Status Report--Retracing the history of the early development of national chronic disease surveillance in Canada and the major role of the Laboratory Centre for Disease Control (LCDC) from 1972 to 2000.

TITRE: Rapport d'étape - Historique des débuts de la surveillance nationale des maladies chroniques au Canada et rôle majeur du Laboratoire de lutte contre la maladie (LLCM) de 1972 à 2000. INTRODUCTION: La surveillance de la santé consiste en l'utilisation systématique et continue de données sur la santé recueillies régulièrement en vue d'orienter les mesures de santé publique en temps opportun. Ce document décrit la création et l'essor des systèmes nationaux de surveillance au Canada et les répercussions de ces systèmes sur la prévention des maladies chroniques et des blessures. En 2008, les auteurs ont commencé à retracer l'historique des débuts de la surveillance nationale des maladies chroniques au Canada, en commençant à 1960, et ils ont poursuivi leur examen jusqu'en 2000. Une publication de 1967 a retracé l'historique de la création du Laboratoire d'hygiène de 1921 à 1967. Notre étude fait suite à cette publication et décrit l'historique de l'établissement de la surveillance nationale des maladies chroniques au Canada, à la fois avant et après la création du Laboratoire de lutte contre la maladie (LCDC).

A reanalysis of cancer mortality in Canadian nuclear workers (1956-1994) based on revised exposure and cohort data.

BACKGROUND: A 15-country study of nuclear workers reported significantly increased radiation-related risks of all cancers excluding leukaemia, with Canadian data a major factor behind the pooled results. We analysed mortality (1956-1994) in the updated Canadian cohort and provided revised risk estimates. METHODS: Employment records were searched to verify and revise exposure data and to restore missing socioeconomic status. Excess relative risks per sievert (ERR/Sv) of recorded radiation dose and 95% confidence intervals (CIs) were estimated using Poisson regression. RESULTS: A significant heterogeneity of the dose-response for solid cancer was identified (P=0.02), with 3088 early (1956-1964) Atomic Energy of Canada Limited (AECL) workers having a significant increase (ERR/Sv=7.87, 95% CI: 1.88, 19.5), and no evidence of radiation risk for 42,228 workers employed by three nuclear power plant companies and post-1964 AECL (ERR/Sv=-1.20, 95% CI: <-1.47, 2.39). Radiation risks of leukaemia were negative in early AECL workers and non-significantly increased in other workers. In analyses with separate terms for tritium and gamma doses, there was no evidence of increased risk from tritium exposure. All workers had mortality lower than the general population. CONCLUSION: Significantly increased risks for early AECL workers are most likely due to incomplete transfer of AECL dose records to the National Dose Registry. Analyses of the remainder of the Canadian nuclear workers (93.2%) provided no evidence of increased risk, but the risk estimate was compatible with estimates that form the basis of radiation protection standards. Study findings suggest that the revised Canadian cohort, with the exclusion of early AECL workers, would likely have an important effect on the 15-country pooled risk estimate of radiation-related risks of all cancer excluding leukaemia by substantially reducing the size of the point estimate and its significance.

Effect of prescribed fire for tick control in California chaparral.

