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Rabies virus (RABV) is a deadly zoonosis that circulates in wild carnivore populations in North America. Intensive management within the USA and Canada has been conducted to control the spread of the raccoon (Procyon lotor) variant of RABV and work towards elimination. We examined RABV occurrence across the northeastern USA and southeastern Québec, Canada during 2008–2018 using a multi-method, dynamic occupancy model. Using a 10 km × 10 km grid overlaid on the landscape, we examined the probability that a grid cell was occupied with RABV and relationships with management activities (oral rabies vaccination (ORV) and trap-vaccinate-release efforts), habitat, neighbour effects and temporal trends. We compared raccoon RABV detection probabilities between different surveillance samples (e.g. animals that are strange acting, road-kill, public health samples). The management of RABV through ORV was found to be the greatest driver in reducing the occurrence of rabies on the landscape. Additionally, RABV occupancy declined further with increasing duration of ORV baiting programmes. Grid cells north of ORV management were at or near elimination ( $\hat{\psi }_{{\rm north}}$ = 0.00, s.e. = 0.15), managed areas had low RABV occupancy ( $\hat{\psi }_{{\rm managed}}$ = 0.20, s.e. = 0.29) and enzootic areas had the highest level of RABV occupancy ( $\hat{\psi }_{{\rm south}}$ = 0.83, s.e. = 0.06). These results provide evidence that past management actions have been being successful at the goals of reducing and controlling the raccoon variant of RABV. At a finer scale we also found that vaccine bait type and bait density impacted RABV occupancy. Detection probabilities varied; samples from strange acting animals and public health had the highest detection rates. Our results support the movement of the ORV zone south within the USA due to high elimination probabilities along the US border with Québec. Additional enhanced rabies surveillance is still needed to ensure elimination is maintained.

With rates of psychiatric illnesses such as depression continuing to rise, additional preclinical models are needed to facilitate translational neuroscience research. In the current study, the raccoon (Procyon lotor) was investigated due to its similarities with primate brains, including comparable proportional neuronal densities, cortical magnification of the forepaw area, and cortical gyrification. Specifically, we report on the cytoarchitectural characteristics of raccoons profiled as high, intermediate, or low solvers in a multi-access problem-solving task. Isotropic fractionation indicated that high-solvers had significantly more cells in the hippocampus (HC) than the other solving groups; further, a nonsignificant trend suggested that this increase in cell profile density was due to increased non-neuronal (e.g., glial) cells. Group differences were not observed in the cellular density of the somatosensory cortex. Thionin-based staining confirmed the presence of von Economo neurons (VENs) in the frontoinsular (FI) cortex, although no impact of solving ability on VEN cell profile density levels was observed. Elongated fusiform cells were quantified in the hippocampus dentate gyrus where high-solvers were observed to have higher levels of this cell type than the other solving groups. In sum, the current findings suggest that varying cytoarchitectural phenotypes contribute to cognitive flexibility. Additional research is necessary to determine the translational value of cytoarchitectural distribution patterns on adaptive behavioral outcomes associated with cognitive performance and mental health. This article is protected by copyright. All rights reserved.

