Raccoons (Procyon lotor) in Germany as potential reservoir species for Lyssaviruses

Raccoons can be found almost everywhere in Germany since their first successful introduction in 1934. Although the animal is a well-known reservoir species for rabies in the USA, during the last European fox rabies epizootic, only a few rabid raccoons were reported from Germany. In recent years, the raccoon population density has increased tremendously, especially in (semi) urban settings. Presently, Germany is free of terrestrial wildlife rabies. To assess the potential risk that the raccoon population in Germany could act as a reservoir species upon reemergence of rabies, the susceptibility of the local raccoon population was investigated. Wild-caught animals were inoculated with the most likely lyssavirus variants to infect the local population. It was shown that the raccoons were fully susceptible for a dog and raccoon rabies virus isolate. Also, five of six raccoons inoculated with a fox rabies virus isolate showed clinical signs. However, none of the raccoons infected with European Bat Lyssavirus type 1 succumbed to rabies; meanwhile, all these raccoons seroconverted. It is concluded that the highest risk for the raccoon population in Germany to become infected with lyssaviruses is through the importation of rabies infected dogs.     

Is the raccoon (<Emphasis Type="Italic">Procyon lotor</Emphasis>) out of control in Europe?

The northern raccoon (Procyon lotor) is an invasive species in Europe and poses a serious threat to indigenous biodiversity and human health. Raccoons can also cause important economic losses. Despite the risks, no comprehensive report on the raccoon status in Europe is available. In this article, I estimate the raccoon invaded range and population trends and evaluate the impacts on native ecosystems and wildlife to (1) determine the raccoon status and discuss the invasion process on a continental scale, (2) identify the threats, and (3) propose guidelines for designing management strategies for raccoon populations. Raccoons have been introduced in Europe since the late 1920s but raccoon population growth and range expansion rates have been increasing since 1970s after a lag period. In addition, recent introductions through the pet trade have resulted in the establishment of new feral raccoon populations. Therefore, the raccoon spatial distribution is a result of multiple introductions and range expansion from the primary raccoon populations in central and eastern Europe over the last 40 years. First individuals escaped from fur farms or were released for hunting. Nowadays, the pet trade is the major introduction pathway. Raccoons have adapted to different environments, even urban and residential areas. In central Europe, the raccoon population has increased by over 300% and grows at exponential rates since the 1990s. The raccoon is out of control in Europe because of increasing population trends, range expansion and no efficient management strategy. However, no impacts have still been reported through an evidence-based approach. There is a pressing need for a long-term management strategy of raccoon populations in Europe.     

Characterization of Major Histocompatibility Complex (MHC) DRB Exon 2 and DRA Exon 3 Fragments in a Primary Terrestrial Rabies Vector (Procyon lotor)

The major histocompatibility complex (MHC) presents a unique system to explore links between genetic diversity and pathogens, as diversity within MHC is maintained in part by pathogen driven selection. While the majority of wildlife MHC studies have investigated species that are of conservation concern, here we characterize MHC variation in a common and broadly distributed species, the North American raccoon (Procyon lotor). Raccoons host an array of broadly distributed wildlife diseases (e.g., canine distemper, parvovirus and raccoon rabies virus) and present important human health risks as they persist in high densities and in close proximity to humans and livestock. To further explore how genetic variation influences the spread and maintenance of disease in raccoons we characterized a fragment of MHC class II DRA exon 3 (250bp) and DRB exon 2 (228 bp). MHC DRA was found to be functionally monomorphic in the 32 individuals screened; whereas DRB exon 2 revealed 66 unique alleles among the 246 individuals screened. Between two and four alleles were observed in each individual suggesting we were amplifying a duplicated DRB locus. Nucleotide differences between DRB alleles ranged from 1 to 36 bp (0.4–15.8% divergence) and translated into 1 to 21 (1.3–27.6% divergence) amino acid differences. We detected a significant excess of nonsynonymous substitutions at the peptide binding region (P = 0.005), indicating that DRB exon 2 in raccoons has been influenced by positive selection. These data will form the basis of continued analyses into the spatial and temporal relationship of the raccoon rabies virus and the immunogenetic response in its primary host.     