Prescribed fire was investigated as a method for controlling ixodid and argasid ticks in chaparral habitats in northern California. Two experimental and two adjacent control plots within a wildlife preserve were monitored for 1 yr postburn. Ticks were collected by flagging vegetation, by CO2-baited pitfall trap, and by live-trapping rodents. Twice as many rodents were caught at control sites compared with burn sites and no dusky-footed woodrats, Neotoma fuscipes Baird, were found in the treatment sites postburn. This species is known to be a reservoir of the agents of Lyme disease, Borrelia burgdorferi sensu stricto Johnson, Schmid, Hyde, Steigerwalt & Brenner, and human granulocytic anaplasmosis, Anaplasma phagocytophilum Dumler, Barbet, Bekker, Dasch, Palmer, Ray, Rikihisa, Rurangirwa. Six ixodid tick species were removed from rodents (Ixodes pacificus Cooley & Kohls, Ixodes jellisoni Cooley & Kohls, Ixodes spinipalpis Hadwen & Nuttall, Ixodes woodi Bishopp, Dermacentor occidentalis Marx, and Dermacentor parumapertus Neumann), two of which transmit bacterial zoonotic agents to people in the far-western United States. There was no decrease in number of ticks per animal trapped at either burn site compared with controls; in fact, the mean number of immature I. pacificus per rodent was significantly higher at one burn site than its control site. Soil refugia may protect ticks from fire-induced mortality; the argasid tick Ornithodoros coriaceus Koch, which lives in soil, was unaffected by the prescribed fire as were I. pacificus and D. occidentalis buried in packets 2.5 cm below ground. We conclude that although prescribed fires in chaparral habitats may diminish local rodent abundance, it does not decrease tick loads on rodents. Furthermore, burning chaparral does not result in a decreased abundance of adult ixodid ticks on vegetation and apparently does not affect argasid or ixodid ticks that are sheltered within soil refugia.

Ability of transstadially infected Ixodes pacificus (Acari: Ixodidae) to transmit West Nile virus to song sparrows or western fence lizards.

The hypothesis that Ixodes pacificus Cooley & Kohls (Acari: Ixodidae) may serve as a reservoir and vector of West Nile virus (family Flaviviridae, genus Flavivirus, WNV) in California was tested by determining the ability of this tick species to become infected with the NY99 strain of WNV while feeding on viremic song sparrows, to maintain the infection transstadially, and then to transmit WNV to recipient naive song sparrows and western fence lizards during the nymphal stage. The percentage of ticks testing positive by reverse transcription-polymerase chain reaction (RT-PCR) decreased from 77% of 35 larvae at day 6 after ticks were transferred to donor song sparrows (day of detachment) to 23% of 35 nymphs at 59 d postinfestation (approximately 19 d after molting to the nymphal stage). However, the percentage of ticks positive by RT-PCR from which infectious virus was recovered by Vero cell assay decreased from 59% on day 6 to 12% on day 59, even though there was no statistically significant decrease in the quantity of RNA within positive ticks. Attempts to improve the sensitivity of plaque assays by blind passage through C6/36 cell cultures were unsuccessful. These data indicated that ticks maintained viral RNA but not necessarily infectious virus over time. Nymphs from larvae that fed on song sparrows with peak viremias ranging from 7.2 to 8.5 log10 plaque-forming units (PFU) per ml were used in transmission attempts. From one to seven RNA-positive nymphal ticks engorged and detached from each of four recipient song sparrows or western fence lizards. Blood samples from sparrows and lizards remained negative, indicating that transmission did not occur. An additional four lizards inoculated with 1,500 PFU of WNV developed moderate viremias, ranging from 4.2 to 5.6 log10 PFU/ml. Our data and data from previous studies collectively indicated that ixodid ticks were not able to experimentally transmit WNV and therefore most likely would not be important vectors in WNV transmission cycles.

Refractoriness of the western fence lizard (Sceloporus occidentalis) to the Lyme disease group spirochete Borrelia bissettii.

The western fence lizard, Sceloporus occidentalis, is refractory to experimental infection with Borrelia burgdorferi sensu stricto, one of several Lyme disease spirochetes pathogenic for humans. Another member of the Lyme disease spirochete complex, Borrelia bissettii, is distributed widely throughout North America and a similar, if not identical, spirochete has been implicated as a human pathogen in southern Europe. To determine the susceptibility of S. occidentalis to B. bissettii, 6 naïve lizards were exposed to the feeding activities of Ixodes pacificus nymphs experimentally infected with this spirochete. None of the lizards developed spirochetemias detectable by polymerase chain reaction for up to 8 wk post-tick feeding, infected nymphs apparently lost their B. bissettii infections within 1-2 wk after engorgement, and xenodiagnostic L. pacificus larvae that co-fed alongside infected nymphs did not acquire and maintain spirochetes. In contrast, 3 of 4 naïve deer mice (Peromyscus maniculatus) exposed similarly to feeding by 1 or more B. bissettii-infected nymphs developed patent infections within 4 wk. These and previous findings suggest that the complement system of S. occidentalis typically destroys B. burgdorferi sensu lato spirochetes present in tissues of attached and feeding I. pacificus nymphs, thereby potentially reducing the probability of transmission of these bacteria to humans or other animals by the resultant adult ticks.

