Oral Rabies Vaccination Program Research Articles

Managing Rabies Cases with a One Health Approach in Various Countries: Literature Review

Introduction: More people die from rabies than from other zoonotic diseases worldwide each year, with an estimated 60,000 deaths attributable to the disease. The management of rabies epidemics in humans and animals is the main goal of the One Health approach's implementation. Reducing the risk of animal-to-human rabies transmission not only lessens the requirement for post-exposure vaccinations but also lessens the financial burden of rabies management in people. Objective: This review aims to provide a general overview and comparison of rabies control programs in various countries using the One Health approach. Method: PRISMA (Preferred Reporting Items for Systematic Reviews and Meta-analyses) is used in the literature review. Using the Science Direct, Scopus, and Pubmed databases covering the years 2012–2022, look up library resources. Result: The results of the literature search with the keywords control, one health approach, and rabies were screened again according to the predetermined inclusion criteria and the publication time range of 2012–2022 until 10 suitable journals were obtained. With financial assistance from the European Union, a number of European nations initiated oral rabies vaccination (ORV) programs in foxes. The vaccine was given in bait form, effectively controlling vulpine rabies. The creation of ORV demonstrates that oral rabies vaccination for wildlife in Serbia was effective and was marked by a rise in the use of vaccine bait as well as an increase in the animal's immunological response. Consequently, until 2018, fewer instances of rabies were documented. Conclusion: With multisectoral support from partners in the government and non-government sectors, implementing One Health is one of the best ways to reduce rabies and guarantee community engagement and knowledge.

Oral Rabies Vaccination of Raccoons (Procyon lotor) across a Development Intensity Gradient in Burlington, Vermont, USA, 2015-2017.

Management of the raccoon rabies virus variant in North America is conducted primarily using oral rabies vaccination (ORV). When a sufficient proportion of the population is vaccinated (∼60%), rabies transmission can be eliminated. To date, ORV programs have successfully controlled and eliminated raccoon rabies in rural areas, but there has been less success in urban areas. We studied the proportions of rabies virus neutralizing antibodies (RVNA) in a raccoon (Procyon lotor) population during a 3-yr ORV trial in developed areas of Burlington, Vermont, US. We used a modified N-mixture model to estimate raccoon abundance, RVNA seroprevalence, and capture rates jointly to examine factors that relate to ORV success to better inform management. We found that raccoon abundance was lower in less-developed areas compared to urban centers. Raccoon RVNA seroprevalence decreased as population abundance increased; it increased as the average age of the population increased. Nontarget opossum (Didelphis virginiana) captures correlated with a decrease in raccoon RVNA seroprevalence in low-development areas, suggesting that they may be competing for baits. The target bait density across the entire study area was 150 baits/km2, but a hand baiting strategy was heavily concentrated on roads, resulting in uneven bait densities within sampling sites (0-484 baits/km2). Uneven bait distribution across the study area may explain low RVNA seroprevalence in some locations. Our results suggest that increases in bait density across the study area may improve RVNA seroprevalence and support annual ORV to account for raccoon population turnover.

In free-roaming dog populations, does egg-based oral rabies vaccination programmes result in improved bait efficacy?

PICO Question In free-roaming dog populations, does an egg-based oral rabies vaccination (ORV) compared with meat and fish based ORVs result in improved bait efficacy? Clinical bottom line Category of research Treatment. Number and type of study designs reviewed Three non-blinded, control trials were critically reviewed. Strength of evidence Weak. Outcomes reported Variables assessed in this Knowledge Summary included the type of bait that stray dogs were most interested in, and whether or not the dog was successfully vaccinated by release of the vaccine sachet into the oral cavity. Conclusion There is weak evidence to show that stray dogs prefer egg-based baits in compared to other bait types, despite egg-based baits allowing for more successful perforation of the vaccine sachet, and hence a higher chance of a successful oral rabies vaccination. How to apply this evidence in practice The application of evidence into practice should take into account multiple factors, not limited to: individual clinical expertise, patient’s circumstances and owners’ values, country, location or clinic where you work, the individual case in front of you, the availability of therapies and resources. Knowledge Summaries are a resource to help reinforce or inform decision making. They do not override the responsibility or judgement of the practitioner to do what is best for the animal in their care.

