Isolation of Anaerobic Dairy Microbes

The pour plate technique was used to isolate the microorganisms [36 ]. For milk samples, 1 mL milk was inoculated into 9 mL of sterile saline solution (S.S.S) whereas, for the solid dairy products, 10 grams were aseptically homogenized in 90 mL of S.S.S. Samples were serially diluted (10 fold) then 1 mL aliquot of each dilution was plated into 9 mL of molten De Man, Rogosa, and Sharpe (MRS) medium agar (Sigma-Aldrich). Inoculated MRS plates were incubated anaerobically at 37°C for 48–72 hrs. in an anaerobic jar (GasPak System—Oxoid, Basingstoke Hampshire, England) using AnaeroPack™-Anaero anaerobic gas generator sachets (Mitsubishi, Japan). Typical colonies were selected according to the morphological characters described by [37 (link)], picked from the plates, and repeatedly sub-cultured to obtain pure isolates.