Isolation of Borrelia burgdorferi from Neotoma fuscipes, Peromyscus maniculatus, Peromyscus boylii, and Ixodes pacificus in Oregon.

The number of Lyme disease cases in Oregon has increased in recent years despite the fact that the pathogen, Borrelia burgdorferi, has never been isolated in the state. Rodent and tick surveys were undertaken in 1997 to isolate and characterize strains of B. burgdorferi from Oregon and to identify potential reservoirs and vectors of Lyme disease. Borrelia burgdorferi was isolated from Neotoma fuscipes, Peromyscus maniculatus, P. boylii, and Ixodes pacificus. Both N. fuscipes and P. maniculatus were infested with I. pacificus and I. spinipalpis. Although I. pacificus infested P. boylii, I. spinipalpis was not found on this rodent, and only 4% of the P. boylii were infected with B. burgdorferi compared with the 19% and 18% infection rates found in N. fuscipes and P. maniculatus, respectively. Variation in the molecular weights of the outer surface proteins A and B were found in these first confirmed isolates of B. burgdorferi from Oregon, as well as truncated forms of outer surface protein B.

Infestation of the southern alligator lizard (Squamata: Anguidae) by Ixodes pacificus (Acari: Ixodidae) and its susceptibility to Borrelia burgdorferi.

To investigate the reservoir potential of the southern alligator lizard, Elgaria multicarinata (Blainville), for the Lyme disease spirochete, Borrelia burgdorferi Johnson, Schmid, Hyde, Steigerwalt & Brenner, 14 lizards were collected from 1 county on each side of the northern Central Valley of California. Seven animals were collected from a Placer County site (Drivers Flat) and a Yolo County site (Cache Creek) where B. burgdorferi had been isolated previously from Ixodes pacificus Cooley & Kohls. Overall, the mean abundance of I. pacificus on all 14 lizards was 34.1 (range, 3-63) for larvae and 11.0 (range, 1-28) for nymphs. In captivity, field-attached I. pacificus larvae and nymphs required, on average, 12.6 (range, 1-37) and 14.4 (range, 5-44) d to feed to repletion, respectively. The prevalence of B. burgdorferi infection in host-seeking I. pacificus nymphs was 1.4% in Cache Creek Canyon and 9.9% in Drivers Flat. Attempts to isolate spirochetes from lizard blood or ticks that had fed on lizards and subsequently molted were unsuccessful as were efforts to cultivate spirochetes in lizard sera. These data suggest that the southern alligator lizard is not a competent reservoir for B. burgdorferi, although it is an important host for I. pacificus subadults.

Ehrlichia phagocytophila genogroup rickettsiae in ixodid ticks from California collected in 1995 and 1996.

A total of 1,246 ixodid ticks collected in 1995 and 1996 from seven California counties were examined for the presence of Ehrlichia phagocytophila genogroup rickettsiae by using a nested PCR technique. Of 1,112 adult Ixodes pacificus Cooley and Kohls ticks tested, nine pools, each containing five ticks, were positive (minimum percentage of ticks harboring detectable ehrlichiae, 0.8%). Positive ticks were limited to four of the seven counties (Sonoma, El Dorado, Santa Cruz, and Orange). In Santa Cruz County, three positive pools were identified at the home of an individual with prior confirmed human granulocytic ehrlichiosis. In El Dorado County, positive ticks were found at sites where cases of granulocytic ehrlichiosis in a horse and a llama had recently occurred. Among 47 nymphal I. pacificus ticks collected in Sonoma County, one positive pool was identified. Fifty-seven adult Dermacentor occidentalis Marx and 30 adult D. variabilis Say ticks, collected chiefly in southern California, were negative. These data, although preliminary, suggest that the prevalence of E. phagocytophila genogroup rickettsiae in ixodid ticks of California may be lower than in cognate vector populations (i.e., I. scapularis Say = I. dammini Spielman, Clifford, Piesman, and Corwin) in the eastern and midwestern United States.

An Ehrlichia strain from a llama (Lama glama) and Llama-associated ticks (Ixodes pacificus).

An ehrlichia was identified in the blood of a diseased llama (lama glama). Sequencing of its 16S rRNA gene showed the ehrlichia to be closely related to members of the Ehrlichia phagocytophila genogroup. The agent was also found in a pool of ticks (Ixodes pacificus) collected at the llama site.

Evidence implicating nymphal Ixodes pacificus (Acari: ixodidae) in the epidemiology of Lyme disease in California.