Retrospective analysis of sero-prevalence and bait uptake estimations in foxes after oral rabies vaccination programmes at European level: Lessons learned and paths forward

Rabies caused by the Classical Rabies Virus (Lyssavirus rabies abbreviated RABV) in the European Union has been close to elimination mainly thanks to Oral Rabies Vaccination (ORV) campaigns targeting wildlife (primarily red foxes). ORV programmes co-financed by the European Commission include a monitoring-component to assess the effectiveness of the ORV campaigns at national level. This assessment is performed by a random collection of red foxes in the vaccinated areas with control of antibodies presence by serological analysis and control of bait uptake by detection of biomarkers (tetracycline incorporated into the baits) in the bones and teeth. ORV programmes aim to a vaccine coverage high enough to immunize (ideally) 70 % of the reservoir population to control the spread of the disease. European Union (EU) programmes that led to almost elimination of rabies on the territory have been traditionally found to have a bait uptake average of 70 % (EU countries; 2010–2020 period) while the seroconversion data showed an average level of 40 % (EU countries; 2010–2020 period). To better understand variations of these indicators, a study was been set up to evaluate the impact of several variables (linked to the vaccination programme itself and linked to environmental conditions) on the bait uptake and the seroconversion rate. Thus, pooling data from several countries provides more powerful statistics and the highest probability of detecting trends. Results of this study advocate the use of a single serological test across the EU since data variation due to the type of test used was higher than variations due to field factors, making the interpretation of monitoring results at EU level challenging. In addition, the results indicates a negative correlation between bait uptake and maximum temperatures reached during ORV campaigns questioning the potential impact of climatic change and associated increase of temperatures on the ORV programmes efficiency. Several hypotheses requesting additional investigation are drawn and discussed in this paper.

Two Sylvatic Rabies Re-Emergences in Central-Eastern Europe over the 2021–2022 Period: An Unprecedented Situation in Recent Years

The implementations of coordinated, standardised, and sustained oral rabies vaccination (ORV) campaigns over large areas have led to the almost elimination of sylvatic rabies from the European Union (EU) territory at the approach of 2020. The annual number of rabies cases reported within EU territory indeed dropped from around 13,000 cases in 1990 to less than 10 cases over the 2017–2019 period. Unfortunately, since 2020, the EU territory has faced two major rabies re-emergence events of rabies lyssavirus in non-flying animals. The first sylvatic rabies outbreak, already described, occurred in 2021 and 2022 in Poland in the Mazowieckie voivodeship, involving the Central Europe variant, while the second one affecting Romania, Hungaria, and Slovakia, increased considerably at the end of 2022 and involved the North East Europe variant only. Thus, Hungary and Slovakia that did not record a single case since 5–7 years, respectively, faced new rabies outbreaks in 2022. This article, therefore, presents these two epidemiological events and discusses the importance and challenges of maintaining ORV programmes and immune belts in the long term, particularly in a complicated context of a pandemic affecting organisation of human societies and of geopolitical conflicts.

Analysis of seroprevalence in target wildlife during the oral rabies vaccination programme in Lithuania

