IN VITRO EVALUATION OF PROBIOTIC ATTRIBUTES OF LACTOBACILLUS SPP ISOLATED FROM DIFFERENT SOURCES Monica Puniya1, Nikhil Kumar, Anita Kumari Garsa, Taufeeq Ahmed, Ravinder Kumar Malik and Anil Kumar Puniya Dairy Microbiology Division, National Dairy Research Institute, Karnal-132001, Haryana. Introduction Genus specific PCR Cell Surface Hydrophobicity “Probiotics are living microorganisms which upon ingestion in sufficient numbers, exert health benefits beyond basic nutrition.” The real concept of using beneficial microorganisms for improved human health is an area of increasing interest for scientific community. The useful effects of probiotics are results of favorable changes in the intestinal health and immune system. The present study was aimed at studying some of the probiotic attributes of lactobacilli isolates for their possible use in controlling the lifestyle related diseases such as obesity. Cell surface hydrophobicity Antibiotic susceptibility Antibiotics C3 C11 9K 12K 17K Amikacin30(AK) R Amoxillin30(AM) S I Ampicillin10(A) Ciprofloxacin10 (CF) Cefuroxime (CU) Cephalokhin(CH) Chloramphenicol (C) Clindamycin(CD) Erythromycin(E) Novobiocin(NV) Kanamycin30(K) Oxytetracylin(O) Primer Sequence: LbLMA 1(5’CTC AAA ACT AAA CAA AGT TTC 3’) and R16-1 (5’CTT GTA CAC ACC GCC CGT CA3’) Methods Acid & Bile tolerance Isolation(different dairy and non-dairy sources) & confirmation of Lactobacillus strains using PCR, catalase test and microbiological examination Screening of Lactobacilli isolates for their survival at different pH (2, 2.5, 6.5) and bile concentrations (0.5%, 1%, 1.5%, 2%) at 0, 1, 2 & 3hrs resp. Acid Tolerance of C3 and C11 Screening of isolates for bile salt hydrolase (BSH) activity Susceptible≥20mm Intermediate≤15-19mm Resistant ≤ 4mm Evaluation of cell surface hydrophobicity , antibiotic susceptibility and anti-bacterial activity Results Bile Tolerance of C3 and C11 Antibiotic Susceptibility Bile salt hydrolase (BSH) assay Conclusions Out of one hundred Lactobacillus isolates, fifteen were exposed for different probiotic attributes & five isolates were found to have good in vitro tolerance and, therefore, can be exploited for their application in functional dairy foods. Overall, these strains may possibly be the probiotics that can be further validated through in vivo studies to elucidate their potential health benefits. Microscopical Examination Gram Staining References Antibacterial Spectrum of Activity FAO/WHO 2002.Guidelines for the evaluation of probiotics in food. Ontario, Canada, USA: Food and Agricultural Organisation. Performance Standards for Antimicrobial Disk Susceptibility tests. 2007. Clinical and Laboratory Standards Institute (CLSI):27(1). ICMR-DBT 2011.Guidelines for evaluation of probiotics in food.New Delhi, India: Indian Council of Medical Research. BSH activity Indicator strain C3 C11 9K 12K 17K Lp P.acedila-ctici Lb 42 ++ +++ Bile Salts C3 C11 9K 12K 17K Lr Sodium deoxycholate (SD) +++ Sodium taurocholate (ST) + Sodium tauroglycocholate (STG) ++ Acknowledgements We gratefully acknowledge DBT, Biocare Women Scientist scheme for funds & Director, NDRI for providing necessary infrastructure & facilities. Antibacterial Spectrum Diameter of zone of inhibition(mm)10-20=+++, 5-10 =++, <5=+ ++: moderate precipitation; +: slight -precipitation; +++: intense precipitation