Enumeration of Lactobacilli in the Fecal Flora of Infant Using Two Different Modified de-Man Rogosa Sharpe Media under Aerobic and Anaerobic Incubation

Regarding the importance of the presence of intestinal lactobacilli and their population in infants, four different treatments were evaluated for Lactobacillus isolation efficiency via reduction in the growth of other groups of bacteria capable of growing on de-Man Rogosa Sharpe (MRS) medium from fecal samples of 11 Iranian infants. MRS-Vancomycin (1 mg L-1) was used as a base medium and application of lactic acid and aerobic incubation of inoculated plates were performed as selective factors. Each fecal sample was cultivated as duplicate on to the base medium with or without lactic acid to reduce the pH to 5.4 ±0.2. Half of the plates were incubated aerobically and the rest of them incubated under 10% CO2 concentration. Total count and Lactobacillus count of all samples were recorded according to the age differences of infants. The counts of false positive colonies were recorded with respect to their cell morphology and gram reaction in all treatments. Anaerobic incubation of lactic acid modified MRS-Vancomycin gave the most Lactobacillus percentage coverage, about 93% among the Lactobacillus positive samples. Using this treatment, the median Lactobacillus count yielded 8.29 log10 cfu g-1 in the younger and 5.70 log10 cfu g-1 in the elder group. It could be concluded that lactic acid might be a proper pH reducing agent when enumeration of lactobacilli from fecal samples is of interest.