Factors affecting carriage and intensity of infection of Calodium hepaticum within Norway rats (Rattus norvegicus) from an urban slum environment in Salvador, Brazil.

Urban slum environments in the tropics are conducive to the proliferation and the spread of rodent-borne zoonotic pathogens to humans. Calodium hepaticum (Brancroft, 1893) is a zoonotic nematode known to infect a variety of mammalian hosts, including humans. Norway rats (Rattus norvegicus) are considered the most important mammalian host of C. hepaticum and are therefore a potentially useful species to inform estimates of the risk to humans living in urban slum environments. There is a lack of studies systematically evaluating the role of demographic and environmental factors that influence both carriage and intensity of infection of C. hepaticum in rodents from urban slum areas within tropical regions. Carriage and the intensity of infection of C. hepaticum were studied in 402 Norway rats over a 2-year period in an urban slum in Salvador, Brazil. Overall, prevalence in Norway rats was 83% (337/402). Independent risk factors for C. hepaticum carriage in R. norvegicus were age and valley of capture. Of those infected the proportion with gross liver involvement (i.e. >75% of the liver affected, a proxy for a high level intensity of infection), was low (8%, 26/337). Sixty soil samples were collected from ten locations to estimate levels of environmental contamination and provide information on the potential risk to humans of contracting C. hepaticum from the environment. Sixty percent (6/10) of the sites were contaminated with C. hepaticum. High carriage levels of C. hepaticum within Norway rats and sub-standard living conditions within slum areas may increase the risk to humans of exposure to the infective eggs of C. hepaticum. This study supports the need for further studies to assess whether humans are becoming infected within this community and whether C. hepaticum is posing a significant risk to human health.

Leptospira in breast tissue and milk of urban Norway rats (Rattus norvegicus).

Leptospirosis is a zoonosis caused by bacteria of the genus Leptospira. The disease is globally distributed and a major public health concern. The Norway rat (Rattus norvegicus) is the main reservoir of the pathogen in urban slums of developing and developed countries. The potential routes of intra-specific leptospire transmission in rats are largely unknown. Herein, we identified pathogenic Leptospira spp. in breast tissue and milk of naturally infected rats. We examined kidney, breast tissue and milk from 24 lactating rats for the presence of leptospires using immunofluorescence, immunohistochemistry, polymerase chain reaction (PCR) and scanning electronic microscopy. All 24 rats had evidence for Leptospira in the kidneys, indicating chronic carriage. The majority of kidney-positive rats had detectable leptospires in milk (18, 75%) and breast tissue (16, 67%), as evidenced by immunofluorescence assay and immunohistochemistry. Four (17%) milk samples and two (8%) breast tissue samples were positive by quantitative real-time PCR. Scanning electron microscopy confirmed the presence of leptospires in breast tissue. No major pathological changes in breast tissue were found. This study, for the first time, identified leptospires in the milk and breast tissue of wild Norway rats, suggesting the possibility of milk-borne transmission of leptospirosis to neonates.

Pre-spillover prevention of emerging zoonotic diseases: what are the targets and what are the tools?

The uneven standards of surveillance, human- or animal-based, for zoonotic diseases or pathogens maintained and transmitted by wildlife H(R)s, or even domestic species, is a global problem, readily apparent even within the United States, where investment in public health, including surveillance systems, has a long and enviable history. As of 2006, there appears to be little scientific, social, or political consensus that animal-based surveillance for zoonoses merits investment in international infrastructure, other than the fledgling efforts with avian influenza, or targeted nontraditional avenues of surveillance and research. National institutions charged with strategic planning for emerging diseases or intentional releases of zoonotic agents have emphasized improving diagnostic capabilities for detecting human infections, modifying the immune status of human or domestic animals through vaccines, producing better antiviral or antibacterial drugs, and enhancing human-based surveillance as an early warning system. With the possible exception of extensive human vaccination, each of these approaches target post-spillover events and none of these avenues of research will have the slightest impact on reducing the risk of additional emergence of viruses or other pathogens from wildlife. Novel schemes of preventing spillover of human pathogens from animal H(R)s can only spring from improving our understanding of the ecological context and biological interactions of pathogen maintenance among H(R)s. Although the benefit derived from investments to improve surveillance and knowledge of zoonotic pathogens circulating among wildlife H(R) populations is uncertain, our experience with HIV and the looming threat of pandemic avian influenza A inform us of the outcomes we can expect by relying on detection of post-spillover events among sentinel humans.