To clarify the role of nymphal versus adult western black-legged ticks (Ixodes pacificus) in the epidemiology of Lyme disease, the seasonal distribution, abundance, and spirochetal infection rates in these stages, and the seasonal occurrence of ticks biting humans and of incident cases of Lyme disease were determined in northern California. Although their seasonal activity periods overlapped for about one-third of the year, nymphs and adults predominated in different seasons, the former from late spring to summer and the latter from fall to early spring. At one site, four (4%) of 100 adults from low vegetation bordering a hardwood forest and 44 (13.6%) of 324 nymphs from leaf litter in the forest were found to contain Borrelia burgdorferi. Biting-collection records revealed that nymphs attach to people more commonly than recognized previously; I. pacificus nymphs comprised 12.5% of 967 ticks of various species and stages and 42% of all nymphs submitted for identification. Attachments by nymphs occurred primarily between April and August, which coincided with the seasonal occurrence of most incident cases of Lyme disease. Collectively, these findings strongly implicate the nymphal stage of I. pacificus as the primary vector of B. burgdorferi to humans in this region.

Distribution and molecular analysis of Lyme disease spirochetes, Borrelia burgdorferi, isolated from ticks throughout California.

Previous studies describing the occurrence and molecular characteristics of Lyme disease spirochetes, Borrelia burgdorferi, from California have been restricted primarily to isolates obtained from the north coastal region of this large and ecologically diverse state. Our objective was to look for and examine B. burdorferi organisms isolated from Ixodes pacificus ticks collected from numerous regions spanning most parts of California where this tick is found. Thirty-one isolates of B. burgdorferi were examined from individual or pooled I. pacificus ticks collected from 25 counties throughout the state. One isolate was obtained from ticks collected at Wawona Campground in Yosemite National Park, documenting the occurrence of the Lyme disease spirochete in an area of intensive human recreational use. One isolate from an Ixodes neotomae tick from an additional county was also examined. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis, immunoblot analysis, agarose gel electrophoresis, Southern blot analysis, and the polymerase chain reaction were used to examine the molecular and genetic determinants of these uncloned, low-passage-number isolates. All of the isolates were identified as B. burgdorferi by their protein profiles and reactivities with monoclonal and polyclonal antibodies, and all the isolates were typed by the polymerase chain reaction as North American-type spirochetes (B. burgdorferi sensu stricto). Although products of the ospAB locus were identified in protein analyses in all of the isolates, several isolates contained deleted forms of this locus that would result in the expression of chimeric OspA-OspB proteins. The analysis of OspC demonstrated that this protein was widely conserved among the isolates but was also quite variable in its molecular mass and the amount of it that was expressed.

Serologic evidence of Yersinia pestis infection in small mammals and bears from a temperate rainforest of north coastal California.

From 1983 to 1985, 463 serum samples from 11 species of mammals in Redwood National Park (RNP) (California, USA) were evaluated for antibodies to Yersinia pestis by the passive hemagglutination method. Yersinia pestis antibodies occurred in serum samples from 25 (36%) of 69 black bears (Ursus americanus), one (50%) of two raccoons (Procyon lotor), five (3%) of 170 dusky-footed woodrats (Neotoma fuscipes), and one (less than 1%) of 118 deer mice (Peromyscus maniculatus). Two hundred seventy-three flea pools, consisting of 14 species of fleas, were collected from small mammals and woodrat nest cups. Viable Y. pestis were not isolated from any of the flea pools. Significant between-year variations in the frequencies of seropositive bear or small mammal sera were not observed. A significantly higher frequency of plague antibodies was observed in bear sera taken during September collections. Frequencies of seropositive bear sera did not vary significantly by sex or age group of bears. Significant differences were not observed in the frequencies of seropositive small mammals by forest habitat type in which they were captured. This is the first report of Y. pestis infection in Redwood National Park, and the first detailed report of Y. pestis activity in a temperate rainforest.

Human and sylvatic Trypanosoma cruzi infection in California.

In August 1982, a 56-year-old woman from Lake Don Pedro, California, developed acute Chagas' disease (American trypanosomiasis). She had not traveled to areas outside the United States with endemic Chagas' disease, she had never received blood transfusions, and she did not use intravenous drugs. Trypanosoma cruzi cultured from the patient's blood had isoenzyme patterns and growth characteristics similar to T. cruzi belonging to zymodeme Z1. Triatoma protracta (a vector of Trypanosoma cruzi) infected with T. cruzi were found near the patient's home, a trypanosome resembling T. cruzi was cultured from the blood of two of 19 ground squirrels (Spermophilus beecheyi), and six of 10 dogs had antibody to T. cruzi. A serosurvey of three groups of California residents revealed antibody to T. cruzi by complement fixation in six of 237 (2.5 per cent) individuals living near the patient and in 12 of 1,706 (0.7 per cent) individuals living in a community 20 miles northeast of the patient's home, but in only one of 637 (0.2 per cent) blood donors from the San Francisco Bay area. This is the first case of indigenously acquired Chagas' disease reported from California and the first case recognized in the United States since 1955. This investigation suggests that transmission of sylvatic Trypanosoma cruzi infection to humans occurs in California but that Chagas' disease in humans is rare.