Modified gam broth

Brain-Heat Infusion (BHI) agar was purchased from Becton Dickinson and Company (Franklin Lakes, NJ). Modified GAM broth and modified GAM agar were purchased from Nissui Pharmaceutical Co., LTD (Tokyo, Japan). Porphyromonas gingivalis ATCC 33277, Fusobacterium nucleatum ATCC 25586, Streptococcus sanguinis ATCC 10556, and Streptococcus mutans ATCC 25175 were purchased from the American Type Culture Collection (Manassas, VA). P. gingivalis and F. nucleatum were grown on GAM broth and GAM agar, and S. sanguinis and S. mutans were grown on BHI broth and BHI agar in an anaerobic chamber with Anaeropack Kenki (Mitsubishi Gas Chemical Company, Tokyo, Japan) at 37 °C.

F. varium Fv113-g1 was isolated from the colon mucus membrane of a patient with UC [15 (link)]. Briefly, the mucus biopsy specimens were first incubated with imipenem (50 μg/ml saline) for 1 hour at 37° C to inactivate extracellular bacteria organisms, and then serially diluted samples were spread on agar plates for anaerobic culture to isolate intracellular bacteria in colon mucus epithelium. Isolated bacteria were examined by the disk diffusion test for imipenem susceptibility (minimal inhibitory concentration less than 4 μg/ml). Fv113-g1 was cultivated in modified GAM broth (Nissui Pharmaceutical Co., Ltd., Tokyo, Japan) or on FM agar (Becton Dickinson, New Jersey, USA) under anaerobic conditions at 37°C.
On the basis of the ATCC product sheets, no sufficient information of pathogenicity for ATCC 8501 and 27725 have been stated; in addition, those isolates are categorized as biosafety level 1, suggesting that those are commensal bacteria as one of human gut flora.