Introduction: conceptualizing and partitioning the emergence process of zoonotic viruses from wildlife to humans.

This introduction provides a telegraphic overview of the processes of zoonotic viral emergence, the intricacies of host-virus interactions, and the distinct role of biological transitions and modifying factors. The process of emergence is conceptualized as two transition stages which are common and required for all disease emergence, (1) human contact with the infectious agent and (2) cross-species transmission of the agent, and two transition stages which are not required for emergence and appear unavailable to many zoonotic pathogens, (3) sustained human-to-human transmission and (4) genetic adaptation to the human host. The latter two transitions are presumably prerequisites for the pandemic emergence of a pathogen. The themes introduced herein are amplified and explored in detail by the contributors to this volume. Each author explores the mechanisms and unique circumstances by which evolution, biology, history, and current context have contrived to drive the emergence of different zoonotic agents by a series of related events; although recognizable similarities exist among the events leading to emergence the details and circumstances are never repetitive.

Animal-based national surveillance for zoonotic disease: quality, limitations, and implications of a model system for monitoring rabies.

Surveillance for zoonotic diseases among wildlife is a research and public health challenge. The inherent limitations posed by the requisite human-animal interactions are often undefined and underappreciated. The national surveillance system for animal rabies in the United States was examined as a model system; reporting of animal rabies is legally mandated, each case of rabies is laboratory confirmed, and data have been consistently collected for more than 50 years. Factors influencing the monthly counts of animal rabies tests reported during 1992-2001 were assessed by univariate and multivariable regression methods. The suitability of passively collected surveillance data for determining the presence or absence of the raccoon-associated variant of rabies within states and within individual counties was assessed by determining critical threshold values from the regression analyses. The size of the human population and total expenditures within a county accounted for 72% and 67%, respectively, of the variance in testing. The annual median number of rabies tests performed was seven for counties without rabies, 22 for counties with non-raccoon rabies, and 34 for counties with raccoon rabies. Active surveillance may be required in locales with sparse human populations when a high degree of confidence in the status of rabies is required.

Persistence of elevated rabies prevention costs following post-epizootic declines in rates of rabies among raccoons (Procyon lotor).

Determining the benefits to cost relationships among different approaches to rabies control and prevention has been hindered by the inherent temporal variability in the dynamics of disease among wildlife reservoir hosts and a tangible and objective measure of the cost of rabies prevention. A major and unavoidable component of rabies prevention programs involves diagnostic testing of animals and the subsequent initiation of appropriate public health responses. The unit cost per negative and positive diagnostic test outcome can be reasonably estimated. This metric when linked to methodologies subdividing the epizootic process into distinct temporal stages provided the requisite detail to estimate benefits derived from rabies control strategies. Oral rabies vaccine (ORV), for prevention of the raccoon-associated variant of rabies, has been distributed in Ohio and adjoining states in an effort to develop an immune barrier to the westward spread of epizootic raccoon rabies. The costs of ORV delivery have been quantified. Herein, the cost structures required to assess the benefits accrued by prevention were developed. A regression model was developed effectively predicting (r2=0.70) the total number of rabies diagnostic tests performed by 53 counties in five northeastern (NE) states from 1992 to 2001. Five temporal stages sufficed to capture the range of variability in the raccoon rabies epizootic process. Unit costs, dollars per diagnostic test outcome, were calculated for negative and positive results from published reports. Ohio counties were matched to NE counties based on similar socioeconomic characters. A "pseudo-epizootic" of raccoon rabies was introduced into Ohio and the costs savings from ORV were derived as the excess costs imposed by epizootic spread throughout the state. At 46 km/year (range modeled, 30-60 km/year), the pseudo epizootic spread, and reached the enzootic stage, in all Ohio counties by year 13 (range modeled, 11-17 years). Cumulative excess costs for Ohio ranged between $11 and $21 million; counties of low socioeconomic status experienced the greatest relative excess costs. The costs for rabies prevention activities reached apices during the epizootic stage of raccoon rabies (2.7-10.8 times baseline) an unforeseen finding indicated elevated costs persisted (1.7-7.2 times baseline) into the enzootic stage.

Assessing the role of long-distance translocation and spatial heterogeneity in the raccoon rabies epidemic in Connecticut.

