Biomedical subjects
R S Lane
Publications and source records attributed to R S Lane.
IgE antibodies in tick bite-induced anaphylaxis.
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Wood rats and kangaroo rats: potential reservoirs of the Lyme disease spirochete in California.
The etiologic agent of Lyme disease, Borrelia burgdorferi Johnson, Schmid, Hyde, Steigerwalt & Brenner, was isolated repeatedly from dusky-footed wood rats, Neotoma fuscipes Baird, and California kangaroo rats, Dipodomys californicus Merriam, in northern California. All animals were collected in a region endemic for Lyme disease but for which the natural reservoir of B. burgdorferi was unknown. Similar attempts to isolate spirochetes from lizards, other species of rodents, jack rabbits, and deer between 1987 and 1991 were unsuccessful. Spirochetes isolated from wood rats and kangaroo rats were antigenically similar to strains of B. burgdorferi that had been isolated previously from the western black-legged tick, Ixodes pacificus Cooley & Kohls, in California. Similar enzootic cycles involving wood rats or kangaroo rats should be sought in other regions of the United States where the reservoirs of this spirochete are unknown.
Lyme disease in California: interrelationship of ixodid ticks (Acari), rodents, and Borrelia burgdorferi.
The association of immature ixodid ticks, several species of rodents, and the Lyme disease spirochete, Borrelia burgdorferi Johnson, Schmid, Hyde, Steigerwalt & Brenner, was studied in two habitats in northern California in spring and summer 1985 and year-round in 1986. A total of 428 rodents were collected from ecotonal chaparral and a woodland-grass-rock outcrop; the former habitat yielded six species, the latter three species. The deer mouse, Peromyscus maniculatus (Wagner), and the piñon mouse, P. truei (Shufeldt), were the dominant species year-round and collectively comprised 78% of rodents captured within chaparral and 87% from the rock outcrop in 1986. In both habitats, rodents were trapped most frequently in winter and spring, and least often in summer and fall. A total of 306 rodent blood films from all six species were assayed for spirochetes by direct immunofluorescence; of these, only one film prepared from P. truei (n = 123 films from 53 individual mice) was found to contain spirochetes. Immature western black-legged ticks, Ixodes pacificus Cooley & Kohls, and Pacific Coast ticks, Dermacentor occidentalis Marx, were collected from each species of rodent. Larvae of I. pacificus infested P. maniculatus and P. truei in low numbers year-round, but nymphs of this tick rarely parasitized these rodents. D. occidentalis larvae infested P. maniculatus and P. truei in spring and particularly in summer; nymphal ticks infested these mice primarily in summer. The efficiency of visual inspection for collecting immatures of these ticks from P. maniculatus ranged from 45 to 69% in spring and summer, whereas the efficiency of a drop-off technique appeared to be 100%. Spirochetes were detected in <1% of D. occidentalis larvae (n = 310) and nymphs (n = 120), and in approximately 4% of I. pacificus larvae (n = 75) derived from these hosts. The potential significance od these findings in the enzootiology of B. burgdorferi is discussed.
Lyme borreliosis: relation of its causative agent to its vectors and hosts in North America and Europe.
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Relationship of Borrelia burgdorferi to its arthropod vectors.
At the IV International Conference on Lyme Borreliosis, a workshop was held to identify the unique development of the Lyme disease spirochete, Borrelia burgdorferi, in its established and suspected arthropod vectors. The following is a summary of the panel's discussions of research aspects concerning relationship(s) of this borrelia to its vectors, and the mode(s) of its transmission to animal hosts.
Clearance of Rh D-positive red cells with monoclonal anti-D.
Two human monoclonal antibodies, one IgG3 and one IgG1, with anti-Rh D specificity, were tested for their ability to clear red cells. Samples of red cells from 12 D-positive subjects were sensitised in vitro with various amounts of antibody, the number of antibody molecules bound to the cells was estimated, and the cells were reinjected into the donor's circulation. Both antibodies mediated clearance but substantially fewer IgG3 than IgG1 antibody molecules were required to produce a given rate of clearance. The IgG3 antibody was slightly more effective than polyclonal anti-D, as judged by comparison with previously published results. Since there is believed to be an association between the rate of red cell clearance by anti-D and the ability of the antibody to suppress D immunisation, the IgG3 monoclonal antibody may prove suitable for the immunoprophylaxis of Rh D haemolytic disease.
