Oral rabies vaccine (ORV) bait uptake by captive striped skunks

Aerial delivery of oral rabies vaccine (ORV) baits has proven effective in large-scale efforts to immunize wildlife against rabies, and in North America this strategy currently is being used to immunize foxes (Urocyon cinereoargenteus and Vulpes vulpes), raccoons (Procyon lotor), and coyotes (Canis latrans). Skunks are also a major reservoir and vector of rabies, but at present oral vaccines for use in skunks are not licensed. Furthermore, given differences in morphology (smaller jaws) and behavior (food handling and consumption), it is unknown if baits currently used in ORV campaigns would be effective for skunks. Because oral vaccine delivery is contingent upon puncture of the vaccine container (VC), baits need to be sufficiently attractive to elicit selection and consumption. Manipulation of the bait to facilitate vaccine ingestion by the target species is a critical element for an effective ORV bait. The objectives of this study were to assess manipulation and consumption of current ORV baits by striped skunks (Mephitis mephitis). We conducted four independent trials with penned animals and various baits to assess bait selection frequency, VC puncture frequency, and consumption. Video recorded trials were used to assess attractiveness of baits and consumption behavior of skunks. Bait characteristics, such as texture, size, and flavor influenced selection and consumption. Fish and chicken flavors were preferred and vaccine containers within selected baits were likely to be punctured. Vaccine ingestion seemed more likely if VCs were directly coated with the bait matrix. To make baits attractive to skunks and to ensure puncture of the VC, modifications to current baits should consider a smaller size, a meat-flavored matrix, a slightly pressurized VC, and a direct coating of matrix on the VC.     

A new flavor-coated sachet bait for delivering oral rabies vaccine to raccoons and coyotes

Research was conducted during 1996-2000 to develop baits for delivering an oral rabies vaccine to raccoons (Procyon lotor) and coyotes (Canis latrans). A bait was sought that: (1) was attractive to the target species, (2) could be distributed by aircraft, (3) was as effective (or more so) than the currently used fish meal polymer bait, and (4) could be produced in large numbers by automated procedures and could be purchased by user groups at substantially lower cost. Ten field trials were conducted to document raccoons' bait flavor preferences, evaluate a new vaccine sachet bait coated with various attractants, and determine if the sachet bait would effectively deliver Raboral V-RG oral rabies vaccine (Merial Limited, Athens, Georgia, USA) to this species. Raccoons preferred fish and crustacean-based flavors over those derived from plant materials. Raccoon visits to tracking stations, frequency of bait removals, and percent of sachets discarded by this species that were emptied of placebo vaccine indicated efficacy of the new bait was equal or superior to the currently used fish meal polymer bait. A field trial conducted in fall 1998 compared aerially distributed vaccine-laden sachet and polymer baits and showed there was no difference between the percent of raccoons from the test and reference areas subsequently found positive for rabies antibody. Four bait trials to determine coyote response to sachet baits were conducted in 1997-98. The propensity for canids to gulp or bolt smaller food items is well known. Thus, a first trial involved offering fish-flavored sachet baits of different sizes to 30 captive coyotes to determine if smaller size baits were more frequently swallowed intact. Two field trials were also conducted in fall 1997 to determine if free-ranging coyotes discriminated among sachet baits coated with different attractants. Finally, Raboral V-RG-laden poultry-flavored sachet baits were aerially dropped and the percent of seropositive coyotes was compared with coyotes from surrounding areas where fish meal polymer vaccine baits had been distributed. Captive coyotes did not swallow sachet baits intact, regardless of size. Bait preference field trials indicated that coyotes preferred poultry, cheese/beef tallow, and fish-flavored sachet baits and that such baits were taken at the same rate as polymer baits. A sample of coyotes from the area baited with vaccine-laden sachet baits had a markedly higher (P = 0.01) seropositivity rate than coyotes from areas where vaccine was distributed in polymer baits. Sachet bait production could be facilitated by automated technology and sachet baits used either as an alternative vaccine delivery device or in combination with the fish meal polymer bait.     