BackgroundRabies vaccination of wildlife carnivores is a powerful tool to prevent, control and eliminate rabies. The presence of neutralizing rabies antibodies in blood is considered a reliable indicator of adequate vaccination. The main purpose of the present study was to analyze the seroprevalence of specific antibodies in target populations of Lithuanian red fox (RF) and raccoon dog (RD) during the oral rabies vaccination (ORV) campaigns during the 2010–2019 period.ResultsOver the ten-year period, 7,261 RF and 2,146 RD sera samples were collected post-mortem in field conditions and tested using a commercial standardized enzyme-linked immunosorbent assay (ELISA) kit in Lithuania. In the ORV spring and autumn vaccination periods, 31.8% (20.3–43.4 95% CI – 95% confidence interval) and 31.7% (21.2–42.1 95% CI) of RF, and 34.1% (22.5–45.7 95% CI) and 34.7% (22.7–46.7 95% CI) of RD sera samples, respectively, were identified as ELISA-positive (seroconversion ≥ 0.5 EU/mL—Equivalent Units per Millilitre). The seroprevalence analysis in adult/ juvenile animal subpopulations indicated that 34.9% (27.2–42.5 95% CI) and 29.2% (20.3–37.9 95% CI) of RF, and 35.6% (25.2–46.0 95% CI) and 30.6% (20.2–40.9 95% CI) of RD sera samples, respectively, were identified as ELISA-positive (seroconversion ≥ 0.5 EU/mL). Statistically strong determinate correlations (r) between the serological results (pos.%) in RF adult/juvenile animal subpopulations (r = 0.937) and between RF and RD positive seroconvert (pos.%) sera samples during the spring vaccinations (r = 0.864) were demonstrated. In different ORV periods, 14–29% of RF and 7–25% of RD sera samples were identified as ELISA-negative (seroconversion < 0.5 EU/mL), but with low (0.125 < 0.49 EU/mL) antibody (Abs) titres.ConclusionsThe 2010–2019 ORV programme has been an effective tool in both RF and RD populations in Lithuania. The rabies-free status of Lithuania was self-declared in 2015 with only three rabies cases identified in buffer zones since then. The percentage of ELISA-positive serum samples (seroconversion ≥ 0.5 EU/mL) during the different periods of vaccination was similar in RF and RD populations—32% and 34% respectively. The identified seroconversion average of 21.5% in RF and 16% in RD sera samples were officially identified as ELISA-negative (seronversion < 0.5 EU/mL), but with low 0.125 < 0.49 EU/mL Abs titres. That low, but positive seroconversion participated in the formation of populations overall immune status and can influence the interpretation of oral vaccination efficacy.

Local rabies transmission and regional spatial coupling in European foxes.

Infectious diseases are often transmitted through local interactions. Yet, both surveillance and control measures are implemented within administrative units. Capturing local transmission processes and spatial coupling between regions from aggregate level data is therefore a technical challenge that can shed light on both theoretical questions and practical decisions. Fox rabies has been eliminated from much of Europe through oral rabies vaccination (ORV) programmes. The European Union (EU) co-finances ORV to maintain rabies freedom in EU member and border states via a cordon sanitaire. Models to capture local transmission dynamics and spatial coupling have immediate application to the planning of these ORV campaigns and to other parts of the world considering oral vaccination. We fitted a hierarchical Bayesian state-space model to data on three decades of fox rabies cases and ORV campaigns from Eastern Germany. Specifically, we find that (i) combining regional spatial coupling and heterogeneous local transmission allows us to capture regional rabies dynamics; (ii) incursions from other regions account for less than 1% of cases, but allow for re-emergence of disease; (iii) herd immunity achieved through bi-annual vaccination campaigns is short-lived due to population turnover. Together, these findings highlight the need for regular and sustained vaccination efforts and our modelling approach can be used to provide strategic guidance for ORV delivery. Moreover, we show that biological understanding can be gained from inference from partially observed data on wildlife disease.

Variation in host home range size decreases rabies vaccination effectiveness by increasing the spatial spread of rabies virus.

Animal movement influences the spatial spread of directly transmitted wildlife disease through host–host contact structure. Wildlife disease hosts vary in home range‐associated foraging and social behaviours, which may increase the spread and intensity of disease outbreaks. The consequences of variation in host home range movement and space use 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 spatiotemporal 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 variation in home range size increases RABV spread and decreases vaccination effectiveness in 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 size for raccoons. We examined how variable home range size influenced the relative effectiveness of three components of oral rabies vaccination (ORV) programmes targeting raccoons—timing and frequency of bait delivery, width of the ORV zone and proportion of hosts immunized.Variability in weekly home range size increased RABV spread rates by 1.2‐fold to 5.2‐fold compared to simulations that assumed a fixed home range size. More variable host home range sizes decreased relative vaccination effectiveness by 71% compared to less variable host home range sizes 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 movement behaviour increases the spatial spread and incidence of RABV. Our vaccination results underscore the importance of prioritizing individual‐level space use and movement data collection to understand wildlife disease dynamics and plan their effective control and elimination.