Spatial heterogeneity and long-distance translocation (LDT) play important roles in the spatio-temporal dynamics and management of emerging infectious diseases and invasive species. We assessed the influence of LDT events on the invasive spread of raccoon rabies through Connecticut. We identified several putative LDT events, and developed a network-model to evaluate whether they became new foci for epidemic spread. LDT was fairly common, but many of the LDTs were isolated events that did not spread. Two putative LDT events did appear to become nascent foci that affected the epidemic in surrounding townships. In evaluating the role of LDT, we simultaneously revisited the problem of spatial heterogeneity. The spread of raccoon rabies is associated with forest cover--rabies moves up to three-times slower through the most heavily forested townships compared with those with less forestation. Forestation also modified the effect of rivers. In the best overall model, rabies did not cross the river separating townships that were heavily forested, and the spread slowed substantially between townships that were lightly forested. Our results suggest that spatial heterogeneity can be used to enhance the effects of rabies control by focusing vaccine bait distribution along rivers in lightly forested areas. LDT events are a concern, but this analysis suggests that at a local scale they can be isolated and managed.

Antibodies to Bartonella species in inner-city intravenous drug users in Baltimore, Md.

BACKGROUND: Bartonella quintana has recently been associated with homeless alcoholic men. Both B quintana and Bartonella henselae have been shown to be opportunistic pathogens of people with acquired immunodeficiency syndrome. The reservoirs and modes of transmission of these infections are incompletely known. OBJECTIVES: To examine serum samples that were taken from inner-city intravenous (IV) drug users for antibodies to Bartonella organisms to determine whether there is an urban transmission cycle for Bartonella species and to examine the demographic and behavioral characteristics of IV drug users to identify possible risk factors for infection with any of these agents. MATERIALS AND METHODS: A serologic survey was conducted, using a convenience sample of serum specimens collected during a study of IV drug use and human immunodeficiency virus infection among 630 inner-city residents in Baltimore, Md. A detailed questionnaire was administered at the initial collection of serum, and additional serum collections and questionnaire updates were made at 6-month intervals. The most recent available serum sample was tested for Bartonella antibody titer by using the indirect immunofluorescent antibody test with 3 antigens: Bartonella elizabethae, B henselae, and B quintana. Univariate and multivariate analyses of selected potential demographic and behavioral risk factors were conducted. RESULTS: Antibodies to Bartonella were highly prevalent in this group; more than 37% of all samples reacted with at least 1 antigen. Overall seroprevalence of antibodies to B elizabethae, B henselae, and B quintana was 33%, 11%, and 10%, respectively. Current IV drug use, frequency of injection, and seronegative human immunodeficiency virus status were significantly associated with Bartonella antibody presence, but these associations varied by analysis. There was a significant inverse association of antibody prevalence to B henselae and B quintana by using CD4+ cell counts among human immunodeficiency virus-seropositive individuals. CONCLUSIONS: Intravenous drug users have an elevated prevalence of antibodies to Bartonella organisms and may be at significant risk of becoming infected. Current IV drug use, high frequency of injection, and seronegative human immunodeficiency virus status are significant risk factors for an increased prevalence of Bartonella antibodies. The current natural histories of Bartonella species are rapidly changing, and mechanisms of transmission remain unknown.

Use of Bartonella antigens for serologic diagnosis of cat-scratch disease at a national referral center.

BACKGROUND: Bartonella henselae (formerly the genus Rochalimaea) has recently been isolated from patients with cat-scratch disease and their cats, and since September 1992 the Centers for Disease Control and Prevention has offered an indirect fluorescent antibody assay for Bartonella-specific antibody. METHODS: Physicians submitted serum samples from patients suspected of having cat-scratch disease or other Bartonella-associated illness and completed a questionnaire that recorded clinical information. Indirect fluorescent antibody assay was performed with the use of antigen derived from three Bartonella species: B henselae, Bartonella quintana, and Bartonella elizabethae. RESULTS: During 16 months, 3088 serum samples were received. The largest numbers of specimens and the highest percentages positive (titer, > or = 64) were observed in the fall and winter. Clinical histories of the first 600 patients for whom serum samples and completed information forms were received were examined in detail; seropositivity was significantly associated with cat contact, cat age of less than 1 year, cat scratch, presence of an inoculation papule, and regional adenopathy. Of 91 patients whose illness met a strict clinical definition of cat-scratch disease, 86 (95%) had titers of 64 or greater to either B henselae or B quintana. A fourfold rise or fall in titer was observed in 87 of 132 patients with paired serum samples. CONCLUSIONS: The indirect fluorescent antibody assay for Bartonella-specific antibody is sensitive for the diagnosis of cat-scratch disease. Redefinition of cat-scratch disease on the basis of cause and use of this assay as a diagnostic criterion is recommended.