Oral baiting is used to deliver vaccines to wildlife to prevent, control, and eliminate infectious diseases. A central challenge is how to spatially distribute baits to maximize encounters by target animal populations, particularly in urban and suburban areas where wildlife such as raccoons (Procyon lotor) are abundant and baits are delivered along roads. Methods from movement ecology that quantify movement and habitat selection could help to optimize baiting strategies by more effectively targeting wildlife populations across space. We developed a spatially explicit, individual-based model of raccoon movement and oral rabies vaccine seroconversion to examine whether and when baiting strategies that match raccoon movement patterns perform better than currently used baiting strategies in an oral rabies vaccination zone in greater Burlington, Vermont, USA. Habitat selection patterns estimated from locally radio-collared raccoons were used to parameterize movement simulations. We then used our simulations to estimate raccoon population rabies seroprevalence under currently used baiting strategies (actual baiting) relative to habitat selection-based baiting strategies (habitat baiting). We conducted simulations on the Burlington landscape and artificial landscapes that varied in heterogeneity relative to Burlington in the proportion and patch size of preferred habitats. We found that the benefits of habitat baiting strongly depended on the magnitude and variability of raccoon habitat selection and the degree of landscape heterogeneity within the baiting area. Habitat baiting improved seroprevalence over actual baiting for raccoons characterized as habitat specialists but not for raccoons that displayed weak habitat selection similar to radiocollared individuals, except when baits were delivered off roads where preferred habitat coverage and complexity was more pronounced. In contrast, in artificial landscapes with either more strongly juxtaposed favored habitats and/or higher proportions of favored habitats, habitat baiting performed better than actual baiting, even when raccoons displayed weak habitat preferences and where baiting was constrained to roads. Our results suggest that habitat selection-based baiting could increase raccoon population seroprevalence in urban-suburban areas, where practical, given the heterogeneity and availability of preferred habitat types in those areas. Our novel simulation approach provides a flexible framework to test alternative baiting strategies in multiclass landscapes to optimize bait-distribution strategies.

OBJECTIVE To evaluate species identification and rabies virus (RABV) characterization among samples from bats submitted for rabies testing in the United States and assess whether a standardized approach to specimen selection for RABV characterization could enhance detection of a sentinel event in virus dissemination among bats. SAMPLE United States public health rabies surveillance system data collected in January 2010 through December 2015. PROCEDURES The number of rabies-tested bats for which species was reported and the number of RABV-positive samples for which virus characterization would likely provide information regarding introduction of novel RABV variants and translocation and host-shift events were calculated. These specimens were designated as specimens of epizootiological importance (SEIs). Additionally, the estimated test load that public health laboratories could expect if all SEIs underwent RABV characterization was determined. RESULTS Species was reported for 74,928 of 160,017 (47%) bats submitted for rabies testing. Identified SEIs were grouped in 3 subcategories, namely nonindigenous bats; bats in southern border states, Florida, Puerto Rico, and the US Virgin Islands; and bats of species that are not commonly found to be inflected with RABV. Annually, 692 (95% CI, 600 to 784) SEIs were identified, of which only 295 (95% CI, 148 to 442) underwent virus characterization. Virus characterization of all SEIs would be expected to increase public health laboratories’ overall test load by 397 (95% CI, 287 to 506) samples each year. CONCLUSIONS AND CLINICAL RELEVANCE Species identification and RABV characterization may aid detection of a sentinel event in bat RABV dissemination. With additional resources, RABV characterization of all SEIs as a standardized approach to testing could contribute to knowledge of circulating bat RABV variants.

ContextDiseases are increasingly contributing to wildlife population declines. Tasmanian devil (Sarcophilus harrisii) populations have locally declined by 82% largely due to the morbidity and mortality associated with two independent transmissible devil facial tumours (DFT1 and DFT2). Toxic baits are often used as a management tool for controlling vertebrate pest populations in Australia, but in other areas of the world oral baits are also used to deliver vaccines or pharmaceuticals to wildlife. Oral vaccine bait products have been distributed for more than 50 years at a landscape scale to protect wildlife from rabies virus and contributed to the elimination of fox rabies from more than ten European nations. An oral bait vaccine to protect against devil facial tumours has been proposed as a management tool to improve the population health, resiliency and fitness of wild Tasmanian devils.AimOur goal was to evaluate the potential use of edible baits as vehicles for vaccine delivery to Tasmanian devils.MethodWe tested placebo versions of baits that are already used in Australia.Key resultsCaptive devils consumed all types of placebo baits but exhibited a preference for ruminant- and fish-based baits. Captive devils also consumed inert capsules inserted into placebo baits. Bait fate trials in the field revealed that baits were generally removed within 24 hours. Tasmanian pademelons (Thylogale billardierii), brushtail possums (Trichosurus vulpecula), and Eastern quolls (Dasyurus viverrinus) were the most common nontarget bait competitors at six private properties in southern Tasmania; wild devils removed approximately 5% of ground baits at these sites. We also evaluated an automated bait dispenser, which reduced nontarget uptake and resulted in over 50% of the baits being removed by devils.ConclusionsThis study demonstrates that captive and wild devils will accept and consume placebo versions of commercial baits. Bait dispensers or modified baits or baiting strategies are needed to increase bait uptake by devils.ImplicationsBait dispensers can be used at a regional scale to deliver baits to Tasmanian devils. These could act as vehicles for preventive or therapeutic vaccines to mitigate the impacts of disease on devil populations.Short summaryThis study aimed to test oral baits as potential vaccine delivery vehicles for Tasmanian devils. Captive and wild devils consumed placebo versions of commercial baits used on mainland Australia. Abundant non-target species, such as brushtail possums, Tasmanian pademelons, and eastern quolls consumed most baits in the wild. Implementation of automated bait dispensers increased bait uptake by devils to over 50% at the same regional field sites.

