This mixture was then ground in a
homogenizer (Stomacher 400-T, Organo) for 60 s to obtain a sample
extract; 9ml of sterilized water was added to 1ml of the sample extract
to make a 1:100 diluent. One milliliter of the 1:100 diluent was
dispensed in five tubes, each containing thioglycollate medium (Eiken
Chemical) heat-dissolved in distilled water and sterilized (hereinafter
referred to as the TGC liquid medium).