ONRAB® oral rabies vaccine is shed from, but does not persist in, captive mammals.

ONRAB® is a human adenovirus rabies glycoprotein recombinant vaccine developed to control rabies in wildlife. To support licensing and widespread use of the vaccine, safety studies are needed to assess its potential residual impact on wildlife populations. We examined the persistence of the ONRAB® vaccine virus in captive rabies vector and non-target mammals. This research complements work on important rabies vector species (raccoon, striped skunk, and red fox) but also adds to previous findings with the addition of some non-target species (Virginia opossum, Norway rats, and cotton rats) and a prolonged period of post vaccination monitoring (41 days). Animals were directly inoculated orally with the vaccine and vaccine shedding was monitored using quantitative real-time PCR applied to oral and rectal swabs. ONRAB® DNA was detected in both oral and rectal swabs from 6 h to 3 days post-inoculation in most animals, followed by a resurgence of shedding between days 17 and 34 in some species. Overall, the duration over which ONRAB® DNA was detectable was shorter for non-target mammals, and by day 41, no animal had detectable DNA in either oral or rectal swabs. All target species, as well as cotton rats and laboratory-bred Norway rats, developed robust humoral immune responses as measured by competitive ELISA, with all individuals being seropositive at day 31. Similarly, opossums showed good response (89% seropositive; 8/9), whereas only one of nine wild caught Norway rats was seropositive at day 31. These results support findings of other safety studies suggesting that ONRAB® does not persist in vector and non-target mammals exposed to the vaccine. As such, we interpret these data to reflect a low risk of adverse effects to wild populations following distribution of ONRAB® to control sylvatic rabies.

Innate immune responses in raccoons after raccoon rabies virus infection.

Zoonotic wildlife diseases pose significant health risks not only to their primary vectors but also to humans and domestic animals. Rabies is a lethal encephalitis caused by rabies virus (RV). This RNA virus can infect a range of terrestrial mammals but each viral variant persists in a particular reservoir host. Active management of these host vectors is needed to minimize the negative impacts of this disease, and an understanding of the immune response to RV infection aids strategies for host vaccination. Current knowledge of immune responses to RV infection comes primarily from rodent models in which an innate immune response triggers activation of several genes and signalling pathways. It is unclear, however, how well rodent models represent the immune response of natural hosts. This study investigates the innate immune response of a primary host, the raccoon, to a peripheral challenge using the raccoon rabies virus (RRV). The extent and temporal course of this response during RRV infection was analysed using genes predicted to be upregulated during infection (IFNs; IFN regulatory factors; IL-6; Toll like receptor-3; TNF receptor). We found that RRV activated components of the innate immune system, with changes in levels of transcripts correlated with presence of viral RNA. Our results suggest that natural reservoirs of rabies may not mimic the immune response triggered in rodent models, highlighting the need for further studies of infection in primary hosts.

Evolution of wildlife rabies control tactics.

The development of tactics for the control of rabies in wildlife species has evolved dramatically during the past few decades in part due to research advances. Historically, rabies control measures primarily involved the culling of target species. However, contemporary advances in the research and development of oral rabies vaccines and delivery systems for wildlife have now made it feasible to treat rabies outbreaks over thousands of square kilometers of habitat. Systems have been developed to control rabies in several of the primary wildlife vectors such as raccoon dogs, red foxes, and raccoons, and rabies has been eliminated from many jurisdictions. However, future research is needed to develop cost-effective and efficacious methods to control rabies in species such as striped skunks as well as in nonterrestrial vectors such as bats. As well, cost-effective rabies management techniques need to be adopted by developing nations so that wildlife rabies control is a realistic and achievable goal globally.

MHC class II DRB diversity in raccoons (Procyon lotor) reveals associations with raccoon rabies virus (Lyssavirus).

