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Borreliosis

Lyme disease or Lyme borreliosis is an infectious tick-borne disease, caused by the Borrelia spirochete, a gram-negative microorganism. more...

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Lyme disease is so named because it is generally believed to have first been observed in and around Old Lyme and Lyme, Connecticut in 1975. Before 1975, elements of Borrelia infection were also known as "tick-borne meningopolyneuritis", Garin-Bujadoux syndrome, Bannwarth syndrome or sheep tick fever. It is transmitted to humans by the bite of infected ticks.

History

The disease was first documented as a skin rash in Europe in 1883. Over the years, researchers there identified additional features of the disease, including an unidentified pathogen, its response to penicillin, the role of the Ixodes tick (wood tick) as its vector, and symptoms that included not only the rash but additional ones that affected the nervous system.

Researchers in the US had been aware of tick infections since the early 1900s. For example, an infection called tick relapsing fever was reported in 1905, and the wood tick, which carries an agent that causes Rocky Mountain spotted fever, was identified soon after. However, the full syndrome now known as Lyme disease, was not identified until a cluster of cases thought to be juvenile rheumatoid arthritis occurred in three towns in southeastern Connecticut, in the United States. Two of these towns, Lyme and Old Lyme, gave the disease its popular name.

In 1982 a novel spirochete was isolated and cultured from the midgut of Ixodes ticks, and subsequently from patients with Lyme disease. The infecting agent was first identified by Jorge Benach, and soon after isolated by Willy Burgdorfer, a scientist at the National Institutes of Health who specialized in the study of spirochete microorganisms. The spirochete was named Borrelia burgdorferi in his honor. Burgdorfer was the partner in the successful effort to culture the spirochete, along with Alan Barbour.

Microbiology

The disease is caused by the parasite Borrelia, which has well over three hundred known genomic strains but is usually cultured as Borrelia burgdorferi, Borrelia afzelii or Borellia garinii. Different Borrelia strains are predominant in Europe and North America.

The disease has been found to be transmitted to humans by the bite of infected Ixodes ticks. Not all ticks carry or can transmit this particular disease. The disease may also, in a few cases, be transmitted by mosquitoes, fleas or blackflies. However, other Borrelia strains (e.g. B. garinii) are probably transmitted this way.

Other tick-borne infections may be transmitted simultaneously with Lyme, including Bartonella, Babesiosis, Ehrlichiosis, and Rickettsia.

Borrelia burgdorferi resembles other spirochetes in that it is a highly specialized, motile, two-membrane, spiral-shaped bacterium which lives primarily as an extracellular pathogen. One of the most striking features of Borrelia burgdorferi as compared with other eubacteria is its unusual genome, which includes a linear chromosome approximately one megabase in size and numerous linear and circular plasmids.

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Borrelia spielmanii erythema migrans, Hungary
From Emerging Infectious Diseases, 11/1/05 by Gabor Foldvari

To the Editor: Lyme disease is the most frequent tickborne human infection in the northern hemisphere. At least 5 species of the Borrelia burgdorferi sensu lato complex, B. burgdorferi sensu stricto, B. afzelii, B. garinii, B. bissettii, and B. lusitaniae, have a pathogenic role in human Lyme disease in central Europe (1-3). A sixth pathogenic strain, A14S, has been isolated from 1 Dutch (4) and 2 German patients with erythema migrans (5). This strain was also detected in 4 questing Ixodes ricinus ticks in Germany (6,7) and 1 in the Czech Republic (8). A14S has recently been described as a new species, B. spielmanii (9); its main reservoir host is probably the garden dormouse (Eliomys quercinus), but B. spielmanii could not be detected in mice or voles. Richter et al. (9) could not find ticks harboring B. spielmanii in 3 of 5 examined areas in Germany. They were present almost exclusively in a single area where the prevalence of infection with this genotype was 15 (6%) of 251. We describe the isolation of this novel Lyme disease spirochete from a human patient with erythema migrans in Hungary.

Since 1999, we have regularly isolated Borrelia burgdorferi sensu lato from skin biopsy specimens of erythema migrans and acrodermatitis chronica atrophicans taken from patients at the Center for Tick-borne Diseases, Budapest, Hungary. To identify the Borrelia species occurring in Hungarian Lyme disease patients, we have started to molecularly analyze cultured isolates that originate from erythema migrans of different patients. DNA was isolated from 8 bacterial pellets by using QIAamp DNA mini kit (Qiagen, Hilden, Germany). Primers BSL-F and BSL-R were used; these amplify an [approximately equal to] 250-bp region of the outer surface protein (osp) A gene from all Lyme disease spirochetes (10). We added 2 [micro]L extracted DNA to a 20-[micro]L reaction mixture composed of 1.0 U HotStartTaq DNA polymerase, 200 [micro]mol/L of each dNTP, 25 pmol of each primer, and 1.5 mmol/L Mg[Cl.sub.2] (HotStartTaq Master Mix, Qiagen). An initial denaturation step at 94[degrees]C for 15 min was followed by 40 cycles of denaturation at 94[degrees]C for 30 s, annealing at 58[degrees]C for 30 s, and extension at 72[degrees]C for 30 s. Final extension was done at 72[degrees]C for 5 min. Amplified DNA was subjected to electrophoresis in a 1.5% agarose gel that was prestained with ethidium bromide and viewed under UV light. After purification, the dideoxy chain termination (Applied Biosystems Division, Foster City, CA, USA) was used for sequencing. Obtained sequences were checked with Chromas v. 1.45 and compared to sequence data available from GenBank by using BLAST (http:// www.ncbi.nlm.nih.gov/BLAST/). New sequences were submitted to GenBank.

