Skip to main content
Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1996 Mar;34(3):524–529. doi: 10.1128/jcm.34.3.524-529.1996

Novel Borrelia burgdorferi isolates from Ixodes scapularis and Ixodes dentatus ticks feeding on humans.

J F Anderson 1, R A Flavell 1, L A Magnarelli 1, S W Barthold 1, F S Kantor 1, R Wallich 1, D H Persing 1, D Mathiesen 1, E Fikrig 1
PMCID: PMC228839  PMID: 8904407

Abstract

Seven cultures of Borrelia burgdorferi differing from strains B31 and ZS7 were identified from among 99 isolates from Ixodes scapularis ticks and from white-footed mice (Peromyscus leucopus) and 1 isolate from an Ixodes dentatus tick. Five of the six novel isolates from I. scapularis and the isolate from I. dentatus were from ticks feeding on humans. The six isolates from I. scapularis lacked OspA and OspB, four possessed an OspD band, and two reacted with an anti-OspC monoclonal antibody. Restriction fragment length polymorphisms of HindIII-digested DNAs from six OspA-negative isolates did not hybridize with radiolabeled ospA or LA88 DNA, and only isolate 46047 hybridized with the pG gene. Fragments similar to those recorded for the standard B. burgdorferi sensu stricto strains B31 and ZS7 were obtained with the fla and the HSP70 genes. Pulsed-field gel electrophoresis patterns of DNA digested with MluI included the specific B. burgdorferi sensu stricto band at 135 kbp for the five OspA-negative isolates from I. scapularis ticks. The six novel isolates apparently lack the 55-kbp plasmid encoding OspA. The pG-containing plasmid may be missing from all but isolate 46047. The isolate from the I. dentatus tick was similar to previous isolates from I. dentatus ticks feeding on rabbits. None of the isolates could be recovered from inoculated C3H/HeNCrlBR or white-footed mice. All isolates reacted with sera from humans with early or late Lyme disease. Our studies demonstrate that these borreliae occur in ticks feeding on humans, and therefore, at least some humans in the northeastern United States are likely being exposed to borreliae other than the classic B31-type strains that have thus far been isolated from humans.