Volatile metabolomic signatures of rabies immunization in two mesocarnivore species.

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.

Evolution of rabies and oral rabies vaccination program for foxes in the EU and in Romania during 2008 - 2018

Decision-makers and risk managers are often called upon to prioritise on and recommend suitable measures to prevent the risk of introduction and spread of pathogens. The main objective of this study was to assess the perceptions of experts in Italy, Switzerland and the United Kingdom concerning the importance, effectiveness, feasibility, costs and acceptability of selected biosecurity measures to prevent the introduction and limit the spread of rabies, blue tongue (BT) and classical swine fever (CSF).After identifying the most relevant measures by the project team, an expert knowledge elicitation was implemented through a questionnaire. After preliminary descriptive analyses, a number of statistical calculations were performed such as weighted medians, Spearman rank correlation tests, Wilcoxon comparison tests and ranking of measures.Three experts from each country completed the questionnaires, one expert for each disease. The mean answer rates for CSF, BT and rabies were 73%, 100% and 99% respectively. “Tracing system for live animal trade” was highlighted as very relevant in all diseases. The implementation of a “restriction zone after a suspicion or confirmation” was also rated as a relevant measure, especially for CSF. We identified generally a small correlation between costs and the other criteria. Among the rabies experts, measures related to “zoonotic risk” were rated highly, supporting the idea of a One Health approach. Disagreement among experts concerned 43 measures for the three pathogens: the debated measures were “control of the wildlife CSF status”, “arthropod-vector control” and “rabies vaccination for domestic animals”.Facing budget restriction, decision-makers need to prioritise their actions and make efficient prevention choices. With this study, we aimed to provide elements for reflection and to inform priority setting. The results can be applied through the implementation of similar surveys or directly from the knowledge already gathered in this study.

Analysis of Iophenoxic Acid Analogues in Small Indian Mongoose (Herpestes Auropunctatus) Sera for Use as an Oral Rabies Vaccination Biological Marker.

The small Indian mongoose (Herpestes auropunctatus) is a reservoir of rabies virus (RABV) in Puerto Rico and comprises over 70% of animal rabies cases reported annually. The control of RABV circulation in wildlife reservoirs is typically accomplished by a strategy of oral rabies vaccination (ORV). Currently no wildlife ORV program exists in Puerto Rico. Research into oral rabies vaccines and various bait types for mongooses has been conducted with promising results. Monitoring the success of ORV relies on estimating bait uptake by target species, which typically involves evaluating a change in RABV neutralizing antibodies (RVNA) post vaccination. This strategy may be difficult to interpret in areas with an active wildlife ORV program or in areas where RABV is enzootic and background levels of RVNA are present in reservoir species. In such situations, a biomarker incorporated with the vaccine or the bait matrix may be useful. We offered 16 captive mongooses placebo ORV baits containing ethyl-iophenoxic acid (et-IPA) in concentrations of 0.4% and 1% inside the bait and 0.14% in the external bait matrix. We also offered 12 captive mongooses ORV baits containing methyl-iophenoxic acid (me-IPA) in concentrations of 0.035%, 0.07% and 0.14% in the external bait matrix. We collected a serum sample prior to bait offering and then weekly for up to eight weeks post offering. We extracted Iophenoxic acids from sera into acetonitrile and quantified using liquid chromatography/mass spectrometry. We analyzed sera for et-IPA or me-IPA by liquid chromatography-mass spectrometry. We found adequate marking ability for at least eight and four weeks for et- and me-IPA, respectively. Both IPA derivatives could be suitable for field evaluation of ORV bait uptake in mongooses. Due to the longevity of the marker in mongoose sera, care must be taken to not confound results by using the same IPA derivative during consecutive evaluations.

Influence of different factors on the costs and benefits of oral vaccination of foxes against rabies.

