Search PubMed⌕ Search

Biomedical subjects

R S Lane

Publications and source records attributed to R S Lane.

At least 37 records · Page 2Linked to original sources

Common ancestry of Borrelia burgdorferi sensu lato strains from North America and Europe.

Ten atypical European Borrelia burgdorferi sensu lato (Borrelia spp. ) strains were genetically characterized, and the diversity was compared to that encountered among related Borrelia spp. from North America. Phylogenetic analyses of a limited region of the genome and of the whole genome extend existing knowledge about borrelial diversity reported earlier in Europe and the United States. Our results accord with the evidence that North American and European strains may have a common ancestry.

Borrelia burgdorferi Group↗

Anti-arthropod saliva antibodies among residents of a community at high risk for Lyme disease in California.

The role of the western black-legged tick (Ixodes pacificus) versus that of other potential arthropod vectors in the epidemiology of Lyme disease was evaluated by determining the prevalence of anti-arthropod saliva antibodies (AASA) among residents (n = 104) of a community at high-risk (CHR). Salivary gland extracts prepared from I. pacificus, the Pacific Coast tick (Dermacentor occidentalis), the western cone-nose bug (Triatoma protracta), and the western tree-hole mosquito (Aedes sierrensis) were used as antigens in an ELISA. Sera from 50 residents of the San Francisco Bay region in northern California and 51 residents of Imperial County in southern California served as comparison groups. The prevalence of AASA ranged from 2% for A. sierrensis to 79% for I. pacificus in study subjects, 0% for D. occidentalis to 36% for I. pacificus among residents of the San Francisco Bay region, and 6% for I. pacificus to 24% for A. sierrensis in residents of Imperial County. The associations between AASA and demographic factors, potential risk factors, probable Lyme disease, and seropositivity for Borrelia burgdorferi were assessed for 85 members of the CHR. Seropositivity for I. pacificus and B. burgdorferi were significantly correlated, the relative risk of seropositivity to B. burgdorferi was about 5 (31% versus 6%) for subjects who were seroreactive to I. pacificus, nearly every individual who was seropositive for B. burgdorferi had elevated levels of antibodies to I. pacificus, and the mean titer for antibodies to I. pacificus was significantly higher for subjects seropositive versus those seronegative for B. burgdorferi. Together, these findings support the widely held belief that I. pacificus is the primary vector of B. burgdorferi for humans in northern California, and they demonstrate the utility of the AASA method as an epidemiologic tool for studying emerging tick-borne infections.

Adolescent↗

Variation in the density of questing Ixodes pacificus (Acari:Ixodidae) nymphs infected with Borrelia burgdorferi at different spatial scales in California.

The density of, and prevalence of infection with Borrelia burgdorferi in, Ixodes pacificus nymphs as well as the density of infected nymphs were compared at 12 properties at a small rural community at high risk for Lyme disease (CHR) and at 12 areas at the University of California Hopland Research and Extension Center (HREC), Mendocino County, California. The mean infection prevalence and density of infected nymphs were 1.7% (range, 0-4.2%) and 0.10 infected nymphs per 100 m2 (range, 0-0.23 per 100 m2) at the HREC, and 12.4% (range, 3.9-41.3%) and 1.83 infected nymphs per 100 m2 (range, 0.29-22.17 per 100 m2) at the CHR. Thus, the mean density of infected nymphs differed 18-fold between CHR and HREC and 76-fold between properties at the CHR. Also, there was up to 10-fold variation in infection prevalence and 16-fold variation in density of infected nymphs between discrete areas within properties at the CHR. The high densities of infected nymphs recorded at the CHR suggest that, despite the low statewide incidence of Lyme disease, the medical community should be alerted that Lyme disease can be highly endemic in rural areas of northwestern California. The prevalence of spirochetal infection was higher for nymphs collected in southern/western, as compared to northern/eastern, exposures at both HREC and CHR. Infection prevalence and nymphal density were negatively associated at the HREC, whereas they tended to be associated positively at the CHR. A positive association was observed between nymphal density and density of infected nymphs when data from CHR and HREC were combined, and when data from the CHR were considered alone, but not for data from the HREC alone.

Animals↗

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.

Animals↗

First isolation and cultivation of Borrelia burgdorferi sensu lato from Missouri.

