BHI-BRAIN HEART INFUSION BROTH

Protocol tips

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	-All the cultures were stored at −80°C in broth supplemented with 20% glycerol. -Cultural characteristics was observed after incubation at 35-37°C for 24-48 hours.

Protocol tips
-All the cultures were stored at −80°C in broth supplemented with 20% glycerol. -Cultural characteristics was observed after incubation at 35-37°C for 24-48 hours.

Publication protocol

For the isolation of lactobacilli, fecal samples were collected from 32 healthy children of age group ranging from 2 to 13 yrs after taking the written informed consent of their parents. The study was approved by the Institutional Human Ethics Committee. The stool sample weighing approximately 1 g was collected in thioglycollate broth (HiMedia laboratories, Mumbai, India) and incubated for 4 h at 37°C in anaerobic jars having 5% carbon dioxide (CO2). Thereafter, 10-fold serial dilutions of the broth were plated onto De Man Rogosa and Sharpe (MRS; HiMedia) agar plates and incubated at 37°C under anaerobic conditions in anaerobic gas jars. Bacterial colonies with different morphologies were selected and preserved in 20% (v/v) glycerol (HiMedia)-containing MRS broth at −80°C. The lactobacilli were identified by Gram-positive staining and catalase-negative phenotype. For experimental purposes, the lactobacilli were cultured in MRS medium from the frozen stocks and propagated twice before use.

The various pathogenic indicator strains used in this study were V. cholerae strain 0139 MTCC 3906, Salmonella enterica Typhimurium MTCC 733, Listeria monocytogenes MTCC 657, Escherichia coli MTCC 119, Shigella flexeri MTCC 1457, V. parahaemolyticus MTCC 451, and Staphylococcus aureus MTCC 96. These strains were procured from Microbial Type Culture Collection, Institute of Microbial Technology, Chandigarh, India. The fungal indicator strain Candida spp. was procured from the Department of Microbiology, Government Medical College, Amritsar. All the pathogenic bacteria were cultured in brain heart infusion (BHI) broth (HiMedia) and Candida spp. was cultured in Sabouroud dextrose broth (HiMedia) at 37°C under aerobic conditions. All the cultures were stored at −80°C in broth supplemented with 20% glycerol.

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