우리 나라 천해양식장 주변의 테트라사이클린 내성균에서 나타나는 tet genes의 분포
Distribution of tet genes in the tetracycline resistant microorganisms isolated from marine-aquatic environments of Korea
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초록
각각의 6종 tet gene을 유사한 크기의 DNA로 cloning한 후 plasmids를 이용하여 multipex PCR을 실시하여 6개의 template를 혼합한 경우까지 모든 경우에서 뚜렷한 특이적 band를 확인할 수 있어 본 방법의 정확성과 분별력 그리고 활용성의 가치를 보다 명확하게 확인할 수 있었다. 확립된 본 기법을 이용하여 양어장 환경에서 채집된 세균총을 분석한 결과, 양식장 주변 해수에서는 6.7%, 저질에서는 24.4% 그리고 net의 부착물질에서는 30.7%의 내성균 빈도를 보였으며, 항생제를 투여한 조피볼락의 장내 세균에서의 내성균 빈도는 2.83%, 항생제를 투여하지 않은 조피볼락은 0.25%의 내성균 빈도를 보여 항생제를 투여한 어류가 항생제를 투여하지 않은 어류에 비하여 높게 나타났다. 분리된 내성균에는 tet (B), tet (C)만을확인할 수 있어 대부분이 두 종류의 내성 유전자에 편중되게 나타남을 알 수 있었다.
Abstract
Expanded potential of multiplex PCR to discriminate six different tet genes in a single reaction was confirmed using the cloned plasmids as targets genes, even applicable for the nucleic acid contained six different tet genes as a pool. Clearly, six different lengths of amplicons after multiplex PCR using the mixture of cloned-plasmids of six different tet genes as templates allowed us to discriminate the specific types of tet genes. With this method, we determined the distribution of tet genes utilized in the microorganisms present in the environments of the marine-aquatic farms of Korea. The frequency of tetracycline (Tc) resistant bacteria present in the seawater, sediment and net was 6.7%, 24.4% and 30.7%, respectively. Interestingly, it was found that these isolates resistant to tetracycline were utilizing relatively restricted tet genes, tet (B) and (C), rather than various tet genes. It is the first report described the proportion of Tc resistant bacteria with analysis of the specific tet genes in the bacteria isolated from the aquatic farms.
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