Isolation of lactate dehydrogenase-elevating viruses from wild house mice and their biological and molecular characterization.

Lactate dehydrogenase-elevating virus (LDV) was first identified as a contaminant of transplantable mouse tumors that were passaged in laboratory mice. It has been assumed that these LDVs originated from LDVs endemic in wild house mouse populations. In order to test this hypothesis and to explore the relationships between LDVs from wild house mice among each other and to those isolated from laboratory mice, we have isolated LDVs from wild house mice and determined their biological and molecular properties. We have screened for LDV tissues of 243 wild house mice that had been caught in various regions of North, Central and South America between 1985 and 1994. We were able to isolate LDVs from the tissues of four mice, three had been caught in Baltimore, MD and one in Montana. We demonstrate that the phenotypic properties (ability to establish a long-term viremic infection, low immunogenicity of the neutralization epitope, high resistance to antibody neutralization and lack of neuropathogenicity) of the four wild house mouse LDVs are identical to those of the primary LDVs isolated from transplantable tumors (LDV-P and LDV-vx), which are distinct from those of the neuropathogenic LDV-C. Furthermore, ORF 5 and ORF 2 and their protein products (the primary envelope glycoprotein VP-3P, and the minor envelope glycoprotein, respectively) of the wild house mouse LDVs were found to be closely related to those of LDV-P and LDV-vx. The LDVs caught in Baltimore, MD were especially closely related to each other, whereas the LDV isolated in Montana was more distantly related, indicating that it had evolved independently. The ectodomain of VP-3P of all four wild house mouse LDVs, like those of LDV-P and LDV-vx, possess the same three polylactosaminoglycan chains, two of which are lacking in the VP-3P ectodomain of LDV-C. These results further strengthen the conclusion that the three polylactosaminoglycan chains are the primary determinants of the phenotypic properties of LDV-P/vx.

Cluster of five children with acute encephalopathy associated with cat-scratch disease in south Florida.

Between August 12 and September 27, 1994, five children in South Florida were hospitalized at a single hospital because of encephalopathy, presenting as status epilepticus, associated with cat-scratch disease (CSD). Diagnoses were confirmed by using an indirect fluorescent antibody test to detect antibody to Bartonella henselae, the causative agent of CSD. These cases represent the first cluster of CSD encephalopathy cases to be recognized in the United States. The patients lived within 7 miles of each other and all reported contact with pet or stray cats before developing regional lymphadenopathy and encephalopathy. All recovered fully. A high proportion of 124 cats from the local area were seropositive (62%) or bacteremic (22%). This study suggests that B. henselae can be associated with geographically focal clusters of CSD encephalitis and should be considered in the evaluation of children with acute encephalopathy.

Mammalian reservoirs and epidemiology of rabies diagnosed in human beings in the United States, 1981-1998.

Between 1981 and 1998, 37 cases of rabies were diagnosed in human beings in the United States. Information directly linking the cause of infection to animal bite was available for only eight of these cases. Indirect incrimination of the vector by analysis of cDNA sequences obtained by reverse transcriptase polymerase chain reaction of samples indicated that for all cases (12/12) believed to have been acquired in foreign countries, variants of the rabies virus (VRVs) associated with dogs (7/12 involved known bite histories) were the cause of the rabies infections. In contrast, VRVs associated with bats (bat-associated VRVs or BAVs) were implicated as the cause of 88% (22/25) of infections believed to have been acquired within the United States (1/22 involved known bite histories). Sequence analyses revealed that a single BAV (Ln/Ps), associated with rabid silver-haired (Lasionycteris noctivagans) and Eastern pipistrelle (Pipistrellus subflavus) bats, was implicated in 73% (16/22) of bat-associated infections. Silver-haired bats are predominantly solitary and migratory. Eastern pipistrelle bats may occur individually or in small clusters. Both species are only infrequently submitted for rabies testing. Unrecognized bites and unique properties of the Ln/Ps BAV may explain its association with the majority of rabies infections in human beings in the United States.

Zoonotic viruses of wildlife: hither from yon.

