Oral vaccination of raccoons (Procyon lotor) with an attenuated (SAD-B19) rabies virus vaccine

Unlike previous reports to the contrary, raccoons (Procyon lotor) were successfully vaccinated against rabies with a liquid SAD-B19 attenuated virus vaccine administered per os and given in vaccine-laden baits. There was neither evidence of vaccine-induced rabies in raccoons nor in a limited safety trial with opossums (Didelphis virginiana) given SAD-B19. Protection from lethal street rabies virus infection was not absolute: only three of nine raccoons given 1 x 10(6.0) TCID/ml were protected versus five of 10 raccoons given 1 x 10(7.0) TCID/ml of SAD-B19 and challenged 4 mo after consumption of vaccine-laden baits. Six of eight raccoons consuming 1 x 10(8.8) TCID/ml of SAD-B19 vaccine in baits survived street rabies virus challenge 2 mo postvaccination. Raccoon survivorship was not wholly dependent upon rabies virus-neutralizing antibody titer on the day of challenge. Vaccinated raccoons demonstrated a prominent anamnestic response within 1 wk following challenge. Surviving raccoons were observed for a minimum of 3 mo following street rabies virus challenge with neither clinical nor pathologic evidence of rabies. The SAD-B19 rabies vaccine administered within baits in captivity appears less effective for raccoons than for its demonstrated efficacy in the immunization of free-ranging foxes (Vulpes vulpes) in Europe.     

Vaccination of free-ranging Pennsylvania raccoons (Procyon lotor) with inactivated rabies vaccine

Thirty-nine free-ranging raccoons (Procyon lotor) in an endemic rabies area of Pennsylvania (USA) were vaccinated with a single intramuscular inoculation of commercial inactivated rabies virus vaccine, 17 June to 23 August 1987. Paired serum samples, pre- and postvaccination, were obtained from eight raccoons and were analyzed in vitro for rabies virus neutralizing antibody using a modified rapid fluorescent focus inhibition test. Seven of eight (88%) recaptured raccoons demonstrated seroconversion within 15 to 26 days of vaccination. At 1 yr postvaccination, three vaccinated raccoons were recaptured and challenged in captivity with street rabies virus, resulting in the death of two of three vaccinates and five of five unvaccinated control raccoons.     

Clinical coccidiosis in raccoons (Procyon lotor)

Clinical coccidiosis was diagnosed in wild-caught and captive raccoons. Eimeria procyonis-like oocysts were seen in 15 of 15 captive raccoons. In 6 of 6 juvenile wild-caught raccoons examined at necropsy, endogenous coccidian stages were seen in the small intestine. Two types of schizonts (large and small) were identified. Large schizonts were up to 110 microm long, contained 10-microm-long merozoites, and were in crypt glandular epithelial cells. Smaller schizonts were 10 microm long, contained 5-microm-long merozoites, and were at the tips of the villi. Only a few gamonts and no oocysts were seen in sections. These stages were thought to be of E. procyonis.     

Experimental inoculation of raccoons (Procyon lotor) with rabies virus of skunk origin.

To determine raccoon (Procyon lotor) susceptibility and serum neutralizing antibody response to a skunk salivary gland rabies virus, raccoons were inoculated with a rabies virus isolated from a naturally-infected striped skunk (Mephitis mephitis). Raccoons were divided into four groups of three animals each. A dilution of the rabies virus suspension, 10(2.4), 10(3.4), or 10(4.8), mouse intracerebral lethal dose50 (MICLD50), was administered into the masseter muscles of each animal. Three negative control animals received only diluent. Saliva and sera were collected on post-inoculation days 35, 63 and 92 for virus isolation and determination of serum neutralizing antibody titer. All animals survived the 92 day observation period and none exhibited the behavioral changes classically associated with clinical rabies virus infections. Rabies virus was not detected in the saliva of any raccoon and two of the three animals receiving the highest inoculum developed serum neutralizing antibodies (SNA). On day 92, a challenge suspension of New York City/Georgia (NYC/GA) strain rabies virus in fox salivary glands (10(3.2) MICLD50) was administered to all 12 raccoons. All animals succumbed to rabies virus except the two animals that had earlier developed SNA. The results of this study provided evidence about the susceptibility of raccoons to a skunk rabies virus and demonstrated that exposed raccoons could survive for at least 92 days following exposure. Furthermore, animals developing SNA under such circumstances were capable of withstanding challenge with rabies virus that was fatal for seronegative raccoons.     

