Prevalence and genetic characterization of a Babesia microti-like species in the North American river otter (Lontra canadensis).

A 4.5-month-old, male, North American river otter (Lontra canadensis) from Athens-Clarke County, Georgia, USA being temporarily housed at a rehabilitation facility, presented with a three-day history of lethargy, anorexia, and severe anemia. Antemortem blood smears revealed intraerythrocytic piroplasms. Supportive care and antiparasitic treatments were initiated, but the animal died three days following presentation. Gross necropsy revealed yellow discoloration of all adipose tissue throughout the carcass and a mildly enlarged, diffusely yellow to pale orange liver. Microscopically, moderate, centrilobular hepatocellular degeneration and necrosis were observed, consistent with hypoxia secondary to apparent hemolytic anemia. Piroplasms were frequently observed in red blood cells in histologic sections. The nearly full-length 18S rRNA gene sequence (1588 bp) was identical to a previously described piroplasm from North American river otters from North Carolina. Phylogenetically, based on the 18S rRNA gene sequence, the otter Babesia sp. was in a sister group with a clade that included several strains of Babesia microti-like species including Babesia sp. from badgers (Meles meles), Babesia vulpes, and Babesia sp. from raccoons (Procyon lotor). To better understand the distribution and genetic variability of this Babesia species, otters from four states in the eastern U.S. and California were tested. Overall, 30 of 57 (53%) otters were positive for Babesia sp. None of four otters from California were positive, but prevalences in eastern states were generally high, 5/9 (55%) in Georgia, 7/14 (50%) in South Carolina, 10/17 (59%) in North Carolina, and 8/13 (62%) in Pennsylvania). Partial 18S rRNA gene sequences from all populations were identical to the clinical case sequence. No Babesia sensu stricto infections were detected. There were six unique COI sequences (937 bp) detected in 18 positive otters. The most common lineage (A) was detected in 12 of 18 (67%) samples from Georgia, North Carolina, South Carolina, and Pennsylvania. Lineage B was found in two otters and the remaining lineage types were found in single otters. These six lineages were 99-99.8% similar to each other and were < 88% similar to related parasites such as B. vulpes, B. microti-like species of raccoons, B. microti, and B. rodhaini. Phylogenetically, the Babesia sp. of otters grouped together in a well-supported clade separate from a sister group including B. vulpes from fox (Vulpes vulpes) and domestic dogs. In conclusion, this report demonstrates that this piroplasm is a potential pathogen of North American river otters and the parasite is widespread in otter populations in the eastern United States.

Distribution of trespass cannabis cultivation and its risk to sensitive forest predators in California and Southern Oregon.

Illegal cannabis cultivation on public lands has emerged as a major threat to wildlife in California and southern Oregon due to the rampant use of pesticides, habitat destruction, and water diversions associated with trespass grow sites. The spatial distribution of cultivation sites, and the factors influencing where they are placed, remain largely unknown due to covert siting practices and limited surveillance funding. We obtained cannabis grow-site locality data from law enforcement agencies and used them to model the potential distribution of cultivation sites in forested regions of California and southern Oregon using maximum entropy (MaxEnt) methods. We mapped the likely distribution of trespass cannabis cultivation sites and identified environmental variables influencing where growers establish their plots to better understand the cumulative impacts of trespass cannabis cultivation on wildlife. We overlaid the resulting grow-site risk maps with habitat distribution maps for three forest species of conservation concern: Pacific fisher (Pekania pennanti), Humboldt marten (Martes caurina humboldtensis), and northern spotted owl (Strix occidentalis caurina). Results indicate that cannabis cultivation is fairly predictably distributed on public lands in low to mid-elevation (~800-1600m) forests and on moderate slopes (~30-60%). Somewhat paradoxically, results also suggest that growers either preferred sites inside of recently disturbed vegetation (especially those burned 8-12 years prior to cultivation) or well outside (>500m) of recent disturbance, perhaps indicating avoidance of open edges. We ground-truthed the model by surveying randomly selected stream courses for cultivation site presence in subsets of the modeling region and found previously undiscovered sites mostly within areas with predicted high likelihood of grow-site occurrence. Moderate to high-likelihood areas of trespass cultivation overlapped with 40 to 48% of modeled habitats of the three sensitive species. For the endangered southern Sierra Nevada fisher population, moderate-high likelihood growing areas overlapped with over 37% of modeled fisher denning habitat and with 100% of annual female fisher home ranges (mean overlap = 48.0% + 27.0 SD; n = 134) in two intensively studied populations on the Sierra National Forest. Locating and reclaiming contaminated cannabis grow sites by removing all environmental contaminants should be a high priority for resource managers.