The emergence of zoonotic viruses maintained by wildlife reservoir hosts is poorly understood. Recent discoveries of Hendra (HENV) and Nipah (NIPV) viruses in Australasia and the emergence of epidemic West Nile virus (WNV) in the United States have added urgency to the study of cross-species transmission. The processes by which zoonotic viruses are transmitted and infect other species are examined as four transitions. Two of these, inter-species contact and cross-species virus transmission (spillover), are essential and sufficient to cause epidemic emergence. Sustained transmission and virus adaptation within the spillover host are transitions not required for virus emergence, but determine the magnitude and scope of subsequent disease outbreaks. Ecologic, anthropogenic, and evolutionary factors modify the probability that viruses complete or move through transitions. As surveillance for wildlife diseases is rare and often outbreak-driven, targeted studies are required to elucidate the means by which important zoonotic viruses are maintained and spillover occurs.

Serologic evidence of hantaviral infections within small mammal communities of Baltimore, Maryland: spatial and temporal patterns and host range.

Serologic evidence was used to investigate the spatial and temporal distribution and host range of hantaviruses in small mammal communities in Baltimore, MD. Immunofluorescent antibody (IFA) reactive to a Baltimore rat isolate of Seoul virus was detected in 44% of 404 Norway rats captured at 4 park or residential locations; 21% of 121 meadow voles captured at 4 park locations possessed significant IFA titers to Prospect Hill virus. Evidence from plaque neutralization assay of rodent sera suggested that Seoul virus and Prospect Hill virus circulated concurrently in voles and rats, respectively, at 1 park. No cross infection of virus between these primary reservoir species was observed, as determined by higher specific neutralizing antibody titers to the characteristic virus for each host species. Only 4% of 449 house mice and 1% of 94 white-footed mice captured at the same sites as the primary host species were seropositive to hantaviruses, despite extensive demonstrated overlap in time and space with the primary host species.

Lymphocytic choriomeningitis virus infection and house mouse (Mus musculus) distribution in urban Baltimore.

Four hundred eighty house mice (Mus musculus) were trapped primarily from urban sites in Baltimore, Maryland from 1984 to 1989 and tested for antibody to lymphocytic choriomeningitis virus (LCMV). The majority of mice (95%) were trapped in residences at two city locations (n = 260), or in an urban park (n = 196); five additional sites were sampled. Overall, 9.0% of the mice were LCMV antibody positive and infected animals were obtained from six of eight sites, including all three of the primary city sites, where the prevalence varied significantly (3.9-13.4%). The location with the highest prevalence was an inner city residential site where positive mice were found significantly clustered within blocks and households. In this location, LCMV antibody prevalence was also significantly correlated with estimates of mouse density within individual blocks. The focal nature of LCMV infection in house mice may result from contact or vertical transmission of virus in conjunction with the highly structured social system of mice, which promotes inbreeding and limited dispersal.

Geographical distribution and age related prevalence of antibody to Hantaan-like virus in rat populations of Baltimore, Maryland, USA.

Recent studies indicate that domestic rats (Rattus norvegicus) from harbor-side areas within port cities of the United States are infected with a Hantaan-like virus. The geographical distribution of seropositive rats may be extremely localized within these urban environments. We surveyed four widely separated residential sites distant from the harbor within Baltimore, Maryland, USA, to determine the geographical distribution and prevalence of antibody to Hantaan-like virus in rats from urban areas of high human population density. Captured rats were weighed and examined for sexual maturity to allow some estimation of age, and their sera were examined by indirect immunofluorescent antibody assay for antibody to Hantaan virus. Seropositive rats were found at all four sites within Baltimore. Increasing antibody prevalence and high titers were associated with increasing rat weight and sexual maturity. Our results show that infection of rats by a Hantaan-like virus is widespread in Baltimore. Antibody in rats may be due to infections acquired during maturation or the delayed seroconversion of rats infected prior to weaning.

Prospective seroepidemiology of hantaviruses and population dynamics of small mammal communities of Baltimore, Maryland.

We used a prospective seroepidemiological study, in conjunction with a mark-release-recapture protocol, to investigate the transmission of hantaviruses in four rodent species from Baltimore, Maryland, from June 1984 to June 1986. A total of 1,208 captures of 762 rodents provided 984 individual blood samples. The antibody prevalence, as determined by frequency of reciprocal indirect fluorescent antibody (IFA) titers greater than or equal to 32, was 33.9% in rats (Rattus norvegicus, n = 466), 28.3% in meadow voles (Microtus pennsylvanicus, n = 67), 1.4% in house mice (Mus musculus, n = 146), and 1.2% in white-footed mice (Peromyscus leucopus, n = 83). Populations of all rodents were maximal during the fall and winter months, but population trends were not clearly associated with periods of virus transmission. The mean incidence of seroconversion to a Hantavirus for rats was 12.06/100 rats/month, but incidence rates could not be established for other species. Rats which seroconverted were generally sexually mature animals, and there was evidence of transmission throughout the year. Animals which seroconverted to a Hantavirus achieved high IFA titers, and remained seropositive for the duration of the study.

