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.     

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).     

The raccoon an emerging rabies host

The geographic distribution of enzootic rabies in raccoons is currently limited to Florida and Georgia. The enzootic area has steadily expanded northward into Georgia during the 1960's. An epizootic in a population of raccoons closely associated with the human residents of a small, off-shore island was investigated. Forty-seven raccoons from the island were confirmed rabies positive by the State Laboratory from January 22 to May 20, 1969. About 10 percent of the raccoons trapped during the peak of the epizootic were positive and all of these had high titers of virus in their salivary glands but were asymptomatic in behavior. Human exposure typically resulted from close approach to a "sick" raccoon. Similar outbreaks may occur elsewhere when humans live in close contact with a semi-domesticated raccoon population. Rabies vaccination of raccoons should be considered as an alternative to population reduction in such situations.     

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.     

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.     

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.     

Adult raccoon survival in an enzootic rabies area of Pennsylvania

Twenty-one adult raccoons (Procyon lotor) were radio-marked on each of two areas in Centre County, Pennsylvania from 17 June to 23 August 1987. Raccoons on Area 1 were vaccinated with a commercial inactivated rabies virus vaccine administered intramuscularly, whereas on Area 2 raccoons were not vaccinated. Survival rates were estimated for three periods: pre-season (23 August to 23 October 1987), harvest season (24 October 1987 to 23 January 1988) and post-season (24 January to 26 March 1988). Kaplan-Meier survival rates (+/- SE) were 1.00 +/- 0.00 for both areas during the pre- and post-season periods. Survival rates during the harvest period were 0.67 +/- 0.11 and 0.69 +/- 0.11 for Area 1 and Area 2, respectively. Survival rates between the two areas were not different (P = 0.929). During 23 August 1987 to 26 March 1988, rabies was not an apparent factor in raccoon survival. Conclusions regarding timing an oral rabies vaccination campaign based upon occurrence of rabies-related mortalities could not be presented because of the lack of obvious rabies mortality. However, our findings, combined with information about immunization, vaccine distribution, and peak periods of raccoon rabies, suggest a late winter or early spring vaccination period would be optimum for reducing the number of raccoons susceptible to rabies.     

A descriptive epidemiological study of raccoon rabies in a rural environment

A recent outbreak of rabies in raccoons, Procyon lotor (L.), in Loudoun County, Virginia (1981-82), prompted a study of the epidemiology of the disease. Parameters studied included the occurrence and movement of the disease over time, sex and age relationships, and behavior patterns of raccoons. During the 18 mo, 427 raccoons were tested, of which 75% were infected with rabies virus. Interpretation of rainfall data and the subsequent spatial occurrence of infected raccoons within the county indicated a cause and effect relationship. The submission rate of female raccoons was greater than that of males. The female raccoons (adult and juvenile) were also found to be infected with the virus more often than the males. Behavior of infected raccoons in a rural environment was similar to those observed in the southeastern United States during earlier epizootics of rabies. The presence of a skunky odor on infected raccoons may be a characteristic of raccoon rabies.     

Landscape modelling spatial bottlenecks: implications for raccoon rabies disease spread

A landscape genetic simulation modelling approach is used to understand factors affecting raccoon rabies disease spread in southern Ontario, Canada. Using the Ontario Rabies Model, we test the hypothesis that landscape configuration (shape of available habitat) affects dispersal, as indicated by genetic structuring. We simulated range expansions of raccoons from New York into vacant landscapes in Ontario, in two areas that differed by the presence or absence of a landscape constriction. Our results provide theoretical evidence that landscape constriction acts as a vicariant bottleneck. We discuss implications for raccoon rabies spread.     

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.     

Exposure time of oral rabies vaccine baits relative to baiting density and raccoon population density

Oral rabies vaccination (ORV) baiting programs for control of raccoon (Procyon lotor) rabies in the USA have been conducted or are in progress in eight states east of the Mississippi River. However, data specific to the relationship between raccoon population density and the minimum density of baits necessary to significantly elevate rabies immunity are few. We used the 22-km2 US National Aeronautics and Space Administration Plum Brook Station (PBS) in Erie County, Ohio, USA, to evaluate the period of exposure for placebo vaccine baits placed at a density of 75 baits/km2 relative to raccoon population density. Our objectives were to 1) estimate raccoon population density within the fragmented forest, old-field, and industrial landscape at PBS: and 2) quantify the time that placebo, Merial RABORAL V-RG vaccine baits were available to raccoons. From August through November 2002 we surveyed raccoon use of PBS along 19.3 km of paved-road transects by using a forward-looking infrared camera mounted inside a vehicle. We used Distance 3.5 software to calculate a probability of detection function by which we estimated raccoon population density from transect data. Estimated population density on PBS decreased from August (33.4 raccoons/km2) through November (13.6 raccoons/km2), yielding a monthly mean of 24.5 raccoons/km2. We also quantified exposure time for ORV baits placed by hand on five 1-km2 grids on PBS from September through October. An average 82.7% (SD = 4.6) of baits were removed within 1 wk of placement. Given raccoon population density, estimates of bait removal and sachet condition, and assuming 22.9% nontarget take, the baiting density of 75/ km2 yielded an average of 3.3 baits consumed per raccoon and the sachet perforated.     

