2013년 생물공학회 추계학술대회, 2013, 02, 217─217

Effect of Protectants on Survivability of Freeze Dried Vibrio Pathogens during Long-term Storage

HyunBae KANG, Na-Ri SHIN, Kwang-Jun Lee

Abstract

Preservation of useful microorganisms in culture collection requires successful long-term storage and reactivation techniques. However, some species like Vibrio often fail to long-term preservation, resulting in loss of them. The aim of this study is therefore to develop a simple and effective method for long-term preservation of Vibrio pathogens including V. vulnificus, V. parahaemolyticus, and V. cholerae. Bacterial cells purely cultured were harvested at the early exponential stage, suspended in 10% skim milk and/or 5% inositol solution as a protectant, and aliquoted in sterilized ampoules. After frozen and dried at -80℃ under vacuum, the ampoules were stored for 2 weeks at 37℃ for accelerated test. Freeze-dried bacterium was observed under the electronic microscope and their survivability was determined by flow cytometry and colony count methods. The data from three species commonly showed great viability (108 CFU ml-1) when cultured in 3% NaCl-LB broth after preserved in 5% inositol solution. These suggest that 5% inositol could be a good candidate as a cryo- and lyo-protectant for Vibrio pathogens and LB broth with 3% NaCl positively helps them to reactivate and proliferate.

• 본 연구는 질병관리본부 연구개발과제(과제번호 2013-NG45006-00) 연구비를 지원받아 수행되었습니다.
• This research was supported by a fund(code 2013-NG45006-00) by Research of Korea Centers for Disease Control and Prevention.