Gifu anaerobic medium agar

Manufactured by Nissui Pharmaceutical

Sourced in Japan

Gifu Anaerobic Medium (GAM) agar is a culture medium used for the isolation and cultivation of anaerobic bacteria. It provides the necessary nutrients and growth conditions for the proliferation of anaerobic microorganisms.

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4 protocols using gifu anaerobic medium agar

Quantifying Gut Microbiome Diversity

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Small intestinal mucosal samples from three jejunal and ileal sites were homogenized and 10-fold diluted in anaerobic diluent (KH₂PO₄ 4.5 g, Na₂HPO₄, 6.0 g, L-cysteine hydrochloride 0.5 g, Tween 80 0.5 g and agar 0.5 g in 1 L of distilled water),^{(14 (link))} and stored at –80°C for later analysis.
The total viable aerobic and facultative anaerobic bacterial counts were determined by culture method on MacConkey agar (Nissui Pharmaceutical Co., Ltd., Tokyo, Japan) and Brain Heart Infusion (BHI) agar (Difco, Detroit, MI) containing 5% horse blood as described previously.^{(16 (link))} The viable obligate anaerobe counts were cultured on Gifu Anaerobic Medium (GAM) agar (Nissui Pharmaceutical Co., Ltd.).

Kurata S., Nakashima T., Osaki T., Uematsu N., Shibamori M., Sakurai K, & Kamiya S. (2014). Rebamipide protects small intestinal mucosal injuries caused by indomethacin by modulating intestinal microbiota and the gene expression in intestinal mucosa in a rat model. Journal of Clinical Biochemistry and Nutrition, 56(1), 20-27.

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Microbiological and Metabolic Analysis of Godo Samples

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Four homemade godo samples were obtained in Towada City. Materials and production process of each sample were shown in Fig. 2.
All samples were brought to the laboratory in a cooler box and stored in the refrigerator.
For microbial count, serial dilutions of godo samples in saline and four kinds of agar media were used. Standard plate count agar (SPC agar; Nissui, Tokyo, Japan) was incubated under aerobic condition at 35°C for 2 days for counting the total aerobic bacteria. The counts of lactic acid bacteria were measured by plate count agar with bromocresol purple (BCP agar; Nissui) with 10 ppm sodium azide and 10 ppm cycloheximide, and yellow colonies were counted after 2-3 days of incubation at 30°C. Gifu anaerobic medium (GAM agar; Nissui) was incubated under anaerobic condition at 35°C for 2 days for counting anaerobic bacteria. Potato dextrose agar (PD agar; Nissui) with 0.01% chloramphenicol was incubated under aerobic condition at 25°C for 5-7 days for counting fungi and yeast.
Biochemical analysis of metabolic products. Concentrations of lactic acid, acetic acid, and ethanol in godo samples were determined enzymatically using F-kit (Roche Diagnostics K.K., Tokyo, Japan) following the manufacturer's instructions.

Shimodate K, & Honda H. (2023). Isolation and identification of psychrotrophic lactic acid bacteria in godo, the traditional fermented soy food in Japan. The Journal of general and applied microbiology, 68(5).

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Bacterial Peptidoglycan Preparation

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Gifu anaerobic medium agar was purchased from Nissui Pharmaceutical Co., Ltd. (Tokyo, Japan). Tryptic soy broth was purchased from Becton Dickinson (Franklin Lakes, NJ, USA). Protease K was purchased from QIAGEN (Hilden, Germany). RNase A was purchased from NIPPON GENE, Co., Ltd. (Tokyo, Japan). DNase was purchased from Promega Corporation (WI, USA). Lysostaphin, lysozyme, methanol, and chloroform were purchased from FUJIFILM Wako Pure Chemical Corporation (Osaka, Japan). Staphylococcus aureus peptidoglycans, Bacillus subtilis peptidoglycans, and Micrococcus luteus peptidoglycans were purchased from Sigma-Aldrich (St. Louis, MO, USA).

Matsumoto Y., Sato E, & Sugita T. (2022). Acute melanization of silkworm hemolymph by peptidoglycans of the human commensal bacterium Cutibacterium acnes. PLoS ONE, 17(9), e0271420.

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Anaerobic Culturing and Chemical Profiling

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Gifu anaerobic medium agar was purchased from Nissui Pharmaceutical Co., Ltd. (Tokyo, Japan). Tryptic soy broth was purchased from Becton Dickinson (Franklin Lakes, NJ, USA). Protease K was purchased from QIAGEN (Hilden, Germany). Propionic acid, acetic acid, lactic acid, and hydrochloric acid were purchased from FUJIFILM Wako Pure Chemical Corporation (Osaka, Japan).

Matsumoto Y., Sato E, & Sugita T. (2024). Acid-treated Staphylococcus aureus induces acute silkworm hemolymph melanization. PLOS ONE, 19(5), e0298502.

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