Effects of Natural Barriers and Habitat on the Western Spread of Raccoon Rabies in Alabama

Abstract: Although domestic animal transmission of rabies has largely been mitigated, the disease remains a concern in both Europe and North America where wildlife transmission has caused epizootics. Raccoon (Procyon lotor) rabies was established in Alabama, USA, in 1975, primarily in the southeastern corner of the state. However, with the exception of isolated events, rabies has not continued to spread westward across the Alabama River. We monitored movements of 100 radiocollared raccoons on 2 sites within hardwood and agriculture habitats in a rabies enzootic area east of the Alabama River, in managed pine habitat area west of the river where rabies sporadically occurs, and in a mixed pine hardwood area outside of the known rabies enzootic area to determine if raccoon movements and habitat use in certain habitat types and the presence of a river may serve as natural barriers preventing the western spread of rabies in Alabama. We also examined raccoon contact rates to determine if they influence disease transmission through static and dynamic interactions. Raccoons in mixed pine-hardwood forest habitats had smaller home ranges and less overlap of ranges compared to the other 3 habitats. However, static interactions between habitats in the use of overlap areas did not differ (F_11,129 = 1.63, P = 0.09). Rabies antibody titers were highest in the managed pine habitat (28%) even prior to oral vaccine bait distributions in spring of 2004 and 2005. Biomarker data from radiocollared and additional raccoons captured after the bait distribution west of the Alabama River demonstrated a low efficacy of the vaccine reaching the small southern raccoons. The combination of the river as a partial barrier, the high percentage of pine forested habitat west of the river, and limited spatial movements of raccoons within these forested habitats appears to have reduced the likelihood of rabies establishing west of the river. Understanding different host-habitat- disease systems is important for successful management of diseases. Based on our results, we recommend that the oral vaccine program continue to use the Alabama River as a partial barrier and baiting be concentrated in the fragmented bottomland hardwood forests and around larger bodies of water where raccoon densities are highest. Success of baiting strategies designed to take advantage of northern raccoon dynamics and habitat use may not be applicable to southern populations.     

Raccoon rabies: the re-emergence of an epizootic in a densely populated area

Investigation of sporadic or isolated cases of rabies was encouraged in the 1950s by the implementation of a national surveillance system for reporting of rabies cases. Low level enzootic disease was recognized in several wild species during this period, including a small focus of rabies in raccoons in Florida. Phylogenetic analysis of the virus transmitted by rabid raccoons and reconstruction of the early history of raccoon rabies in the United States suggests that rabies was present in these animals long before its recognition by the public health community and that the emergence of raccoon rabies as a major threat to public health was the result of several natural and human-mediated events. In recent years, reports of raccoon-human contact and rabies in raccoons have increased as changes in human demographics and land development have concentrated raccoon populations in urban and suburban settings. In addition to the natural spread of rabies from Florida to raccoon populations in Georgia, Alabama, South Carolina and North Carolina, a separate focus in the northeastern states is believed to have been caused by the translocation of raccoons from the southeast to Virginia and West Virginia. Oral vaccination of wild species may be an important adjunct to traditional rabies control measures in the management of this disease in the future.     

New insights on the geographical origins of the Caribbean raccoons

The introduction of species outside their natural range is one of the major threats to biodiversity and has often been identified as a menace to agricultural production and human health. The raccoon is recognized as a globally invasive species. However, several populations in the Caribbean were long considered native and endemic species. Although previous genetic studies have shown that raccoons from the islands of the West Indies belong to the northern raccoon Procyon lotor, the history and origin of these introductions remain poorly known. In this study, we investigated the geographical origin of Caribbean raccoon populations using newly available molecular genetic data. We used haplotype network analyses of two mitochondrial markers, Cytochrome b and Control Region, with new sequences and those from GenBank. We also specifically investigated the origin of the endangered endemic Cozumel raccoon, Procyon pygmaeus, by re-analyzing data. Our results confirmed that all Caribbean raccoon populations belong to the northern raccoon. Bahamian populations originated from two different sources in Florida, and the Lesser Antilles raccoons seem to originate from northern regions of the native range. In addition, our results question the taxonomic status of the Cozumel raccoon, as currently available genetic data support a conspecific status with the northern raccoon. These results have important implications in the context of conservation and ecosystem management. Identifying origins of introduced populations and understanding the history of their introductions will facilitate studies on the impact of the raccoon on insular ecosystems.     