The oral rabies vaccination (ORV) programme is the most widespread vaccination programme that is implemented in the European Union and targets wildlife diseases. The size of the project requires significant financial resources, so a cost-benefit analysis is necessary to establish the economic value of the project. We summarized all ORV costs during the period 2011-2016 in Croatia, calculated all established benefits and presented the cost-benefit ratio (CBR). Additionally, we analysed all components included in the ORV and rabies control programme (surveillance, preventive human treatment and vaccination of dogs) to find possibilities to increase the benefits. According to our results, in the period 2011-2016, the CBR was only 0.05, and the majority of the cost was derived from the preventive vaccination of dogs (72.3%). With the implementation of 2- or 3-year vaccination intervals, the CBR can be increased to 1.46 or 1.92, respectively, confirming positive economic value. This study shows the importance of analysing all rabies control and eradication components based on the specific characteristics of a particular country to determine the factors that can be modified to potentially improve the benefits of ORV.

Optimizing spatial and seasonal deployment of vaccination campaigns to eliminate wildlife rabies.

Understanding how the spatial deployment of interventions affects elimination time horizons and potential for disease re-emergence has broad application to control programmes targeting human, animal and plant pathogens. We previously developed an epidemiological model that captures the main features of rabies spread and the impacts of vaccination based on detailed records of fox rabies in eastern Germany during the implementation of an oral rabies vaccination (ORV) programme. Here, we use simulations from this fitted model to determine the best vaccination strategy, in terms of spatial placement and timing of ORV efforts, for three epidemiological scenarios representative of current situations in Europe. We found that consecutive and comprehensive twice-yearly vaccinations across all regions rapidly controlled and eliminated rabies and that the autumn campaigns had the greater impact on increasing the probability of elimination. This appears to result from the need to maintain sufficient herd immunity in the face of large birth pulses, as autumn vaccinations reach susceptible juveniles and therefore a larger proportion of the population than spring vaccinations. Incomplete vaccination compromised time to elimination requiring the same or more vaccination effort to meet similar timelines. Our results have important practical implications that could inform policies for rabies containment and elimination in Europe and elsewhere. This article is part of the theme issue 'Modelling infectious disease outbreaks in humans, animals and plants: epidemic forecasting and control'. This theme issue is linked with the earlier issue 'Modelling infectious disease outbreaks in humans, animals and plants: approaches and important themes'.

Spatial ecology of urban striped skunks (Mephitis mephitis) in the Northern Great Plains: a framework for future oral rabies vaccination programs

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.

Challenges to controlling rabies in skunk populations using oral rabies vaccination: A review.

Controlling rabies in skunk populations is an important public health concern in many parts of the United States due to the potential for skunk rabies outbreaks in urban centres and the possible role for skunks in raccoon rabies variant circulation. Oral rabies vaccination (ORV) programmes have supported wildlife rabies control efforts globally but using ORV to control rabies in skunk populations has proven more challenging than with other target species, like foxes, coyotes and raccoons. A review of published studies found that some ORV constructs are immunogenic in skunks and protect against virulent rabies virus challenges, especially when delivered by direct installation into the oral cavity. However, in field ORV programmes using currently available vaccine-bait formats and distribution methods targeting other rabies reservoir species, skunks often fail to seroconvert. Field effectiveness of ORV in skunks appears to be limited by poor bait uptake or inadequate ingestion of vaccine rather than from poor vaccine efficacy. Observations of captive skunks revealed vaccine spillage when handling and biting into baits such that modification of bait formats might improve field effectiveness. In addition, a dose-response relationship between bait distribution density and post-baiting seroconversion among skunks was observed across the limited number of field studies. Additional research is needed to identify opportunities to modify ORV baits and distribution strategies to improve the viability of ORV as a rabies control strategy in skunks.

EVALUATION OF ORAL RABIES VACCINATION: PROTECTION AGAINST RABIES IN WILD CAUGHT RACCOONS ( PROCYON LOTOR).