ContextEthyl-Iophenoxic acid (Et-IPA) has been widely used as a bait biomarker to determine oral bait consumption by vertebrate wildlife species. Oral bait vaccines have been delivered to numerous wildlife species to protect them from disease. The Tasmanian devil (Sarcophilis harrisii), the largest extant carnivorous marsupial species, is threatened by the transmissible cancers known as devil facial tumour disease (DFTD). Development of a protective DFTD vaccine is underway, and an oral bait has been proposed to deliver the vaccine in the wild. The bait delivery system requires a biomarker that can be detected for several months post-consumption in Tasmanian devils.AimTo determine the suitability of Et-IPA as a bait biomarker in the Tasmanian devil.MethodTwo Tasmanian devils were fed 50 mg Et-IPA (4.5 to 7.1 mg Et-IPA/kg bodyweight). Liquid chromatography with tandem mass spectrometry (LC-MS/-MS) was used to directly measure Et-IPA in baseline serum samples and samples collected on days 1, 14, 26 and 56 post-baiting.Key resultBoth devils retained serum Et-IPA concentrations at two orders of magnitude above negative control sera when this study concluded.ConclusionsEt-IPA is a useful bait biomarker for Tasmanian devils and can be included in future DFTD bait vaccine field trials to determine bait vaccine uptake.

Oral baiting is used to deliver vaccines to wildlife to prevent, control, and eliminate infectious diseases. A central challenge is how to spatially distribute baits to maximize encounters by target animal populations, particularly in urban and suburban areas where wildlife like raccoons (Procyon lotor) are abundant and baits are delivered along roads. Methods from movement ecology that quantify movement and habitat selection could help to optimize baiting strategies by more effectively targeting wildlife populations across space. We developed a spatially explicit, individual-based model of raccoon movement and oral rabies vaccine seroconversion to examine whether and when baiting strategies that match raccoon movement patterns perform better than currently employed baiting strategies in an oral rabies vaccination zone in greater Burlington, Vermont, USA. Habitat selection patterns estimated from locally radio-collared raccoons were used to parameterize movement simulations. We then used our simulations to estimate raccoon population rabies seroprevalence under currently used baiting strategies (actual baiting) relative to habitat selection-based baiting strategies (habitat baiting). We conducted simulations on the Burlington landscape and artificial landscapes that varied in heterogeneity relative to Burlington in the proportion and patch size of preferred habitats. We found that the benefits of habitat baiting strongly depended on the magnitude and variability of raccoon habitat selection and the degree of landscape heterogeneity within the baiting area. Habitat baiting improved seroprevalence over actual baiting for raccoons characterized as habitat specialists but not for raccoons that displayed weak habitat selection similar to radio-collared individuals—except when baits were delivered off roads where preferred habitat coverage and complexity was more pronounced. In contrast, in artificial landscapes with either more strongly juxtaposed favored habitats and/or higher proportions of favored habitats, habitat baiting performed better than actual baiting, even when raccoons displayed weak habitat preferences and where baiting was constrained to roads. Our results suggest that habitat selection-based baiting could increase raccoon population seroprevalence in urban-suburban areas, where practical, given the heterogeneity and availability of preferred habitat types in those areas. Our novel simulation approach provides a flexible framework to test alternative baiting strategies in multiclass landscapes to optimize bait distribution strategies.

