Fermentation of non-sterilized fish biomass with a mixed culture of film-forming yeasts and lactobacilli and its effect on innate and adaptive immunity in mice
Section snippets
Culture medium
Medium for each microbe culture was prepared after a textbook (11). MRS (Man, Rogosa, and Sharpe) medium was used for LAB culture, YG (Yeast extract-glucose) medium for yeast culture, and PCA (Plate count agar) medium for general bacteria culture. The composition of each medium is shown in Table 1. After preparation, media were sterilized by heating (121 °C, 10 min). In the case of solidification, two percent of agar was added to each media.
Peptone [Bacto peptone (Difco)] and yeast extract were
Cultivation characteristics of C. ethanolica
After preculturing in YPD2 medium at 27 °C for 24 h with slight shaking (40 rpm), 35 ml of precultured C. ethanolica broth was inoculated to 700 ml YPD13 medium in a 1-L cylindrical vessel, and cultivated at 27 °C for 220 h. After 24 h of cultivation, some ring-shaped masses of yeasts were observed, and by 72 h, a thick film layer had formed. After 72 h, the film layer was almost constant in a thickness. During an entire period of cultivation, ethanol concentration, sugar concentration and pH
Acknowledgments
We appreciate Dr. Hideo Tsukamoto, Ms Yoshiko Shinozaki, and Ms Kayoko Iwao, the Education and Research Support Center, Tokai University, for their valuable advice and technical support in this study. We also appreciate Mr. Satoru Serizawa, Cooperative Numazu Suisan Kaihatsu Center, for his kindly supply of fish biomass with valuable information. Special thanks also to Ms Kimiyo Shibata, Ms Takako Kato, and Mr. Ryota Seki for their technical support in this study.
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Present address: Yamazaki Baking Co. Ltd, Kanagawa 244-8525, Japan.
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Present address: Maruzen Food Industry Co. Ltd, Shizuoka 410-1312, Japan.