Isolation of spirochetes from Japanese ixodid ticks, Ixodes tanuki, Ixodes turdus, and Ixodes columnae

Abstract

Spirochaetes were isolated from midgut tissues of ticks and cultured in BSK medium. These were: Hk501 from an adult of Ixodes tanuki fed on a raccoon dog (Nyctereutes procyonoides albus) in Hokkaido, Japan; Iw501 from an unfed female of I. tanuki in Iwate; Ya501 from an adult of I. turdus fed on the bulbul Hypsipetes amaurotis in Yamagata Prefecture; Ac502 from an adult of I. turdus on H. amaurotis in Aichi Prefecture; and AM501 from an unfed nymph of I. columnae [see N. Takada & H. Fujita (1992) Journal of the Acarological Society of Japan, 1: 37-44] collected by flagging vegetation in Aomori Prefecture. Whole-cell lysates of the spirochaetes were subjected to SDS-PAGE and Western blotting using various monoclonal antibodies as probes. None of these 5 isolates were identical to the North American B31 strain of Borrelia burgdorferi in comparison with protein profiles. The epitope for MAb H9724 was present in the 41-kDa flagellin protein of all isolates examined. Both Hk501 and Iw501 possessed the 32-kDa OspA protein reactive with MAb H5332. MAb H5332 also reacted with the 33-kDa protein of Ya501. The reactivities of MAbs confirmed that the isolates were strains of B. burgdorferi. The 2 isolates from I. tanuki were almost identical with each other in their protein compositions.