Rabies lyssavirus (RABV) is enzootic in raccoons across the eastern United States. Intensive management of RABV by oral rabies vaccination (ORV) has prevented its spread westward and shown evidence of local elimination in raccoon populations of the northeastern US. The USDA, Wildlife Services, National Rabies Management Program (NRMP) collaborates with other agencies to implement broad-scale ORV and conducts extensive monitoring to measure the effectiveness of the management. Enhanced Rabies Surveillance (ERS) was initiated during 2005 and updated in 2016 to direct surveillance efforts toward higher-value specimens by assigning points to different methods of encountering specimens for collection (strange-acting, roadkill, surveillance-trapped, etc., specimen point values ranged from 1 to 15). We used the 2016–2019 data to re-evaluate the point values using a dynamic occupancy model. Additionally, we used ERS data from 2012–2015 and 2016–2019 to examine the impact that the point system had on surveillance data. Implementation of a point system increased positivity rates among specimens by 64%, indicating a substantial increase in the efficiency of the ERS to detect wildlife rabies. Our re-evaluation found that most points accurately reflect the value of the surveillance specimens. The notable exception was that samples from animals found dead were considerably more valuable for rabies detection than originally considered (original points = 5, new points = 20). This work demonstrates how specimen prioritization strategies can be used to refine and improve ERS in support of wildlife rabies management.

A necessary component of elimination programmes for wildlife disease is effective surveillance. The ability to distinguish between disease freedom and non‐detection can mean the difference between a successful elimination campaign and new epizootics. Understanding the contribution of different surveillance methods helps to optimize and better allocate effort and develop more effective surveillance programmes. We evaluated the probability of rabies virus elimination (disease freedom) in an enzootic area with active management using dynamic occupancy modelling of 10 years of raccoon rabies virus (RABV) surveillance data (2006–2015) collected from three states in the eastern United States. We estimated detection probability of RABV cases for each surveillance method (e.g. strange acting reports, roadkill, surveillance‐trapped animals, nuisance animals and public health samples) used by the USDA National Rabies Management Program. Strange acting, found dead and public health animals were the most likely to detect RABV when it was present, and generally detectability was higher in fall–winter compared to spring–summer. Found dead animals in fall–winter had the highest detection at 0.33 (95% CI: 0.20, 0.48). Nuisance animals had the lowest detection probabilities (~0.02). Areas with oral rabies vaccination (ORV) management had reduced occurrence probability compared to enzootic areas without ORV management. RABV occurrence was positively associated with deciduous and mixed forests and medium to high developed areas, which are also areas with higher raccoon (Procyon lotor) densities. By combining occupancy and detection estimates we can create a probability of elimination surface that can be updated seasonally to provide guidance on areas managed for wildlife disease. Synthesis and applications. Wildlife disease surveillance is often comprised of a combination of targeted and convenience‐based methods. Using a multi‐method analytical approach allows us to compare the relative strengths of these methods, providing guidance on resource allocation for surveillance actions. Applying this multi‐method approach in conjunction with dynamic occupancy analyses better informs management decisions by understanding ecological drivers of disease occurrence.

