생명과학회지 (Journal of Life Science)

  • 제23권9호
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  • Pages.1133-1139
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  • 2013
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  • 1225-9918(pISSN)
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  • 2287-3406(eISSN)
한국생명과학회 (Korean Society of Life Science)
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Serratia marcescens 검출을 위한 PCR 기법 개발 및 돼지정액 유래균주에 대한 항생제 감수성 양상

Specific Detection of Serratia marcescens Based on a PCR Assay and Antimicrobial Susceptibility of S. marcescens Isolated from Boar Semen

  • 정지아 (농림축산검역본부 세균질병과) ;
  • 김애란 (농림축산검역본부 세균질병과) ;
  • 서병주 (농림축산검역본부 세균질병과) ;
  • 정석찬 (농림축산검역본부 세균질병과) ;
  • 김인철 (국립축산과학원 양돈과) ;
  • 정기화 (경남과학기술대학교 동물소재공학과) ;
  • 정병열 (농림축산검역본부 세균질병과)
  • Jung, Ji-A (Bacteriology Division, Animal and Plant Quarantine Agency) ;
  • Kim, Aeran (Bacteriology Division, Animal and Plant Quarantine Agency) ;
  • Seo, Byoung Joo (Bacteriology Division, Animal and Plant Quarantine Agency) ;
  • Jung, Suk Chan (Bacteriology Division, Animal and Plant Quarantine Agency) ;
  • Kim, In Cheul (Swine Science Division, National Institute of Animal Science, Rural Development Administration) ;
  • Chung, Ki Hwa (Department of Animal Resources Technology, Gyeongnam National University of Science and Technology) ;
  • Jung, Byeong Yeal (Bacteriology Division, Animal and Plant Quarantine Agency)
  • 투고 : 2013.07.10
  • 심사 : 2013.09.09
  • 발행 : 2013.09.30
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초록

돼지 원정액의 채취나 희석정액의 제조과정 중 세균오염이 많이 발생하는데, 이는 정자활력의 감소뿐 아니라 모돈의 수태율 저하 등을 유발한다. 특히 Serratia marcescens는 환경에 널리 존재하며 비위생적으로 제조된 정액에 많이 분리되고 있다. 본 연구에서는 S. marcescens의 신속한 동정을 위하여 PCR 기법을 개발하였으며, 국내 돼지정액 유래 S. marcescens을 이용하여 최소생육억제농도(MIC) 등을 조사하고 유효 항생제를 선발하고자 하였다. 개발 PCR 기법은 S. marcescens에서만 306 bp의 특이 유전자 증폭산물을 형성하였으며, 기타 돼지정액에서 분리 보고된 균주나 Serratia 속균에서는 유전자 증폭 산물이 형성되지 않아 특이성이 인정되었다. PCR 기법의 민감도는 S. marcescens에서 추출된 DNA $50pg/{\mu}l$까지 검출이 가능하였다. 디스크확산법에 의한 국내 돼지정액 유래 S. marcescens의 항생제 감수성을 조사한 결과, gentamicin, ceftiofur, neomycin 등에서 80% 이상의 높은 감수성을 보였다. 한편 ceftiofur, enrofloxacin, gentamicin, neomycin의 $MIC_{90}$는 각각 8, 8, 8, $16{\mu}g/ml$로 나타났다. 따라서 개발된 PCR 기법은 S. marcescens를 동정하는 유용한 방법이며, gentamicin 등 선발된 항생제는 S. marcescens에 의한 정액 내 세균오염을 관리하기 위한 희석제용 항생제로 추천된다.

During the collection of boar semen, bacterial contamination usually occurs. The contamination has deleterious effects both on semen quality and on sow fertility. The majority of contaminants are gram-negative bacteria, especially Serratia marcescens. In this study, we developed a PCR assay for the identification of S. marcescens targeting the luxS gene (GenBank no. EF164926). S. marcescens yielded a specific 306 bp PCR product. However, no amplification was observed in the other strains tested. The detection limit of PCR was $50pg/{\mu}l$ of template DNA of S. marcescens. The antimicrobial susceptibility patterns of S. marcescens isolated from boar semen were tested using the disk diffusion method. Gentamicin, ceftiofur, florfenicol, and neomycin showed high sensitivity in this test. The minimum inhibitory concentration (MIC) was also determined by the broth microdilution method. The $MIC_{90}$ values of ceftiofur, enrofloxacin, gentamicin, and neomycin were 8, 8, 8, and $16{\mu}g/ml$, respectively. These results indicate that PCR amplification of the luxS gene is a reliable and effective method for the identification of S. marcescens and that ceftiofur, enrofloxacin, gentamicin, and neomycin are effective semen extenders for controlling S. marcescens.

키워드

참고문헌

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피인용 문헌

  1. Detection of Insect Pathogen Serratia marcescens in Protaetia brevitarsis seulensis (Kolbe) from Korea vol.28, pp.2, 2014, https://doi.org/10.7852/ijie.2014.28.2.25