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An enzyme-linked immunosorbent assay was developed to detect antibodies to Borrelia burgdorferi in cottontail rabbits captured in Millbrook, N.Y., and New York, N.Y. Five antigenically variable strains of B. burgdorferi were analyzed to determine the variability of serologic test results. In analyses of 79 serum samples, seropositivity ranged from 56% for a strain cultured from kidney tissues of a cottontail rabbit to 68% for a strain isolated from a larva of Ixodes dentatus, a tick that parasitized a cottontail rabbit. There were false-positive results when reference rabbit antisera to B. hermsii and Treponema pallidum were screened against B. burgdorferi. Cross-reactivity with antisera to Leptospira interrogans serovars was less pronounced. Western blot (immunoblot) analyses revealed reactivities of test sera to two or more surface or subsurface proteins of B. burgdorferi with approximate molecular masses of 18, 25 to 27, 34, 36, 41, and 59 kilodaltons. Cottontail rabbits respond immunologically to B. burgdorferi, but the observed variations in serologic test results should not be a limitation in field and laboratory investigations of Lyme borreliosis.

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Spirochetes were isolated from 71 subadult Ixodes dentatus removed from cottontail rabbits captured in Millbrook, N.Y., and in New York, N.Y. Spirochetes were also cultured from kidney tissues of six rabbits. While all isolates reacted with monoclonal antibody H9724, which identifies the spirochetes as borreliae, more than half did not bind with antibody H5332 and even fewer reacted with H3TS, both of which were produced to outer surface protein A of Borrelia burgdorferi. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis protein profiles of three isolates differed from one another and from all previously characterized B. burgdorferi strains from humans, ticks, and wildlife in North America. The 12 periplasmic flagella that originated subterminally from each pointed end of a rabbit Borellia isolate contrasted with the 11 or fewer flagella for B. burgdorferi reported previously from North America. Although DNA homology and restriction endonuclease analysis also revealed differences among a rabbit kidney isolate, an I. dentatus isolate, and B. burgdorferi B31, similarities were sufficient to lead us to conclude that the borreliae in rabbits and I. dentatus are B. burgdorferi. Enzyme-linked immunosorbent assay titers of sera from humans with diagnosed Lyme disease to rabbit tick B. burgdorferi were often similar to one another and to those recorded for a reference B. burgdorferi strain.

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A previously undescribed Borrelia burgdorferi antigenic variant was isolated from each of four Ixodes dammini larvae removed from white-footed mice, Peromyscus leucopus, captured in Millbrook, N.Y. This site is in the northern range of the known distribution of the tick in the northeastern United States. The molecular weights of approximately 32,500 and 35,500 for outer surface A and outer surface B proteins, respectively, were distinctly higher than those for previously characterized isolates from North American ticks, humans, and wild mammals. A prominent low-molecular-weight protein of about 23,500 was also present. All four isolates infected Syrian hamsters and retained their antigenicity after passage through these rodent hosts. Serum samples from patients with Lyme disease tended to have immunoglobulin M, immunoglobulin G, and immunoglobulin antibodies to Connecticut B. burgdorferi 2591 at titers slightly higher than or equal to those recorded for an antigenically different strain of B. burgdorferi from Millbrook, N.Y.

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An enzyme-linked immunosorbent assay (ELISA) and indirect fluorescent-antibody test were used to detect antibodies to Borrelia burgdorferi, the causative agent of Lyme disease, in Peromyscus leucopus (white-footed mouse). Of the 661 mice captured in Connecticut, Rhode Island, and New York during 1980 and 1983 to 1987, 166 (25.1%) had antibodies to B. burgdorferi by ELISA. Comparative analyses of 210 serum specimens, collected in areas where Lyme disease is endemic, revealed a threefold difference in sensitivity between the ELISA (38.1% positive) and the indirect fluorescent-antibody method (12.4%). Although prevalence of seropositive P. leucopus was highest during June, elevated amounts of antibody (1:1,280 to 1:2,560) were detected in mice that harbored spirochetes during all seasons. Being reservoirs for B. burgdorferi, these rodents are suitable for monitoring spirochete infections at foci and should be included in field evaluations of control programs aimed at suppressing Lyme disease.

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