Oral rabies vaccination (ORV) is an effective tactic for wildlife rabies control, particularly for containment of disease spread along epizootic fronts. As part of the continuing evaluation of the ORV program in free-ranging raccoons ( Procyon lotor) in the US, 37 raccoons from ORV-baited areas in Pennsylvania were live-trapped and transferred to captivity to evaluate protection against rabies in animals with varying levels of existing neutralizing antibodies, expressed in international units per milliliter (IU/mL). Among the 37 raccoons at the date of capture, 24% (9/37) of raccoons were seronegative (<0.05 IU/mL), 22% (8/37) were low positive (≥0.05-0.11 IU/mL), 27% (10/37) were medium positive (>0.11-<0.5 IU/mL), and 27% (10/37) were high positive (≥0.5 IU/mL). Raccoons were held for 86-199 d between the date of capture and rabies virus challenge. At challenge, 68% (25/37) raccoons were seronegative. The overall survival rate among challenged animals was 46% (17/37). Based on the antibody titers at the time of challenge, survivorship was 24% (6/25) among seronegative animals, 100% (4/4) among low positive animals, 83% (5/6) among medium positive animals, and 100% (2/2) among high positive animals. Evidence of high-titer seroconversion after vaccination is a good surrogate indicator of rabies survival; however, survival rates of approximately 45% (15/35) were found among raccoons with detectable titers below 0.5 IU/mL. In contrast, any detectable titer at the time of challenge (>3 mo after vaccination) appeared to be a surrogate indicator of survival. Overall, we illustrated significant differences in the value of specific titers as surrogates for survival based on the timing of measurement relative to vaccination. However, survivorship was generally greater than 45% among animals with any detectable titer regardless of the timing of measurement. These findings suggest that lower titer cutoffs may represent a valid approach to measuring immunization coverage within ORV management zones, balancing both sensitivity and specificity for estimating herd immunity.

First four Oral Rabies Vaccination campaigns of the red foxes in Greece: Evaluating factors and assessment

Following the last animal rabies outbreak in Greece in 2012, Oral Rabies Vaccination (ORV) campaigns of red foxes (Vulpes vulpes) were conducted in order to halt the spread of the disease, as widely and effectively have also been implemented in other countries. The present study aims to report the main outcomes following the first Greek ORV campaigns during autumn 2013, 2014, 2015 and spring 2016, to assess their effectiveness and to investigate factors potentially related to their success. Blood samples, mandible bones and teeth, derived by 452 foxes, were tested for rabies antibody titration, animal age determination and tetracycline (TTC) detection. The laboratory results obtained were statistically analyzed. High seroprevalence and TTC detection rates were obtained following the autumn campaigns studied, while these rates were significantly reduced following the spring campaign. The year or the season of the vaccination campaign, the estimated age group of the animal and the geographical Regional Unit (RU), where the animal was hunted, were identified as important factors. On the contrary, no significance could be ascertained for TTC detection based on exclusively previous uptake, use of filter paper, blood sample type and quality, as well as sex of animal. Based on the monitoring results achieved, the first ORV campaigns conducted in the country can be generally considered to be satisfactory. No positives cases were detected since May 2014. Seasonal, geographical parameters and factors related to fox ecology may interfere with monitoring results and should be always considered when planning future ORV programs.

FLAVOR PREFERENCE AND EFFICACY OF VARIABLE DOSE ONTARIO RABIES VACCINE BAIT (ONRAB) DELIVERY IN STRIPED SKUNKS (MEPHITIS MEPHITIS)

In North America, terrestrial wildlife rabies control is achieved by oral rabies vaccination programs that principally target mesocarnivores. Success at rabies control in striped skunks ( Mephitis mephitis) has been more limited and may require additional enhancements to existing bait products or novel bait designs and attractants. We evaluated preference among captive striped skunks for six different flavors of placebo Ontario Rabies Vaccine Bait (ONRAB®) "Ultralite" Baits (Artemis Technologies, Guelph, Ontario, Canada). Different doses and delivery methods of ONRAB vaccine were tested for efficacy in a subsequent experiment with the same skunks. Cheese-, egg-, and chicken-flavored baits were preferred over plain-flavored baits, but a strong preference for a singular flavor was not observed. Vaccine efficacy of 80-100% was observed among skunks challenged at 335 d postvaccination across a log range of doses tested by a direct instillation into the oral cavity route, respectively (109.3-1010.2 median tissue culture infective doses), in contrast to more-limited efficacy by bait delivery. Our results extended the duration of ONRAB vaccine efficacy in skunks and suggested that there may be limited flexibility to alter vaccine titer and volume in novel bait designs for skunks.