Sylvatic maintenance of Borrelia burgdorferi (Spirochaetales) in Northern California: untangling the web of transmission.

Lyme borreliosis is associated with several genospecies of Borrelia burgdorferi sensu lato (s.l.) (Spirochaetales), but human disease has been associated only with Borrelia burgdorferi sensu stricto (s.s.) Johnson, Schmid, Hyde, Steigerwalt & Brenner in the western United States. Restriction fragment length polymorphism (RFLP) analysis of rrf-rrl amplicons from 124 tick and mammalian isolates from various habitats yielded 13 RFLP patterns. Of these patterns, six were patterns previously associated either with Borrelia bissettii Postic, Marti Ras, Lane, Hendson & Baranton or Borrelia burgdorferi s.s., and the remaining seven patterns belonged to diverse and previously uncharacterized Borrelia spp. Uncharacterized Borrelia spp. were cultured most frequently from Ixodes spinipalpis Hadwen & Nuttall and California kangaroo rats, Dipodomys californicus Merriam, inhabiting grasslands, and B. bissettii from I. spinipalpis and dusky-footed woodrats, Neotoma fuscipes Baird, associated with oak woodlands or chaparral. B. burgdorferi s.s. typically was isolated from host-seeking Ixodes pacificus Cooley & Kohls collected in dense oak woodlands, woodland-grass, or redwood forests. Although some isolates of B. burgdorferi s.s. were cultured from woodrats, there was no clear association of this human pathogen with any vertebrate host. These findings, along with recent evidence indicating that the western gray squirrel, Sciurus griseus Ord, may be an important reservoir of B. burgdorferi s.s. in Californian oak woodlands, suggest that our earlier hypothesis implicating an enzootic cycle involving woodrats and I. spinipalpis is insufficient to account for observed patterns of infection in nature.

Geographical distribution patterns and habitat suitability models for presence of host-seeking ixodid ticks in dense woodlands of Mendocino County, California.

We used drag sampling to examine the geographical distribution patterns of ixodid ticks engaging in open (non-nidicolous) host-seeking behavior in dense woodland habitats of the climatically and ecologically diverse Mendocino County in north coastal California. The findings based on this sampling methodology reflect risk of human exposure to host-seeking ticks rather than the true distribution of the ticks. Drag sampling in 78 sites yielded 7,860 nymphal or adult Ixodes pacificus Cooley & Kohls, 220 Dermacentor occidentalis Marx, 150 Ixodes spinipalpis Hadwen & Nuttall, 15 Hemaphysalis leporispalustris (Packard), 12 Ixodes angustus Neumann, 12 Ixodes auritulus Neumann, and a single Dermacentor variabilis (Say). I. pacificus, which is the primary vector of the Lyme disease spirochete Borrelia burgdorferi to humans in California, occurred in all 78 sites examined. D. occidentalis, another tick species commonly biting humans in California, and H. leporispalustris typically were encountered in oak-associated woodlands in the central or eastern parts of the county. In contrast, three species of Ixodes ticks (I. angustus, I. auritulus, and I. spinipalpis) most commonly were found questing openly in woodlands with redwood present in the western part of the county. I. angustus and I. spinipalpis are occasional human biters and known experimental vectors of B. burgdorferi. Our study represents the first collection of large numbers of openly host-seeking I. spinipalpis ticks. Univariate tests of associations between presence of ticks (D. occidentalis, H. leporispalustris, I. angustus, I. auritulus, or I. spinipalpis) and environmental geographical information systems-remote sensing (GIS/ RS)-based data indicated that elevation, number of growing degree-days, and tasseled cap brightness, greenness, and wetness are especially useful predictors of presence of openly hostseeking ticks. Combinations of the above-mentioned GIS/RS-based data yielded significant logistic regression models for habitat suitability of host-seeking ticks for all five above-mentioned species. The model equations were used to create spatial surfaces of predicted presence of suitable habitat for openly host-seeking ticks in Mendocino County dense woodlands.