Raccoon (Procyon lotor) in Iberia: Status update and suitable habitats for an invasive carnivore

Raccoons are American carnivores, considered invasive across several countries worldwide, especially in Europe. In the Iberian Peninsula, previous studies on raccoons documented several breeding populations in Spain a decade ago and only two confirmed records from isolated individuals in Portugal. Given the need for updating its Iberian distribution and identifying suitable areas with higher invasion risk, we compiled presence records from established breeding populations and isolated individuals. By using a Maxent approach based on breeding records, we forecasted the suitable habitats in Iberia with higher invasion risk for raccoons and identified the related environmental drivers. Overall, we collected 1039 records of raccoon presence throughout the Iberian Peninsula, including 980 records from established breeding populations. Their origin is probably linked to escapes from captivity. Climatic conditions, linked to both drier and wetter environments, and proximity to water bodies were the main predictors of suitable areas for raccoon’s expansion from the currently established breeding nuclei in Iberia. The forecasted high probability areas showed a wide, but fragmented distribution concentrated on four main areas: central, central-north, central-east, and north-west Iberia. NW Portugal seems to be the area with higher invasion risk in the country, although field surveys showed no evidence of raccoon presence yet. However, there are several records in Spain near the Portuguese border, comprising isolated individuals and breeding populations. Therefore, it is crucial to ensure regular monitoring of areas with high invasion risk, particularly those near facilities with captive raccoons that often act as a source of feral individuals, to assure early detection and effective control for the expansion of this invasive carnivore.     

Raccoon Social Networks and the Potential for Disease Transmission

Raccoons are an important vector of rabies and other pathogens. The degree to which these pathogens can spread through a raccoon population should be closely linked to association rates between individual raccoons. Most studies of raccoon sociality have found patterns consistent with low levels of social connectivity within populations, thus the likelihood of direct pathogen transmission between raccoons is theoretically low. We used proximity detecting collars and social network metrics to calculate the degree of social connectivity in an urban raccoon population for purposes of estimating potential pathogen spread. In contrast to previous assumptions, raccoon social association networks were highly connected, and all individuals were connected to one large social network during 15 out of 18 months of study. However, these metrics may overestimate the potential for a pathogen to spread through a population, as many of the social connections were based on relatively short contact periods. To more closely reflect varying probabilities of pathogen spread, we censored the raccoon social networks based on the total amount of time spent in close proximity between two individuals per month. As this time criteria for censoring the social networks increased from one to thirty minutes, corresponding measures of network connectivity declined. These findings demonstrate that raccoon populations are much more tightly connected than would have been predicted based on previous studies, but also point out that additional research is needed to calculate more precise transmission probabilities by infected individuals, and determine how disease infection changes normal social behaviors.     

Cryptosporidium sp. skunk genotype in wild raccoons (Procyon lotor) naturally infected with Baylisascaris procyonis from Central Germany

Cryptosporidium spp. are apicomplexan parasites of public health concern. They are one of the main causes of intestinal diseases in humans and animals. Contaminated water is among the main sources of infection for humans and mammals. Raccoons are an introduced species in Germany. They are anthropogenic adapters with a natural affinity for water bodies. We collected samples from wild raccoons in the Federal States of Saxony and Thuringia, Central Germany. Through molecular genotyping, we found Cryptosporidium sp. skunk genotype in one raccoon from Saxony (1/24) and in one animal from Thuringia (1/27). Both raccoons were also infected with the zoonotic nematode Baylisascaris procyonis. This is the first report of co-infection with these two parasites in raccoons from Germany. Our study highlights the potential of these animals as carriers of zoonotic pathogens. Since raccoons can thrive in human settlements, this study provides data that can be used as a baseline for preventive programs.     

The genetic structure of raccoon introduced in Central Europe reflects multiple invasion pathways

Invasions of non-native species are of great concern as they have a devastating impact on native biodiversity and can also affect the economy of a region. Multiple introductions in several locations of a new range greatly promote the success of non-native species. The raccoon (Procyon lotor) is an omnivore whose native distribution extends from southern Canada to Panama. It has been successfully introduced in many European countries. We examined the microsatellite and mitochondrial diversity of raccoon populations in Central Europe (Germany, Poland, and Czech Republic) in order to determine their introduction sources and pathways as well as the factors affecting genetic structure in this invasive species. We found low diversity of the mtDNA control region and moderate diversity of microsatellite markers. Raccoon showed three hierarchic levels of genetic structure which separate at different levels sampled from Czech Republic, Germany and raccoon inhabiting two different habitats in Poland. In Poland the raccoon population was established through migration from Germany to Czech Republic. Analysis of the intensity of migration between two different habitat types indicated source-sink dynamics in the Polish populations of raccoons. Our results confirm the high intensity of the raccoon invasion in Central Europe and point to specific measures needed as part of an effective management strategy.     