Hemoparasites of raccoons (Procyon lotor) in Florida

Four hemoparasite species (Babesia lotori, Trypanosoma cruzi, Dirofilaria tenuis and Mansonella llewellyni) were found in raccoons (Procyon lotor) collected from 1972 to 1974 in Duval (n = 14) and Collier (n = 170) counties, Florida (USA). Trypanosoma cruzi was found in thin blood smears from one raccoon at each locality. The prevalence of B. lotori was 79% and 80% in samples taken in December 1973 in Collier and Duval counties, respectively. No patent infections by B. lotori were detected in raccoons collected in Collier County in December 1972, but 42% of the raccoons examined in September 1973 were infected. In Collier County there were no significant differences in the prevalence of B. lotori by host sex or age. In Duval County, overall D. tenuis prevalence was 7%, whereas that of M. llewellyni was 14%; the latter species was not found in Collier County. Adult raccoons had a significantly greater prevalence of D. tenuis (32%) than did subadults and juveniles (7%), and male raccoons showed a significantly greater prevalence (51%) than did females (8%).     

Architecture of the masticatory apparatus in eastern raccoons (Procyon lotor lotor)

The structure and function of the masticatory apparatus of raccoons resemble those found in carnivores. In this study, the architecture of the skull, dentition, and masticatory apparatus is described, and a model is proposed that suggests a mechanism used by raccoons to reduce different foods. The model suggests that jaw movements are similar to those of cats, the posterior regions of the superficial and deep parts of the temporalis and the anterior region of the medial pterygoid generate horizontal jaw movements, and the anterior portions of the superficial and deep temporalis as well as portions of the masseteric complex generate vertical closing movement. The distributions of slow, fast fatigable, and fast fatigue-resistant fibers for the temporalis and masseteric complex are related to the possible actions of these muscles during mastication, as are the regional cross-sectional areas of the masticatory muscles.     

Yohimbine reversal of ketamine-xylazine immobilization of raccoons (Procyon lotor)

Six adult raccoons (Procyon lotor) were sedated with a combination of ketamine hydrochloride (KH) at 10 mg/kg body weight and xylazine hydrochloride (XH) at 2 mg/kg body weight intramuscularly (i.m.). Twenty min after the KH-XH combination was given, yohimbine hydrochloride (YH) at either 0.1 mg/kg (Trial 1) or 0.2 mg/kg (Trial 2) body weight or a saline control (Trial 3) was administered intravenously (i.v.). The time to arousal, time to sternal recumbency and time to walking were recorded. These times were significantly shortened after YH administration [e.g., mean time to walking (MTW) at 0.2 mg/kg YH = 23.7 min] as compared to the saline controls (MTW = 108.8 min). Heart and respiratory rates both increased after YH administration, while body temperature remained constant. A fourth trial was performed using a higher ratio of KH to XH (45:1 rather than 5:1) to mimic sedation as performed in the field. The mean time to arousal (MTA) and MTW in this trial (1.3 and 23.7 min, respectively) were significantly shorter than controls and similar to YH trials performed after immobilization with 5:1 KH-XH. Yohimbine hydrochloride may be useful in field studies that require sedation of raccoons using KH-XH combinations.     

Field immobilization of raccoons (Procyon lotor) with Telazol and xylazine

The effectiveness of tiletamine plus zolazepam (Telazol) and xylazine was evaluated as an immobilizing combination for raccoons (Procyon lotor). Fifteen raccoons were injected intramuscularly with a 3:2 mixture of Telazol (3.2+/-0.6 mg/kg [mean+/-SD]) and xylazine (2.1+/-0.4 mg/kg) at Pictured Rocks National Lakeshore, Michigan, USA, during May-October, 2001-03. Mean induction time was 4.8+/-3.8 min; mean recovery time was 128.5+/-48.4 min. No linear relationships were found between the amount (mg/kg) of Telazol-xylazine injected and induction (r2 = 0.06, P = 0.40) or recovery times (r2 = 0.01, P = 0.78). Mean heart rate, respiratory rate, and body temperature declined through 20 min after induction (P< 0.05). No mortality occurred and no short-term adverse effects were observed in recaptured individuals. I conclude that a 3:2 mixture of Telazol-xylazine is a safe and effective immobilizing agent for raccoons when conducting nonsurgical field procedures. Immobilizing raccoons with Telazol at 3 mg/kg and xylazine at 2 mg/kg should provide up to 60 min of handling time and usually allow full recovery in about 120 min.     