Acarologic risk of exposure to Borrelia burgdorferi spirochaetes: long-term evaluations in north-western California, with implications for Lyme borreliosis risk-assessment models.

Over a 5-year period (1997-2001) the population densities of Ixodes pacificus Cooley & Kohls (Acari: Ixodidae) nymphs infected with spirochaetes of Borrelia burgdorferi sensu lato (s.l.) were evaluated in areas of 2000 ha at two localities (CHR, nine sites; HREC, seven sites) 25 km apart in Mendocino County, north-western California. The 5-year median density of infected nymphs was significantly higher at CHR than at HREC (0.51 vs. 0.09 per 100 m(2) and site-specific yearly densities exceeding one infected nymph per 100 m2 were 10-fold more likely to occur at CHR than at HREC. The importance of long-term data in acarologic risk assessment was demonstrated by significantly higher median yearly densities of infected nymphs at CHR from 1997 to 1999, whereas both areas had similar densities during 2000-2001. Overall, the causative agent of Lyme borreliosis in North America, B. burgdorferi Johnson et al. sensu stricto (s.s.) accounted for 76% of 46 genetically characterized B. burgdorferi s.l. infections from I. pacificus nymphs. Tremendous variability in acarologic risk was recorded within both areas: yearly densities of infected nymphs varied 11-97-fold between sites at CHR and 8-30-fold at HREC. Part of this variation could be explained by environmental traits, most notably deer usage. However, correlations between environmental factors and density of infected nymphs (for CHR and HREC combined) did not necessarily apply when these areas were considered separately. Thus, a Lyme borreliosis ecology model developed in one of these areas needs testing in the other area.

Seasonal activity patterns of Ixodes pacificus nymphs in relation to climatic conditions.

In western North America, the tick Ixodes pacificus Cooley & Kohls (Acari: Ixodidae) is the primary vector to humans and domestic animals of the disease agents causing Lyme disease and granulocytic ehrlichiosis. We examined the seasonal activity patterns of I. pacificus nymphs over a 4-year period, including the wet and cold El Niño winter/spring of 1998, in a dry oak/madrone woodland, and for one year in a cooler and moister redwood/tanoak woodland in Mendocino County, California. Linear regressions were used to estimate when nymphal densities first exceeded and then fell below 25, 50 and 75% of the recorded yearly peak densities. In oak/madrone woodland, nymphs typically were active by mid-March, reached 50% of their yearly peak densities in early to mid-April, peaked by early May, fell below 50% of their peak densities by early to mid-June, and were absent by late July to mid-August. The lengths of the periods with nymphal densities exceeding 50 and 75% of the recorded yearly peaks in oak/madrone woodland were associated positively with rainfall and negatively with maximum air temperatures during April-May. Moreover, nymphal numbers typically reached 50% of their peak 10-15 days later, remained at levels above 50% of the peak 1.3-1.5 times longer, and started declining 4-6 weeks later under cooler, moister climatic conditions (oak/madrone woodland in 1998 and redwood/tanoak woodland in 2000) relative to warmer, drier conditions (oak/madrone woodland in 2000-2001). In oak/madrone woodland, nymphal densities typically started to decline when mean maximum daily air temperatures exceeded 23 degrees C. Nymphal densities were higher in dry oak/madrone relative to moist redwood/tanoak woodland from mid-March to late May 2000, similar in both habitat types in early June, but higher in redwood/tanoak woodland from late June onwards. We conclude that large-scale studies of the density of I. pacificus nymphs in California need to consider spatial variation in the length of nymphal activity periods and select temporal sampling regimens that yield representative data for all included habitat types.