Control of rabies in Slovenia

Red foxes (Vulpes vulpes) are the main reservoir of rabies in Slovenia, whereas cases of rabies in other wildlife species occur sporadically. In 1995, a program of oral vaccination of wildlife in Slovenia was initiated; baits with oral vaccine were distributed by air at a density of 20 baits/km(2). During 1995, when the oral vaccination program was started, 1,089 cases of rabies (including both wild and domestic animals) were reported. Five years later (1999), only six positive animals were detected among 1,195 tested (0.5%). Despite an increase in bait density (25 baits/km(2)) during the years 2000 and 2001, reported rabies cases increased to 115 and 135, respectively. In 2003, following initiation of a new bait-dropping strategy, which incorporated perpendicular rather than parallel flight lines, the number of rabies cases decreased to eight.     

Comparing ONRAB® AND RABORAL V-RG® oral rabies vaccine field performance in raccoons and striped skunks, New Brunswick, Canada, and Maine, USA

Control of rabies in mesocarnivore reservoirs through oral rabies vaccination (ORV) requires an effective vaccine bait. Oral rabies vaccine performance in the field may be affected by a variety of factors, including vaccine bait density and distribution pattern, habitat, target species population density, and the availability of competing foods. A field study in which these covariates were restricted as much as possible was conducted along the international border of the state of Maine (ME), USA, and the province of New Brunswick (NB), Canada, to compare the performance of two oral rabies vaccines in raccoons (Procyon lotor) and striped skunks (Mephitis mephitis). RABORAL V-RG(?) (vaccinia-rabies glycoprotein recombinant oral vaccine in fishmeal-coated sachet) or ONRAB(?) (adenovirus-rabies glycoprotein recombinant oral vaccine in Ultralite bait matrix) were distributed in ME and NB, respectively, by fixed-wing aircraft at a density of 75 baits/km(2) along parallel flight lines spaced 1.0 km apart. Sera were collected from live-trapped raccoons and skunks 5-7 wk post-ORV and assayed to determine antibody prevalence in each area. Duplicate serum samples were provided blind to two different laboratories for analyses by rabies virus serum neutralization assays (at both laboratories) and a competitive enzyme-linked immunosorbent assay (at one laboratory). There was no significant difference in the proportion of antibody-positive animals determined by the three serologic methods, nor was there a significant difference between ONRAB and RABORAL V-RG in the proportion of antibody-positive striped skunks observed post-ORV. In contrast, the proportion of antibody-positive raccoons was significantly higher in the ONRAB- versus the RABORAL V-RG-baited areas (74% vs. 30%; χ(2)=89.977, df=5, P<0.0001). These data support that ONRAB may serve as an effective tool for raccoon rabies control.     

Prevalence of tetracycline and rabies virus antibody in raccoons, skunks, and foxes following aerial distribution of V-RG baits to control raccoon rabies in Ontario, Canada

More than 3.6 million baits containing a recombinant vaccinia virus-rabies glycoprotein (V-RG) oral rabies vaccine were aerially or hand-distributed during 1999-2006 in an approximate 4,000-9,000 km(2) area of eastern Ontario, Canada, as part of a multitactic approach to control the raccoon variant of rabies. The efficacy of the program was assessed through the collection and testing of > 6,900 animals for bait acceptance and rabies virus-specific antibodies. Raccoon acceptance of rabies vaccine baits was significantly greater (71-83% ) in areas baited at a density of 150 baits/km(2) compared to areas baited at 75 baits/km(2) (26-58% ), and more raccoons consumed vaccine baits in areas baited with a flight line spacing of 0.75 km (45.3% [321/708]) than with a spacing of 1.5 km (33.8% [108/320]). In addition, greater numbers of raccoons consumed vaccine baits during a drop in September (52.7% [213/404]) as opposed to a June bait drop (34.6% [216/624]). Seropositivity rates for raccoons ranged between 7% and 28% in areas baited at 75/km(2) and 10% to 27% in areas baited at 150/km(2) with statistical differences varying among years and treatments. The last case of raccoon-variant rabies reported in Ontario was in September 2005. The control of raccoon rabies in Ontario has resulted in an estimated $6M to $10 M Cdn annual savings in rabies-associated costs.     