Human Seroprevalence of Tick-Borne Anaplasma phagocytophilum, Borrelia burgdorferi, and Rickettsia Species in Northern California.

There is a paucity of data on human exposure to tick-borne pathogens in the western United States. This study reports prevalence of antibodies against three clinically important tick-borne pathogens (Borrelia burgdorferi, Anaplasma phagocytophilum, and Rickettsia spp.) among 249 people in five counties in northern California. Individuals from Humboldt County were recruited and answered a questionnaire to assess risk of exposure to tick-borne pathogens. Samples from other counties were obtained from a blood bank and were anonymized. Seventeen (6.8%) samples were seropositive for antibodies against at least one pathogen: five for A. phagocytophilum, eight for B. burgdorferi, and four for Rickettsia spp. Women and people aged 26-35 had higher seroprevalence compared to other demographic groups. Santa Cruz County had no seropositive individuals, northern Central Valley counties had three seropositive individuals (all against A. phagocytophilum), and Humboldt County had 14 (all three pathogens), a significant, four-fold elevated risk of exposure. The Humboldt County questionnaire revealed that a bird feeder in the yard was statistically associated with exposure to ticks, and lifetime number of tick bites was associated with increasing age, time watching wildlife, and time hiking. Three-quarters of respondents were concerned about tick-associated disease, 81.0% reported experiencing tick bites, and 39.0% of those bitten reported a tick-borne disease symptom, including skin lesions (76.4%), muscle aches (49.1%), joint pain (25.5%), or fever (23.6%). Despite high levels of concern, many individuals who had been bitten by a tick were not tested for a tick-borne pathogen, including those with consistent symptoms. We highlight the need for further research and dissemination of information to residents and physicians in Northern California regarding tick-associated disease, so that appropriate medical attention can be rapidly sought and administered.

Endemic Skunk amdoparvovirus in free-ranging striped skunks (Mephitis mephitis) in California.

The genus Amdoparvovirus includes the newly discovered skunk amdoparvovirus and the well-characterized Aleutian disease virus which causes significant health impacts in farmed mink worldwide. In 2010-2013, an outbreak of fatal amdoparvovirus-associated disease was documented in free-ranging striped skunks (Mephitis mephitis) from the San Francisco Bay Area of California. To characterize the geographic distribution, earliest occurrence and abundance of this virus, as well as possible impacts on sympatric mustelids of conservation concern, we tested blood samples from skunks throughout California and fishers (Pekania pennanti) from northern California for amdoparvovirus DNA. Amdoparvovirus DNA was detected in 64.8% of sampled skunks (140/216), and test-positive skunks were distributed widely throughout the state, from as far north as Humboldt County and south to San Diego County. The first test-positive skunks were detected from 2004, prior to the 2010-2013 outbreak. No significant spatial or temporal clustering of infection was detected. Although healthy and clinically ill animals tested positive for amdoparvovirus DNA, histopathologic evaluation of a subset from clinically ill skunks indicated that positive PCR results were associated with pneumonia as well as there being more than one inflammatory type lesion. None of 38 fishers were PCR-positive. Given the widespread geographic distribution and lack of a clear epizootic centre, our results suggest the presence of an endemic skunk-associated amdoparvovirus strain or species. However, if the virus is not host-specific, skunks' ubiquitous presence across rural and urban habitats may pose a risk to susceptible domestic and wild species including mustelids of conservation concern such as fishers and Pacific martens (Martes caurina).

Prevalence, distribution, and diversity of cryptic piroplasm infections in raccoons from selected areas of the United States and Canada.

