Transferable R plasmid of Streptococci Ioslation from Diseased Olive Flounder (Paralichthys olivaceus) in Jeju

제주도 양식넙치병어에서 분리된 연쇄상구균의 약제내성 전이성 plasmid

  • Kim, Jong-Hun (Department of Aqualife Medicine, Chonam National University) ;
  • Lee, Chang-Hoon (Jeju Fisheries Research Institute, National Fisheries Research & Development Institute) ;
  • Kim, Eun-Heui (Department of Aqualife Medicine, Chonam National University)
  • 김종훈 (전남대학교 수산생명의학과) ;
  • 이창훈 (국립수산과학원 제주수산연구소) ;
  • 김은희 (전남대학교 수산생명의학과)
  • Published : 2006.12.01

Abstract

Seventy-five streptococci were isolated from diseased olive flounder, Paralichthys olivaceus in Jeju. Their drug susceptibility and transferable multiple drug resistance were characterized. All isolates were resistant to flumequine (AR) and oxolinic acid (OA) and 26 isolates (34.7%) showed 4~6 multiple resistance of ampicillin (ABPC), AR, doxycycline (DOXY), erythromycin (EM), norfloxacin(NOR), OA and oxytetracycline (OTC) in various combinations. pST9 of a transferable R plasmid was detected from a multiple drug resistance strain, Streptococcus sp., ST9 originated from diseased flounder in Jeju, previously. We performed DNA hybridization to know the distribution of plasmid with the same DNA structure as pST9 in streptococci. Thirteen out of 60 isolates analyzed were positive in colony DNA hybridization and the part of bacteria isolated from raw meal was also hybridized with pST9. It suggested that raw meal is one of the origin of the resistance plasmid and R plasmid with DNA structure differing from pST9 is also involving in multiple drug resistance of the streptococci. In conjugation experiment, we found transferable R plasmid carrying OTC, DOXY and/or EM resistance determinant in the 13 resistance strains. all of the streptococci carrying the transferable R plasmid were similar in RAPD patterns. However, pST -type R plasmid was rare in S. iniae most frequently appearing in flounder farm.

제주도내 양식넙치 병어에서 분리된 연쇄상구균 75균주의 항균제에 대한 감수성 경향 및 내성균 출현빈도를 조사하고 다재내성 균주로부터 전이성 R plasmid를 검출하였다. 넙치 병어에서 분리된 모든 연쇄상구균은 flumequine (AR)과 oxolinic acid (OA)에 대하여 저항성이었으며, 그 중 30균주 (41%)는 ampicillin (ABPC), doxycycline (DOXY), erythromycin (EM), norfloxacine (NFLX), oxyteracycline (OTC)의 약제에 대하여 다양한 조합으로 동시내성을 나타내었다. AR과 OA를 포함하여 4~6약제에 대하여 다재내성을 보인 균주는 26주였다. 연쇄상구균의 다재내성 전이에 관여하는 R plasmid인 pST9와 DNA구조가 유사한 plasmid의 분포를 알아보기 위하여 60 분리 균주에 대한 colony 혼성화를 실시하였다. 그 결과 13 분리균에서 혼성화 양성반응이 나타났으며 생사료의 원료로 사용되는 양치와 고등어에서도 양성반응이 나타남으로써 동일 DNA구조의 plasmid가 분포함을 시사하였다. 혼성화 양성반응을 보인 13균주로부터 전이성 R plasmid를 검출하기 위하여 conjugation을 실시하였다. OTC, DOXY, EM에 대한 항균제 내성 marker인 Otc, Doxy, Em은 R plasmid에 의하여 수용균인 Streptococcus sp.로 전이가 이루어졌다. 또한 이들 전이성 R plasmid를 보유하고 있는 연쇄상구균들이 동일 분류군에 속하는지를 알아보기 위하여 RAPD pattern을 분석하였다. 내성균주들의 RAPD pattern은 모두 유사하였으며, Streptococcus iniae type의 RAPD pattern (Kim and Kim, 2003)은 나타나지 않았다. 그러므로 pST9와 동일 DNA 구조의 R plasmid는 S. iniae에서의 출현빈도가 매우 낮을 것으로 예상되었다.

Keywords

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