Five Borrelia burgdorferi sensu lato isolates from Missouri are described. This represents the first report and characterization of such isolates from that state. The isolates were obtained from either Ixodes dentatus or Amblyomma americanum ticks that had been feeding on cottontail rabbits (Sylvilagus floridanus) from a farm in Bollinger County, Mo., where a human case of Lyme disease had been reported. All isolates were screened immunologically by indirect immunofluorescence by using monoclonal antibodies to B. burgdorferi-specific outer surface protein A (OspA) (antibodies H3TS and H5332), B. burgdorferi-specific OspB (antibody H6831), Borrelia (genus)-specific antiflagellin (antibody H9724), and Borrelia hermsii-specific antibody (antibody H9826). Analysis of the isolates also involved a comparison of their protein profiles by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Finally, the isolates were analyzed by PCR with six pairs of primers known to amplify selected DNA target sequences specifically found in the reference strain B. burgdorferi B-31. Although some genetic variability was detected among the five isolates as well as between them and the B-31 strain, enough similarities were found to classify them as B. burgdorferi sensu lato.

Animals↗

Expanded diversity among Californian borrelia isolates and description of Borrelia bissettii sp. nov. (formerly Borrelia group DN127).

Up to now, the only species in the complex Borrelia burgdorferi sensu lato known to cause Lyme borreliosis in the United States has been B. burgdorferi sensu stricto. However, some atypical strains closely related to the previously designated genomic group DN127 have been isolated in the United States, mostly in California. To explore the diversity of B. burgdorferi sensu lato group DN127, we analyzed the nucleotide sequences of the rrf-rrl intergenic spacer regions from 19 atypical strains (18 from California and one from New York) and 13 North American B. burgdorferi sensu stricto strains (6 from California). The spacer region sequences from the entire B. burgdorferi sensu lato complex available in data banks were used for comparison. Phylogenetic analysis of sequences shows that the main species of the B. burgdorferi sensu lato complex (B. afzelii, B. garinii, B. andersonii, B. japonica, B. burgdorferi sensu stricto, B. valaisiana, and B. lusitaniae) each form a coherent cluster. A heterogeneous group comprising strains belonging to the previously designated group DN127 clustered separately from B. burgdorferi sensu stricto. Within this cluster, the deep branches expressing the distances between the rrf-rrl sequences reflect a high level of divergence. This unexpected diversity contrasts with the monomorphism exhibited by B. burgdorferi sensu stricto. To clarify the taxonomic status of this highly heterogeneous group, analysis of the rrs sequences of selected strains chosen from deeply separated branches was performed. The results show that these strains significantly diverge at a level that is compatible with several distinct genomic groups. We conclude that the taxonomy and phylogeny of North American B. burgdorferi sensu lato should be reevaluated. For now, we propose that the genomic group DN127 should be referred to as a new species, B. bissettii sp. nov., and that other related but distinct strains, which require further characterization, be referred to as Borrelia spp.

Borrelia↗

Borreliacidal factor in the blood of the western fence lizard (Sceloporus occidentalis).

In some populations of the western black-legged tick, Ixodes pacificus, the prevalence of infection with Lyme disease spirochetes (Borrelia burgdorferi) in nymphal ticks exceeds those in adult ticks by 3-4-fold. Experiments were conducted to determine if the reduced spirochetal prevalence in adult ticks is due to the presence of anti-borrelial antibodies or to another borreliacidal factor in the blood of the western fence lizard, Sceloporus occidentalis, a primary host of subadult I. pacificus, or to loss of spirochetes as nymphal ticks molt to the adult stage. Ten lizards were each exposed to the feeding activities of 10 nymphs having a 78% prevalence of B. burgdorferi infection. Five of the lizards had been hyperimmunized first with 10(8) heat-killed spirochetes and 5 were seronegative to B. burgdorferi. After repletion and the transstadial molt, none of 62 resultant adult ticks from both groups of lizards was found to contain spirochetes. In contrast, 11 of 20 (55%) infected nymphs that had fed on 4 preimmune rabbits passed spirochetes to adult ticks. Taken together, these findings demonstrate that host immunoglobulins and the transstadial molt by themselves are not necessary for eliminating B. burgdorferi from infected nymphal ticks. A novel in vitro assay revealed that nearly all spirochetes placed in plasma or sera from lizards died in less than 1 hr, whereas many spirochetes injected into mouse plasma or sera survived for 72 hr. When spirochetes were put in lizard sera that had been preheated (100 C for 10 min) and allowed to cool, survival was extended to 72 hr. We conclude that the blood of S. occidentalis contains a thermolabile, borreliacidal factor, probably a protein, that destroys spirochetes in the midgut diverticula of feeding I. pacificus nymphs.

Animals↗

Role of small mammals in the ecology of Borrelia burgdorferi in a peri-urban park in north coastal California.