Demonstration of rabies virus-specific antibody in the sera of free-ranging Iowa raccoons (Procyon lotor).

Between 1984 and 1988, a study was conducted to evaluate the frequency of rabies virus neutralizing antibodies in raccoons (Procyon lotor) in two counties in Iowa. Nine hundred eighty five raccoons were trapped and tagged in Guthrie and Cerro Gordo counties during the spring, summer and fall of each year. Sex, age and weight were recorded for each animal and a blood sample was collected. Serum samples were tested for the presence of serum neutralizing antibodies (SNA) by the rapid fluorescent focus inhibition test (RFFIT), mouse serum neutralization test (MSN), and an indirect fluorescent antibody (IFA) technique for detecting immunoglobulin G. Fifty-one raccoons (5%) were found to have SNA by the RFFIT. Thirty-six serum samples (24 with RFFIT antibody titer greater than 3.0, and 12 less than 3.0) were also tested by the MSN, with results correlating well with the RFFIT results (r = 0.86, P less than 0.01, Kappa = 0.93). In 35 raccoons with SNA by the RFFIT, six individuals had immunoglobulin G binding activity by the IFA test. These results provided serologic evidence of exposure of raccoons to rabies virus in an area free of enzootic raccoon rabies.     

Susceptibility of raccoons (Procyon lotor) to infection with Mycobacterium bovis

Tuberculosis due to Mycobacterium bovis infection is endemic in white-tailed deer (Odocoileus virginianus) in the northeastern portion of the lower Michigan peninsula (USA). Various wild carnivores and omnivores, including raccoons (Procyon lotor), are infected with M. bovis within the endemic area. To investigate the pathogenesis of tuberculosis in raccoons and the likelihood of M. bovis transmission from infected raccoons to other susceptible hosts, we experimentally inoculated raccoons with single oral doses of M. bovis (ranging from 30 to 1.7 x 10(5) colony forming units [CFU]), five daily oral doses of M. bovis (ranging from 10 to 1 x 10(5) CFU), or a single intravenous (i.v.) dose of 1 x 10(5) CFU of M. bovis, from November 1998 through December 2000. Granulomatous lesions consistent with tuberculosis, or tissue colonization with M. bovis, were seen in one of five raccoons in the single low oral dose group, one of five raccoons in the multiple low oral dose group, two of five raccoons in the multiple medium oral dose group, five of five raccoons in the multiple high oral dose group, and five of five raccoons in the i.v. inoculated group. In oral inoculated raccoons, lesions were most common in the tracheobronchial and mesenteric lymph nodes and lung. Excretion of M. bovis in saliva or nasal secretions was noted in all i.v. inoculated raccoons and two of five multiple low oral dose raccoons. Mycobacterium bovis was not isolated from urine or feces from any experimentally inoculated raccoons. The need for multiple large oral doses to establish infection, and the low number of orally inoculated raccoons that excreted M. bovis in nasal secretions or saliva, suggest that wide-spread tuberculosis among raccoons is unlikely.     

Sarcocystis in muscles of raccoons (Procyon lotor L.)

Sarcocysts of Sarcocystis sp. were found in 26 (50%) of 52 raccoons (Procyon lotor) from Ohio, Pennsylvania, Florida, and Maryland. Although only 4 (7.7%) of 52 cardiac muscles specimens were found to contain sarcocysts, 25% to 36.5% of tongue, diaphragm, masseter muscle, and esophagus specimens were found infected. By light microscopy, sarcocyst walls were less than 3 micron thick and had no conspicuous projections; interior septa were indistinct. By transmission electron microscopy, sarcocyst walls had short (mean = 2.7 micron), villus-like protrusions; thin septa were seen within the sarcocysts. The raccoon may be an intermediate host for a Sarcocystis sp. that completes its life cycle in an unidentified, wild carnivore.     

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.     

