There is increasing scientific evidence and commercial interest for using probiotics for eliminating and handling of specific diseases. Probiotics can be evaluated for its role and performance against isolated pathogens from contaminating sources. The present work reports on invitro antimicrobial activity of commercial selected probiotics against pathogenic microbe Vibrio parahaemolyticus. The work also describes cytotoxic activities using MTT assay and adherence studies of selected probiotics. Results for the studies showed maximum zone of inhibition 13.66±0.46mm in probiotic enteroplus,12.33±0.93mm in lactobacillus (NCIM2056) and 10.66±0.93mm in Avant Bact. Cytotoxicity was expressed as IC50(µg/ml) values, observed on CaCO cell lines for different probiotics. Avant Bact showed a IC50 value of 104.7745, Lactobacillus (NCIM2056) a value of 58.13223 and Enteroplus a value of 50.09716. These values expressed different safety aspects of probiotics used for study. Finally the adherence study was done to check probiotic colonizing capacity. The probiotics showed varied adherence capacity against caco cell lines. Enteroplus has % adhesion of 10.25±0.74, Avant Bact. 7.25±0.82 and Lactobacillus (NCIM2056) 7.5±1.12. In conclusion antimicrobial results show importance of probiotics to be used against specific gastro intestinal diseases. Cytotoxicity determines safety aspects of probiotics and adherence study determines probiotic as a promising candidate for in vivo studies.