Modeling Raccoon (Procyon lotor) Habitat Connectivity to Identify Potential Corridors for Rabies Spread.

The United States Department of Agriculture (USDA), Animal and Plant Health Inspection Service (APHIS), Wildlife Services National Rabies Management Program has conducted cooperative oral rabies vaccination (ORV) programs since 1997. Understanding the eco-epidemiology of raccoon (Procyon lotor) variant rabies (raccoon rabies) is critical to successful management. Pine (Pinus spp.)-dominated landscapes generally support low relative raccoon densities that may inhibit rabies spread. However, confounding landscape features, such as wetlands and human development, represent potentially elevated risk corridors for rabies spread, possibly imperiling enhanced rabies surveillance and ORV planning. Raccoon habitat suitability in pine-dominated landscapes in Massachusetts, Florida, and Alabama was modeled by the maximum entropy (Maxent) procedure using raccoon presence, and landscape and environmental data. Replicated (n = 100/state) bootstrapped Maxent models based on raccoon sampling locations from 2012–2014 indicated that soil type was the most influential variable in Alabama (permutation importance PI = 38.3), which, based on its relation to landcover type and resource distribution and abundance, was unsurprising. Precipitation (PI = 46.9) and temperature (PI = 52.1) were the most important variables in Massachusetts and Florida, but these possibly spurious results require further investigation. The Alabama Maxent probability surface map was ingested into Circuitscape for conductance visualizations of potential areas of habitat connectivity. Incorporating these and future results into raccoon rabies containment and elimination strategies could result in significant cost-savings for rabies management here and elsewhere.

Progress towards Bait Station Integration into Oral Rabies Vaccination Programs in the United States: Field Trials in Massachusetts and Florida.

Bait stations for distribution of oral rabies vaccine baits are designed for rabies management in highly-developed areas where traditional distribution of oral rabies vaccine baits may be difficult. As part of national efforts to contain and eliminate the raccoon (Procyon lotor) variant of the rabies virus (raccoon rabies) in the eastern United States, the United States Department of Agriculture, Animal and Plant Health Inspection Service, Wildlife Services program, distributed vaccine baits by bait stations experimentally and operationally in Massachusetts during 2006-present, and in Florida during 2009–2015. In Massachusetts, a rabies virus-neutralizing antibody (RVNA) response of 42.1% for raccoons captured in areas baited with high density bait stations during 2011–2015 was achieved, compared with 46.2% in areas baited by hand, suggesting the continuation of this as a strategy for the oral rabies vaccination (ORV) program there, and for similar locations. Non-target competition for vaccine baits is problematic, regardless of distribution method. In Massachusetts, bait station visitation rates for targeted raccoons and non-target opossums (Didelphis virginiana) were similar (1.18:1) during 2006–2009 (p > 0.05). Bait station modifications for reducing non-target uptake were tested, and in Massachusetts, reduced non-target bait access was achieved with two design alternatives (p < 0.001). However, no difference was noted between the control and these two alternative designs in Florida. Due to ongoing trials of new vaccines and baits, the bait station performance of an adenovirus rabies glycoprotein recombinant vaccine bait, ONRAB® bait (Artemis Technologies, Guelph, ON, Canada) and a vaccinia-rabies glycoprotein recombinant vaccine bait, RABORAL V-RG®bait (Merial Limited, Athens, GA, USA), was compared. While uptake of the ONRAB bait was greater in Massachusetts (p < 0.001) in this limited trial, both types performed equally well in Florida. Since bait station tampering or theft as well as potential human bait contacts has been problematic, performance of camouflaged versus unpainted white bait stations was analyzed in terms of internal temperatures and maintaining a stable bait storage environment. In Massachusetts, camouflaged bait station interiors did not reach higher average temperatures than plain white bait stations in partially- or fully-shaded locations, while in Florida, camouflaged bait stations were significantly warmer in light exposure categories (p < 0.05). As ORV operations expand into more heavily-urbanized areas, bait stations will be increasingly important for vaccine bait distribution, and continued refinements in the strategy will be key to that success.