Land-use effects on prevalence of raccoon roundworm (Baylisascaris procyonis)

The raccoon (Procyon lotor) is the definitive host of Baylisascaris procyonis, a large intestinal roundworm that is zoonotic and can result in fatal or severe central nervous system disease in young children. Prevalence of infection among raccoon populations often is high, and in the midwestern United States, B. procyonis has been reported in 68-82% of raccoons. Raccoon populations have increased in response to changes in human land use, and often reach higher densities in urban and suburban landscapes than rural landscapes. However, shifts in foraging behavior among urban raccoons could impact the transmission of B. procyonis if small vertebrate intermediate hosts are not a significant part of the raccoon diet. The objective of this study was to compare prevalence of B. procyonis infection between urban and rural raccoon populations on a regional scale. Necropsy was done on 204 raccoons collected from September through February during 2000-2005 from seven states across the Midwest (regional sample). Baylisascaris procyonis was found in 54% of examined raccoons. Prevalence differed between land-use types (chi2=11.56, df=1, P=0.0007), and was higher among animals collected from rural locations (65%) than those collected in urban locations (41%). Intensity of infection also differed (F=5.52, df=1, P=0.02), with rural raccoons having greater worm burdens (x=29.63+/-36.42) than urban raccoons (x=13.85+/-18.47). Despite high densities of raccoons in urban landscapes, fewer urban raccoons were infected with B. procyonis, suggesting decreased dependence on intermediate hosts as a food source. This possible explanation was supported by a similar trend in prevalence among subsamples of raccoons collected from three Chicago-area populations (local samples) with differing levels of urbanization, population densities, and foraging behavior that had been intensively monitored during 1995-2002. Decreased transmission of B. procyonis in urban landscapes may be due to decreased predation of intermediate hosts, and contact of juvenile raccoons with B. procyonis eggs may be an important factor in maintaining infections within such populations.     

Frequency of deposition and location of Baylisascaris procyonis eggs in raccoon feces

Baylisascaris procyonis is a large ascarid nematode found in the small intestine of raccoons (Procyon lotor). Infection with larvae of B. procyonis can produce visceral, ocular, and neural larval migrans in humans. Infected raccoons can shed millions of eggs a day in their feces. However, it is unknown whether eggs are consistently shed or whether eggs occur at irregular intervals by the population of female nematodes within a host. We trapped, infected, and collected daily fecal samples from 11 raccoons maintained in captivity. Eggs from B. procyonis were obtained from anterior, central, and posterior sections of raccoon feces, isolated by flotation, and quantified under 100× magnification. Naturally infected raccoons were collected and used as a comparison with the experimentally infected group. All raccoons in the experimental group (n=11) became infected with B. procyonis after consuming one infected mouse. Additionally, differential egg deposition rates were observed among individual raccoons from the experimental and naturally infected groups. Mean number of eggs per gram of feces (means±SE) was 16,563±4,321, which was less than previously reported for the species. However, no differences (F(2,30)=0.84, P=0.45) were noted in mean number of eggs per gram of feces among fecal sections. Wildlife biologists, veterinarians, health officials, and researchers of B. procyonis should collect daily fecal samples for a minimum of 3 days before identifying a raccoon as negative for B. procyonis infection. However, it does not matter where within the fecal matter the sample is obtained.     

First outbreak of Baylisascaris procyonis larva migrans in rabbits in Japan

A syndrome of progressive neurological signs was noticed in rabbits (Oryctolagus cuniculus) kept in a small wildlife park in mid-July 2000. Three out of 12 common raccoons (Procyon lotor) kept in this park were infected with Baylisascaris procyonis, and the larvae were found from affected rabbits. This outbreak is the first proven B. procyonis larva migrans in Japan, and the potential risk of serious zoonosis by this ascarid species should be considered by pet owners, veterinarians, physicians and public health authorities in this country as in North America where raccoons are endemic.     

Survey of raccoons on Key Largo, Florida, USA, for Baylisascaris procyonis

Numbers of the endangered Key Largo woodrat (KLWR; Neotoma floridana smalli) have been declining for at least 25 yr. The raccoon (Procyon lotor) roundworm, Baylisascaris procyonis, has been found to have an adverse effect on the survival of Alleghany woodrats (N. magister). High densities of raccoons can exacerbate this problem by increasing the amount of feces containing viable eggs of B. procyonis available to woodrats. In 2002, 64 fecal samples were collected and examined for eggs of B. procyonis from >32 raccoons within the KLWR's known range on Key Largo, Florida, USA. All samples were negative for eggs of B. procyonis. Raccoon density in this area was approximately 0.62 raccoons/ha. Despite this high density of raccoons, B. procyonis does not appear to be a threat to the KLWR population.     