The role of small mammals other than woodrats in the enzootiology of the Lyme disease spirochete, Borrelia burgorferi, was assessed in the peri-urban park. Mammals were collected monthly from September through to April. Following tick removal, the animals were tested for B. burgdorferi by culture of ear-punch biopsies. Larvae and nymphs that were intermediate in morphology between Ixodes spinipalpis and Ixodes neotomae occurred on several species of rodents (Peromyscus truei, Peromyscus californicus, Microtus californicus, Rattus rattus and Reithrodontomys megalotis) and the brush rabbit (Sylvilagus bachmani). Morphometric analyses of these I. spinipalpis-like ticks and the offspring from two I. neotomae females from the site suggest that I. neotomae may bo conspecific with I. spinipalpis. Borrelia burgdorferi was isolated from eight out of 109 (7.3%), three out of 16 (18.8%), two out of 38 (5.3%) and two out of six (33.3%) P. truei, P. maniculatus, M. californicus and R. rattus, respectively. One bush rabbit yielded the first isolate of B. burgdorferi from a lagomorph in western North America. This isolate and three others derived from unfed I. spinipalpis-like nymphs failed to produce infection when inoculated intradermally into 11-12 P. maniculatus each. Likewise, no spirochetes were detected in 420 Ixodes pacificus nymphs derived from larvae fed on animals inoculated with these isolates. An additional isolate, derived from an I. spinipalpis-like nymph, was recovered by ear-punch biopsies from five our of 12 (42%) needle-inoculated P. maniculatus. However, spirochetes were not detected in 20 I. pacificus nymphs fed as larvae on each of five mice (two infected and three uninfected) inoculated with this isolate. We conclude that brush rabbits and several species of rodents besides woodrats may contribute to the maintenance of B. burgdorferi because they harbour the spirochete and are fed upon by competent enzootic vectors.

Animals↗

Genetic heterogeneity of Borrelia burgdorferi in the United States.

To examine in detail Borrelia burgdorferi strain diversity in the United States, 186 isolates from human, tick, and rodent sources were analyzed from multiple distinct geographic regions of the United States and abroad. Strains were characterized by genomic macrorestriction analysis and ospA and 23S rDNA gene sequencing followed by phylogenetic analysis. Results indicate that spirochetal isolates from the United States fall into two major divisions and nine or more subdivisions; human isolates fell into five of these subdivisions. Greater genetic diversity was observed among B. burgdorferi isolates from moderate climatic regions, consistent with increased tick vector and reservoir diversity. All of the Borrelia isolates were reactive by ospA polymerase chain reaction except for Borrelia hermsii controls and several tick isolates from the Northeast, which were shown to lack the 49-kb plasmid encoding outer surface protein A (OspA). The data suggest that US B. burgdorferi isolates demonstrate substantial genetic heterogeneity, with regional differences in spirochete populations.

Animals↗

Population genetic structure of Ixodes pacificus (Acari:Ixodidae) using allozymes.

Genetic analysis of the population structure of the western blacklegged tick, Ixodes pacificus Cooley & Kohls, was conducted using allozymes. This vector tick transmits the Lyme disease spirochete, Borrelia burgdorferi Johnson, Schmid, Hyde, Steigerwalt & Brenner, in the far-western United States. It ranges from British Columbia to Baja California and disjunct populations are present in Oregon, Nevada, Utah, and Arizona. Host-seeking adult ticks were collected from vegetation across the range of the species and were partially fed on rabbits prior to analysis. Twelve putative loci were resolved using starch gel electrophoresis. One locus, glucose-6-phosphate isomerase, formed an apparent north/south latitudinal cline and showed significant geographic structure. None of the remaining loci exhibited much genetic differentiation. Estimates of gene flow were high relative to other arthropods. Isolation-by-distance analysis suggests a recent and rapid range expansion. We conclude that the overall lack of differentiation is due high rates of gene flow.

Alleles↗

Taxonomic status of Ixodes neotomae and I. spinipalpis (Acari: Ixodidae) based on mitochondrial DNA evidence.

Ixodes spinipalpis Hadwen & Nuttall and I. neotomae Cooley are enzootic vectors of Borrelia burgdorferi Johnson, Schmid, Hyde, Steigerwalt & Brenner in western North America. The taxa overlap in host preference, habitat, and morphology. Mitochondrial DNA was compared between the taxa to test for reproductive isolation. A 300-bp region of the mitochondrial 16S ribosomal DNA gene was amplified in 28 specimens of I. neotomae and 149 specimens of I. spinipalpis. These products were screened for sequence variation using single-strand conformation polymorphism analysis, and 9 haplotypes were detected. Haplotype frequencies varied between taxa; however, Shannon diversity analysis indicated that most variation arose among collections within each taxon, and no unique haplotypes characterized either one. Phylogenetic analysis of 18 sequences, representing a replicate of each of the 9 haplotypes, was performed with Ixodes pacificus Cooley & Kohls and Ixodes jellisoni Cooley & Kohls as outgroups. Strong monophyletic support was found for a clade containing I. neotomae and I spinipalpis and within this clade no lineages comprised exclusively either taxon. These results argue against treatment of I. neotomae and I. spinipalpis as distinct species.