Pseudorabies: experimental studies in raccoons with different virus strains

Raccoons (Procyon lotor) were infected by the nasal/oral route with as little as 10(2) plaque forming units (PFU) of pseudorabies virus (PrV). There was no apparent difference in the susceptibility of raccoons to infection with either of two virulent field strains or with the naturally avirulent K strain which has been used in modified live virus vaccines. Each of these three viruses was transmitted by contact to uninfected raccoons. All raccoons that were infected with virulent field strains died; however only two of 11 (18%) raccoons that were infected with the K strain died. One of four raccoons that survived infection with the K strain survived superinfection with virulent virus. This finding was significant because it could be a mechanism by which virulent PrV can be introduced and persist in the raccoon population. The possibility of this event occurring is increasing because of the widespread prevalence of PrV and the use of modified live virus vaccines for controlling clinical pseudorabies in swine. Virus neutralizing activity was found in five of 47 serums collected from raccoons that were trapped in PrV endemic areas. This observation implies that a herpesvirus, possibly PrV, may be present in the wild raccoon population.     

MHC class II DRB diversity in raccoons (<Emphasis Type="Italic">Procyon lotor</Emphasis>) 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.     

Age,but not Sex or Genetic Relatedness,Shapes Raccoon Dominance Patterns

Raccoons are generally regarded as solitary, yet several studies have found that raccoons frequently form social affiliations. One benefit to sociality in many mammal species is that relatives and close associates can form coalitions against third parties during agonistic encounters. We tested whether raccoon dominance patterns were influenced by age, sex, genetic relatedness, and association patterns at two anthropogenic feeding stations in an urban forest. We found that genetic relatedness had no significant effect on patterns of agonism at one of the feeding stations. At the second feeding station, raccoons were more likely to act aggressively toward close relatives, which is opposite of the predicted pattern. However, when we controlled for the number of times raccoons arrived at feeding stations in close proximity, the effect of relatedness on dominance patterns was not significant at either feeding station. These results suggest that relatedness plays little or no role in shaping dominance patterns of raccoons. Older raccoons were ranked significantly higher in the dominance hierarchies regardless of sex. This pattern leads us to conclude that age is the primary factor driving the outcome of aggressive interactions in raccoons at our study site. Despite frequent social interactions at the study site, the patterns of raccoon dominance more closely resemble patterns found in solitary animals. To confirm the generality of these results and to better understand the evolution of raccoon social behavior, similar studies need to be undertaken in other raccoon populations.     

Spatial dynamics and molecular ecology of North American rabies

Rabies, caused by a single-stranded RNA virus, is arguably the most important viral zoonotic disease worldwide. Although endemic throughout many regions for millennia, rabies is also undergoing epidemic expansion, often quite rapid, among wildlife populations across regions of Europe and North America. A current rabies epizootic in North America is largely attributable to the accidental introduction of a particularly well-adapted virus variant into a naive raccoon population along the Virginia/West Virginia border in the mid-1970s. We have used the extant database on the spatial and temporal occurrence of rabid raccoons across the eastern United States to construct predictive models of disease spread and have tied patterns of emergence to local environmental variables, genetic heterogeneity, and host specificity. Rabies will continue to be a remarkable model system for exploring basic issues in the temporal and spatial dynamics of expanding infectious diseases and examining ties between disease population ecology and evolutionary genetics at both micro- and macro-evolutionary time scales.     

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.     

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.     

