution in tissues of infected animals to identify possible mechanisms of transmission. SKAV was detected in 72.5% (37/51) skunks and was present at high rates at all locations tested across North America. Analysis of the complete genomic sequence of 29 strains showed a clear geographic segregation, frequent recombination and marked differences in the evolutionary dynamics of the major structural (VP2) and non-structural (NS1) proteins. NS1 was characterized by a higher variability and a higher percentage of positively selected codons. This could indicate that antibody-mediated enhancement of infection occurs in SKAV, an infection strategy that may be conserved across amdoparvoviruses. Finally, in situ hybridization revealed virus in epithelium of the gastrointestinal tract, urinary tract and skin, indicating that viral transmission could occur via oronasal, faecal and/or urinary secretions, as well as from skin and hair. The endemicity of SKAV over large geographic distances and its high genetic diversity suggest a long-term virus-host association. Persistent shedding and high environmental stability likely contribute to efficient viral spread, simultaneously offering opportunities for cross-species transmission with consequent risk to sympatric species, including domestic animals and wildlife.

The New Face of Human Rabies in Mexico, What's Next After Eradicating Rabies in Dogs.

In pre-Hispanic Mexico, dogs were not identified as an important source of rabies. We know from codexes and chronicles of the conquerors that at that time, rabies cases in humans and domestic animals were caused by local wildlife species such as bats. Canine-rabies virus variant seems to have arrived with Europeans. The first documented case of canine rabies in the Americas is found in Mexico in the Annals of the Holy Inquisition (16th century). During Mexico's independence, cases were frequently reported. In the 19th century the first attempts to control human rabies were made through sanitary measures such as elimination of rabid dogs and applying postexposure vaccination. During the first half of the 20th century, the efficacy of canine vaccination to prevent human rabies was established. However, in Mexico, despite reports of numerous human cases (>70/year), canine vaccination did not have enough coverage. It was only during the 1990s that Mexico made a serious commitment to eliminate dog-transmitted human rabies. Since the beginning, vaccination campaigns have been free and massive. Coverage increased from 7,100,000 doses in 1990 to more than 18,000,000 since 2017. This culminated in the elimination of dog-mediated human rabies cases since 2006. Subsequently, the epidemiology of rabies had changed. Nowadays, it is wildlife species (mainly bats and skunks) that are the source of human rabies. As a mega-biodiverse country Mexico has numerous wildlife species with potential to transmit rabies virus. Thus it is paramount to remain vigilant with respect to canine vaccination campaigns and to promote rabies research in wildlife.

IN SITU HYBRIDIZATION AND VIRUS CHARACTERIZATION OF SKUNK ADENOVIRUS IN NORTH AMERICAN WILDLIFE REVEALS MULTISYSTEMIC INFECTIONS IN A BROAD RANGE OF HOSTS.

Skunk adenovirus-1 (SkAdV-1) has been reported infecting several North American wildlife species; however, lesions associated with disease have not yet been completely characterized, particularly in porcupines. We describe and characterize the tissue distribution and lesions associated with SkAdV-1 infection in 24 wildlife diagnostic cases submitted between 2015 and 2020, including 16 North American porcupines (Erethizon dorsatum), three striped skunks (Mephitis mephitis), and five raccoons (Procyon lotor), which constitute a new host species. The most common lesion in all species was severe necrotizing bronchopneumonia with (n=12) or without (n=10) interstitial involvement. Intranuclear inclusion bodies were common in respiratory epithelium (n=21) and less often in renal tubular (n=6) and biliary epithelium (n=1). Several cases (n=4) had secondary bacterial infections, including Bordetella bronchiseptica, Pasteurella multocida, and Streptococcus zooepidemicus. In situ hybridization in porcupine (n=6), raccoon (n=1), and skunk (n=1) revealed SkAdV-1 DNA in multiple tissue types, including lung, trachea, turbinates, liver, kidney, lymph node, and brain, and multiple cell types including epithelial, endothelial, and mesothelial cells. These findings were consistent across species. Comparison of viral genomes from a porcupine and a raccoon with that originally isolated from a skunk demonstrated DNA point mutations affecting several viral genes, including the fiber protein gene. Our findings show the spectrum of disease associated with SkAdV-1 infection in a broad host range of wildlife species.