Animals↗

Emerging bacterial zoonotic and vector-borne diseases. Ecological and epidemiological factors.

Among the etiologic agents of emerging infectious diseases are several bacterial organisms that naturally reside in animal and arthropod hosts. The most compelling emerging bacterial zoonotic and vector-borne diseases in the United States are Lyme disease; a Southern erythema migrans-like illness; human monocytic ehrlichiosis; human granulocytic ehrlichiosis; a novel cat flea-associated typhus group rickettsiosis; bartonelloses of immunocompetent and immunocompromised persons, particularly with AIDS; and sylvatic plague. Some of these antimicrobial-treatable infections are life threatening. During the acute stage of illness when antimicrobial agents are most effective, the flulike clinical signs and symptoms and available laboratory tests frequently do not point to a particular diagnosis. Epidemiological factors determined by the ecology of the bacteria are often the most useful diagnostic clues. The recognition of these evolving problems emphasizes the need for development of better laboratory diagnostic methods, for surveillance for and tracking of disease, and for continued research into factors contributing to transmission of the organisms. The continual appearance of previously unidentified bacterial infections requires prospective national strategies for timely recognition of the syndrome, identification of the agent, establishment of criteria and methods for diagnosis, optimization of the treatment regimen, and determination of successful approaches to prevention and control.

Animals↗

Field and laboratory studies on the timing of oviposition and hatching of the western black-legged tick, Ixodes pacificus (Acari: Ixodidae).

The timing of oviposition and hatching of Ixodes pacificus was investigated in the field and at constant temperatures in the laboratory. Replete females held at temperatures between 9 and 29 degrees C began depositing eggs a mean of 9-70 days after drop off. Egg masses held between 12 and 25 degrees C commenced hatching 25-178 days after the onset of oviposition. Eggs held at 9 or 29 degrees C did not hatch. The lower temperature thresholds for development (LTD) for oviposition and hatching were 6.5 and 9 degrees C, respectively. The number of degree days required for oviposition and hatching was 173 and 588, respectively. Replete females placed in the field on 2 December through to 8 March deposited eggs from 2 February through to 24 April; the eggs commenced hatching between 2 July and 21 August. Unfed larvae from two of 20 egg masses survived through the winter and fed readily when exposed to deer mice (Peromyscus maniculatus) on 22 April. Replete larvae were returned to the field and moulted between 9 and 21 August. Larvae exposed to deer mice in August, 4 weeks after hatching, also fed readily. Although further studies are needed to clarify the timing of nymphal development, the present study suggests that I. pacificus requires more than 1 year to complete its life cycle.

Animals↗

Ixodes (Ixodes) jellisoni and I. (I.) neotomae (Acari:Ixodidae): descriptions of the immature stages from California.

Nymphal and larval stages of Ixodes (Ixodes) jellisoni Cooley & Kohls and I. (I.) neotomae Cooley are described for the first time. These 2 tick species occur only in the western United States, predominantly in California. The primary host for I. jellisoni is the California kangaroo rat, Dipodomys californicus (Merriam); that for I. neotomae is the dusky-footed woodrat, Neotoma fuscipes Baird. The etiologic agent of Lyme disease Borrelia burgdorferi Johnson, Schmidt, Hyde, Steigerwalt & Brenner has recently been isolated from both tick species, and I. neotomae was proven a competent enzootic vector of the Lyme disease spirochete.

Animals↗

Evaluation of permethrin-impregnated cotton balls as potential nesting material to control ectoparasites of woodrats in California.

