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Antibacterial effects of natural products on Vibrio vulnificus growth in seawater and shellfish

바닷물과 조개에서 패혈증비브리오균 증식에 대한 천연물의 항균 효과

  • Hong, Yeongjin (Department of Microbiology, Chonnam National University Medical School) ;
  • Ryu, Phil Youl (Department of Microbiology, Chonnam National University Medical School) ;
  • Kim, Seung Yong (College of Pharmacy, Chonnam National University) ;
  • Yeom, Kyung Min (College of Pharmacy, Chonnam National University) ;
  • Park, Jung Up (College of Pharmacy, Chonnam National University) ;
  • Jo, Se Jin (College of Pharmacy, Chonnam National University) ;
  • Guo, Rui Hong (College of Pharmacy, Chonnam National University) ;
  • Tran, The Bach (Institute of Ecology and Biological Resources (IEBR), Vietnam Academy of Science and Technology (VAST)) ;
  • Jin, Hang (Institute of Medicinal Plants, Yunnan Academy of Agricultural Sciences (YAAS)) ;
  • Kim, Young Ran (College of Pharmacy, Chonnam National University)
  • Received : 2019.10.28
  • Accepted : 2020.02.04
  • Published : 2020.02.29

Abstract

Vibrio vulnificus, an opportunistic pathogen, causes septicaemia when raw shellfish and fish are eaten by patients with hepatic diseases or reduced immunity. In this study, we evaluated inhibitory effects of some natural products on V. vulnificus growth using 96-well microplate assay. We found that Phyllanthus emblica L., Rosa chinensis Jacq., Rosa rugose Thub., and Chukrasia tabularis A. Juss. significantly inhibited V. vulnificus growth in Luria-Bertani (LB) broth. Among these four extracts, the inhibition diameter of Chukrasia tabularis was 16.00 ± 0.58 mm in disc diffusion assay on V. vulnificus growth. In addition, these four natural products protected HeLa cells from V. vulnificus-induced cytotoxicity. A cocktail containing these four products showed an inhibitory effect on V. vulnificus growth in seawater and shellfish by reducing its growth by 75.7% and 97%, respectively. These results suggest that these four natural products are safe and effective natural antimicrobial candidates to prevent V. vulnificus infection.

본 연구에서는 패혈증비브리오균에 대한 PEM, RCM, RRM과 CTM 메탄올 추출물의 항균 활성을 평가하였다. 96-well microplates를 사용한 액체 배양법 실험 결과 4가지 천연물은 200 ㎍/mL 농도에서 80%이상의 항균 효과를 나타내었다. 또한 페이퍼디스크 확산법을 사용하여 항균 활성을 평가한 결과 4가지 천연물은 20, 40 mg/disc의 농도에서 농도 의존적으로 패혈증비브리오균의 성장을 억제하였다. 다음으로 4가지 천연물의 안전성을 평가하기 위해 MTS assay를 통해 HeLa 세포에 대한 독성을 확인한 결과 100, 200 ㎍/mL의 농도에서 세포 독성을 나타내지 않았다. 또한 LDH assay를 사용하여 패혈증비브리오균의 HeLa 세포에 대한 세포막 손상 억제 효과를 평가한 결과 PEM, RCM, RRM과 CTM 메탄올 추출물은 200 ㎍/mL 농도에서 세포막 손상억제 효과를 나타내었다. 다음으로 바닷물과 조개를 사용한 수족관 모델에서 4가지 천연물의 메탄올 추출물 혼합액 200 ㎍/mL 농도는 colony 생성을 현저하게 감소시켜 패혈증비브리오균의 증식을 억제 하는 결과를 나타내었다. 이와 같은 결과를 통해 4가지 천연물의 메탄올 추출물은 바닷물과 해산물에 존재하는 패혈증비브리오균에 대해 뛰어난 항균효과를 나타내면서 인체에는 무해한 천연소독제로 응용할 수 있을 것으로 판단된다.

Keywords

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