The order Piroplasmida contains a diverse group of intracellular parasites, many of which can cause significant disease in humans, domestic animals, and wildlife. Two piroplasm species have been reported from raccoons (Procyon lotor), Babesia lotori (Babesia sensu stricto clade) and a species related to Babesia microti (called B. microti-like sp.). The goal of this study was to investigate prevalence, distribution, and diversity of Babesia in raccoons. We tested raccoons from selected regions in the United States and Canada for the presence of Babesia sensu stricto and Babesia microti-like sp. piroplasms. Infections of Babesia microti-like sp. were found in nearly all locations sampled, often with high prevalence, while Babesia sensu stricto infections had higher prevalence in the Southeastern United States (20-45% prevalence). Co-infections with both Babesia sp. were common. Sequencing of the partial 18S rRNA and cytochrome oxidase subunit 1 (cox1) genes led to the discovery of two new Babesia species, both found in several locations in the eastern and western United States. One novel Babesia sensu stricto sp. was most similar to Babesia gibsoni while the other Babesia species was present in the 'western piroplasm' group and was related to Babesia conradae. Phylogenetic analysis of the cox1 sequences indicated possible eastern and western genetic variants for the three Babesia sensu stricto species. Additional analyses are needed to characterize these novel species; however, this study indicates there are now at least four species of piroplasms infecting raccoons in the United States and Canada (Babesia microti-like sp., Babesia lotori, a novel Babesia sensu stricto sp., a novel western Babesia sp.) and a possible fifth species (Babesia sensu stricto) in raccoons in Japan.

Grass is not always greener: rodenticide exposure of a threatened species near marijuana growing operations.

OBJECTIVE: Marijuana (Cannabis spp.) growing operations (MGO) in California have increased substantially since the mid-1990s. One environmental side-effect of MGOs is the extensive use of anticoagulant rodenticides (AR) to prevent damage to marijuana plants caused by wild rodents. In association with a long-term demographic study, we report on an observation of brodifacoum AR exposure in a threatened species, the northern spotted owl (Strix occidentalis caurina), found freshly dead within 669-1347 m of at least seven active MGOs. RESULTS: Liver and blood samples from the dead northern spotted owl were tested for 12 rodenticides. Brodifacoum was the only rodenticide detected in the liver (33.3-36.3 ng/g) and blood (0.48-0.54 ng/ml). Based on necropsy results, it was unclear what role brodifacoum had in the death of this bird. However, fatal AR poisoning has been previously reported in owls with relatively low levels of brodifacoum residues in the liver. One likely mechanism of AR transmission from MGOs to northern spotted owls in California is through ingestion of AR contaminated prey that frequent MGOs. The proliferation of MGOs with their use of ARs in forested landscapes used by northern spotted owls may pose an additional stressor for this threatened species.

Diversity of rickettsiae in a rural community in northern California.

Far northern California forests are highly biodiverse in wildlife reservoirs and arthropod vectors that may propagate rickettsial pathogens in nature. The proximity of small rural communities to these forests puts people and domestic animals at risk of vector-borne infection due to spillover from wildlife. The current study was conducted to document exposure to rickettsial pathogens in people and domestic animals in a rural community, and identify which rickettsiae are present in sylvatic and peri-domestic environments near this community. Blood samples from people, domestic animals (dogs, cats, and horses) and wild carnivores were tested for Rickettsia spp. antibodies and DNA (people and domestic animals only) by serology and real time (RT)-PCR, respectively. Ectoparasites were collected from dogs, wild carnivores and from vegetation by flagging, and tested for Rickettsia spp. DNA by RT-PCR. DNA sequencing of the rickettsial 17kDa protein gene or the ompA gene was used for species identification. Despite a seroprevalence of 3% in people, 42% in dogs, 79% in cats, 33% in gray foxes, and 83% in bobcats, RT-PCR on blood was consistently negative, likely because the sensitivity of this test is low, as Rickettsia spp. do not often circulate in high numbers in the blood. Rickettsia spp. DNA was found in four flea species collected from bobcats and Ctenocephalides felis collected from domestic dogs. All amplicons sequenced from fleas were R. felis. Ixodes pacificus collected by flagging were commonly infected with a Rickettsia sp. endosymbiont. Rickettsia rhipicephali DNA was found in Dermacentor variabilis from dogs, black bears, a gray fox, and a D. occidentalis collected by flagging. Dermacentor variabilis from dogs and black bears also contained R. montanensis DNA. Multiple Rickettsia spp. (including species with zoonotic and pathogenic potential) were found among human biting arthropod vectors of both wild and domestic carnivores and on flags. Knowledge of the diversity of Rickettsia spp. that are present within arthropod vectors to which people and domestic animals are exposed is an essential first step is making an accurate diagnosis and in better understanding the epidemiology of these potential pathogens. Within-host and vector interaction among these species may play a role in spillover into human and domestic animals.