Life cycle of Ixodes pacificus (Acari: Ixodidae): timing of developmental processes under field and laboratory conditions.

The developmental timing of Ixodes pacficus Cooley & Kohls, the primary vector of the Lyme disease spirochete and the agent of human granulocytic ehrlichiosis in the far-western United States, was determined under field and laboratory conditions. During their seasonal peaks of abundance, each of the three parasitic stages of L. pacificus, both fed and unfed, was placed inside silk-screen packets. These packets were apportioned between four topographic exposures of two hilltop sites in northwestern California. The sites differed in vegetational composition and elevation: the first (elevation 390 m) was dominated by woodland-grass, the second (elevation 914 m) by chaparral. The timing of oviposition, larval eclosion, molting, and mortality were recorded in the field every 2-3 wk for 2.5 yr. Microenvironmental temperatures were measured on all four exposures at both sites. Accelerated developmental rates of all three stages were correlated with warmer soil temperatures and the time of placement in the field. In the laboratory, replete female I. pacificus maintained under uniform environmental conditions sustained constant preovipositional and prehatch periods independent of date-of-feeding. In the field, all unfed stages survived through one active feeding season with most larvae and nymphs remaining in behavioral diapause between late summer and early spring. No life stage survived through two active feeding periods which suggests that cohorts do not overlap. We concluded that I. pacificus takes a minimum of 3 yr to complete its life cycle in northwestern California.

Birds and their ticks in northwestern California: minimal contribution to Borrelia burgdorferi enzootiology.

Birds and their attendant ticks were surveyed for infection with the Lyme disease spirochete Borrelia burgdorferi, in chaparral and woodland-grass habitats in northwestern California from March to July, 1998 to 1999. In total, 234 birds were captured and recaptured (15%); nearly 2.5 times more birds were captured in chaparral than in woodland-grass. Overall, 34 species representing 15 families were collected during this study; of these, 24 species were caught in chaparral, 19 in woodland-grass, and 9 in both vegetational types. The most frequently captured birds were sage sparrows (Amphispiza belli) in chaparral, and American robins (Turdus migratorius) and oak titmice (Baelophus inornatus) in woodland-grass. Birds hosted 35 Ixodes pacificus (15 larvae, 20 nymphs) and 9 Haemaphysalis leporispalustris (3 larvae, 5 nymphs, 1 adult) ticks, of which 32 were removed from chaparral birds and 12 from woodland birds. The prevalence of tick infestation was 13% (21/167) in chaparral and 5% (3/67) in woodland-grass, but the relative and mean tick intensities of 0.19 and 1.5 for chaparral birds, and 0.18 and 4.0 for woodland birds, respectively, did not differ significantly by habitat. Spirochetes were not detected in either bird-blood or tick-tissue samples when tested by culture, immunofluorescence, or Giemsa-staining. In contrast, over 90% (86/94) of western fence lizards (Sceloporus occidentalis) collected in June or July were infested with an average of 6.9 and 8.9 immature I. pacificus in chaparral and woodland-grass, respectively. We conclude that birds contribute little to the enzootiology of B. burgdorferi in chaparral and woodland-grass habitats in northwestern California because of their limited parasitism by tick vectors and lack of detectable spirochetemias.

Field trial of systemically delivered arthropod development-inhibitor (fluazuron) used to control woodrat fleas (Siphonaptera: Ceratophyllidae) and ticks (Acari: Ixodidae).