In North America, the raccoon rabies virus (RRV) is an endemic wildlife disease which causes acute encephalopathies and is a strong selective force on raccoons (Procyon lotor), with estimates of ∼85% of the population succumbing to the disease when epizootic. RRV is regarded as a lethal disease if untreated; therefore, no evolutionary response would be expected of raccoon populations. However, variable immune responses to RRV have been observed in raccoons indicating a potential for evolutionary adaptation. Studies of variation within the immunologically important major histocompatibility complex (MHC) have revealed relationships between MHC alleles and diseases in humans and other wildlife species. This enhances our understanding of how hosts and pathogens adapt and co-evolve. In this study, we used RRV as a model system to study host-pathogen interaction in raccoons from a challenge study and from four wild populations that differ in exposure times and viral lineages. We investigated the potential role of Prlo-DRB polymorphism in relation to susceptibility/resistance to RRV in 113 RRV positive and 143 RRV negative raccoons. Six alleles were found to be associated with RRV negative status and five alleles with RRV positive animals. We found variable patterns of MHC associations given the relative number of selective RRV sweeps in the studied regions and correlations between MHC diversity and RRV lineages. The allelic associations established provide insight into how the genetic variation of raccoons may affect the disease outcome and this can be used to examine similar associations between other rabies variants and their hosts.

Stray dog population health in Jodhpur, India in the wake of an animal birth control (ABC) program.

Our objectives were to (1) estimate the prevalence of various health indices in the stray dog population in Jodhpur, India and (2) determine if there was an association between an animal birth control (ABC) program and the prevalence of these health indices in this population. A prevalence survey of 323 sexually intact stray dogs >3 months caught from the streets of Jodhpur from September to November, 2005 indicated that low body condition score (70%), skin conditions (69%) and tick infestation (68%) were the most common health problems in this population. An observational study of 888 stray dogs on the streets of Jodhpur from March to April, 2006 revealed that sterilized dogs were more likely to have a higher body condition score (BCS) than sexually intact dogs when controlling for age, based on a multinomial regression model. However, sterilized dogs were more likely to have a skin condition than sexually intact dogs, based on a logistic regression model. Our observations of the surgical/kennel facility indicated that an effective tick control program was needed. Additionally, the current parasite control protocol at the kennel/shelter facility was inadequate to treat mange, a contact-transmitted skin disease. This is the first study to evaluate the associations between an ABC program and stray dog health, apart from rabies.

Stray dog population demographics in Jodhpur, India following a population control/rabies vaccination program.

Animal Birth Control (ABC) is a program by which stray dogs are sterilized and vaccinated against rabies with the aim of controlling both dog population size and rabies. Population size and demographics of stray dogs were measured before and after implementation of an ABC program in Jodhpur, India. Dog population size declined (p<0.05) in three of five areas surveyed, showed a decreasing trend (p>0.05) in 1 area, and remained stable in 1 area between 2005 and 2007. By 2007, 61.8-86.5% of the free-roaming dog population was surgically sterilized and vaccinated for rabies in the areas surveyed. In March-May, 2007, adults comprised 80-96% of the free-roaming dog population, while subadults and puppies comprised 0-18 and 0-4%, respectively. The male:female ratio among dogs>3 months old was 1.4:1. A population demographic model predicted that at the current level of sterilization/rabies vaccination, vaccination coverage would remain above 70%, and the dog population would decrease by 69% reaching stability after 13-18 years. A surgical sterilization coverage under 40% would maintain the dog population at current levels.

Assessing reproductive patterns and disorders in free-ranging dogs in Jodhpur, India to optimize a population control program.

The objectives were to test the hypothesis that estrus and pregnancy are seasonal in free-ranging female dogs (>3 mo old) in Jodhpur, India, and to determine litter size, and the prevalence of fetal resorption in this population. The prevalence of estrus and pregnancy was determined in 5400 free-ranging bitches (trapped and released) at the time of ovariohysterectomy. In a separate study, the uteri and ovaries of 246 free-ranging bitches were examined to determine litter size and fetal resorption. The bitches exhibited seasonal estrus and pregnancy (P < 0.00001), with a higher percentage of bitches in estrus or pregnant during the late monsoon season (September to November) compared to the other three seasons. The mean litter size based on embryo/fetal counts was 4.6 (95% CI = 4.0-5.3; n = 40) and based upon placental site counts was 4.4 (95% CI = 3.9-4.8; n = 105). Prevalence of fetal resorption was 32.6% (95% CI = 20.5-47.5; n = 43) with a mean of 2.8 resorptions per litter in those with at least one resorption (95% CI = 1.8-3.8; n = 14). This was the first study to estimate previous litter size of non-pregnant, free-ranging dogs based upon placental sites. Litter size data from this study will be used in a population demographic model to predict the long-term impact of animal birth control (ABC) on the free-ranging dog population in Jodhpur. Increasing the efforts to surgically sterilize bitches prior to the time of year of peak pregnancy or whelping will help maximize the impact of an ABC program on the Jodhpur free-ranging dog population.