Patterns of Natural and Human-Caused Mortality Factors of a Rare Forest Carnivore, the Fisher (Pekania pennanti) in California.

Wildlife populations of conservation concern are limited in distribution, population size and persistence by various factors, including mortality. The fisher (Pekania pennanti), a North American mid-sized carnivore whose range in the western Pacific United States has retracted considerably in the past century, was proposed for threatened status protection in late 2014 under the United States Endangered Species Act by the United States Fish and Wildlife Service in its West Coast Distinct Population Segment. We investigated mortality in 167 fishers from two genetically and geographically distinct sub-populations in California within this West Coast Distinct Population Segment using a combination of gross necropsy, histology, toxicology and molecular methods. Overall, predation (70%), natural disease (16%), toxicant poisoning (10%) and, less commonly, vehicular strike (2%) and other anthropogenic causes (2%) were causes of mortality observed. We documented both an increase in mortality to (57% increase) and exposure (6%) from pesticides in fishers in just the past three years, highlighting further that toxicants from marijuana cultivation still pose a threat. Additionally, exposure to multiple rodenticides significantly increased the likelihood of mortality from rodenticide poisoning. Poisoning was significantly more common in male than female fishers and was 7 times more likely than disease to kill males. Based on necropsy findings, suspected causes of mortality based on field evidence alone tended to underestimate the frequency of disease-related mortalities. This study is the first comprehensive investigation of mortality causes of fishers and provides essential information to assist in the conservation of this species.

High Time for Conservation: Adding the Environment to the Debate on Marijuana Liberalization.

The liberalization of marijuana policies, including the legalization of medical and recreational marijuana, is sweeping the United States and other countries. Marijuana cultivation can have significant negative collateral effects on the environment that are often unknown or overlooked. Focusing on the state of California, where by some estimates 60%-70% of the marijuana consumed in the United States is grown, we argue that (a) the environmental harm caused by marijuana cultivation merits a direct policy response, (b) current approaches to governing the environmental effects are inadequate, and

Canine distemper in an isolated population of fishers (Martes pennanti) from California.

Four fishers (Martes pennanti) from an insular population in the southern Sierra Nevada Mountains, California, USA died as a consequence of an infection with canine distemper virus (CDV) in 2009. Three fishers were found in close temporal and spatial relationship; the fourth fisher died 4 mo later at a 70 km distance from the initial group. Gross lesions were restricted to hyperkeratosis of periocular skin and ulceration of footpads. All animals had necrotizing bronchitis and bronchiolitis with syncytia and intracytoplasmic inclusion bodies. Inclusion bodies were abundant in the epithelia of urinary bladder and epididymis but were infrequent in the renal pelvis and the female genital epithelia. No histopathologic or immunohistochemical evidence for virus spread to the central nervous system was found. One fisher had encephalitis caused by Sarcocystis neurona and another had severe head trauma as a consequence of predation. The H gene nucleotide sequence of the virus isolates from the first three fishers was identical and was 99.6% identical to the isolate from the fourth fisher. Phylogenetically, the isolates clustered with other North American isolates separate from classical European wildlife lineage strains. These data suggest that the European wildlife lineage might consist of two separate subgroups that are genetically distinct and endemic in different geographic regions. The source of infection as well as pertinent transmission routes remained unclear. This is the first report of CDV in fishers and underscores the significance of CDV as a pathogen of management concern.

Anticoagulant rodenticides on our public and community lands: spatial distribution of exposure and poisoning of a rare forest carnivore.

