Anaerobic Bacterium FS03 Conditioned Medium

Conditioned medium (FS03CM) was prepared from an anaerobic bacterium, FS03, which is closely related to Terrisporobacter spp., and had been isolated from soil in a previous study [12 (link)] following the method described by Pahalagedara, Jauregui, Maclean, Altermann, Flint, Palmer, Brightwell and Gupta [27 (link)]. Briefly, colonies of isolate FS03 grown on sheep blood agar (SBA) were used to inoculate the cooked meat glucose starch (CMGS) medium (45 mL) (Fort Richard Laboratories, New Zealand) supplemented with yeast extract (0.0005%), hemin (0.1%), and vitamin K (1%), and incubated inside a 35 °C anaerobic chamber for 48 h. After the incubation period, the culture was centrifuged at 10,000× g for 40 min at 4 °C, and the supernatant was filter-sterilized using a 0.22 µm polyvinylidene fluoride syringe filter (Millipore, Ireland). The sterile conditioned media was aliquoted and stored frozen at −20 °C until use.