Potential resource competition between an invasive mammal and native birds: overlap in tree cavity preferences of feral raccoons and Ural owls

Invasive mammals include good tree climbers that use tree cavities for resting and nesting. Tree cavities are important but limited resources in most forests; thus, some invasive mammals can be serious competitors for native cavity-using species, especially cavity-nesting birds. Despite the potential impact, such inter-class competition has rarely been considered. We examined the possibility of resource competition for tree cavities between the invasive raccoon Procyon lotor and the native Ural owl Strix uralensis. Both species are nocturnal and use tree cavities during daytime. We assessed an overlap in cavity use for both species as an indication of potential competition by monitoring 341 cavities during their breeding season in a natural park in Hokkaido, Japan. Of 341 potentially available cavities, raccoons and Ural owls used 37 and 32, respectively. The characteristics of 58 cavities used by raccoons or owls were compared to 49 random cavities to determine if they selected cavities with certain characteristics. As predicted from a large amount of tree cavities and a low raccoon density in this managed forest, we did not find direct evidence of competition, such as physical interaction, intrusion to cavities, or habitat segregation. Cavity types used by both species overlapped considerably in terms of height, entrance size, depth, and other characteristics: their habitats were widely overlapped. Further, in four cavities, one species was replaced by the other. Given the similar habitat requirements, the invasive raccoon could be a potential competitor for Ural owl when raccoon density increases and/or cavity availability decreases, which is the case for many forests in Japan. This study suggests that potential threats of resource competition among not only closely but also distantly related taxa should be taken into consideration when studying the impacts of invasive species.     

Life cycle of Sarcocystis neurona in its natural intermediate host,the raccoon,Procyon lotor

Sarcocystis neurona causes encephalomyelitis in many species of mammals and is the most important cause of neurologic disease in the horse. Its complete life cycle is unknown, particularly its development and localization in the intermediate host. Recently, the raccoon (Procyon lotor) was recognized as a natural intermediate host of S. neurona. In the present study, migration and development of S. neurona was studied in 10 raccoons that were fed S. neurona sporocysts from experimentally infected opossums; 4 raccoons served as controls. Raccoons were examined at necropsy 1, 3, 5, 7, 10, 14, 15, 22, 37, and 77 days after feeding on sporocysts (DAFS). Tissue sections of most of the organs were studied histologically and reacted with anti-S. neurona-specific polyclonal rabbit serum in an immunohistochemical test. Parasitemia was demonstrated in peripheral blood of raccoons 3 and 5 DAFS. Individual zoites were seen in histologic sections of intestines of raccoons euthanized 1, 3, and 5 DAFS. Schizonts and merozoites were seen in many tissues 7 to 22 DAFS, particularly in the brain. Sarcocysts were seen in raccoons killed 22 DAFS. Sarcocysts at 22 DAFS were immature and seen only in skeletal muscle. Mature sarcocysts were seen in all skeletal samples, particularly in the tongue of the raccoon 77 DAFS; these sarcocysts were infective to laboratory-raised opossums. This is the first report of the complete development of S. neurona schizonts and sarcocysts in a natural intermediate host.     

Ecology of wildlife rabies in Europe

• 1 The number of wildlife rabies cases has increased in Europe in recent years. We review the epizootiology of wildlife rabies in Europe, paying special attention to recent changes to the situation of two important vector species: the red fox and the raccoon dog. Red fox Vulpes vulpes has been the main vector of rabies since 1945, but the number and proportion of raccoon dog Nyctereutes procyonoides cases has rapidly increased during the past few years, particularly in north‐eastern Europe.
• 2 The transmission rate (average number of susceptible animals infected by each rabid animal) is critical for rabies spread and is partly determined by population density. Both raccoon dogs and foxes live in pairs. Foxes also live in family groups. Pairs and groups share their territories. Home range size usually correlates negatively with population density. Fox home ranges are 50–1500 ha, those of raccoon dogs 150–700 ha. The threshold value for rabies spread among foxes is estimated to be 0.63 individuals/km². Although fox density in eastern and northern Europe may be lower than this, the pooled density of foxes and raccoon dogs exceeds the threshold density.
• 3 Animal movements, especially dispersal of young, pose a risk for rabies spread. Although the likelihood of an epizootic is highest where fox and raccoon dog densities are highest, rabies may spread fastest where population densities are lower, because dispersal distances tend to correlate negatively with population density.
• 4 Oral vaccinations have been more effective in rabies control than culling foxes. Where two vector species exist, vaccination should be conducted twice a year, because most raccoon dogs disperse in autumn but some foxes do not disperse before mid‐ or late winter.
• 5 New rabies models, based on two vector species and their interaction, and which take into account the hibernation period of raccoon dogs, are needed for north‐eastern Europe.