Full Text

The Full Text of this article is available as a PDF (326.8 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Anderson J. F., Magnarelli L. A. Epizootiology of Lyme disease and methods of cultivating Borrelia burgdorferi. Ann N Y Acad Sci. 1992 Jun 16;653:52–63. doi: 10.1111/j.1749-6632.1992.tb19629.x. [DOI] [PubMed] [Google Scholar]
  2. Anderson J. F., Magnarelli L. A., LeFebvre R. B., Andreadis T. G., McAninch J. B., Perng G. C., Johnson R. C. Antigenically variable Borrelia burgdorferi isolated from cottontail rabbits and Ixodes dentatus in rural and urban areas. J Clin Microbiol. 1989 Jan;27(1):13–20. doi: 10.1128/jcm.27.1.13-20.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Anderson J. F., Magnarelli L. A., McAninch J. B. New Borrelia burgdorferi antigenic variant isolated from Ixodes dammini from upstate New York. J Clin Microbiol. 1988 Oct;26(10):2209–2212. doi: 10.1128/jcm.26.10.2209-2212.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Baranton G., Postic D., Saint Girons I., Boerlin P., Piffaretti J. C., Assous M., Grimont P. A. Delineation of Borrelia burgdorferi sensu stricto, Borrelia garinii sp. nov., and group VS461 associated with Lyme borreliosis. Int J Syst Bacteriol. 1992 Jul;42(3):378–383. doi: 10.1099/00207713-42-3-378. [DOI] [PubMed] [Google Scholar]
  5. Barbour A. G., Hayes S. F., Heiland R. A., Schrumpf M. E., Tessier S. L. A Borrelia-specific monoclonal antibody binds to a flagellar epitope. Infect Immun. 1986 May;52(2):549–554. doi: 10.1128/iai.52.2.549-554.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Barbour A. G., Heiland R. A., Howe T. R. Heterogeneity of major proteins in Lyme disease borreliae: a molecular analysis of North American and European isolates. J Infect Dis. 1985 Sep;152(3):478–484. doi: 10.1093/infdis/152.3.478. [DOI] [PubMed] [Google Scholar]
  7. Barbour A. G. Isolation and cultivation of Lyme disease spirochetes. Yale J Biol Med. 1984 Jul-Aug;57(4):521–525. [PMC free article] [PubMed] [Google Scholar]
  8. Barbour A. G., Tessier S. L., Todd W. J. Lyme disease spirochetes and ixodid tick spirochetes share a common surface antigenic determinant defined by a monoclonal antibody. Infect Immun. 1983 Aug;41(2):795–804. doi: 10.1128/iai.41.2.795-804.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Belfaiza J., Postic D., Bellenger E., Baranton G., Girons I. S. Genomic fingerprinting of Borrelia burgdorferi sensu lato by pulsed-field gel electrophoresis. J Clin Microbiol. 1993 Nov;31(11):2873–2877. doi: 10.1128/jcm.31.11.2873-2877.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Benach J. L., Bosler E. M., Hanrahan J. P., Coleman J. L., Habicht G. S., Bast T. F., Cameron D. J., Ziegler J. L., Barbour A. G., Burgdorfer W. Spirochetes isolated from the blood of two patients with Lyme disease. N Engl J Med. 1983 Mar 31;308(13):740–742. doi: 10.1056/NEJM198303313081302. [DOI] [PubMed] [Google Scholar]
  11. Berger B. W., Johnson R. C., Kodner C., Coleman L. Cultivation of Borrelia burgdorferi from erythema migrans lesions and perilesional skin. J Clin Microbiol. 1992 Feb;30(2):359–361. doi: 10.1128/jcm.30.2.359-361.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Bissett M. L., Hill W. Characterization of Borrelia burgdorferi strains isolated from Ixodes pacificus ticks in California. J Clin Microbiol. 1987 Dec;25(12):2296–2301. doi: 10.1128/jcm.25.12.2296-2301.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Brown R. N., Lane R. S. Lyme disease in California: a novel enzootic transmission cycle of Borrelia burgdorferi. Science. 1992 Jun 5;256(5062):1439–1442. doi: 10.1126/science.1604318. [DOI] [PubMed] [Google Scholar]
  14. Burgdorfer W., Barbour A. G., Hayes S. F., Benach J. L., Grunwaldt E., Davis J. P. Lyme disease-a tick-borne spirochetosis? Science. 1982 Jun 18;216(4552):1317–1319. doi: 10.1126/science.7043737. [DOI] [PubMed] [Google Scholar]
  15. Canica M. M., Nato F., du Merle L., Mazie J. C., Baranton G., Postic D. Monoclonal antibodies for identification of Borrelia afzelii sp. nov. associated with late cutaneous manifestations of Lyme borreliosis. Scand J Infect Dis. 1993;25(4):441–448. doi: 10.3109/00365549309008525. [DOI] [PubMed] [Google Scholar]
  16. Fikrig E., Tao H., Kantor F. S., Barthold S. W., Flavell R. A. Evasion of protective immunity by Borrelia burgdorferi by truncation of outer surface protein B. Proc Natl Acad Sci U S A. 1993 May 1;90(9):4092–4096. doi: 10.1073/pnas.90.9.4092. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Johnson R. C., Marek N., Kodner C. Infection of Syrian hamsters with Lyme disease spirochetes. J Clin Microbiol. 1984 Dec;20(6):1099–1101. doi: 10.1128/jcm.20.6.1099-1101.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Johnson S. E., Klein G. C., Schmid G. P., Bowen G. S., Feeley J. C., Schulze T. Lyme disease: a selective medium for isolation of the suspected etiological agent, a spirochete. J Clin Microbiol. 1984 Jan;19(1):81–82. doi: 10.1128/jcm.19.1.81-82.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Lane R. S., Pascocello J. A. Antigenic characteristics of Borrelia burgdorferi isolates from ixodid ticks in California. J Clin Microbiol. 1989 Oct;27(10):2344–2349. doi: 10.1128/jcm.27.10.2344-2349.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Magnarelli L. A., Anderson J. F. Enzyme-linked immunosorbent assays for the detection of class-specific immunoglobulins to Borrelia burgdorferi. Am J Epidemiol. 1988 Apr;127(4):818–825. doi: 10.1093/oxfordjournals.aje.a114864. [DOI] [PubMed] [Google Scholar]