Few studies have investigated the ecology of urban striped skunks (Mephitis mephitis) despite their role as a primary rabies vector species paired with an ability to thrive in these landscapes. Information on home range, nightly movements, and habitat selection, is important for rabies management planning regarding the placement of oral rabies vaccine (ORV) baits and for management of skunk populations more generally. Our aim was to obtain baseline ecological information with an emphasis on spatial ecology of urban striped skunks in the Northern Great Plains region that is lacking in the literature. We used radio telemetry to track 22 (4 M, 18 F) skunks during September 2016 to November 2016 and March 2017 to November 2017. Size of home range using kernel density estimation with smoothing by least squares cross validation identified that male skunks ( $$ \overline{x} $$ = 4.36 km2, SE ± 0.79) had larger home ranges than females ( $$ \overline{x} $$ = 1.79 km2, SE ± 0.24). Female skunk home ranges varied by season with the largest home ranges detected during summer 2017. We found no evidence that use-availability was associated with habitat type or season among female skunks. Female skunks were more likely to be found closer to water; however, we found no association of use with road characteristics and geographic location within the study area. Targeting ORV efforts in areas near water may be warranted, but considering differences in habitat selection for skunk studies in urban areas, we caution that ORV baiting programs may not be one-size-fits-all, and a framework for effective bait placement would benefit from studies specific to the management area of interest.

Innovative problem solving, repeated innovation, learning, and inhibitory control are cognitive abilities commonly regarded as important components of behaviorally flexible species. Animals exhibiting these cognitive abilities may be more likely to adapt to the unique demands of living in novel and rapidly changing environments, such as urbanized landscapes. Raccoons (Procyon lotor) are an abundant, generalist species frequently found in urban habitats, and are capable of innovative problem solving, which makes them an ideal species to assess their behavioral flexibility. We gave 20 captive raccoons a multi-access puzzle box to investigate which behavioral and cognitive mechanisms enable the generation of innovative and flexible behaviors in this species. Over two-thirds of raccoons tested were not only capable of innovative problem solving, but displayed repeated innovation by solving more than one solution on the multi-access puzzle box and demonstrated that they learned multiple solutions to a novel problem. Although we found no relationship between our measure of inhibitory control and a raccoon's ability to exhibit repeated innovations, we did find a positive relationship between the diversity of behaviors that an individual exhibited when interacting with the problem and the number of solution types that they solved. We identified other predictors of problem-solving performance, including neophobia and persistence. Finally, we examine the implications of our results in the context of the cognitive-buffer hypothesis and consider whether the widespread success of an adaptive generalist carnivore could be due in part to having these cognitive and behavioral traits.

The arctic fox variant of the rabies virus (RABV) is enzootic in the circumpolar north. Reports of abortive RABV exposures motivated a retrospective analysis of sera from 41 arctic foxes (Vulpes lagopus) captured at Karrak Lake in Nunavut, Canada, during 2011-15. Estimated RABV antibody prevalence among foxes was 15% (95% confidence interval, 7-28%).

The Ontario Rabies Vaccine (ONRAB) is a human adenovirus rabies glycoprotein recombinant oral vaccine immunogenic for small Indian mongooses when delivered by direct instillation into the oral cavity. We offered Ultralite baits containing ~1.8 mL 109.5 TCID50 ONRAB oral rabies vaccine to 18 mongooses, while 6 mongooses were offered identical baits in placebo form. We collected sera from individual mongooses at days 0, 14 and 30 post vaccination (pv) and quantified rabies virus neutralizing antibodies (RVNA) using the rapid fluorescent focus inhibition test, with titers greater than or equal to 0.1 IU/mL considered positive. All study subjects were RVNA negative prior to bait offering. Bait consumption was variable: all 6 sham and 13 of 18 (72%) treatment animals consumed/punctured the baits offered. By day 30 pv, RVNA were detected among 11 of 13 (84.6%) of treatment mongooses that consumed/punctured baits, whereas sham-vaccinated mongooses remained RVNA negative throughout the study. We conclude ONRAB is immunogenic for mongooses by Ultralite bait delivery, although the bait design may need further optimization.

