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.     

Comparison of suburban vaccine distribution strategies to control raccoon rabies

Helicopters and hand baiting are commonly used to distribute vaccine-laden baits to help control raccoon (Procyon lotor) rabies in suburban landscapes, but these techniques may be labor intensive, costly, or unavailable in some areas. We tested conventional baiting strategies against polyvinyl-chloride (PVC) bait stations in Erie County (New York, USA) during July and August 2003-05. Hand, helicopter, and bait station treatments were randomly assigned to six 25-km(2) suburban study sites. To estimate the proportion of raccoons that ingested baits, tooth and blood samples from 954 raccoons were collected and examined for tetracycline biomarker and rabies-neutralizing antibodies, respectively. Overall, 38% (358/954) of the raccoons in Erie County tested positive for tetracycline; 16% (155/954) tested seropositive for rabies virus. Year of study significantly impacted biomarker prevalence; fewer raccoons tested positive for tetracycline in 2004. Probability of seropositivity increased with raccoon age. No statistically significant differences existed between baiting strategies and frequencies of biomarker and antibody-positive raccoons across all years combined. Thus, bait stations could be used as part of an integrated rabies control strategy.     

Oral rabies vaccination of a northern Ohio raccoon population: relevance of population density and prebait serology

Ohio's oral rabies vaccination (ORV) program was established to prevent the westward spread of the raccoon (Procyon lotor) rabies virus (Lyssavirus, Rhabdoviridae) in Ohio, USA. The program, which targets raccoons, distributes vaccine-bait units (VBU) at a target density of 75 units/km2. Few studies have examined the relationship of VBU density and target population density to the prevalence of rabies virus-neutralizing antibodies (RVNA). We conducted experimental VBU distributions in August 2003 and August 2004, 150 km west of the ORV zone where there was no history of raccoon rabies. We measured change in RVNA titers in blood collected from live-trapped raccoons before and after VBU distributions. A closed population mark-recapture estimate of the size of the target population was 91 raccoons/km2, compared to the realized VBU distribution density of 70 units/km2. Surprisingly, 41% of 37 serum samples were RVNA-positive (>or=0.05 IU/ml) before VBU distribution in 2003, but all titers were <0.25 IU/ml. Although viable VBUs were distributed in August 2003, only 21% of 315 samples were RVNA-positive before VBU distribution in 2004, but 9% had titers>or=0.25 IU/ml. Tetracycline (biomarker in bait) prevalence in teeth indicated that 57% of raccoons ingested VBUs after distribution in 2003, and 54% ingested VBUs after distribution in 2004. However, only 8% and 11% of sera were positive for RVNA (>or=0.05 IU/ml) after VBU distribution in 2003 and 2004, respectively. Only 4-5% of sera collected after bait distribution had titers>or=0.25 IU/ml each year. The standard distribution density of 75 VBUs/km2 was insufficient to produce a population-wide immunoprotective response against rabies infection in our high-density target population. Presence of RVNA in a presumed na?ve population before baiting demonstrates that estimating prevalence of RVNA after oral rabies vaccination can be problematic without knowledge of background titers and seasonal changes in prevalence of RVNA before and after baiting.     

Evaluation and significance of tetracycline stability in rabies vaccine baits

Tetracycline is widely used as a biomarker for bait consumption by wildlife; tetracycline is incorporated into bones and teeth and can be detected by fluorescence microscopy several weeks postconsumption. During 2003, the United States Department of Agriculture distributed more than 10 million tetracycline-containing rabies-vaccine baits to control the spread of wildlife vectored rabies to humans, pets, and livestock. To estimate the percentage of target species consuming the baits, raccoons and skunks were collected in baited areas and teeth were analyzed for the presence of the biomarker. Several incidents of low biomarker detection rates prompted an investigation of the stability of the biomarker in the baits. Baits were collected at several points along the manufacturing and distribution chain. Baits were analyzed for free and polymer-bound tetracycline and the less active isomer epitetracycline. Results indicated that a portion of the tetracycline was converted to epitetracycline. Additionally, significant quantities of both compounds were trapped in the polymer, which is homogeneously distributed throughout the bait. The results of this study suggest that approximately 40% of the target quantity of tetracycline was unavailable for absorption. This situation could contribute to low biomarker detection rates and suggests that formulation modification should be considered.     

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.     

