Fecal Microbiome Analysis in Rats

On days 0, 7, 14 and 21, a pooled fecal sample from the rats in each subgroup was taken. Just after collection, each subgroup’s fecal sample was weighed and homogenized in sterile phosphate buffer saline (PBS), and then, tenfold serial dilution was made in PBS. Aerobic and anaerobic microorganisms were cultured in Plate Count Agar (Merck, Germany). Lactic acid bacteria (LAB) were cultured in MRS (de Man, Rogosa, Sharpe) agar (Merck, Germany). Coliforms were cultured in VRBL (Violet Red Bile Lactose) agar (Merck, Germany). LAB and anaerobic bacteria were incubated at anaerobic condition. All plates were incubated at 37°C for 48 h. The microflora enumeration was expressed as CFU/gr of feces. In order to count bacterial spores in fecal samples, the homogenized samples in PBS were put into a water bath at 80°C for 10 min, and then cooled immediately to help the spore germination. Then the samples were serially diluted in PBS and cultured in Plate Count Agar (Merck, Germany). The plates were incubated at 37°C for 48 h. The spore enumeration was expressed as spores/gr. All the samples were cultured in duplicate (17 (link), 19 ).