Epizootiology of an outbreak of cerebrospinal nematodiasis in cottontail rabbits and woodchucks

An epizootic of cerebrospinal nematodiasis in cottontail rabbits (Sylvilagus floridanus) and woodchucks (Marmota monax) caused by Baylisascaris procyonis larvae followed the establishment of an ascarid-infected raccoon (Procyon lotor) population in a woodlot. Five of seven raccoons examined from the woodlot harbored ascarids, with one heavily infected animal shedding approximately 27,500 eggs per gram of feces. A laboratory-reared cottontail rabbit developed neurologic disease due to larval migration 80 days after infection with B. procyonis eggs from the raccoons.     

Gnathostoma procyonis from south Georgia and north Florida raccoons

From 2004 to 2006, 511 raccoons collected by the United States Department of Agriculture Wildlife Services as part of a study to evaluate the effects of mesomammalian predator removal on bobwhite-quail reproduction, were examined for the presence of Gnathostoma procyonis, a nematode that parasitizes the stomach of its definitive host, the raccoon. One hundred forty-one raccoons (27.6%) contained G. procyonis, with males being equally infected (27.5%) as females (27.8%). Mean intensity did not differ significantly between sexes (males-mean = 2.88, range 1-17; females-mean = 2.78, range 1-10) or between collection sites. A significant seasonal prevalence of G. procyonis was noted with a peak in March (43.3%, n = 120) and a minimum in September (5.4%, n = 37). Both collection sites exhibited significant seasonal decreases in G. procyonis, but were not significantly different from each other. There was no cumulative seasonal pattern in mean intensity of G. procyonis, and no seasonal pattern at either collection site. Host weight was not related to either G. procyonis prevalence or mean intensity. Host removal did not have an apparent effect on prevalence or mean intensity of G. procyonis.     

Genetic polymorphism of Baylisascaris procyonis in host infrapopulations and component populations in the Central USA

Baylisascaris procyonis is a nematode of significant concern to public and domestic animal health as well as wildlife management. The population genetics of B. procyonis is poorly understood. To gain insights into patterns of genetic diversity within (infrapopulation level) and among (component population level) raccoon (Procyon lotor) hosts, and specifically to assess the relative importance of indirect and direct transmission of the parasite for explaining observed population structure, we collected 69 B. procyonis from 17 wild raccoons inhabiting five counties in Missouri and Arkansas, USA. Informative regions of mitochondrial (CO1, CO2) and nuclear (28S, ITS2) genes were amplified and the distribution and genetic variability of these genes were assessed within and across raccoons. Concatenation of the CO1 and CO2 mtDNA sequences resulted in 5 unique haplotypes, with haplotype diversity 0.456?±?0.068. The most common haplotype occurred in 94% of raccoons and 72.5% of B. procyonis. Sequences for 28S rDNA revealed four unique nuclear genotypes, the most common found in 100% of raccoons and 82.6% of B. procyonis. ITS2 genotypes were assessed using fragment analysis, and there was a 1:1 correspondence between 28S and ITS-2 genotypes. Infrapopulation variation in haplotypes and genotypes was high and virtually all hosts infected with multiple sequenced nematodes also harbored multiple haplotypes and genotypes. There was a positive relationship between the size of the analyzed infrapopulation (i.e., the number of nematodes analyzed) and the number of haplotypes identified in an individual. Collectively this work emphasizes the importance of indirect transmission in the lifecycle to this parasite.     

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%).     

Global Distribution of Bartonella Infections in Domestic Bovine and Characterization of Bartonella bovis Strains Using Multi-Locus Sequence Typing

Bartonella bovis is commonly detected in cattle. One B. bovis strain was recently isolated from a cow with endocarditis in the USA, suggesting its role as an animal pathogen. In the present study, we investigated bartonella infections in 893 cattle from five countries (Kenya, Thailand, Japan, Georgia, and Guatemala) and 103 water buffaloes from Thailand to compare the prevalence of the infection among different regions and different bovid hosts. We developed a multi-locus sequence typing (MLST) scheme based on nine loci (16S rRNA, gltA, ftsZ, groEL, nuoG, ribC, rpoB, ssrA, and ITS) to compare genetic divergence of B. bovis strains, including 26 representatives from the present study and two previously described reference strains (one from French cows and another from a cow with endocarditis in the USA). Bartonella bacteria were cultured in 6.8% (7/103) of water buffaloes from Thailand; all were B. bovis. The prevalence of bartonella infections in cattle varied tremendously across the investigated regions. In Japan, Kenya, and the Mestia district of Georgia, cattle were free from the infection; in Thailand, Guatemala, and the Dusheti and Marneuli districts of Georgia, cattle were infected with prevalences of 10–90%. The Bartonella isolates from cattle belonged to three species: B. bovis (n=165), B. chomelii (n=9), and B. schoenbuchensis (n=1), with the latter two species found in Georgia only. MLST analysis suggested genetic variations among the 28 analyzed B. bovis strains, which fall into 3 lineages (I, II, and III). Lineages I and II were found in cattle while lineage III was restricted to water buffaloes. The majority of strains (17/28), together with the strain causing endocarditis in a cow in the USA, belonged to lineage I. Further investigations are needed to determine whether B. bovis causes disease in bovids.     