Salmonella in raccoons (Procyon lotor) in southern Ontario, Canada

Numerous serotypes of Salmonella have been detected in a variety of wild animals, including raccoons (Procyon lotor). Raccoons are common, mid-size omnivores that live in close association with people in urban and rural areas in Ontario. Although raccoons are known to shed Salmonella, little is known about their potential long-term role in maintaining Salmonella infections. We sampled feces from raccoons in three areas of Ontario: one primarily urban site around Niagara, one primarily rural site north of Guelph, and the grounds of the Toronto Zoo, in 2007 to identify which serotypes of Salmonella were commonly shed by raccoons in southern Ontario. In addition, we conducted a longitudinal study at the Toronto Zoo site to determine if raccoons remain persistently infected with Salmonella. Salmonella was found in 45% of samples. The prevalence of Salmonella in raccoon feces ranged from 27% at the rural site to 65% at the urban site. We detected 16 serotypes of Salmonella in 83 positive samples. The most common serotype detected in raccoons from the rural and zoo sites was Salmonella enterica serotype Typhimurium, whereas Salmonella Newport was detected most commonly in the urban site. Only one raccoon of 11 that were captured in four or more consecutive trapping sessions shed the same Salmonella serotype for two consecutive months, suggesting that raccoons regularly acquire new Salmonella serotypes from the environment.     

Development of 14 multiplexed microsatellite loci for raccoons Procyon lotor

In order to complement ecological information with genetic data we isolated and characterized 14 polymorphic microsatellite markers from raccoons (Procyon lotor). Three multiplexed panels comprising the loci were developed and 29 individuals from a contiguous habitat patch in northern Indiana, USA were genotyped. The number of alleles per locus ranged from four to 18, and overall heterozygosities ranged from 0.31 to 1.00. One locus was identified as possibly being X‐linked, since males appeared to be hemizygous. Data generated using these markers will be used to further our understanding of small‐scale raccoon population dynamics in a highly fragmented landscape.     

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.     

Proposed field evaluation of a rabies recombinant vaccine for raccoons (Procyon lotor): site selection, target species characteristics, and placebo baiting trials

Prior to a limited field application of an orally-administered vaccinia-rabies glycoprotein (V-RG) recombinant virus vaccine for wildlife, background data were obtained for the proposed site on Parramore Island, Virginia (USA). Mammalian target and nontarget species, potentially at risk for exposure to vaccine were inventoried. Placebo baiting trials with a fishmeal polymer bait resulted in high bait disturbance (88 to 100%), primarily by raccoons (Procyon lotor), with infrequent visitation and no evidence of bait consumption by deer (Odocoileus virginianus), small mammals or avian species. Definitive bait acceptance rates by raccoons (indicative of bait ingestion) were difficult to accurately determine based exclusively on premolar and vibrissae samples collected antemortem from live-trapped raccoons for tetracycline and rhodamine B biomarker analyses, respectively. Bait acceptance rate was more accurately determined during a pilot baiting trial conducted on North Island, South Carolina, when mandibles (postmortem samples) were examined for tetracycline incorporation. Parasitologic findings in raccoons on Parramore Island included Hepatozoan procyonis, Phagicola angrense and Physaloptera rara and a variety of incidental microscopic lesions, and provided baseline pathological data for comparison subsequent to V-RG vaccine application. A population density estimate of one raccoon/2.7 ha was calculated using mark-recapture data for comparison after vaccine deployment. Limited reproductive data, including estimates of pregnancy rates by palpation, the number of live kits/litter live-trapped with previously pregnant raccoons or observed in the dens of radio-collared raccoons, was gathered to assess the effect of proposed oral vaccination with V-RG vaccine. Home ranges were assessed by radio-telemetry of 15 raccoons; all radio-collared raccoons currently reside on Parramore Island.(ABSTRACT TRUNCATED AT 250 WORDS)     

Genetic evidence for multiple introduction events of raccoons (Procyon lotor) in Spain

The common raccoon (Procyon lotor) is endemic to Central and North America, although non-native populations have become established around the world. In Spain, growing evidence of the introduction of raccoons has been reported across the country in the last decade, especially in Central Spain where the largest population is thought to occur. We used mitochondrial and microsatellite DNA data to investigate the genetics of invasive raccoons in Central Spain and to infer: the number of introduction events, the number of founders and the genetic variability of the introduced populations compared to a native population. We found that at least two introduction events have occurred along the Jarama and Henares Rivers in Central Spain, which currently constitute two genetically differentiated subpopulations. In both localities the number of effective founders from a native population was estimated as 2–4 individuals. These newly founded populations have expanded and show evidence of incipient contact and reproduction between them. This may allow for an increase in the genetic variability and adaptive potential of the population(s), possibly increasing the difficulty of controlling this invasive species. Our results reveal the ability to longitudinally monitor the genetics of the raccoon range expansion and emphasize the urgent need to control the pet trade of potentially invasive species.     

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.