Anticoagulant rodenticide (AR) poisoning has emerged as a significant concern for conservation and management of non-target wildlife. The purpose for these toxicants is to suppress pest populations in agricultural or urban settings. The potential of direct and indirect exposures and illicit use of ARs on public and community forest lands have recently raised concern for fishers (Martes pennanti), a candidate for listing under the federal Endangered Species Act in the Pacific states. In an investigation of threats to fisher population persistence in the two isolated California populations, we investigate the magnitude of this previously undocumented threat to fishers, we tested 58 carcasses for the presence and quantification of ARs, conducted spatial analysis of exposed fishers in an effort to identify potential point sources of AR, and identified fishers that died directly due to AR poisoning. We found 46 of 58 (79%) fishers exposed to an AR with 96% of those individuals having been exposed to one or more second-generation AR compounds. No spatial clustering of AR exposure was detected and the spatial distribution of exposure suggests that AR contamination is widespread within the fisher's range in California, which encompasses mostly public forest and park lands Additionally, we diagnosed four fisher deaths, including a lactating female, that were directly attributed to AR toxicosis and documented the first neonatal or milk transfer of an AR to an altricial fisher kit. These ARs, which some are acutely toxic, pose both a direct mortality or fitness risk to fishers, and a significant indirect risk to these isolated populations. Future research should be directed towards investigating risks to prey populations fishers are dependent on, exposure in other rare forest carnivores, and potential AR point sources such as illegal marijuana cultivation in the range of fishers on California public lands.

Prevalence to Toxoplasma gondii and Sarcocystis spp. in a reintroduced fisher (Martes pennanti) population in Pennsylvania.

Understanding the role of disease in population regulation is important to the conservation of wildlife. We evaluated the prevalence of Toxoplasma gondii exposure and Sarcocystis spp. infection in 46 road-killed and accidentally trapper-killed fisher (Martes pennanti) carcasses collected and stored at -20 C by the Pennsylvania Game Commission from February 2002 to October 2008. Blood samples were assayed for T. gondii antibodies using the modified agglutination test (MAT, 1 : 25) and an indirect immunofluorescent antibody test (IFAT, 1 : 128). For genetic analysis, DNA samples were extracted from thoracic and pelvic limb skeletal muscle from each carcass to test for Sarcocystis spp. using 18s-rRNA PCR primers. Antibodies to T. gondii were found in 100% (38 of 38) of the fishers tested by MAT and in 71% (32 of 45) of the fishers tested by IFAT. PCR analysis revealed that 83% (38 of 46) of the fishers were positive for Sarcocystis spp. Sequence analysis of 7 randomly chosen amplicons revealed the fisher sarcocysts had a 98.3% to 99.1% identity to several avian Sarcocystis spp. sequences in GenBank. Data from our study suggest that a high percentage of fishers in Pennsylvania have been exposed to T. gondii and are infected with Sarcocystis spp.

Pilot immunization of mice infected with an equine strain of Corynebacterium pseudotuberculosis.

This pilot study evaluated protection of an equine autogenous bacterin-toxoid vaccine against Corynebacterium pseudotuberculosis infection. Twenty-four BALB/c mice were inoculated with two doses of bacterin-toxoid vaccine or two injections of a placebo. Clinical, microbiologic, and pathologic outcomes were assessed after intradermal infection with one of two equine-origin C. pseudotuberculosis strains. Mice receiving bacterin-toxoid from fast-growing C. pseudotuberculosis showed significant protection from challenge infection, as evidenced by a higher survival rate, fewer gross and histopathologic lesions, and lower bacterial levels on culture. Successful protection via a vaccine against equine internal abscesses might provide supplementary management options against an important, potentially fatal disease.

Effectiveness of rapid diagnostic tests to assess pathogens of fishers (Martes pennanti) and gray foxes (Urocyon cinereoargenteus).

