Lactobacilli isolated from various sources were identified on the basis of 16S–23S rRNA gene intergenic region amplification and subsequent sequencing of the smaller intergenic region. An in vitro analysis of probiotic properties including binding, ability to tolerate different concentrations of bile, survival in acidic buffer and antimicrobial activity of four different isolates and two standard strains (Lactobacillus plantarum American Type Culture Collection (ATCC) 8014 and L. rhamnosus GG (LGG)) was carried out. The ability of each isolate to stimulate Caco-2 cells, human peripheral blood mononuclear cells (PBMC) and THP-1 cells resulting in immunomodulation of these cells was analysed. Isolates L. rhamnosus CS25 and L. delbrueckii M and standard strain ATCC 8014 showed broad antimicrobial activity, and isolates CS25 (percentage of survival 6·9 % at pH 2·5, 5·1 % at pH 2·0) and L. plantarum CS23 (5·7 % at pH 2·5, 4·9 % at pH 2·0) have shown good tolerance to acidic pH. Isolate CS23 showed a good survival (14 %) after 2 h incubation in de Man, Rogosa and Sharpe (MRS) medium containing 3 % bile salts. Isolates CS23, CS25 and L. fermentum ASt1 could stimulate Caco-2 cells, human PBMC and THP-1 cells for a strong and varied immunomodulatory response in these cells. Though LGG showed poor antimicrobial activity as well as bile and acid tolerance, it was found to be the best binding strain tested. Child faecal isolate CS23 from the present study showed high binding ability (seventeen bacteria/Caco-2), high tolerance to acidic pH and bile salts and significant immunomodulation; therefore it is a good potential probiotic candidate.
(Received May 26 2009)
(Revised November 19 2009)
(Accepted December 03 2009)
(Online publication January 14 2010)
Abbreviations: ATCC, American Type Culture Collection; cfu, colony-forming units; LGG, Lactobacillus rhamnosus GG; MRS, de Man, Rogosa and Sharpe; PBMC, peripheral blood mononuclear cells; RPMI, Roswell Park Memorial Institute