The dusky-footed woodrat, Neotoma fuscipes Baird is a natural reservoir of the Lyme disease spirochete, Borrelia burgdorferi Johnson, Schmid, Hyde, Steigerwalt & Brenner, in California. To investigate the potential of host-targeted insecticide to control the tick vectors of B. burgdorferi, permethrin-impregnated or untreated cotton balls were distributed in metal cylinders as potential nesting material adjacent to 95 woodrat houses in chaparral-covered rangeland. Laboratory experiments demonstrated that adult woodrats would enter the cylinders and construct nests from permethrin-treated or untreated cotton. The residual concentration of permethrin did not vary significantly during an 11-mo period and remained > 60% of the registered insecticidal formulation (7.5% [AI] by cotton weight). The abundance of 4 species of ticks (Ixodes neotomae Cooley; the western blacklegged tick I. pacificus Cooley & Kohls; I. woodi Bishopp; and the Pacific Coast tick, Dermacentor occidentalis Marx) infesting woodrats was similar in the treatment and control areas. Although > 90% of the cotton disappeared from the metal cylinders in both areas, examination of 8 active woodrat houses revealed that small amounts of cotton had been incorporated into the nest cups of only 25%. In contrast, the abundance of the flea Orchopeas sexdentatus (Baker) decreased significantly in the treatment area only. Spirochetes were not detected in 168 adult O. sexdentatus fleas that had fed on spirochetemic woodrats, which demonstrates that this flea is an inefficient host of B. burgdorferi. We conclude that the use of permethrin-impregnated cotton as potential nesting material is ineffective for controlling ticks associated with the dusky-footed woodrat in brushlands, but this methodology may be useful for reducing populations of sylvatic fleas.

Animals↗

Comparison of infectivities of six tick-derived isolates of Borrelia burgdorferi for rodents and ticks.

The infectivity and dissemination to the skin of six isolates of Borrelia burgdorferi were evaluated by inoculating them into groups of deer mice (Peromyscus maniculatus), hamsters, and Swiss Webster mice. Rodent infection was assayed by culture of ear punch biopsy specimens taken at 4, 8, and 12 weeks postinoculation (p.i.). Spirochetes were detected in biopsy specimens from individuals of all three host species that had been inoculated with four isolates (CA3, CA4, CA7, and CA8). Ear punch biopsy specimens taken from Swiss Webster mice at 12 weeks p.i. yielded an additional reisolate (CA2), even though these animals did not seroconvert. The remaining isolate (CA9) was not recovered from any host. However, two deer mice and all hamsters and Swiss Webster mice inoculated with CA9 seroconverted. All six isolates were of low infectivity to ticks when inoculated intramuscularly into hosts. Only 4 (1.6%) of 250 Ixodes pacificus larvae acquired and transstadially maintained infection from hosts inoculated intramuscularly. Infectivity of three isolates for ticks also was tested in Swiss Webster mice injected intradermally. The mean prevalences of infection in xenodiagnostic ticks fed on these mice at 4 weeks p.i. were 47.9, 1.2, and 2.2% for isolates CA4, CA7, and CA8, respectively. The mean prevalences of infection for ticks fed on the same mice at 12 weeks p.i. were 36.4, 11.8, and 20.4%, respectively. Such differences in the infectivity and rate of dissemination of individual isolates of B. burgdorferi should be considered during studies of reservoir and vector competence.

Animals↗

Reservoir competence of four chaparral-dwelling rodents for Borrelia burgdorferi in California.

Aspects of the reservoir competence of four rodents for the Lyme disease spirochete, Borrelia burgdorferi, were evaluated in California. Rodents were live-trapped and ear-punch biopsies were cultured during each season. A second set of biopsies was cultured from representative individuals after 2-3 weeks of captivity and the results of culturing biopsies taken on both dates were compared with the results of feeding Ixodes pacificus larvae on hosts xenodiagnostically. The prevalence of infections did not differ significantly between dusky-footed woodrats (Neotoma fuscipes) and California kangaroo rats (Dipodomys californicus) nor among seasons. Combined results of the three tests showed that 85.7% of dusky-footed woodrats (n = 21) and 78.6% of California kangaroo rats (n = 14) were infected with B. burgdorferi. In contrast, only 22.2% of brush mice (Peromyscus boylii) (n = 14) and 7.1% of pinyon mice (P. truei) (n = 9) were infected. The sensitivity of culturing ear-punch biopsies as an assay for borrelial infection was significantly greater when biopsies were taken after a short period of captivity (0.89) rather than on the day of capture (0.52). Tick xenodiagnosis, in which I. pacificus was used as the vector, revealed borrelial infections in 90.3% of infected rodents. Spirochetes were observed in 37.7% of 239, 45.2% of 155, 60.0% of 10, and 7.1% of 14 cultures of nymphal I. pacificus fed as larvae on naturally infected woodrats, kangaroo rats, brush mice, and a pinyon mouse, respectively. The mean prevalence of infection in xenodiagnostic ticks varied significantly among host species with a greater proportion of ticks infected while feeding on woodrats and kangaroo rats than on mice. This study reconfirms previous reports that implicated woodrats and kangaroo rats as reservoirs of B. burgdorferi in California.

Animals↗