  21. Magnarelli L. A., Anderson J. F., Johnson R. C. Cross-reactivity in serological tests for Lyme disease and other spirochetal infections. J Infect Dis. 1987 Jul;156(1):183–188. doi: 10.1093/infdis/156.1.183. [DOI] [PubMed] [Google Scholar]
  22. Magnarelli L. A., Anderson J. F., Johnson R. C., Nadelman R. B., Wormser G. P. Comparison of different strains of Borrelia burgdorferi sensu lato used as antigens in enzyme-linked immunosorbent assays. J Clin Microbiol. 1994 May;32(5):1154–1158. doi: 10.1128/jcm.32.5.1154-1158.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Magnarelli L. A., Miller J. N., Anderson J. F., Riviere G. R. Cross-reactivity of nonspecific treponemal antibody in serologic tests for Lyme disease. J Clin Microbiol. 1990 Jun;28(6):1276–1279. doi: 10.1128/jcm.28.6.1276-1279.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Marconi R. T., Liveris D., Schwartz I. Identification of novel insertion elements, restriction fragment length polymorphism patterns, and discontinuous 23S rRNA in Lyme disease spirochetes: phylogenetic analyses of rRNA genes and their intergenic spacers in Borrelia japonica sp. nov. and genomic group 21038 (Borrelia andersonii sp. nov.) isolates. J Clin Microbiol. 1995 Sep;33(9):2427–2434. doi: 10.1128/jcm.33.9.2427-2434.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Nadelman R. B., Pavia C. S., Magnarelli L. A., Wormser G. P. Isolation of Borrelia burgdorferi from the blood of seven patients with Lyme disease. Am J Med. 1990 Jan;88(1):21–26. doi: 10.1016/0002-9343(90)90122-t. [DOI] [PubMed] [Google Scholar]
  26. Norris S. J., Carter C. J., Howell J. K., Barbour A. G. Low-passage-associated proteins of Borrelia burgdorferi B31: characterization and molecular cloning of OspD, a surface-exposed, plasmid-encoded lipoprotein. Infect Immun. 1992 Nov;60(11):4662–4672. doi: 10.1128/iai.60.11.4662-4672.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Oliver J. H., Jr, Owsley M. R., Hutcheson H. J., James A. M., Chen C., Irby W. S., Dotson E. M., McLain D. K. Conspecificity of the ticks Ixodes scapularis and I. dammini (Acari: Ixodidae). J Med Entomol. 1993 Jan;30(1):54–63. doi: 10.1093/jmedent/30.1.54. [DOI] [PubMed] [Google Scholar]
  28. Persing D. H., Rutledge B. J., Rys P. N., Podzorski D. S., Mitchell P. D., Reed K. D., Liu B., Fikrig E., Malawista S. E. Target imbalance: disparity of Borrelia burgdorferi genetic material in synovial fluid from Lyme arthritis patients. J Infect Dis. 1994 Mar;169(3):668–672. doi: 10.1093/infdis/169.3.668. [DOI] [PubMed] [Google Scholar]
  29. Rawlings J. A., Fournier P. V., Teltow G. J. Isolation of Borrelia spirochetes from patients in Texas. J Clin Microbiol. 1987 Jul;25(7):1148–1150. doi: 10.1128/jcm.25.7.1148-1150.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Sears J. E., Fikrig E., Nakagawa T. Y., Deponte K., Marcantonio N., Kantor F. S., Flavell R. A. Molecular mapping of Osp-A mediated immunity against Borrelia burgdorferi, the agent of Lyme disease. J Immunol. 1991 Sep 15;147(6):1995–2000. [PubMed] [Google Scholar]
  31. Steere A. C., Grodzicki R. L., Kornblatt A. N., Craft J. E., Barbour A. G., Burgdorfer W., Schmid G. P., Johnson E., Malawista S. E. The spirochetal etiology of Lyme disease. N Engl J Med. 1983 Mar 31;308(13):733–740. doi: 10.1056/NEJM198303313081301. [DOI] [PubMed] [Google Scholar]
  32. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Wallich R., Brenner C., Kramer M. D., Simon M. M. Molecular cloning and immunological characterization of a novel linear-plasmid-encoded gene, pG, of Borrelia burgdorferi expressed only in vivo. Infect Immun. 1995 Sep;63(9):3327–3335. doi: 10.1128/iai.63.9.3327-3335.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Wallich R., Helmes C., Schaible U. E., Lobet Y., Moter S. E., Kramer M. D., Simon M. M. Evaluation of genetic divergence among Borrelia burgdorferi isolates by use of OspA, fla, HSP60, and HSP70 gene probes. Infect Immun. 1992 Nov;60(11):4856–4866. doi: 10.1128/iai.60.11.4856-4866.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Wilske B., Preac-Mursic V., Göbel U. B., Graf B., Jauris S., Soutschek E., Schwab E., Zumstein G. An OspA serotyping system for Borrelia burgdorferi based on reactivity with monoclonal antibodies and OspA sequence analysis. J Clin Microbiol. 1993 Feb;31(2):340–350. doi: 10.1128/jcm.31.2.340-350.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Wilske B., Preac-Mursic V., Schierz G., Busch K. V. Immunochemical and immunological analysis of European Borrelia burgdorferi strains. Zentralbl Bakteriol Mikrobiol Hyg A. 1986 Dec;263(1-2):92–102. doi: 10.1016/s0176-6724(86)80108-0. [DOI] [PubMed] [Google Scholar]
  37. Wilske B., Preac-Mursic V., Schierz G., Kühbeck R., Barbour A. G., Kramer M. Antigenic variability of Borrelia burgdorferi. Ann N Y Acad Sci. 1988;539:126–143. doi: 10.1111/j.1749-6632.1988.tb31846.x. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Clinical Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES