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강원도 양식 연어과 어류에서 분리된 에로모나스 종의 유전학적 동정

Genetic identification of Aeromonas species using a housekeeping gene, rpoD, in cultured salmonid fishes in Gangwon-Do

  • 임종원 (강릉원주대학교 해양생물공학과) ;
  • 구본형 (강원수산질병관리원) ;
  • 김광일 (동해수산연구소 양식산업과) ;
  • 정현도 (부경대학교 수산생명의학과) ;
  • 홍수희 (강릉원주대학교 해양생물공학과)
  • Lim, Jongwon (Department of Marine Biotechnology, Gangneung-Wonju National University) ;
  • Koo, Bonhyeong (Kangwon Fisheries disease control center) ;
  • Kim, Kwang Il (Aquaculture Industry Division, East Sea Fisheries Research Institute) ;
  • Jeong, Hyun Do (Department of Aquatic Life Medicine, Pukyong National University) ;
  • Hong, Suhee (Department of Marine Biotechnology, Gangneung-Wonju National University)
  • 투고 : 2017.11.14
  • 심사 : 2017.12.05
  • 발행 : 2017.12.30

초록

현재 양식장에서는 Aeromonad를 비롯한 다양한 병원균에 의한 전염병으로 인해 많은 경제적 손실을 겪고 있다. 연어과 어류뿐만 아니라 담수 및 해수어류에도 치명적인 감염을 야기하는 Aeromonas 종은 적어도 26종 이상이 보고되어왔으며, 전염병을 유발하는 유비쿼터스 세균이다. Aeromonas 종을 확인하기 위해 16S rDNA 및 하우스 키핑 유전자의 핵산 서열을 기반으로 한 분자생물학적 기술이 사용될 수 있다. 본 연구에서 Aeromonas 종은 강원도 16개 양식장의 연어과 어류로부터 분리되었으며 Aeromonad의 16S rDNA와 하우스 키핑 유전자의 서열, 즉 RNA polymerase sigma factor ${\sigma}^{70}$ (rpoD) 또는 DNA gyrase subunit B (gyrB)를 기반으로 계통 발생 학적으로 동정했다. 그 결과 대서양 연어 (Salmo salar), 은연어 (Oncorhynchus keta), 산천어 (Oncorhynchus masou masou), 무지개송어 (Oncorhynchus mykiss)에서 96 개의 균주가 수집되었으며, 36개의 균주가 16S rDNA 분석에 의해 Aeromonas 속으로 확인되었다. 확인된 Aeromonas 속 균주는 rpoD 또는 gyrB 유전자 서열을 기반으로 추가 분석되어 Aeromonas salmonicida (24 균주), A. sobria (10 균주), A. media (1 균주) 및 A. popoffii (1 균주)로 검출되었으며, 이 것은 Aeromonas salmonicida가 강원도의 연어과 어류에서 주요 감염균임을 나타낸다. 또 하우스 키핑 유전자의 서열에 기초한 Aeromonas 종의 계통발생학적 동정은 16S rDNA 서열보다 더 정확하다는 것이 또한 증명되었다.

At the present, fish farms are suffering a lot of economic losses due to infectious diseases caused by various pathogens including aeromonad. Aeromonad is ubiquitous bacteria that causes infectious diseases. At least 26 species in the genus Aeromonas have been reported to cause fatal infections not only in salmonid fishes, but also in other freshwater and seawater fishes. Molecular techniques based on nucleic acid sequences of 16S rDNA and housekeeping genes can be used to identify the Aeromonas species. In this study, The genus Aeromonas was isolated from salmonid fishes of sixteen fish farms in Gangwon-Do, Korea and phylogenetically identified based on the sequences of 16S rDNA and housekeeping genes for Aeromonad, i.e. RNA polymerase sigma factor ${\sigma}^{70}$ (rpoD) or DNA gyrase subunit B (gyrB). Consequently, 96 strains were collected from Atlantic salmon (Salmo salar), coho salmon (Oncorhynchus kisutch), masou salmon (Oncorhynchus masou) and rainbow trout (Oncorhynchus mykiss), and 36 isolates were identified as the genus Aeromonas by 16S rDNA analysis. Thirty six Aeromonad isolates were further analysed based on rpoD or gyrB gene sequences and found Aeromonas salmonicida (24 isolates), A. sobria (10 isolates), A. media (1 isolates) and A. popoffii (1 isolates), indicating that A. salmonicida is a main infectious bacteria in Salmonid fishes in Gangwon-Do. It was also proved that the phylogenetic identification of Aeromonas species based on the sequences of housekeeping gene is more precise than the 16S rDNA sequence.

키워드

참고문헌

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