Distribution and Identification of Halophilic Bacteria in Solar Salts Produced during Entire Manufacturing Process

천일염 생산공정별 미생물 분포 조사 및 호염미생물 동정

  • Na, Jong-Min (Functional Food & Nutrition Division, Department of Agrofood Resources, National Academy of Agricultural Sciences (NAAS), RDA) ;
  • Kang, Min-Seung (Functional Food & Nutrition Division, Department of Agrofood Resources, National Academy of Agricultural Sciences (NAAS), RDA) ;
  • Kim, Jin-Hyo (Chemical Safety Division, National Academy of Agricultural Sciences(NAAS), RDA) ;
  • Jin, Yong-Xie (Functional Food & Nutrition Division, Department of Agrofood Resources, National Academy of Agricultural Sciences (NAAS), RDA) ;
  • Je, Jeong-Hwan (Functional Food & Nutrition Division, Department of Agrofood Resources, National Academy of Agricultural Sciences (NAAS), RDA) ;
  • Kim, Jung-Bong (Functional Food & Nutrition Division, Department of Agrofood Resources, National Academy of Agricultural Sciences (NAAS), RDA) ;
  • Cho, Young-Sook (Functional Food & Nutrition Division, Department of Agrofood Resources, National Academy of Agricultural Sciences (NAAS), RDA) ;
  • Kim, Jae-Hyun (Functional Food & Nutrition Division, Department of Agrofood Resources, National Academy of Agricultural Sciences (NAAS), RDA) ;
  • Kim, So-Young (Functional Food & Nutrition Division, Department of Agrofood Resources, National Academy of Agricultural Sciences (NAAS), RDA)
  • 나종민 (농촌진흥청 국립농업과학원 농식품자원부 기능성식품과) ;
  • 강민승 (농촌진흥청 국립농업과학원 농식품자원부 기능성식품과) ;
  • 김진효 (농촌진흥청 국립농업과학원 농산물안전성부 유해화학과) ;
  • 김영섭 (농촌진흥청 국립농업과학원 농식품자원부 기능성식품과) ;
  • 제정환 (농촌진흥청 국립농업과학원 농식품자원부 기능성식품과) ;
  • 김정봉 (농촌진흥청 국립농업과학원 농식품자원부 기능성식품과) ;
  • 조영숙 (농촌진흥청 국립농업과학원 농식품자원부 기능성식품과) ;
  • 김재현 (농촌진흥청 국립농업과학원 농식품자원부 기능성식품과) ;
  • 김소영 (농촌진흥청 국립농업과학원 농식품자원부 기능성식품과)
  • Received : 2010.10.05
  • Accepted : 2011.05.25
  • Published : 2011.06.28

Abstract

In this study, we determined the changes in microbial numbers in solar salts according to the manufacturing process and storage duration. The salt samples were harvested from salt farms in Shinan (area 2) and Yeonggwang (area 1). They were serially diluted ten-fold and then placed on 4 kinds of cultivable media (mannitol salt agar, eosin methylene blue, plate count agar, and trypticase soy agar). After incubation, we obtained 62 halophilic isolates from the salt samples. Coliform and general bacteria were not detected in all salt samples. By 16S rRNA sequencing analysis, we found 12 kinds of halophilic bacteria belonging to the genera Halobacillus, Halomonas, Bacillus, Idiomarina, Marinobacter, Pseudoalteromonas, Vibrio, Salinivibrio, Virgibacillus, Alteromonas, Staphylococcus and some un-known stains. In our study, we discovered two novel species that have a 16S rDNA sequence similarity below 97%.

우리나라 전남지역에서 생산되는 천일염의 생산공정별 미생물 분포 조사 및 배양 분리법을 이용하여 호염미생물을 분리하여 동정하는 것을 이번 연구의 목표로 삼았다. 천일염 시료는 생산지를 고려하여 육지 염전인 영광군 한 지역과 해안 염전 지역인 신안군 두 군데에서 생산공정별로 세분화하여 총 28개 분석시료를 수집하여 사용하였으며, 이들 시료를 십진 희석하여 4종류의 미생물 분리용 배지에 도말, 배양한 후 나타난 집락(colony)을 계수하였다. 그 결과 천일염 생산 공정 중 대장균 등의 식품 오염지표 미생물은 검출되지 않았지만, 저장수에서 증발지 단계로 진행되면서 $1.1{\times}10^3{\sim}1.8{\times}10^5$ CFU/g의 호염성 미생물들이 검출되었다. 그러나 이후 단계인 함수저장고, 결정지에서는 미생물 수가 점차적으로 감소되었으며, 소금저장소에 보관된 천일염 시료에서는 미생물이 전혀 검출되지 않았다. 이들 분리균들은 형태학적 특성에 따라 무작위로 62개 집락을 선정하여 분리하였다. 순수 분리된 미생물들은 PCR 기법을 통해 16S rRNA 유전자의 염기서열을 분석한 후, 기존에 보고된 미생물 유전자 database와 비교함으로써 12속의 천일염 유래 호염균들의 존재를 확인하였다. 이번 연구 결과 천일염 생산 단계에서 분리된 halophilic bacteria은 Halobacillus, Halomonas, Bacillus, Idiomarina, Marinobacter, Pseudoalteromonas, Vibrio, Salinivibrio, Virgibacillus, Alteromonas, Staphylococcus 및 un-known 등이다.

Keywords

References

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