An orally delivered arthropod development-inhibitory (fluazuron) was evaluated for its potential to reduce the number of flea and tick vectors found on the dusky-footed woodrat Neotoma fuscipes Baird, a reservoir host important in disease enzootiology in northern California. Pigmented bait cubes containing fluazuron were distributed monthly to woodrat nests in a chaparral habitat for 1 yr. When compared with control woodrats, the numbers of fleas [primarily Orchopeas sexdentatus (Baker)] on treated woodrats were significantly reduced 3-4 mo after initial application, and remained so for the duration of the application period. By contrast, tick numbers were not significantly reduced on treated woodrats. After the cessation of treatments, flea indices remained lower on treated animals for up to 2 mo after application. Approximately 93% of woodrats captured in the treatment area excreted pigmented feces and 93% of distributed bait cubes were removed by woodrats, which indicates that the bait cube formulation and delivery system were highly effective. Bait cubes also were attractive to small rodents and ground-frequenting birds. The results of this study suggest that a monthly application program of fluazuron delivered by bait cube is effective in reducing woodrat flea-burdens, but is not effective, at least in the short-term, in controlling ticks.

Acarologic risk of exposure to emerging tick-borne bacterial pathogens in a semirural community in northern California.

An acarologic study was conducted in a semirural community in northern California to determine the relative abundance of, and the prevalence of infection with, three emerging bacterial pathogens in the western black-legged tick (Ixodes pacificus). These included the agents causing Lyme disease (Borrelia burgdorferi), human granulocytic ehrlichiosis [Ehrlichia phagocytophila (formerly Ehrlichia equi)], and human monocytic ehrlichiosis (Ehrlichia chaffeensis). The study area in Sonoma County consisted of two properties each with four residents and an uninhabited adjacent comparison area. Six of the eight residents had been either physician-diagnosed or serodiagnosed previously with Lyme disease, and, of these, one also had been serodiagnosed with human monocytic ehrlichiosis. Direct immunofluorescent/culture assays and bacterial species-specific polymerase chain reaction assays were used to test whole ticks individually for presence of B. burgdorferi and Ehrlichia spp., respectively. Overall, 6.5% of the nymphal (n = 589) and 1.6% of the adult ticks (n = 318) from the same generational cohort were found to contain B. burgdorferi. In contrast, none of 465 nymphs and 9.9% of 202 adults were infected with E. phagocytophila. Excised tissues from another 95 adult ticks yielded a comparable E. phagocytophila infection prevalence of 13.7%. E. chaffeensis was not detected in either nymphal or adult ticks. Using a combination of culture and polymerase chain reaction assays, coinfection of I. pacificus adults with B. burgdorferi and E. phagocytophila was demonstrated for the first time. The marked disparity in the infection prevalence of these pathogens in nymphal and adult ticks suggests that their maintenance cycles are inherently different.

Nymphs of the western black-legged tick (Ixodes pacificus) collected from tree trunks in woodland-grass habitat.

Nymphs of the western black-legged tick, Ixodes pacificus, were found on the trunks of trees during spring and summer in northwestern California. In a woodland-grass habitat, large- and medium-sized (> 130 cm and 80-130 cm in circumference, respectively), moss-covered oak (Quercus spp.) trees supported ticks significantly more often than trees without these characteristics. Additionally, trees with basal leaf-litter and lacking shade (at time of sampling) were significantly associated with the presence of ticks. Mean tick-burdens were > 1 for all oaks (1.06), all trees with basal leaf-litter (1.05), and all trees of large-circumference (1.19); 0.79 ticks per tree were collected over the entire study. Moss reduced the surface temperature of trees by a mean of 1.9 degrees C (range of 1.6-5.0 degrees C) and increased relative humidity by up to 2.5% from the ambient. These microclimatic changes, along with the presence of refugia in bark and western fence lizard (Sceloporus occidentalis) hosts on the lower-most surface of trees, likely accounted for ticks questing on the trunks. Although of undetermined epidemiological significance, the presence of host-seeking I. pacificus nymphs on tree trunks may shed light on the relation of abiotic and biotic factors to the life history of this important vector of disease.

Prevalence and abundance of Ixodes pacificus immatures (Acari: Ixodidae) infesting western fence lizards (Sceloporus occidentalis) in northern California: temporal trends and environmental correlates.