Development of a bait and baiting system for delivery of oral rabies vaccine to free-ranging African wild dogs (Lycaon pictus)

The objective of the study was to develop a bait and baiting system capable of delivering one effective dose of oral rabies vaccine to each member of a free-ranging African wild dog (Lycaon pictus) pack. Trials were conducted between June and October 2000. The results of cafeteria-style bait preference trials testing seven candidate baits in captive wild dogs revealed a significant preference for chicken heads (June trials: P = 0.023, September trials: P = 0.021). Trials using a topical biomarker (rhodamine B) showed that chicken head baits were sufficiently chewed on most occasions to rupture the vaccine container. Free-ranging wild dogs and young pups ingested chicken head baits. Significant dominance of bait intake by a single individual was seen in four of six study packs and in the three packs in which an alpha pair could be distinguished, the dominant feeder was an alpha animal. Pattern of bait distribution and degree of satiation had no effect on pack coverage (proportion of pack ingesting at least one bait). Pack coverage was significantly related to trial number (r = 0.71, P < 0.001), with pack coverage increasing with increased exposure of the pack to the baits. During 46 hr of diurnal observations of free-ranging wild dogs only two baits were lost to non-target species. A baiting system for the oral vaccination of captive and free-ranging wild dogs is proposed.     

Spatio-temporal Use of Oral Rabies Vaccines in Fox Rabies Elimination Programmes in Europe

In Europe, the elimination of wildlife rabies using oral rabies vaccination [ORV] of foxes for more than 30 years has been a success story. Since a comprehensive review on the scope of the different oral rabies vaccine baits distributed across Europe has not been available yet, we evaluated the use of different vaccine baits over the entire period of ORV [1978–2014]. Our findings provide valuable insights into the complexity of ORV programs in terms of vaccine related issues. More than 10 oral vaccines against rabies were used over the past four decades. Depending on many factors, the extent to which oral rabies virus vaccines were used varied considerably resulting in huge differences in the number of vaccine doses disseminated in ORV campaigns as well as in large spatial and temporal overlaps. Although vaccine virus strains derived from the SAD rabies virus isolate were the most widely used, the success of ORV campaigns in Europe cannot be assigned to a single oral rabies virus vaccine alone. Rather, the successful elimination of fox rabies is the result of an interaction of different key components of ORV campaigns, i.e. vaccine strain, vaccine bait and strategy of distribution.     

Wild carnivore acceptance of baits for delivery of liquid rabies vaccine

A series of experiments are described on the acceptance, by red foxes (Vulpes vulpes) and other species, of two types of vaccine-baits intended to deliver liquid rabies vaccine. The baits consisted of a cube of sponge coated in a mixture of tallow and wax, or a plastic blister-pack embedded in tallow. All baits contained tetracycline as a biological marking agent: examination of thin sections of carnivore canines under an ultraviolet microscope revealed a fluorescent line of tetracycline if an individual had eaten baits. Baits were dropped from fixed-wing aircraft flying about 100 m above ground at approximately 130 km/h. Flight lines followed the edges of woodlots midway between parallel roads. Baits were dropped at one/sec, resulting in one bait/36 m on the ground, or 17 to 25 baits per km2. Crows (Corvus brachyrhynchos) removed many baits, but did not appear to lower the percent of the fox population which took bait. Dropping baits only into corn and woodland to conceal baits, to reduce depredation by crows, reduced acceptance by foxes. Acceptance by foxes ranged between 37 and 68%. Meat added as an attractant did not raise acceptance. Presence, absence, color and perforations of plastic bags did not alter bait acceptance. Dispersal by juvenile foxes probably lowered the estimates of bait acceptance. It took 7 to 17 days for 80% (n = 330) of foxes to eat their first bait. The rapidity with which foxes picked up their first bait appeared more affected by unknown characteristics of years or study areas than by experimental variables. Skunks (Mephitis mephitis) and raccoons (Procyon lotor) also ate these baits, but acceptance was lower. Small mammals contacted baits, but rarely contacted the vaccine, which had the potential for vaccine-induced rabies in some species. Aerial distribution of baits was more cost-effective than ground distribution as practiced in Europe. This system has potential for field control of rabies, although higher acceptance will be desirable.     