Nucleotide sequence variability in the nontranscribed spacer of the rRNA locus in the oyster parasite Perkinsus marinus.

We examined the sequence variability of the nontranscribed spacer (NTS) and internal-transcribed spacer (ITS1 and ITS2) domains of the rRNA locus of Perkinsus marinus from Maryland, Florida, and Louisiana. The sequence of P. marinus DNA including the 5S rRNA, NTS, small subunit (SSU) rRNA, ITSI, and ITS2 regions confirmed their contiguity in the rRNA locus and revealed differences at 28 positions with the SSU rRNA sequences published earlier. The 307-bp polymerase chain reaction (PCR)-amplified fragments from the NTS domain of the various P. marinus isolates revealed the presence of 2 distinct sequences, designated as types I and II, that differed at 6 defined nucleotide positions. Based on these differences, nested PCR and restriction enzyme digests were used to distinguish between the 2 types. Sequences of the ITS1 and ITS2 domains of samples from either NTS type I (n = 3) or type II (n = 3) showed no variation and were identical to published sequences. Frequencies of the P. marinus NTS sequence types I and II in infected oysters varied with the geographic origin of the samples. All Maryland samples examined (n = 19) corresponded to the NTS type I sequence, the type II was the most frequent in the Florida samples (n = 17), and both types were about equally represented in the Louisiana samples (n = 19), with both sequence types found in individual oyster specimens. Although it has been suggested that P. marinus is diploid, it remains to be determined if both NTS sequence types can be present in a single P. marinus trophozoite.     

Helminths of the raccoon (Procyon lotor) in western Kentucky

Seventy raccoons (Procyon lotor) from western Kentucky were examined for helminths from December 1985 through May 1986. Twenty-three species of helminths were collected including 10 species of Trematoda (Brachylaima virginiana, Euryhelmis squamula, Eurytrema procyonis, Fibricola cratera, Gyrosoma singulare, Maritreminoides nettae, Mesostephanus appendiculatoides, Metagonimoides oregonensis, Paragonimus kellicotti, Pharyngostomoides procyonis), 2 species of Cestoda (Atriotaenia procyonis, Mesocestoides variabilis), 10 species of Nematoda (Arthrocephalus lotoris, Baylisascaris procyonis, Capillaria putorii, C. plica, Crenosoma goblei, Dracunculus insignis, Gnathostoma procyonis, Molineus barbatus, Physaloptera rara, Trichinella spiralis), and 1 species of Acanthocephala (Macracanthorhynchus ingens). A mean of 6.4 (3-11) helminth species per host was recorded. Fibricola cratera, Atriotaenia procyonis, Mesocestoides variabilis, Arthrocephalus lotoris, Capillaria plica, Dracunculus insignis, Molineus barbatus, and Physaloptera rara were ubiquitous parasites of the raccoon, whereas specific nidi were observed for Eurytrema procyonis, Gyrosoma singulare, Paragonimus kellicotti, Baylisascaris procyonis, Trichinella spiralis, and Macracanthorhyncus ingens. With an overall prevalence of 10% or higher, 15 of the 23 helminth species were considered common parasites of the raccoon in western Kentucky. When the 10% prevalence rate was applied within geographical quadrants to correct for the presence of nidi it was found that 18 of the 23 helminth species were common and 5 were regarded as rare parasites of the raccoon. Two species of nematodes, T. spiralis and B. procyonis, displayed a markedly higher prevalence in male raccoons.     

Seroprevalence of Trypanosoma cruzi in raccoons from South Carolina and Georgia

Trypanosoma cruzi, the causative agent of American trypanosomiasis or Chagas' disease, is of both medical and veterinary importance as is evidenced by chronic phase myocarditis in humans and dogs. Further, T. cruzi has been reported from over 20 species of wildlife reservoir hosts in the USA, with raccoons (Procyon lotor) and opossums (Didelphis virginiana) being the most common. Whereas previous studies on T. cruzi in raccoons have included only culture and direct examination of blood, the indirect immunofluorescent antibody test (IFAT) was used in the current study to detect anti-T. cruzi antibodies in the serum of raccoons. Of 221 raccoons trapped at 13 sites representing the five physiographic regions of South Carolina plus five sites in the Piedmont region of Georgia (from April 1997 to February 2000), 104 (47%) were seropositive. A higher seroprevalence in raccoons was observed in the coastal regions, with seroprevalence in the Lower Coastal Plain South (61%) being significantly higher than that in the Foothills (37%), Piedmont (42%), and Upper Coastal Plain (40%) regions. However, at a seroprevalence of 52%, the Lower Coastal Plain North was not significantly different from any other region. Although more female raccoons were infected than males, no statistical difference in prevalence was observed between sexes. The high seroprevalence of T. cruzi in raccoons, together with a few reports of wildlife isolates being infective for other wildlife species and domestic/laboratory animals, suggests that risk of T. cruzi infection may be higher than previously suspected.     

