Identification of Borrelia burgdorferi sensu stricto, Borrelia garinii and Borrelia afzelii in Ixodes ricinus Ticks from Southern Bohemia Using Monoclonal Antibodies

doi:10.1016/S0934-8840(00)80005-5

Zentralblatt für Bakteriologie

Volume 289, Issue 8, January 2000, Pages 797-806

https://doi.org/10.1016/S0934-8840(00)80005-5 Get rights and content

Summary

The prevalence of Borrelia burgdorferi sensu lato (s.l.) in 761 Ixodes ricinus ticks collected in different localities of southern Bohemia was investigated. The minimum infection rate of nymphs, males and females as determined by dark-field microscopy was 4.8 % in 1997. The total B. burgdorferi prevalence determined in ticks in 1998 was 11.04 %. A significant local variability of the prevalence of B. burgdorferi was not found. Nine isolates were identified at a genospecies level by immunoblotting with prepared monoclonal antibodies (Mabs). The most prevalent genospecies in ticks was B. burgdorferi sensu stricto (s. s.) (6 strains). Two tick isolates reacted with Mabs specific to both B. garinii and B. afzelii and one isolate was detected as a mixed culture of B. burgdorferi s. s. and B. garinii. Considerable variability in protein composition among the strains obtained was found to exist. Five B. burgdorferi genospecies-specific Mabs were prepared and used for the identification of spirochaete isolates. Mabs 9B8 and 6C5 reacted with OspA and OspB proteins of B. burgdorferi s. s., while 4D7 and 1D11 recognised OspA protein of B. garinii and 9D2 specifically reacted with the OspB protein of B. afzelii. In addition, Mab 5H4 reacted with flagellin of all the above-mentioned genospecies of B. burgdorferi s.l.

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Cited by (37)

Selected phenotypic features of BR91, a unique spirochaetal strain isolated from the Culex pipiens mosquito
2014, Microbiological Research
Citation Excerpt :
However, spirochaetes in high cell density survived longer at 28 °C. For spirochaete primary isolation attempts, cultivation medium is generally supplemented with antibiotics such as phosphomycin, 5-fluorouracil, trimethoprim, sulfamethoxazole, gentamicin, kanamycin, neomycin and rifampicin to suppress the growth of contaminating agents (Cacciapuoti et al. 1995; Oliver et al. 1998; Strle et al. 1995; Štěpánová-Tresová et al. 1999; Tresová et al. 1998). We found that strain BR91 was sensitive to low concentration of trimethoprim (10 μg/mL), while B. burgdorferi s.l. tolerated significantly higher concentration of this antibiotic (Morshed et al. 1993).
Growth temperature range, resistance to selective antibiotics, activities of 23 enzymes, protein fingerprints and fatty acids composition of the spirochaetal strain BR91, isolated from the Culex pipiens mosquito, were tested. The spirochaetes were grown in BSK-H Complete liquid medium. The optimal in vitro growth temperature of the strain was 33 °C. Strain BR91 was sensitive to trimethoprim, nalidixic acid, 5-fluorouracil, and tolerated phosphomycin. The strain produced acid and alkaline phosphatase, esterase (C4), esterase–lipase (C8), leucine arylamidase, naphthol-AS-BI-phosphohydrolase and α-fucosidase. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) assay revealed several major proteins in the size range of 13–16 kDa, 22–30 kDa and 37–131 kDa. Fatty acid methyl ester (FAME) analysis showed that C_14:0, C_16:0, C_18:1 ω9c and summed feature 5 (C_18:2 ω6,9c and/or C_18:0 anteiso) are major fatty acids. This study highlights certain phenotypic differences between strain BR91 and the Lyme disease spirochaete Borrelia burgdorferi, and supports the hypothesis that strain BR91 represents a unique taxonomical entity in a system of spirochaetal species.
IrAM-An α<inf>2</inf>-macroglobulin from the hard tick Ixodes ricinus: Characterization and function in phagocytosis of a potential pathogen Chryseobacterium indologenes
2009, Developmental and Comparative Immunology
The universal protease inhibitors of the α₂-macroglobulin (α₂M) family are evolutionarily conserved constituents of innate immunity, presumably because they guard organisms against undesired proteolytic attacks of a different origin. Here, we determined the primary structure of α₂-macroglobulin from the hard tick Ixodes ricinus (IrAM) by sequencing of overlapping PCR products. Predicted disulfide and glycosylation patterns, post-translational cleavage and alternative splicing within its ‘bait region’ demonstrate that IrAM is closely related to the α₂-macroglobulin from the soft tick Ornithodoros moubata. The IrAM message is expressed in all tick developmental stages and tissues, except for the gut, and the protein was detected to be mainly present in the hemolymph. Silencing of IrAM by dsRNA interference markedly reduced the phagocytosis of a potential pathogen, Chryseobacterium indologenes, by tick hemocytes both in vitro and in vivo. In contrast, phagocytosis of the Lyme disease spirochete Borrelia burgdorferi or a commensal bacteria Staphylococcus xylosus was not affected by the IrAM knock-down. Similar results were obtained upon deactivation of all thioester proteins in tick hemolymph by methylamine. We have further demonstrated that phagocytosis of C. indologenes is dependent on an active metalloprotease secreted by the bacteria. These data indicate that interaction of tick α₂-macroglobulin with a protease of an invading pathogen is linked with cellular immune response.
Tailored Functionalization of Plasmonic Ag/C:H:N:O Nanocomposite for Sensitive and Selective Detection of the Lyme Disease Pathogen
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2021, Frontiers in Immunology
Design of a broadly reactive Lyme disease vaccine
2020, npj Vaccines