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.     

Bait delivery for oral rabies vaccine to gray foxes

Rabies is a widespread zoonotic disease that has reached epizootic proportions in gray foxes (Urocyon cinereoargenteus) in central Texas. Because each species of carnivore has different food preferences and foraging strategies, it is essential that the efficacy of a bait delivery program be examined for gray foxes prior to an oral vaccination program being attempted. Field trials were conducted to determine bait preferences of free-ranging gray foxes to selected baits and odor attractants. Baits consisted of polymer cubes made of either dog food meal or fish meal, and a wax-lard cake that was enhanced with marshmallow flavoring. Attractants added to baits exuded sulfurous, fatty, cheesy, or sweet odors and flavors. During 3,589 operable bait station nights, gray fox visitation and bait uptake rates were 9.2% and 8.3%, respectively. Gray foxes exhibited no preference in bait uptake rates between bait and odor attractant combinations. Gray foxes exhibited no difference in cumulative bait uptake rates between onroad and offroad sites; however, the uptake rate by raccoons was significantly greater for baits placed on roads than for baits randomly placed. Raccoons were the major non-target species competing for baits, being attributed with 73% of the total uptake. Visitation and bait uptake rates by raccoons significantly increased after a 7-day lethal removal of raccoons (n = 37) from the study area. Random distribution of baits is recommended; it reduced bait uptake by non-target species without adversely affecting uptake by gray foxes.     

A baiting system for delivery of an oral plague vaccine to black-tailed prairie dogs

Laboratory and field studies were conducted between July and October 1999 to identify bait preference, biomarker efficacy, and bait acceptance rates for delivering an oral plague vaccine to black-tailed prairie dogs (Cynomys ludovicianus). Twenty juvenile captive prairie dogs were offered alfalfa baits containing either alfalfa, alfalfa and 5% molasses, or alfalfa, 5% molasses and 4% salt. Based on the results of these trials we selected a bait containing alfalfa, 7% molasses, and 1% salt for field trials to determine bait acceptance rates by free-ranging animals. The biomarkers DuPont Blue dye, iophenoxic acid, and tetracycline hydrochloride were orally administered to captive prairie dogs to determine their efficacy. Only tetracycline proved effective as a biomarker. Two field trials were conducted at separate prairie dog colonies located at the Buffalo Gap National Grassland (Pennington County, South Dakota, USA). In Trial 1, three baits containing tetracycline were distributed around each active burrow entrance and an additional bait was placed inside the burrow (1,276 baits total). In Trial 2, baits were distributed at the same density per burrow as Trial 1, but along transects spaced 10 m apart (1,744 baits total). Trapping began 3 days after bait distribution, and 30 prairie dogs then were captured at each site to determine the percentage of animals marked. In Trial 1, 67% of the prairie dogs captured had tetracycline deposits indicative of bait consumption. In Trial 2, 83% of the prairie dogs had ingested a bait. Approximately 15% of the animals in both trials ate more than one bait. Fleas (Opisocrostis hirsutus) were found on 64 of 70 (91%) of the prairie dogs captured during this study.     

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.     

The elimination of raccoon rabies from Wolfe Island, Ontario: animal density and movements

During 1996 to 1998, an average of 52% to 55% of the raccoon (Procyon lotor) population on Wolfe Island, Ontario was vaccinated against rabies during proactive trap-vaccinate-release (TVR) operations. However, during 1999, the percent of the population vaccinated declined to 39% and an outbreak (6 cases) of raccoon rabies occurred on the island from December 1999 to January 2000. The raccoon population on Wolfe Island declined dramatically (71% reduction) from 1,067 raccoons (mean density = 8.4/km(2) [6.4-12.4, 95% CI]) during 1999 to 305 raccoons (mean density = 2.4/km(2) [0.87-4.1, 95% CI]) in the spring of 2000. Raccoon density immediately following the outbreak was significantly lower in cells with rabies cases, suggesting that rabies had a negative effect on population size. However, raccoon density had doubled by 1 yr following the outbreak. Movement of raccoons on Wolfe Island was as great as 24 km. Male raccoons moved greater distances than females. Movements to surrounding islands were also noted for raccoons ear tagged on Wolfe Island which indicates the island could serve as a focus for greater geographic rabies spread. Point infection control (PIC) during 2000, TVR during 2001-02, and the aerial distribution of Vaccinia-Rabies Glycoprotein (V-RG) baits during 2000 and 2003-05 were used to eliminate rabies from Wolfe Island. No cases have been detected since late January 2000 (to February 2007).     