Evaluation of attractant flavours for use in oral vaccine baits for badgers (<Emphasis Type="Italic">Meles meles</Emphasis>)

European badgers (Meles meles) are a wildlife reservoir for Mycobacterium bovis infection (tuberculosis) in Ireland and the UK and are implicated in the transmission of infection to livestock. Vaccination of badgers with the human BCG vaccine (Bacille Calmette Guerin) is considered as an important strategy to reduce the burden of disease in this species, and a pragmatic approach is likely to involve oral vaccination. In this study, we evaluated nine different flavours for use as attractants in a prototype oral vaccine bait for European badgers (M. meles): aniseed, apple, cocoa powder, carob powder, curry, fish, garlic, peanut and strawberry. The bait matrix was composed of a natural lipid formulation, developed as a vehicle for oral vaccination against tuberculosis in wildlife. A ‘food for work’ paradigm was employed during the trials to ensure the animals were actively seeking the baits. The trials showed carob and cocoa powders were equally attractive and more attractive than any of the other candidates. Carob and cocoa show potential as bait attractants for badgers and might form part of a novel vaccine delivery system.     

Acceptance of baits for delivery of oral rabies vaccine to raccoons.

During 1993-96 the Ontario Ministry of Natural Resources (Canada) implemented a research project to evaluate the efficacy of three candidate baits to deliver oral rabies vaccine to wild raccoons (Procyon lotor). Extensive field testing revealed that raccoon acceptance of Sugar-Vanilla baits (SV) at densities of 200/km2 and 400/km2, hand-placed in urban habitats of Scarborough (Ontario) during 1993, was 74% and 82%, respectively. Raccoon density in those areas averaged 11/km2. Aerial placement of SV baits in rural habitats in Barrie (Ontario) during 1993 and 1994, yielded raccoon acceptance levels of 58% with a density of 100 baits/km2, 59% at 75 baits/km2, and 47% at 50 baits/km2. Raccoon acceptance of SV baits was significantly lower in areas baited at the density of 50/km2. Acceptance of Cheese baits (CH) at a density of 75 baits/km2 was 52%. During 1996 trials in Barrie, modified SV baits with blister packs protruding through the matrix yielded raccoon acceptance values of 51% at a bait density of 54/km2, whereas acceptance of regular SV baits was 39% at a density of 51 baits/km2. Pooling of bait acceptance data for all years revealed that bait acceptance was highest for adult male raccoons. Raccoon density in rural habitats (Barrie, Ontario) where the studies took place, averaged 11-13/km2. Puncture and impact testing of blister packs in baits suggested that they would adequately serve as a vehicle to contain oral rabies vaccine for delivery to raccoons via baits.     

Acceptance of simulated oral rabies vaccine baits by urban raccoons

In summer 1986, a study was conducted to evaluate raccoon (Procyon lotor) acceptance of oral baits that could be used for rabies vaccination. One thousand wax-coated sponge bait cubes were filled with 5 mg of a seromarker (iophenoxic acid), placed in polyethylene bags, and hand-distributed in an 80 ha area within an urban National Park in Washington, D. C. (USA). After 3 wk, target and nontarget animals were trapped and blood samples collected to evaluate bait uptake. Thirty-three of 52 (63%) raccoons had elevated blood iodine levels indicating they had eaten at least one bait, 13 (25%) were negative, and six (12%) had marginal values. These results indicate that sponge baits hand-placed at a density of 12.4/ha can reach a significant proportion of an urban raccoon population. Implications for oral rabies vaccination of raccoons are discussed.     