Wildlife managers often need to assess the current health status of wildlife communities before implementation of management actions involving surveillance, reintroductions, or translocations. We estimated the sensitivity and specificity of a commercially available domestic canine rapid diagnostic antigen test for canine parvovirus and a rapid enzyme-linked immunosorbent assay for the detection of antibodies toward Anaplasma phagocytophilum on populations of fishers (Martes pennanti) and sympatric gray foxes (Urocyon cinereoargenteus). Eighty-two fecal samples from 66 fishers and 16 gray foxes were tested with both SNAP((R)) PARVO rapid diagnostic test (RDT) and a nested polymerase chain reaction (PCR). Whole blood samples from 23 fishers and 53 gray foxes were tested with both SNAP 4Dx RDT and immunofluorescence assays (IFAs). The SNAP PARVO RDT detected no parvovirus, whereas PCR detected the virus in 17 samples. Eleven samples were positive using the SNAP 4Dx RDT, whereas 46 samples tested by IFA were positive for A. phagocytophilum. Both RDTs had low sensitivity and poor test agreement. These findings clearly demonstrate the importance of validating RDTs developed for domesticated animals before using them for wildlife populations.

Seroprevalence of Trypanosoma cruzi among eleven potential reservoir species from six states across the southern United States.

Trypanosoma cruzi, the causative agent of Chagas' disease, is a substantial public health concern in Latin America. Although rare in humans and domestic animals in the United States, T. cruzi is commonly detected in some wildlife species, most commonly raccoons (Procyon lotor) and Virginia opossums (Didelphis virginiana). To increase our understanding of the reservoir host species range and geographic distribution, 11 species of mammals from six states spanning the known range of T. cruzi (Arizona, California, Florida, Georgia, Missouri, and Virginia) were tested for antibodies to T. cruzi using indirect immunofluorescent antibody testing. In addition, culture isolation attempts were conducted on a limited number of animals from Georgia and Florida. Evidence of T. cruzi was found in every state except California; however, low numbers of known reservoirs were tested in California. In general, the highest seroprevalence rates were found in raccoons (0-68%) and opossums (17-52%), but antibodies to T. cruzi were also detected in small numbers of striped skunks (Mephitis mephitis) from Arizona and Georgia, bobcats (Lynx rufus) from Georgia, two coyotes (Canis latrans) from Georgia and Virginia, and a ringtail (Bassariscus astutus) from Arizona. Culture-based prevalence rates for raccoons were significantly greater than those for opossums; however, seroprevalences of raccoons and opossums from several geographic locations in Georgia and Florida were not different, indicating that exposure rates of these two species are similar within these areas. For both raccoons and opossums, seroprevalence was significantly higher in females than in males. No difference was detected in seroprevalence between adults and juveniles and between animals caught in urban and rural locations. Our results indicate that T. cruzi prevalence varies by host species, host characteristics, and geographic region and provides data to guide future studies on the natural history of T. cruzi in the United States.

Ecology of Anaplasma phagocytophilum infection in gray foxes (Urocyon cinereoargenteus) in northwestern California.

Although granulocytic anaplasmosis, caused by infection of Anaplasma phagocytophilum, is an emerging human and domestic animal disease, the ecology and natural history of the parasite is not well understood. Gray foxes (Urocyon cinereoargenteus) are relatively common, occasionally peri-urban mesocarnivores whose geographic distribution overlaps the reported distribution of granulocytic anaplasmosis in humans and domestic animals in North America. We evaluated the potential of foxes as hosts and reservoirs of A. phagocytophilum in both urban and backcountry habitats of the Hoopa Valley Indian Reservation, Humboldt County, California, USA. We trapped 54 individual foxes and had 16 recaptures for a total of 70 fox samples between June 2003 and October 2004 in delineated urban and backcountry zones. We collected 296 adult and 145 nymphal ticks from the 70 captured foxes including 193 Ixodes pacificus, 149 Ixodes texanus, 98 Dermacentor variabilis, and one Dermacentor occidentalis. There were seasonal differences in tick intensities, with most I. pacificus adults occurring in winter and spring (P < 0.001), most I. texanus nymphs in spring (P = 0.03), and most D. variabilis adults in spring and summer (P = 0.01). Thirty-six (51%) of the 70 fox sera had antibodies against A. phagocytophilum, with a higher (P = 0.24) prevalence in backcountry foxes (16 of 23) than in urban-zone foxes (12 of 31). Six (9%) of 70 fox samples were polymerase chain reaction-positive for A. phagocytophilum. Twenty-eight (31%) of 90 domestic dogs sampled from vaccine clinics within the study area were seropositive for A. phagocytophilum. There was a significant difference in prevalence between dogs and backcountry foxes (70%), but no differences were found between dogs and urban foxes (39%). We propose that gray foxes are a good sentinel species for A. phagocytophilum infections in northwestern California.