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)     

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.     

Molecular characterization of Histomonas meleagridis and other parabasalids in the United States using the 5.8S, ITS-1, and ITS-2 rRNA regions

Extracted DNA from 28 Histomonas meleagridis -infected avian tissue samples from multiple hosts and geographic locations was analyzed for variation in the 5.8S rRNA and the flanking internal transcribed spacer regions (ITS 1 and ITS 2). Samples were amplified by polymerase chain reaction, sequenced, and compared with known sequences from GenBank accessions of H. meleagridis and other related protozoa. The analyses revealed significant genetic variation within H. meleagridis sequences and suggested the possibility of multiple genotypes within the samples or a possible misdiagnosis. Related protozoa found in some samples were mostly identified as Tetratrichomonas spp. However, 1 sample had a 93% identity to Simplicimonas similis , a newly described organism, suggesting the possibility of a new pathogen in poultry. A phylogenetic tree analyzing the 5.8S and flanking ITS regions was inconclusive and we were unable to resolve all H. meleagridis into a single grouping. In contrast, a tree constructed only on the 5.8S rRNA grouped all but 1 H. meleagridis sample into 1 clade, including GenBank accessions submitted from Europe. This suggests that the 5.8S region alone is more reliable in identifying H. meleagridis than are the combined 5.8S and flanking ITS regions. There was no correlation between genotypes and host species or geographic location, suggesting that H. meleagridis moves freely between multiple avian species in the sampled regions.     

Changes in transmission of Baylisascaris procyonis to intermediate hosts as a function of spatial scale

Physical changes in landscapes alter the abundance and distribution of species. Higher-order effects can occur when changes in ecological processes result in altered interspecific interactions and subsequent changes in a species' abundance or persistence. Baylisascaris procyonis , a roundworm parasite of raccoons ( Procyon lotor ), is pathogenic to numerous small vertebrates that serve as intermediate hosts, including white-footed mice ( Peromyscus leucopus ). Raccoons have responded positively to agriculturally induced changes in landscapes, with potential consequences for intermediate hosts of B . procyonis . We examined white-footed mice from a homogeneous, predominantly forested landscape in south-central Indiana and a heterogeneous, predominantly agricultural landscape in northwestern Indiana for presence of larval B . procyonis . We compared prevalence of infection, intensity of infection, and average number of larvae per mouse between the landscapes, and among forest patches within the highly fragmented agricultural landscape. Prevalence, intensity of infection, and average number of larvae were significantly higher in the highly fragmented landscape. Within the agricultural landscape, regression models predicted probability of infection, intensity of infection, and average number of larvae per mouse per patch as functions of forest patch area and isolation. We conclude that positive responses of raccoons to agriculturally induced fragmentation of forests have resulted in increased encounter rates between white-footed mice and infective stages of B . procyonis between and within landscapes, resulting in increased transmission of the parasite to intermediate hosts.     

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.     

Sequence conservation in the internal transcribed spacers and 5.8S ribosomal RNA of parasitic scuticociliates Miamiensis avidus (Ciliophora, Scuticociliatia)

The scuticociliate Miamiensis avidus is a histophagous parasite that causes high mortality in cultured marine fishes, with clinical signs of severe ulcers and hemorrhages in the skeletal muscle. The internal transcribed spacer (ITS) region, which is widely used in taxonomy and molecular phylogeny because of a high degree of variation, was compared for 21 cloned strains of M. avidus (Ciliophora, Scuticociliatia). These strains were isolated from olive flounder Paralichthys olivaceus, ridged-eye flounder Pleuronichthys cornutus and spotted knifejaw Oplegnathus fasciatus in Korea and Japan. The ITS1 (140 bp), ITS2 (236 bp) and 5.8S (119 bp) regions from 21 strains were identical, indicating that these regions are highly conserved in M. avidus. Phylogenic analysis of ITS2 shows that the ciliate should be included in the Philasterida with a close relationship to Pseudocohnilembus hargisi. This study exhibits the first detailed analysis of the ITS1, 5.8 S and ITS2 rRNA regions of M. avidus.     

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.