Host-seeking behavior of adult Ixodes pacificus (Acari: Ixodidae) as determined by flagging vegetation.
Diurnal host-seeking by adults of the western black-legged tick, Ixodes pacificus Cooley & Kohls in relation to vegetative habitat type and meterological factors was investigated at a coastal and an inland site in northern California. Host-seeking behavior and relative tick abundance were determined with a tick drag method from late fall to midwinter 1987-1988. At the inland site, tick abundance usually was significantly greater in chaparral-grassland ecotones than in adjoining dense chaparral on the south-facing slope of a mountaintop, whereas both of these vegetative types produced significantly fewer ticks on a north slope compared with a contiguous south-facing slope. There was no evidence for an association between tick abundance and plant species within ecotonal chaparral. Multiple regression analyses revealed that tick abundance in ecotonal chaparral at the inland site and in grassland at the coastal site was not associated consistently with either ambient temperature or relative humidity. Compared with the inland site, ticks were considerably more abundant at the coastal site where sampling occasionally yielded more than 100 ticks per 100 drag samples. The incidence of infection with Borrelia burgdorferi Johnson, Schmid, Hyde, Steigerwalt & Brenner, the etiologic agent of Lyme borreliosis, in adult ticks collected in the morning (1.5%) versus afternoon (2.2%), and in male (1.3%) versus female ticks (2.5%), at the coastal site in winter was similar.
Survey of birds and lizards for ixodid ticks (Acari) and spirochetal infection in northern California.
A total of 138 birds (24 species) was captured in an oak woodland between December 1988 and June 1989 at the University of California, Sierra Foothill Range Field Station, Yuba County, Calif. Ticks were not found on 71 birds captured between December 1988 and March 1989. Five subadult Ixodes pacificus Cooley & Kohls were removed from 3 of 67 birds caught between April and June 1989. These three birds, an orange-crowned warbler (Vermivora celata (Say], a lazuli bunting (Passerina amoena (Say], and a chipping sparrow (Spizella passerina (Bechstein], represent new host records for I. pacificus in California. Tissues from two ticks and thick blood films prepared from 126 birds tested negative for spirochetes by direct immunofluorescence (DI). A total of 172 larval and 197 nymphal I. pacificus was removed from 15 of 16 western fence lizards (Sceloporus occidentalis Baird & Girard) caught between April and June 1989 in the same location as were birds. Thick blood films prepared from all 16 lizards and tissue smears from 334 of the ticks (143 larvae and 191 nymphs) were DI test-negative for spirochetes. One (1.1%) of 93 adult I. pacificus collected at the bird-lizard capture site in February 1989 was infected with spirochetes that resembled B. burgdorferi.
A simple plasma anticoagulant-exchange method to increase the recovery of factor VIII in therapeutic concentrates.
Donor blood, primarily anticoagulated by acid citrate dextrose formula A (ACD-A), was separated by means of the HemaScience Autopheresis C plasmapheresis device. The citrated plasma was collected directly into a solution of heparin and calcium chloride to achieve a final plasma-ionised calcium concentration of approximately 2 mM, and a heparin concentration of 1.0 IU/ml. Heparin at this concentration provided adequate anticoagulation, and did not result in insoluble cryoprecipitates. Three pairs of donor-matched 4-kg plasma pools (anticoagulant-exchanged variant and ACD-A-anticoagulated control) were constructed and subsequently fractionated to an intermediate stage. The mean recovery of factor VIII from 3 anticoagulant-exchanged pools (394 IU/kg) was 23% greater than the mean recovery from the matched control pools (319 IU/kg). This increased recovery was not achieved at the expense of specific activity.
DNA and protein analyses of tick-derived isolates of Borrelia burgdorferi from California.