Six sequences (DQ007298, DQ007299, DQ007300, DQ007301, DQ007302, DQ007303) showed 100% homology to B. afzelii Khab 625 strain (AY502599). One (DQ007297) of the remaining 2 samples showed 99.6% similarity with B. burgdorferi B31 (AE000790), and the other (AY995900) showed 99.21% similarity with B. spielmanii (AF102057).

The patient whose culture showed B. spielmanii was a 42-year-old woman with a homogenous erythema migrans, diagnosed on September 24, 1999. The erythema was 10 cm in diameter on the front surface of the knee at the first visit (see online Figure, available at http://www. cdc.gov/ncidod/EID/vol11no11/050542-G.htm). The immunoglobulin M (IgM) and IgG Borrelia immunoblot that applied B. afzelii (ACA1) antigen was negative in serum drawn on the seventh day after the appearance of erythema migrans. The patient did not remember a tick bite and had not traveled abroad during the previous 6 months. She complained of an "extremely unusual," intense, serous nasal discharge that started 3 weeks before the appearance of erythema migrans and of a moderate headache; both disappeared spontaneously 2 weeks before treatment.

Our results show at least 3 distinct species or B. burgdorferi sensu lato in Hungary. In addition to B. burgdorferi sensu stricto and B. afzelii, known throughout Europe, we detected the recently described species B. spiel manii among randomly selected samples. Together with 2 previous publications (4,5), our observation also suggest that B. spielmanii has a pathogenic role in human Lyme disease. Although B. spielmanii is distributed more focally than other species of the B. burgdorferi sensu lato complex (9), it occurs from the Netherlands through Germany and Czech Republic to Hungary (4,5,7,8).

Acknowledgment

We acknowledge the advice and the positive control samples provided by Martin J. Kenny.

Gabor Foldvari, * Robert Farkas, * and Andras Lakos ([dagger])

* St. Istvan University Faculty of Veterinary Science, Budapest, Hungary; and ([dagger]) Center for Tick-borne Disease, Budapest, Hungary

Address for correspondence: Andras Lakos, Center for Tick-borne Diseases, Visegradi u. 14. H-1132 Budapest, Hungary; fax: 36-1-329-3898; email: alakos@t-online.hu

References

(1.) Postie D, Ras NM, Lane RS, Hendson M, Baranton G. Expanded diversity among Californian Borrelia isolates and description of Borrelia bissettii sp. nov. (formerly Borrelia group DN127). J Clin Microbiol. 1998;36:3497-504.

(2.) Maraspin V, Cimperman J, Lotric-Furlan S, Ruzic-Sabljic E, Jurca T, Pieken RN, et al. Solitary borrelial lymphocytoma in adult patients. Wien Klin Wochenschr. 2002;114:515-23.

(3.) Collares-Pereira M, Couceiro S, Franca I, Kurtenbaeh K, Schafer SM, Vitorino L, et al. First isolation of Borrelia lusitaniae from a human patient. J Clin Microbiol. 2004;42:1313-8.

(4.) Wang G, van Dam AP, Dankert J. Phenotypic and genetic characterization of a novel Borrelia burgdorferi sensu lato isolate from a patient with Lyme borreliosis. J Clin Microbiol. 1999;37:3025-8.

(5.) Fingerle V, Michel H, Schulte-Spechtel U, Gottner G, Hizo-Teufel C, Hofmann H, et al. A14S--a new Borrelia burgdorferi s.l. genospecies as relevant cause of human disease [abstract]. Int J Med Microbiol. 2004;294(Suppl 1):207.

(6.) Rauter C, Oehme R, Dietrich I, Engele M, Hartung T. Distribution of clinically relevant Borrelia genospecies in ticks assessed by a novel, single-run real-time PCR. J Clin Microbiol. 2002;40:36-43.

(7.) Michel HB, Wilske B, Hettche G, Gottner G, Heimerl C, Reischl U, et al. An ospA-polymerase chain reaction/restriction fragment length polymorphism-based method for sensitive detection and reliable differentiation of all European Borrelia burgdorferi sensu lato species and OspA types. Med Microbiol Immunol. 2003;193:219-26.

(8.) Derdakova M, Beati L, Pet'ko B, Stanko M, Fish D. Genetic variability within Borrelia burgdorferi sensu lato genospecies established by PCR-single-strand conformation polymorphism analysis of the rrfA-rrlB intergenic spacer in Ixodes ricinus ticks from the Czech Republic. Appl Environ Microbiol. 2003;69:509-16.

(9.) Richter D, Schlee DB, Allgower R, Matuschka FR. Relationships of a novel Lyme disease spirochete, Borrelia spielmani sp. nov., with its hosts in central Europe. Appl Environ Microbiol. 2004;70:6414-9.

(10.) Demaerschalck I, Benmessaoud A, Dekesel, M, Hoyois B, Lobet Y, Hoet P, et al. Simultaneous presence of different Borrelia burgdorferi genospecies in biological fluids of Lyme-disease patients. J Clin Microbiol. 1995;33:602-8.

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