Safety studies on an adenovirus recombinant vaccine for rabies (AdRG1.3-ONRAB) in target and non-target species.

A replication-competent human adenovirus vector in which the rabies virus glycoprotein gene was inserted (AdRG1.3-ONRAB) was given by direct instillation into the oral cavity to representatives of three wildlife vector species of concern in Ontario (red fox, raccoon and striped skunk) and to a variety of non-target wildlife species, domestic and laboratory species. Despite use of a relatively high dose of vaccine, no untoward clinical signs were observed. Subsequent to vaccine exposure, detection of vaccine virus in lung, spleen, intestine, liver, kidney and brain of each animal was attempted using an ONRAB-specific assay combining PCR with Southern blotting (PCR-SB). Of the 1280 tissue samples obtained from vaccinates or contact animals, 18 (1.4%) were found to be PCR-SB positive. Virus isolation attempts were performed utilizing cell culture for all PCR-SB positive tissues and a selection of PCR-SB negative tissues. Histological examination performed on all PCR-SB positive tissues failed to identify lesions attributed to the vaccine. A quantitative real-time PCR was used to determine the excretion of the vaccine in feces and in the oral cavity with 0.8% of oral swabs and 6.8% of fecal specimens found to be positive. The low rates of recovery of vaccine virus from tissues, feces and the oral cavity suggest that the likelihood of ONRAB causing a negative impact on wildlife species is unlikely.

Multiple movement modes by large herbivores at multiple spatiotemporal scales.

Recent theory suggests that animals should switch facultatively among canonical movement modes as a complex function of internal state, landscape characteristics, motion capacity, and navigational capacity. We tested the generality of this paradigm for free-ranging elk (Cervus elaphus) over 5 orders of magnitude in time (minutes to years) and space (meters to 100 km). At the coarsest spatiotemporal scale, elk shifted from a dispersive to a home-ranging phase over the course of 1-3 years after introduction into a novel environment. At intermediate spatiotemporal scales, elk continued to alternate between movement modes. During the dispersive phase, elk alternated between encamped and exploratory modes, possibly linked to changes in motivational goals from foraging to social bonding. During the home-ranging phase, elk movements were characterized by a complex interplay between attraction to preferred habitat types and memory of previous movements across the home-range. At the finest temporal and spatial scale, elk used area-restricted search while browsing, interspersed with less sinuous paths when not browsing. Encountering a patch of high-quality food plants triggered the switch from one mode to the next, creating biphasic movement dynamics that were reinforced by local resource heterogeneity. These patterns suggest that multiphasic structure is fundamental to the movement patterns of elk at all temporal and spatial scales tested.

Socially informed random walks: incorporating group dynamics into models of population spread and growth.

Simple correlated random walk (CRW) models are rarely sufficient to describe movement of animals over more than the shortest time scales. However, CRW approaches can be used to model more complex animal movement trajectories by assuming individuals move in one of several different behavioural or movement states, each characterized by a different CRW. The spatial and social context an individual experiences may influence the proportion of time spent in different movement states, with subsequent effects on its spatial distribution, survival and fecundity. While methods to study habitat influences on animal movement have been previously developed, social influences have been largely neglected. Here, we fit a 'socially informed' movement model to data from a population of over 100 elk (Cervus canadensis) reintroduced into a new environment, radio-collared and subsequently tracked over a 4-year period. The analysis shows how elk move further when they are solitary than when they are grouped and incur a higher rate of mortality the further they move away from the release area. We use the model to show how the spatial distribution and growth rate of the population depend on the balance of fission and fusion processes governing the group structure of the population. The results are briefly discussed with respect to the design of species reintroduction programmes.

Evidence of Parelaphostrongylus tenuis infections in free-ranging elk (Cervus elaphus) in southern Ontario.

The antemortem detection of a Parelaphostrongylus tenuis infection in a free-ranging wild elk (Cervus elaphus) in southern Ontario is documented. Postmortems on other free-ranging elk that died during 2000-2005 indicated that 59% (17/29) were infected with P. tenuis, based on presence of lesions in the brain.

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).

Rabies in vaccinated raccoons from Ontario, Canada.

From 1999 to 2006, 132 cases of raccoon rabies, caused by the raccoon variant of rabies virus, were confirmed in eastern Ontario, Canada. Trap-vaccinate-release (TVR) and point infection control (PIC) programs were implemented to control the disease; 43,014 raccoons (Procyon lotor) were vaccinated against rabies by injection (Imrab3) during that period. Two vaccinated raccoons were diagnosed with rabies at 6 mo and at 2 wk postvaccination. One may have been due to a vaccination failure. The other was likely due to the animal being in the late stages of incubation for rabies at the time of vaccination. This information will be useful to wildlife rehabilitators and agencies that hold raccoons in captivity in that a vaccinated raccoon is not necessarily immune to rabies.

Porcupine quills in raccoons as an indicator of rabies, distemper, or both diseases: disease management implications.

A relationship was detected between the presence of embedded porcupine quills and the diagnosis of rabies in raccoons in eastern Canada during 1999-2004. No relationship was found between the presence of quills in raccoons and the diagnosis of canine distemper. Raccoons with embedded quills should be submitted for rabies testing.

Behavior, movements, and demographics of rabid raccoons in Ontario, Canada: management implications.

During 1999-2003, 127 cases of raccoon variant rabies were reported in raccoons (Procyon lotor) and striped skunks (Mephitis mephitis) in Ontario, Canada. Raccoons accounted for 98% (125/127) of the reported cases with behaviors/conditions including aggression, fighting with dogs, ataxia, vocalizations, appearance of being sick, and the presence of porcupine (Erethizon dorsatum) quills. Seventy-eight percent of the rabid raccoons were adults. Juveniles were underrepresented (22%) compared with the adult/juvenile ratios found in nonrabid Ontario raccoon populations. Of the known aged raccoons, 83% were < or = 3 yr of age, and 22% of the rabid adult female raccoons had evidence of having had a litter during the year in which they were found to be rabid. The majority of rabid raccoons were reported during the fall, winter, and spring, suggesting a relationship between raccoon behavioral activities such as denning and breeding and the timing of rabies outbreaks. Multiple cases of raccoon rabies occurred at several barns, suggesting that those structures serve as focal points of rabies transmission as a result of denning activities. Movements of five rabid raccoons (range 1,564-4,143 m) were not different from movements of nonrabid raccoons in Ontario. Sixty-six percent of the rabid animals were submitted by government staff, stressing the importance of those agencies in rabies control and surveillance operations. Increased knowledge of the behaviors of rabid raccoons should assist in the development of management strategies for rabies.

Estimating the incubation period of raccoon rabies: a time-space clustering approach.

We used a time-space clustering approach to estimate the incubation period of raccoon rabies in the wild using data from the 1999-2001 invasion of raccoon rabies into eastern Ontario from northern New York State. The time differences and geographical distances between all possible pairs of rabies cases were computed, classified and assembled into a time-space matrix. The rows of that matrix represented differences in cases in weeks and the columns represent distances between cases in kilometers and the values in the cells of the matrix represent the counts of cases at specific time and distance intervals. There was a significant cluster of pairs 5 weeks apart with apparent harmonics at additional 5-week intervals. These results are explained by assuming the incubation period of raccoon rabies had a mode of 5 weeks. The time clusters appeared consistently at distance intervals of 5 km. We discuss the possibility that the spatial intervals were influenced by the 5 km radius of the point infection control depopulation process used in 1999 and the 10-15 km radial areas used in 2000. With the practical limits of those radii, there was an intensive effort to eliminate raccoons. Our procedure is easy to implement and provides an estimate of the shape of the distribution of incubation periods for raccoon rabies.