Nine isolates of Borrelia burgdorferi from ixodid ticks collected in northern California were characterized. Restriction endonuclease analysis, pulsed-field gel electrophoresis, and Western blot (immunoblot) analysis were used in this study. Four isolates were very similar to each other. The others shared some similarities but were classified as having unique genotypes. A strain from an Ixodes neotomae tick displayed the greatest genetic and antigenic diversity when compared to the isolates collected from Ixodes pacificus ticks. A computerized library based on DNA banding patterns of the isolates by restriction enzyme analysis is also reported. This library was created by using a scanning laser densitometer.
Interlaboratory and intralaboratory comparisons of indirect immunofluorescence assays for serodiagnosis of Lyme disease.
A conventional indirect immunofluorescence assay (IFA) and an anticomplement indirect immunofluorescence assay (ACIF) for detecting serum antibodies to Borrelia burgdorferi in humans were evaluated during a prevalence survey in northern California. Sera obtained from 119 current or former residents of an area in which Lyme disease is endemic were split and tested by the IFA in two laboratories and the ACIF in a third. The seropositivity rate ranged from 15 to 20% with 88 to 93% agreement among laboratories. Interlaboratory agreement was statistically highly significant in each of the three pairwise comparisons and was positively associated with clinical manifestations of Lyme disease. Intralaboratory agreement ranged from 93 to 96% in two laboratories and was also statistically highly significant. Immunoblotting confirmed 100 of 101 of the nondiscrepant immunofluorescence test results and likewise was positively correlated with the degree of interlaboratory agreement. The ACIF was found to be a highly specific test (100% specificity) with a much lower cutoff titer (1:8) than the conventional IFA (determined to be 1:128 or 1:256 in two laboratories) for detecting antibodies to B. burgdorferi. It also appeared to be more sensitive (80 versus 68%) than the IFA as determined by comparative immunoblotting, though the absolute sensitivity of the ACIF for serodiagnosis of early Lyme disease has yet to be determined. Significant serologic cross-reactivity was demonstrated between B. burgdorferi, Borrelia coriaceae, and Borrelia hermsii by the IFA, which may confound spirochetal serosurveys in California where all three spirochetes are known to coexist.
Susceptibility of the western fence lizard (Sceloporus occidentalis) to the Lyme borreliosis spirochete (Borrelia burgdorferi).
Attempts to infect juvenile and adult western fence lizards (Sceloporus occidentalis) with the Lyme borreliosis spirochete (Borrelia burgdorferi) were largely unsuccessful. Spirochetes could not be isolated from the blood and various tissues of 14 lizards 21-32 days after they had been inoculated ip (n = 8) or sc (n = 6) with 10(6) or 10(8) B. burgdorferi representing 3 tick isolates, although 1 lizard apparently developed a transitory spirochetemia lasting 2 days. Similarly, spirochetes could not be detected in the blood or tissues of 5 lizards fed upon by 2- greater than 8 infected larvae or nymphs of the western black-legged tick (Ixodes pacificus). Sixty-five blood samples from 59 lizards in an endemic area and various tissues from 20 of the same lizards were also assayed for B. burgdorferi with negative results. The implications of these findings for the maintenance of this spirochete in natural foci are discussed.
Treatment of clothing with a permethrin spray for personal protection against the western black-legged tick, Ixodes pacificus (Acari: Ixodidae).
The synthetic pyrethroid, permethrin, when applied to clothing with a pressurized spray at an application rate estimated previously to be 4 micrograms a.i./cm2, was found to be 100% effective for personal protection against all three parasitic stages of the western black-legged tick, Ixodes pacificus Cooley and Kohls. This tick has been implicated as the primary vector of the Lyme disease spirochete (Borrelia burgdorferi) to humans in the far-western United States. Periods of exposure to permethrin-treated cloth as brief as 10 and 45 seconds incapacitated 100% of subadult and adult ticks, respectively, within 1-3 h post-treatment. Under field conditions, permethrin apparently does not repel questing adult ticks, though 100% of ticks recovered from treated clothing after exposures as brief as less than or equal to 15 seconds were moribund 1 h later.
Lyme disease in California: interrelationship of Ixodes pacificus (Acari: Ixodidae), the western fence lizard (Sceloporus occidentalis), and Borrelia burgdorferi.
