Improved Isolation of Uncultured Anaerobic Bacteria using Medium Prepared with Separate Sterilization of Agar and Phosphate

Souichiro Kato; Mia Terashima; Ayano Yama; Megumi Sato; Wataru Kitagawa; Kosei Kawasaki; Yoichi Kamagata

doi:10.1264/jsme2.ME19060

Short Communication

Improved Isolation of Uncultured Anaerobic Bacteria using Medium Prepared with Separate Sterilization of Agar and Phosphate

Souichiro Kato, Mia Terashima, Ayano Yama, Megumi Sato, Wataru Kitagawa, Kosei Kawasaki, Yoichi Kamagata

Author information

Souichiro Kato
Bioproduction Research Institute, National Institute of Advanced Industrial Science and Technology (AIST)
Division of Applied Bioscience, Graduate School of Agriculture, Hokkaido University
Mia Terashima
Bioproduction Research Institute, National Institute of Advanced Industrial Science and Technology (AIST)
Institute of Low Temperature Science, Hokkaido University
Ayano Yama
Bioproduction Research Institute, National Institute of Advanced Industrial Science and Technology (AIST)
Hokkaido High-Technology College
Megumi Sato
Bioproduction Research Institute, National Institute of Advanced Industrial Science and Technology (AIST)
Hokkaido High-Technology College
Wataru Kitagawa
Bioproduction Research Institute, National Institute of Advanced Industrial Science and Technology (AIST)
Division of Applied Bioscience, Graduate School of Agriculture, Hokkaido University
Computational Bio Big Data Open Innovation Laboratory (CBBD-OIL), AIST
Kosei Kawasaki
Bioproduction Research Institute, National Institute of Advanced Industrial Science and Technology (AIST)
Yoichi Kamagata
Bioproduction Research Institute, National Institute of Advanced Industrial Science and Technology (AIST)
Division of Applied Bioscience, Graduate School of Agriculture, Hokkaido University
Bioproduction Research Institute, AIST

Keywords: culturability, anaerobic bacteria, wastewater treatment plant, agar medium preparation, phosphate

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Supplementary material

2020 Volume 35 Issue 1 Article ID: ME19060

DOI https://doi.org/10.1264/jsme2.ME19060

Details

Abstract

We previously demonstrated that a simple modification in the preparation of agar media, i.e., autoclaving phosphate and agar separately (termed the “PS protocol”), improved the culturability of aerobic microorganisms by reducing the generation of reactive oxygen species. We herein investigated the effects of the PS protocol on the cultivation of anaerobic microorganisms using sludge from a wastewater treatment system as a microbial source. The application of the PS protocol increased colony numbers and the frequency of phylogenetically novel isolates under aerobic, nitrate reduction, and fermentation conditions. The PS protocol is useful for isolating both aerobic and anaerobic microorganisms.

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