Oral vaccination against raccoon rabies: landscape heterogeneity and timing of distribution influence wildlife contact rates with the ONRAB vaccine bait

Aerial distribution of oral vaccine baits is one of the available strategies for controlling the spread of infectious wildlife diseases. This technique has commonly been used to control rabies in wild carnivores and, together with other techniques, was used to immunize wild populations of raccoons (Procyon lotor) and striped skunks (Mephitis mephitis) after the detection of the first rabid raccoon in the province of Quebec, Canada, in 2006. Vaccine bait distribution was conducted over large areas where agricultural land is dominant but interspersed with residual forest patches. Our objective was to evaluate the effect of habitat (forest vs. agricultural crops) in space and time on the contact rate between wildlife and the ONRAB(?) vaccine bait, a recent alternative to the V-RG(?). Four transects of eight vaccine baits each were installed parallel to, and at different distances from, the forest's edge (under forest cover, at field-forest edge, and at 50 and 200 m from forest edge in agricultural crops) at three sites composed of various crop types interspersed with forest patches. This experiment was conducted during three periods (late spring, 1-7 June; summer, 27 July-2 August; and fall, 24-30 October) in 2009. Contact rates with vaccine baits were monitored for 7 days in each period to evaluate the potential temporal variations generated within the habitat types. Contact rates with ONRAB vaccine baits were highest under forest cover and in the fall. Of 13 species observed in proximity to the vaccine baits, raccoons were the most frequent (49.5%, n=55 visits). Our study underlines the importance of taking into account landscape heterogeneity and timing of distribution when planning the distribution of vaccine baits to control rabies in raccoons.     

Formulation and evaluation of baits for oral rabies vaccination of raccoons (Procyon lotor)

Captive raccoons were offered a variety of vaccine containers and bait components in a series of three-choice tests. Paraffin wax ampules were the most readily accepted vaccine container. Preferred bait components included corn and shellfish oils, deep fried corn meal batter, and egg, apple and buttermilk flavorings. These results, together with factors including ease of bait formulation, cost, and suitability for field use, were used to develop an experimental delivery system for an oral rabies vaccine. The developed system was composed of a polyurethane sleeve (1.5 x 5.5 cm) dipped in a commercial food batter mix together with corn meal, milk and egg. The sleeve was deep fried in corn oil and a 2.0 ml ampule containing a recombinant rabies vaccine was then inserted into the sleeve bait. These baits were presented to 10 captive raccoons. Nine of the 10 animals developed high levels of rabies virus neutralizing antibodies. Field tests are needed to determine if the delivery system developed also is effective for wild raccoons.     

Oral rabies vaccine contact by raccoons and nontarget species in a field trial in Florida

Rabies is enzootic in raccoons (Procyon lotor) in the eastern United States. Oral vaccination of free-ranging raccoons against rabies has the potential to control the disease in a principal reservoir and reduce the risk of rabies exposure among domestic animals and humans. Free-ranging animal contact with baits containing a vaccinia virus recombinant vaccine expressing the rabies glycoprotein gene (V-RG) was monitored in Pinellas County (Florida, USA) from February through May 1997. Bait contact was assessed with 423 tracking plate nights; conducted in four land use zones: single residential, multiple residential, industrial-commercial, and undeveloped. The undeveloped land use zone was further described by six vegetation communities: mangrove swamp, red maple swamp, beach dune, pine forest, mixed oak hammock, and cabbage palm hammock. Seven animal taxa contacted the baited tracking plates across the four land use zones: raccoons, opossums (Didelphis virginiana), cats (Felis catus), dogs (Canis familiaris), rabbits (Sylvilagus sp.), unidentified rodents, and birds. A total of 252/413 (61%) of the baits was contacted by animals; 95 (38%) of these were specifically by the raccoon, the target species. Overall bait contact by all animals was significantly different among the four land use zones, being highest in the undeveloped zone (82%) and lowest in the industrial-commercial zone (34%). Bait contact by raccoons also was significantly different among the undeveloped and pooled urban zones. Among the six vegetation communities, bait contact by all animals was significantly different ranging from 95% in the mangrove to 50% in the cabbage palm hammock. Among the four vegetation communities tested, bait contact by raccoons also was significantly different.     

Prerequisites for oral immunization of free-ranging raccoons (Procyon lotor) with a recombinant rabies virus vaccine: study site ecology and bait system development.