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Zentralblatt für Bakteriologie

Summary

References (22)

Rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein dye binding

Analyt. Biochem.

Nitrocellulose membrane as an antigen or antibody carrier for screening hybridoma cultures

J. Immunol. Meth.

Prevalence of Borrelia burgdorferi in Ixodes ricinus ticks in Berlin (West)

Zbl. Bakt. Hyg. A

Identification of Borrelia burgdorferi sensu lato tick isolates from Slovakia by PCR typing with 16S rRNA primers

Zbl. Bakt.

Reactivity of European and American isolates of Borrelia burgdorferi with different monoclonal antibodies by means of a microimmunoblot technique

Zbl. Bakt.

Delineation of Borrelia burgdorferi sensu stricto, Borrelia garinii sp. nov., and group VS461 associated with Lyme borreliosis

Int. J. Syst. Bact.

Comparison in the immunological properties of Borrelia burgdorferi isolates from Ixodes ricinus derived from three endemic areas in Switzerland

Epidemiol. Infect.

Investigation of haematophagous artropods for borreliae-summarized data, 1988–1996

Folic Parasitol.

Variations in the occurrence of Borrelia in the ticks, Ixodes ricinus (L.) in the Olomouc District

Cesk. Epidemiol. Mikrobiol. Immunol.

Continuous cultures of fused cells secreting antibody of predefined specificity

Nature (London)

Cleveage of structural proteins during the assembly of the head of bacteriophage T4

Nature (London)

Cited by (37)

Selected phenotypic features of BR91, a unique spirochaetal strain isolated from the Culex pipiens mosquito

IrAM-An α<inf>2</inf>-macroglobulin from the hard tick Ixodes ricinus: Characterization and function in phagocytosis of a potential pathogen Chryseobacterium indologenes

Tailored Functionalization of Plasmonic Ag/C:H:N:O Nanocomposite for Sensitive and Selective Detection of the Lyme Disease Pathogen

Ixodes ricinus salivary serpin iripin-8 inhibits the intrinsic pathway of coagulation and complement

Identification of Tick Ixodes ricinus Midgut Genes Differentially Expressed During the Transmission of Borrelia afzelii Spirochetes Using a Transcriptomic Approach

Design of a broadly reactive Lyme disease vaccine