Spatial patterns of neutral and functional genetic variations reveal patterns of local adaptation in raccoon (Procyon lotor) populations exposed to raccoon rabies

Local adaptation is necessary for population survival and depends on the interplay between responses to selective forces and demographic processes that introduce or retain adaptive and maladaptive attributes. Host–parasite systems are dynamic, varying in space and time, where both host and parasites must adapt to their ever‐changing environment in order to survive. We investigated patterns of local adaptation in raccoon populations with varying temporal exposure to the raccoon rabies virus (RRV). RRV infects approximately 85% of the population when epizootic and has been presumed to be completely lethal once contracted; however, disease challenge experiments and varying spatial patterns of RRV spread suggest some level of immunity may exist. We first assessed patterns of local adaptation in raccoon populations along the eastern seaboard of North America by contrasting spatial patterns of neutral (microsatellite loci) and functional, major histocompatibility complex (MHC) genetic diversity and structure. We explored variation of MHC allele frequencies in the light of temporal population exposure to RRV (0–60 years) and specific RRV strains in infected raccoons. Our results revealed high levels of MHC variation (66 DRB exon 2 alleles) and pronounced genetic structure relative to neutral microsatellite loci, indicative of local adaptation. We found a positive association linking MHC genetic diversity and temporal RRV exposure, but no association with susceptibility and resistance to RRV strains. These results have implications for landscape epidemiology studies seeking to predict the spread of RRV and present an example of how population demographics influence the degree to which populations adapt to local selective pressures.     

Predictive spatial dynamics and strategic planning for raccoon rabies emergence in Ohio

Rabies is an important public health concern in North America because of recent epidemics of a rabies virus variant associated with raccoons. The costs associated with surveillance, diagnostic testing, and post-exposure treatment of humans exposed to rabies have fostered coordinated efforts to control rabies spread by distributing an oral rabies vaccine to wild raccoons. Authorities have tried to contain westward expansion of the epidemic front of raccoon-associated rabies via a vaccine corridor established in counties of eastern Ohio, western Pennsylvania, and West Virginia. Although sporadic cases of rabies have been identified in Ohio since oral rabies vaccine distribution in 1998, the first evidence of a significant breach in this vaccine corridor was not detected until 2004 in Lake County, Ohio. Herein, we forecast the spatial spread of rabies in Ohio from this breach using a stochastic spatial model that was first developed for exploratory data analysis in Connecticut and next used to successfully hind-cast wave-front dynamics of rabies spread across New York. The projections, based on expansion from the Lake County breach, are strongly affected by the spread of rabies by rare, but unpredictable long-distance translocation of rabid raccoons; rabies may traverse central Ohio at a rate 2.5-fold greater than previously analyzed wildlife epidemics. Using prior estimates of the impact of local heterogeneities on wave-front propagation and of the time lag between surveillance-based detection of an initial rabies case to full-blown epidemic, specific regions within the state are identified for vaccine delivery and expanded surveillance effort.     

Elimination of rabies from red foxes in eastern Ontario

The province of Ontario (Canada) reported more laboratory confirmed rabid animals than any other state or province in Canada or the USA from 1958-91, with the exception of 1960-62. More than 95% of those cases occurred in the southern 10% of Ontario (approximately 100,000 km2), the region with the highest human population density and greatest agricultural activity. Rabies posed an expensive threat to human health and significant costs to the agricultural economy. The rabies variant originated in arctic foxes: the main vector in southern Ontario was the red fox (Vulpes vulpes), with lesser involvement of the striped skunk (Mephitis mephitis). The Ontario Ministry of Natural Resources began a 5 yr experiment in 1989 to eliminate terrestrial rabies from a approximately 30,000 km2 study area in the eastern end of southern Ontario. Baits containing oral rabies vaccine were dropped annually in the study area at a density of 20 baits/km2 from 1989-95. That continued 2 yr beyond the original 5 yr plan. The experiment was successful in eliminating the arctic fox variant of rabies from the whole area. In the 1980's, an average of 235 rabid foxes per year were reported in the study area. None have been reported since 1993. Cases of fox rabies in other species also disappeared. In 1995, the last bovine and companion animal cases were reported and in 1996 the last rabid skunk occurred. Only bat variants of rabies were present until 1999, when the raccoon variant entered from New York (USA). The success of this experiment led to an expansion of the program to all of southern Ontario in 1994. Persistence of terrestrial rabies, and ease of elimination, appeared to vary geographically, and probably over time. Ecological factors which enhance or reduce the long term survival of rabies in wild foxes are poorly understood.     

Investigations into rabies II

Antonie van Leeuwenhoek -     

Oral vaccination of skunks with raccoon poxvirus recombinants expressing the rabies glycoprotein or the nucleoprotein.

Twenty nine skunks (Mephitis mephitis) were vaccinated orally with raccoon poxvirus (RCN) recombinants: 10 with a recombinant expressing the rabies virus glycoprotein (RCNRG), 10 with RCNRG mixed with a recombinant expressing the rabies virus nucleoprotein (RCNRN) and nine with RCN alone. Rabies virus neutralizing antibodies were detected in six of the 20 skunks; five skunks (three given RCNRG, two given a mixture of recombinants) survived a rabies challenge that was lethal for nine skunks vaccinated with RCN alone.