A model baiting system suitable for the delivery of an oral rabies vaccine to free-ranging raccoons (Procyon lotor) was developed and tested on barrier islands in South Carolina (USA). Features of barrier island physiography and ecology were studied relative to selective bait deployment and site biosecurity. Capture-mark-recapture data were obtained from 228 raccoons. Raccoon density estimates, using a modified census assessment technique, were one raccoon per 1.8 to 2.7 ha. Mean (+/- SE) and range home area estimates of radio-collared raccoons were 84 (+/- 15.6) ha (27 to 176 ha) by a minimum convex polygon method and 138 (+/- 22.8) ha (43 to 241 ha), by a harmonic mean transformation method. Habitat utilization determinations of radio-collared raccoons were conducted to identify study areas to potentially maximize selectivity of bait towards raccoons and to reduce the absolute number of baits deployed. Island raccoons showed a habitat preference for maritime forest, maritime shrub and marsh areas. Additionally, there was no evidence of inter-island or mainland exchange of ear-tagged or radio-collared raccoons. A disease and mortality survey was conducted to identify baseline pathology and incidental lesions in the target raccoon population, prior to actual vaccination initiation. Thirty-eight percent of 30 clinically suspect raccoons sampled had intracytoplasmic eosinophilic inclusions diagnostic of canine distemper; no other lesions suggestive of viral etiologies were found. Serological surveys for raccoon poxvirus and rabies virus antibodies were negative. Antibody titers to canine adenovirus 1 and 2 indicated a moderate level of exposure (approximately 10 to 16%) in the raccoon population. Overall, 93 to 100% of placebo baits were consistently disturbed by 7 days post-bait deployment, and bait acceptance rates by raccoons ranged from 49 to 85%, by using a modular systems approach to select the optimum combination of bait attractant, biomarker, matrix, density, and distribution. These results suggest that a large proportion (up to 85%) of a free-ranging island raccoon population can be selectively and safely targeted, marked and monitored utilizing a proposed oral bait delivery system for recombinant or other rabies vaccines.     

A study of techniques for the distribution of oral rabies vaccine to wild raccoon populations

This study evaluates a technique for delivering an oral rabies vaccine to wild raccoon (Procyon lotor) populations. Various baits and attractants were first tested on caged raccoons and baiting trials were then conducted in two distinct physiographic regions of Virginia (USA), the coastal plain and the Piedmont plateau. Raccoon population density studies preceded the field trials. Each polyurethane sponge bait distributed contained approximately 200 mg tetracycline as a tissue biomarker, and was presented in an outer bag with a fish-based attractant. Baits were frozen until used and distributed from an aeroplane throughout two 4-km2 sites in each region. One site received 450 baits/km2 and the other 120 baits/km2. Postbaiting evaluation included the direct observation of baits in the field and the examination of teeth and bone from trapped and hunted animals for evidence of the biomarker. Between 30% and 73% of the captured animals showed evidence of bait consumption. The proportion of animals with evidence of bait uptake changed when areas adjacent to the actual baiting site were included. The percentage of animals taking baits was not related to the density of baits that were distributed.     

Experience with antirabies vaccination of foxes using the oral route coordinated among several European countries and perspectives on the use of recombinant vaccinia-rabies virus

Campaigns of fox vaccination against rabies were carried out in Belgium, grand-duchy of Luxembourg and France in September 1986, June and September 1987. The SAD B19 attenuated strain of rabies virus, contained in baits (Tübingen baits) was used as vaccine. Baits were distributed at a range density of 11 to 15 baits per km2. First results are very encouraging. A recombinant vaccinia virus harbouring the rabies virus glycoprotein gene has been developed. This recombinant virus can be given to the fox by the oral route and protects it against rabies virus challenge; it is also innocuous for the fox and other non-target European species. A first trial of fox vaccination against rabies using this recombinant vaccinia-rabies virus has been carried out in Belgium, on a military domain, in October 1987.     