The relationship of immature western black-legged ticks, Ixodes pacificus Cooley and Kohls, to the western fence lizard, Sceloporus occidentalis Baird and Girard, and to the Lyme disease spirochete, Borrelia burgdorferi, was investigated in chaparral and woodland-grass habitats in northern California from 1984 to 1986. Immature ticks were found on lizards in spring and summer, but the prevalence and abundance of ticks on this host were considerably greater in spring. The peak of larval abundance preceded that of nymphs by several weeks, but there was considerable seasonal overlap between these parasitic stages. Larvae and nymphs attached primarily to the lateral nuchal pockets of lizards in chaparral (99.5%) and woodland-grass (91.8%). The numbers of larvae infesting lizards in spring fit the negative binomial distribution in woodland-grass but not in chaparral; insufficient data precluded similar analyses for nymphs. Tick loads did not differ significantly with respect to age or gender of the lizard. Spirochetal infection rates (range, 0-3.7%) in I. pacificus immatures were comparable in both habitats and were similar to those reported previously for adults of this tick. Overall, 1 (0.9%) of 117 larvae and 10 (1.8%) of 552 nymphs were infected with spirochetes resembling B. burgdorferi. Spirochetes were not observed in blood smears prepared from 261 wild-caught lizards, including five lizards fed upon by infected ticks at the time of collection. These and other findings suggest that S. occidentalis, although an important host of I. pacificus immatures, may be less important as a source for infecting ticks with B. burgdorferi.
Severely heated therapeutic factor VIII concentrate of high specific activity.
A new method for the manufacture of a heated factor VIII concentrate of high specific activity (2-6 IU factor VIII:C/mg protein) has been developed. Addition of heparin to cryoprecipitate extract at acid pH precipitated fibrinogen and fibronectin. Factor VIII was then recovered from the supernatant by precipitation with glycine and sodium chloride. After re-solution and desalting on Sephadex G-25, the concentrate was sterile-filtered and lyophilised. The dried product was stable to heating in the final container at 80 degrees C for 72 h. Data from 25 consecutive batches of concentrate prepared from 1,200-1,500 kg plasma pools are presented. The mean final yield of heated product was 190 IU factor VIII:C/kg plasma. The concentrate has been found to be safe and effective in clinical use.
Antigenic characteristics of Borrelia burgdorferi isolates from ixodid ticks in California.
Twenty (1.4%) of 1,421 adult Ixodes pacificus ticks and 2 (20%) of 10 adult Ixodes neotomae ticks collected in five counties of northern California were found to contain spirochetes by direct immunofluorescence examination of their tissues with a polyvalent conjugate. Borreliae isolated from the tissues of nine of these ticks (I. pacificus, 8; I. neotomae, 1) were identified as Borrelia burgdorferi with specific monoclonal antibodies and characterized further by polyacrylamide gel electrophoresis and Western blot (immunoblot) analyses. The isolate from I. neotomae was the first to be characterized from a tick other than I. pacificus in western North America. All strains were relatively homogeneous with respect to the kind of OspA proteins they produced, whereas they were heterogeneous with regard to their OspB proteins and to several low-molecular-weight proteins in the 21,500-to-24,000 region. Significant phenotypic variation was observed among isolates obtained within and between populations of I. pacificus. This investigation nearly doubles the number of isolates of B. burgdorferi that have been characterized from ixodid ticks in the far western United States.
Lagomorphs as sentinels for surveillance of borreliosis in the far western United States.
Brush rabbits (Sylvilagus bachmani) and black-tailed jackrabbits (Lepus californicus) from California (USA) were assayed for antibodies to Borrelia burgdorferi, the etiologic agent of Lyme borreliosis. Significant antibody titers were detected in 90% (range, 67 to 100%) of brush rabbits from four of six localities, and in 90% of jackrabbits from a single locality, in northern California. One of the populations of brush rabbits that did not yield seropositive individuals inhabited an oceanic island devoid of any other terrestrial mammal, whereas the other population was located on an isolated flood plain bordering San Francisco Bay. Absorption tests using B. burgdorferi as antigen revealed that antibodies detected in both species of lagomorphs were directed against borreliae. These findings reinforce the earlier suggestion that lagomorphs may be useful as sentinel animals for surveillance of borreliosis in the far western United States.