Since the 1990s, oral rabies vaccination (ORV) has been used successfully to halt the westward spread of the raccoon rabies virus (RV) variant from the eastern continental USA. Elimination of raccoon RV from the eastern USA has proven challenging across targeted raccoon (Procyon lotor) and striped skunk (Mephitis mephitis) populations impacted by raccoon RV. Field trial evaluations of the Ontario Rabies Vaccine Bait (ONRAB) were initiated to expand ORV products available to meet the rabies management goal of raccoon RV elimination. This study describes the continuation of a 2011 trial in West Virginia. Our objective was to evaluate raccoon and skunk response to ORV occurring in West Virginia for an additional two years (2012&ndash, 2013) at 75 baits/km2 followed by three years (2014&ndash, 2016) of evaluation at 300 baits/km2. We measured the change in rabies virus-neutralizing antibody (RVNA) seroprevalence in targeted wildlife populations by comparing levels pre- and post-ORV during each year of study. The increase in bait density from 75/km2 to 300/km2 corresponded to an increase in average post-ORV seroprevalence for raccoon and skunk populations. Raccoon population RVNA levels increased from 53% (300/565, 95% CI: 50&ndash, 57%) to 82.0% (596/727, 95% CI: 79&ndash, 85%) during this study, and skunk population RVNA levels increased from 11% (8/72, 95% CI: 6&ndash, 20%) to 39% (51/130, 95% CI: 31&ndash, 48%). The RVNA seroprevalence pre-ORV demonstrated an increasing trend across study years for both bait densities and species, indicating that multiple years of ORV may be necessary to achieve and maintain RVNA seroprevalence in target wildlife populations for the control and elimination of raccoon RV in the eastern USA.

Background Rabies is a viral disease of animals and people causing fatal encephalomyelitis if left untreated. Although effective pre- and post-exposure vaccines exist, they are not widely available in many endemic countries within Africa. Since many individuals in these countries remain at risk of infection, post-exposure healthcare-seeking behaviors are crucial in preventing infection and warrant examination. Methodology A rabies knowledge, attitudes, and practices survey was conducted at 24 geographically diverse sites in Uganda during 2013 to capture information on knowledge concerning the disease, response to potential exposure events, and vaccination practices. Characteristics of the surveyed population and of the canine-bite victim sub-population were described. Post-exposure healthcare-seeking behaviors of canine-bite victims were examined and compared to the related healthcare-seeking attitudes of non-bite victim respondents. Wealth scores were calculated for each household, rabies knowledge was scored for each non-bitten survey respondent, and rabies exposure risk was scored for each bite victim. Logistic regression was used to determine the independent associations between different variables and healthcare-seeking behaviors among canine-bite victims as well as attitudes of non-bitten study respondents. Results A total of 798 households were interviewed, capturing 100 canine-bite victims and a bite incidence of 2.3 per 100 person-years. Over half of bite victims actively sought medical treatment (56%), though very few received rabies post-exposure prophylaxis (3%). Bite victims who did not know or report the closest location where PEP could be received were less likely to seek medical care (p = 0.05). Respondents who did not report having been bitten by a dog with higher knowledge scores were more likely to respond that they would both seek medical care (p = 0.00) and receive PEP (p = 0.06) after a potential rabies exposure event. Conclusions There was varying discordance between what respondents who did not report having been bitten by a dog said they would do if bitten by a dog when compared to the behaviors exhibited by canine-bite victims captured in the KAP survey. Bite victims seldom elected to wash their wound or receive PEP. Having lower rabies knowledge was a barrier to theoretically seeking care and receiving PEP among not bitten respondents, indicating a need for effective and robust educational programs in the country.

Preventing zoonotic emergence from bats requires integrative research

The COVID-19 pandemic highlights the substantial public health, economic, and societal consequences of virus spillover from a wildlife reservoir. Widespread human transmission of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) also presents a new set of challenges when considering viral spillover from people to naïve wildlife and other animal populations. The establishment of new wildlife reservoirs for SARS-CoV-2 would further complicate public health control measures and could lead to wildlife health and conservation impacts. Given the likely bat origin of SARS-CoV-2 and related beta-coronaviruses (β-CoVs), free-ranging bats are a key group of concern for spillover from humans back to wildlife. Here, we review the diversity and natural host range of β-CoVs in bats and examine the risk of humans inadvertently infecting free-ranging bats with SARS-CoV-2. Our review of the global distribution and host range of β-CoV evolutionary lineages suggests that 40+ species of temperate-zone North American bats could be immunologically naïve and susceptible to infection by SARS-CoV-2. We highlight an urgent need to proactively connect the wellbeing of human and wildlife health during the current pandemic and to implement new tools to continue wildlife research while avoiding potentially severe health and conservation impacts of SARS-CoV-2 "spilling back" into free-ranging bat populations.