Evaluating seasonal bait delivery to badgers using rhodamine B

In the UK and Ireland, research on the control of bovine tuberculosis in badgers includes the development of a palatable bait for oral delivery of a vaccine and a means of its deployment in the field. In the present study, we carried out field deployment of bait according to the established method of bait marking in early spring and early summer to compare the effects of seasonality on bait uptake rates. All baits contained rhodamine B (RhB) which was subsequently detected in the hair and whiskers of captured badgers. During the 8 days of bait feeding at 14 badger setts, 99% of baits deployed in spring, and 100% of those deployed in summer were removed. The presence of RhB in captured badgers indicated high rates of uptake amongst adult badgers in spring (93%) and summer (98%). Only cubs captured in summer showed evidence of having taken bait (91%). Between 67% and 100% of each social group was estimated to have taken bait. The detection of RhB in 96% of badgers captured at outlier setts, where bait was not fed, suggested that deployment at main setts alone may be sufficient to target a relatively high proportion of the badger population. The number of baits deployed per marked badger suggested that a similar level of uptake might be achievable using fewer baits. The results clearly demonstrate the potential value of the bait-marking methodology for delivering vaccine baits to badgers during spring and summer, but that deployment in early summer is necessary to target cubs.     

Use of Rhodamine B to Reveal Patterns of Interspecific Competition and Bait Acceptance in Raccoons

Abstract: Passive treatment of raccoons (Procyon lotor) through distribution of vaccine-laden baits recently has emerged as a potential solution to address health and economic conflicts associated with raccoon rabies and may have applications in the management of other pathogens carried by raccoons if frequent bait deployments are used. Consumption of baits by nontarget species reduces the efficiency in which baits can be used to manage wildlife disease, although no study has explicitly evaluated the influence of bait competitor density on the ability to treat raccoons. Our objectives were to use the biomarker Rhodamine B (RB) to 1) evaluate patterns of raccoon bait acceptance as a function of competition with Virginia opossums (Didelphis virginiana), the dominant bait competitor; 2) characterize attributes of opossum bait acceptance to improve efficiency of raccoon treatment; and 3) evaluate the effect of repeated bait exposure on rates of bait acceptance as may be required in the management of wildlife disease issues beyond rabies. Identifying bait consumption by individuals based on the presence of an RB mark in a sample of whiskers, we used logistic regression to model raccoon and opossum bait acceptance as a function of bait availability, previous exposure to baits, demographic attributes, and an index of time spent in the baited area (residency index). For both raccoons and opossums, the best measure of bait availability was the variable number of baits per opossum. The most parsimonious logistic regression model for raccoon bait acceptance included the variables baits per opossum, exposure history, and residency index. The strength of the variable baits per opossum relative to competing measures of bait availability indicated bait consumption by opossums significantly limited the ability to treat raccoons. The most parsimonious model for opossum acceptance was composed of the variables baits per opossum, sex, weight, residency index, baits per opossum X sex, and weight X sex. Patterns of opossum bait acceptance likely were driven by effects of bait availability and sex-dependent differences in movement. Our results call attention to the importance of bait competition in limiting the ability to effectively treat raccoon populations through distribution of baits and suggest managers incorporate information on density of bait competitors, particularly opossums, in allocation of baits.     

Complete Genome and Molecular Epidemiological Data Infer the Maintenance of Rabies among Kudu (Tragelaphus strepsiceros) in Namibia

Rabies in kudu is unique to Namibia and two major peaks in the epizootic have occurred since it was first noted in 1977. Due to the large numbers of kudu that were affected, it was suspected that horizontal transmission of rabies occurs among kudu and that rabies was being maintained independently within the Namibian kudu population – separate from canid cycles, despite geographic overlap. In this study, it was our aim to show, through phylogenetic analyses, that rabies was being maintained independently within the Namibian kudu population. We also tested, through complete genome sequencing of four rabies virus isolates from jackal and kudu, whether specific mutations occurred in the virus genome due to host adaptation. We found the separate grouping of all rabies isolates from kudu to those of any other canid species in Namibia, suggesting that rabies was being maintained independently in kudu. Additionally, we noted several mutations unique to isolates from kudu, suggesting that these mutations may be due to the adaptation of rabies to a new host. In conclusion, we show clear evidence that rabies is being maintained independently in the Namibian kudu population – a unique phenomenon with ecological and economic impacts.