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.     

Epidemiology of raccoon rabies in Virginia, 1984 to 1989.

Geographical and temporal trends in reports of rabid raccoons (Procyon lotor) in Virginia were summarized for 1984 to 1989; 3,256 raccoons were submitted for rabies testing, of which 1,053 (32.3%) had rabies. Both the absolute number of rabid raccoons and the percent of rabid raccoons (number rabid divided by number submitted) were examined for seasonal and yearly trends. Geographically, the epidemic moved eastward and southward in the state. The seasonal trend showed bimodal peaks in late winter and early fall and a seasonal low in summer. The percent of rabies positive raccoons peaked 1 mo earlier than the absolute number of rabies positive raccoons. The peak in the number of rabies positive raccoons occurred in 1987, while the percent of rabies positive raccoons peaked in 1986. These trends were used to recommend timing and placement of oral vaccine as one strategy to control raccoon rabies in wildlife.     

Safety of lyophilized SAG2 oral rabies vaccine in collared lemmings

Fifteen collared lemmings (Dicrostonyx groenlandicus) were exposed to a lyophilized oral rabies vaccine designed to immunize wild carnivore populations. No animals contracted rabies from the vaccine as determined by the absence of clinical signs after 37 days and lack of rabies virus in brain tissue determined by the fluorescent antibody (FA) test. These results suggest that collared lemmings would not contract rabies if they ingested this lyophilized vaccine in the wild during bait vaccination programs for arctic foxes (Alopex lagopus).     

Immunization of arctic foxes (Alopex lagopus) with oral rabies vaccine

Arctic foxes (Alopex lagopus) were successfully immunized against rabies using an orally-administered, liquid SAD-BHK21 live virus vaccine in a sausage bait. Immunization was determined by serologic response and by resistance to challenge with an arctic rabies virus strain. Virus was not shed in saliva following oral vaccination, indicating that arctic foxes would not infect other foxes after ingesting this vaccine. High antibody levels were present in all experimental foxes 2 wk following initial vaccination. A booster vaccination at 56 wk induced a significant serologic response within 1 wk, suggesting an anamnestic response but titers began to decline within 8 wk in most foxes. Foxes were observed for 16 mo following the challenge and exhibited no symptoms of rabies. The SAD-BHK21 rabies vaccine in a sausage bait system has a strong potential for vaccinating wild populations of arctic fox.     

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.     

Differential permeability of rivers to raccoon gene flow corresponds to rabies incidence in Ontario, Canada

The correlation of landscape features with genetic discontinuities reveals barriers to dispersal that can contribute to understanding present and future spread of wildlife diseases. This knowledge can then be used for targeting control efforts. The impact of natural barriers on raccoon dispersal was assessed through genetic analysis of samples from two regions, Niagara ( N  = 666) and St. Lawrence ( N  = 802). These areas are transected by major rivers and are at the northern front of a raccoon rabies epizootic. Genetic clusters were identified in each region using Bayesian clustering algorithms. In the Niagara region, two clusters were identified corresponding to either side of the Niagara River. For the St. Lawrence region, spatially congruent clusters were not identified, despite the presence of the intervening St. Lawrence River. These genetic data are consistent with raccoon rabies incidence data where rabies has been detected across the St. Lawrence River in Ontario while no cases have been detected in Ontario across the Niagara River. This is despite expectations of rabies incidence in Niagara before the St. Lawrence based on the progression of rabies from New York. The results from the two regions suggest different permeabilities to raccoons between New York and Ontario that may be attributed to the rivers. However, other factors have also been explored that could contribute to this difference between these study sites including the shape of the landscape and resource distribution.     

保存温度和时间对狂犬病疫苗效力稳定性的影响

对人用精制狂犬病疫苗和浓缩狂犬疫苗分析在－33℃,2－8℃,25℃,37℃保存0－6个月进行效力的稳定性观察,结果表明随存放时间及温度增加,两者的生物活性均有不同程度的降低。在2－8℃条件下疫苗的效力明显比其它温度稳定（P＜0．02）;精制疫苗与浓缩疫苗比较,其效力有更稳定的趋势。