Zoonotic Bartonella species in fleas collected on gray foxes (Urocyon cinereoargenteus).

Bartonella spp. are fastidious, gram-negative, rod-shaped bacteria and are usually vector-borne. However, the vector has not been definitively identified for many recently described species. In northern California, gray foxes (Urocyon cinereoargenteus) are infected with two zoonotic Bartonella species, B. rochalimae and B. vinsonii subsp. berkhoffii. Fleas (range 1-8 fleas per fox) were collected from 22 (41.5%) of 54 gray foxes from urban and backcountry zones near Hoopa, California. The flea species were determined, and DNA was individually extracted to establish the Bartonella species harbored by these fleas. Of the 108 fleas collected, 99 (92%) were identified as Pulex simulans. Overall, 39% (42/108) of the fleas were polymerase chain reaction (PCR)-positive for Bartonella, with B. rochalimae and B. vinsonii subsp. berkhoffii identified in 34 (81%) and 8 (19%) of the PCR-positive fleas, respectively. There was no difference between the prevalence of Bartonella spp. in P. simulans for the urban and backcountry zones. Fourteen (64%) of the 22 foxes were Bartonella bacteremic at one or more of the capture dates. In 10 instances, both the foxes and the fleas collected from them at the same blood collection were Bartonella-positive. B. rochalimae was the predominant species identified in both foxes and fleas. The competency of Pulex fleas as a vector of B. rochalimae has not been confirmed and will need to be demonstrated experimentally. Pulex spp. fleas readily feed on humans and may represent a source of human exposure to zoonotic species of Bartonella.

Infective endocarditis in a dog and the phylogenetic relationship of the associated "Bartonella rochalimae" strain with isolates from dogs, gray foxes, and a human.

The first case of canine endocarditis caused by "Bartonella rochalimae" is reported. By PCR-restriction fragment length polymorphism, sequence, and phylogenetic analyses, Bartonella isolates from a dog with endocarditis, 22 gray foxes, and three dogs, described as B. clarridgeiae like, were confirmed to belong to the new species "B. rochalimae," suggesting canids as the natural reservoir.

Gray foxes (Urocyon cinereoargenteus) as a potential reservoir of a Bartonella clarridgeiae-like bacterium and domestic dogs as part of a sentinel system for surveillance of zoonotic arthropod-borne pathogens in northern California.

Two species of Bartonella, a novel Bartonella clarridgeiae-like bacterium and B. vinsonii subsp. berkhoffii, were isolated from rural dogs and gray foxes in northern California. A novel B. clarridgeiae-like species was isolated from 3 (1.7%) of 182 dogs and 22 (42%) of 53 gray foxes, while B. vinsonii subsp. berkhoffii was isolated from 1 dog (0.5%) and 5 gray foxes (9.4%). PCR and DNA sequence analyses of the citrate synthase (gltA) gene and the 16S-23S intergenic spacer region suggested that strains infecting dogs and gray foxes were identical. Fifty-four dogs (29%) and 48 gray foxes (89%) had reciprocal titers of antibodies against Bartonella spp. of > or =64. The high prevalence of bacteremia and seroreactivity to Bartonella spp. in gray foxes suggests that they may act as a reservoir species for the B. clarridgeiae-like species in this region. Domestic dogs were also tested for other arthropod-borne infectious agents. Fifty-one dogs (28%) were positive for Dirofilaria immitis antigen, seventy-four (40%) were seroreactive to Anaplasma phagocytophilum, and five (2.7%) were seropositive for Yersinia pestis. Fourteen dogs (7.6%) were PCR positive for A. phagocytophilum. Polytomous logistic regression models were used to assess the association of Bartonella antibody titer categories with potential risk factors and the presence of other vector-borne agents in domestic dogs. Older dogs were more likely to be seroreactive to Bartonella spp. There was no association between the exposure of dogs to Bartonella and the exposure of dogs to A. phagocytophilum in this study.