Introduction: The Tasmanian devil (Sarcophilus harrisii) is the largest extant carnivorous marsupial. Since 1996, its population has declined by 77% primarily due to a clonal transmissible tumor, known as devil facial tumor (DFT1) disease. In 2014, a second transmissible devil facial tumor (DFT2) was discovered. DFT1 and DFT2 are nearly 100% fatal.Areas covered: We review DFT control approaches and propose a rabies-style oral bait vaccine (OBV) platform for DFTs. This approach has an extensive safety record and was a primary tool in large-scale rabies virus elimination from wild carnivores across diverse landscapes. Like rabies virus, DFTs are transmitted by oral contact, so immunizing the oral cavity and stimulating resident memory cells could be advantageous. Additionally, exposing infected devils that already have tumors to OBVs could serve as an oncolytic virus immunotherapy. The primary challenges may be identifying appropriate DFT-specific antigens and optimization of field delivery methods.Expert opinion: DFT2 is currently found on a peninsula in southern Tasmania, so an OBV that could eliminate DFT2 should be the priority for this vaccine approach. Translation of an OBV approach to control DFTs will be challenging, but the approach is feasible for combatting ongoing and future disease threats.

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 (

Rabies is a zoonotic disease caused by infection with rabies virus, which circulates naturally in several wild carnivore and bat reservoirs in the United States (US). The most important reservoir in the US from an animal and public health perspective is the raccoon (Procyon lotor). To prevent the westward expansion of a significant raccoon rabies epizootic along the eastern seaboard, an operational control program implementing oral rabies vaccination (ORV) has existed in the US since the 1990s. Recently, two vaccine efficacy studies conducted with raccoons and striped skunks (Mephitis mephitis) provided the opportunity to determine if volatile fecal metabolites might be used to non-invasively monitor ORV programs and/or predict virus protection for these species. The volatile metabolome is a rich source of information that may significantly contribute to our understanding of disease and infection. Fecal samples were collected at multiple time points from raccoons and striped skunks subjected to oral treatment with rabies vaccine (or sham). Intramuscular challenge with a lethal dose of rabies virus was used to determine protection status at six (raccoons) and 11 (skunks) months post-vaccination. In addition to fecal samples, blood was collected at various time points to permit quantitative assessment of rabies antibody responses arising from immunization. Feces were analyzed by headspace gas chromatography with mass spectrometric detection and the chromatographic responses were grouped according to cluster analysis. Cluster scores were subjected to multivariate analyses of variance (MANOVA) to determine if fecal volatiles may hold a signal of immunization status. Multiple regression was then used to build models of the measured immune responses based on the metabolomic data. MANOVA results identified one cluster associated with protective status of skunks and one cluster associated with protective status of raccoons. Regression models demonstrated considerably greater success in predicting rabies antibody responses in both species. This is the first study to link volatile compounds with measures of adaptive immunity and provides further evidence that the volatile metabolome holds great promise for contributing to our understanding of disease and infections. The volatile metabolome may be an important resource for monitoring rabies immunization in raccoons and striped skunks., Author summary Permanent damage to ecosystems and direct transmission to humans (zoonoses) are two serious consequences of infectious wildlife diseases. Rabies is one such zoonotic disease of concern. Recently, two rabies vaccine efficacy studies were conducted with raccoons (Procyon lotor) and striped skunks (Mephitis mephitis). These studies allowed us the opportunity to investigate the presence of volatile signals of rabies immunization coded in fecal odors. These signals, if sensitive and specific, could permit non-invasive monitoring of rabies vaccination in wildlife populations. Semi-quantitative analyses of fecal volatiles and quantitative assessment of rabies binding antibodies (rVBA; which represent humoral immune responses to the vaccine or virus) were made at multiple time points. Regression models predicting rVBA responses using volatile fecal metabolites demonstrated predictive qualities in both species. This is the first study to link volatile compounds with adaptive immunity and provides further evidence that the volatile metabolome holds great promise for contributing to our understanding of wildlife diseases.