The prevalence and abundance of immature Ixodes pacificus ticks on western fence lizards (Sceloporus occidentalis) were examined in relation to time of year, host attributes (i.e., age, gender, and presence or absence of blood parasites), and 5 environmental characteristics, including topographic exposure and ground cover substrate, over a 2-year period in northern California. Lizards were infested with subadult ticks from early March until late July or early August, with peak median numbers of larvae and nymphs recorded in late April and early May of both years. Peak larval and nymphal abundances differed between years. The overall ratio of larvae to nymphs on adult male lizards was low, ranging from 0.80 in 1999 to 2.41 in 2000. Such intensive feeding of nymphs versus larvae on these lizards, which are reservoir-incompetent for Borrelia burgdorferi spirochetes, may explain previous observations of decreasing spirochetal infection prevalence from the nymphal to adult stage in northwestern California. Adult male lizards were more likely to be infested with nymphs and harbored greater abundances of larvae and nymphs than adult females. Lizards uninfected with blood parasites had more nymphs than infected lizards. The measured environmental characteristics could explain only a small percentage of the total variation observed in larval prevalence (22%) and in larval and nymphal abundance (12 and 3%, respectively).

Density and spatial distribution of Ixodes pacificus (Acari: Ixodidae) in two recreational areas in north coastal California.

The density and distribution of Ixodes pacificus was assessed at 2 parks in north coastal California. The density of I. pacificus adults and nymphs varied significantly between years, trails, and sides of trails. Adult ticks occurred on vegetation along sun-exposed trails in January through March, their density (0-1.93 per 20 m) correlated with brush density, trail width, and presence of an uphill slope. Nymphs (0.06-5.10 per 20 m) occurred in leaf litter along shaded trails in May-July. Adult I. pacificus were rare at picnic sites (0.00-0.24 per 20 m), but nymphal densities (0.93-2.37 per 20 m) were comparable with those along some shaded trails. The prevalence of Borrelia burgdorferi in ticks (2.8% overall) did not differ significantly between locations, years, or stages. We conclude that the risk of acquiring Lyme disease in these sites is low, but varies among trails, seasons, and years.

Genetic characteristics of Borrelia coriaceae isolates from the soft tick Ornithodoros coriaceus (Acari: Argasidae).

Two Borrelia isolates (CA434 and CA435) cultured from the soft tick Ornithodoros coriaceus were analyzed by contour-clamped homogeneous electric field gel electrophoresis of unrestricted and ApaI-restricted DNA, standard electrophoresis of BamHI- and HindIII-restricted DNA, Southern hybridization, restriction fragment length polymorphism and sequencing of the 16S rRNA gene, and amplification of the 5S-23S intergenic spacer region. These isolates were compared with Borrelia coriaceae type strain Co53, B. burgdorferi sensu stricto strain CA4, and the relapsing-fever spirochete B. parkeri (undesignated). The 16S rRNA region of CA434 and CA435 differed from that of B. coriaceae type strain Co53 by the presence of 1 base (C) at position 367 (GenBank accession no. U42286). The linear plasmid profile of CA434 was similar to that of Co53, and the ApaI, BamHI, and HindIII restriction fingerprints of the total cellular DNA of CA434 and Co53 were similar. In contrast, CA435 differed somewhat from CA434 and Co53, which demonstrates that B. coriaceae is genetically diverse. Southern hybridization showed that the DNAs of CA434 and CA435 hybridized strongly with the digoxigenin-labeled DNA of Co53. Low homology was found between the DNA of Co53 and that of B. parkeri. The 16S rRNA sequence of B. parkeri was identical to previously published results for B. parkeri strain M3001 (GenBank accession number U42296). CA434 and CA435 represent only the second and third isolates of B. coriaceae obtained from any source since its initial isolation from an O. coriaceus tick in 1985. All three B. coriaceae isolates were derived from adult ticks collected from the same locality in northwestern California. Difficulties encountered in detecting B. coriaceae in, and isolating this spirochete from, the tissues of O. coriaceus are discussed. The lack of concordance between different detection or isolation methods suggests that reliance upon a single technique may grossly underestimate the true prevalence of spirochetal infection in wild-caught O. coriaceus ticks.