Serologic evidence of rickettsialpox (Rickettsia akari) infection among intravenous drug users in inner-city Baltimore, Maryland.

We tested single serum samples from 631 intravenous (i.v.) drug users from inner-city Baltimore, Maryland for serologic evidence of exposure to spotted fever group rickettsiae. A total of 102 (16%) individuals had titers > or = 64 to Rickettsia rickettsii by an indirect immunofluorescence assay. Confirmation that infection was caused by R. akari was obtained by cross-adsorption studies on a subset of serum samples that consistently resulted in higher titers to R. akari than to R. rickettsii. Current i.v. drug use, increased frequency of injection, and shooting gallery use were significant risk factors for presence of group-specific antibodies reactive with R. rickettsii. There was a significant inverse association with the presence of antibodies reactive to R. rickettsii and antibodies reactive to the human immunodeficiency virus. This study suggests that i.v. drug users are at an increased risk for R. akari infections. Clinicians should be aware of rickettsialpox, as well as other zoonotic diseases of the urban environment, when treating i.v. drug users for any acute febrile illness of undetermined etiology.

Evidence of rodent-associated Bartonella and Rickettsia infections among intravenous drug users from Central and East Harlem, New York City.

We tested serum samples collected in 1997 and 1998 from a cohort of 204 injection drug users (IDUs) recruited from Central and East Harlem, New York City, New York, for antibodies reactive with seven rickettsial or Bartonella spp. antigens. Rodent-associated Bartonella elizabethae and Rickettsia akari were the primary etiologic agents of interest. The testing panel also included Bartonella henselae, Bartonella quintana, Rickettsia prowazekii, Rickettsia rickettsii, and Rickettsia typhi. The highest prevalence of seroreactive serum samples (46%) was found with B. elizabethae antigens; 10% of the samples reacted with B. henselae antigens, while 2% reacted with B. quintana antigens. Reactivity to the latter two antigens was likely due to cross-reactivity with B. elizabethae antigens in most instances. Among the spotted fever group rickettsiae, 18 (9%) samples reacted with R. akari, including 10 samples (5%) that also reacted with R. rickettsii. Cross-adsorption studies demonstrated that most of the spotted fever group rickettsiae antibodies were due to R. akari infections. Among the typhus group rickettsiae, 5 samples reacted weakly to R. prowazekii antigens, and no samples reacted with R. typhi antigens. These findings suggest that Harlem IDUs are commonly exposed to two rodent-associated zoonotic agents. Further study of IDU populations may help elucidate transmission cycles of these agents in inner cities where higher levels of transmission occur.

Persistent infection in Neotoma fuscipes (Muridae: Sigmodontinae) with Ehrlichia phagocytophila sensu lato.

Dusky-footed wood rats (Neotoma fuscipes Baird) and two species of Peromyscus mice (P. maniculatus Wagner and P. truei Shufeldt) were collected over a 16-month period from three sites in Sonoma County, California. Blood was collected from 93 wood rats and 177 mice and serum or plasma was tested for seroreactivity with Ehrlichia phagocytophila sensu lato (also known as the human granulocytic ehrlichiosis agent). Thirty-five (37.6%) wood rats and 15 (8.5%) mice were seropositive. Positive Neotoma serology by site ranged from 9.4% to 62.1%. Polymerase chain reaction (PCR) testing for the Ehrlichia groESL heat shock operon was performed on all the seropositive and selected seronegative wood rats; 24 (68.6%) seropositive animals were PCR positive. Two seroconversions and no seroreversions were detected among 18 of the seropositive wood rats that were recaptured and tested multiple times (range = 2-6). Fourteen (77.8%) of the 18 were also PCR positive with six of these positive at every testing point (range = 2-6). One wood rat remained serologically and PCR positive in six specimens collected over a 14-month period. One male of 84 questing adult Ixodes pacificus Cooley & Kohls collected was PCR-positive for E. phagocytophila. Borrelia burgdorferi, the agent of Lyme disease, was cultured from ear punch biopsies from six of seven E. phagocytophila seropositive and one of four seronegative wood rats.