In the US, rabies lyssavirus (RABV) only circulates in wildlife species and the most significant reservoir from a public and animal health perspective is the raccoon (Procyon lotor). Management of wildlife rabies relies principally on oral rabies vaccination (ORV) strategies using vaccine-laden bait delivery to free-ranging target hosts, in order to reduce the susceptible population to prevent the spread of and eliminate RABV circulation. Our objective was to evaluate efficacy of the Ontario Rabies Vaccine Bait (ONRAB) against a lethal RABV challenge in captive raccoons. Sham or live vaccine baits were offered to 50 raccoons and efficacy was evaluated in 46, split into two trials of 17 and 29 raccoons. Raccoons were challenged with a lethal dose of RABV 180 days post-vaccination and observed for 90 days post-infection. Raccoon bait interactions were assigned increasing integer scores for approach, oral manipulation, puncture, and consumption behaviors. Higher bait interaction scores were observed in the fall compared to the spring trial, indicating that more raccoons consumed baits in the fall. Although animal age did not explain variation in bait interaction scores, the geometric mean rabies virus antibody titers among juvenile vaccinates were higher than adults at all pre-challenge time points. The prevented fraction associated with ONRAB delivery was 0.73 (8/11, 95% CI 0.39–0.94) in the spring trial and 0.91 (21/23, 95% CI 0.72–0.99) in the fall trial. All sham-vaccinated raccoons (12/12) succumbed to rabies infection, in contrast to 15% (5/34) mortality among vaccinated raccoons. Our results indicate a high efficacy of ONRAB bait vaccination in protecting adult and juvenile raccoons against RABV infection for a minimum of six months. These data complement experimental field trials that have also demonstrated the potential of ONRAB for the control and prevention of RABV circulation in free-ranging raccoon populations in the US.

The small Indian mongoose (

Animal movement influences the spatial spread of wildlife infectious diseases through host-host contact structure and hence pathogen transmission. Wildlife disease hosts vary in characteristics related to pathogen transmission, which can increase the spread and intensity of disease outbreaks. The consequences of home range size variation on wildlife disease dynamics are poorly understood, but could help to predict disease spread and determine more effective disease management strategies.We developed a spatially-explicit individual-based model to examine the effect of variation in host home range size on the spatial spread rate, persistence, and incidence of rabies virus (RABV) in raccoons (Procyon lotor). We tested the hypothesis that host home range area variation decreases vaccination effectiveness in wildlife host populations following pathogen invasion into a vaccination zone.We simulated raccoon demography and RABV dynamics across a range of magnitudes and variances in weekly home range radius distributions for raccoons, and compared results to conditions that assumed a fixed host home range area. We examined how variable host home range radius distributions influenced the relative effectiveness of three components of orally-baited raccoon RABV vaccination (ORV) programs—timing and frequency of bait delivery, width of the zone where ORV baits were delivered, and proportion of hosts immunized.Variability in home range radius distributions increased RABV spread rates by 1.2 - 5.2-fold compared to simulations with fixed radii. More variable host home range radius distributions decreased relative vaccination effectiveness by 71% compared to less variable host home range radius distributions under conventional vaccination conditions. We found that vaccination timing was more influential for vaccination effectiveness than vaccination frequency or vaccination zone width.Our results suggest that variation in wildlife home range exploration increases the spatial spread and incidence of wildlife disease. Our vaccination results underscore the importance of prioritizing individual-level space use data collection to understand the dynamics of wildlife diseases and plan their effective control and elimination.