The present study aimed to analyze the colonization potential of indigenous probiotic strains and to assess their effects on physiology of Caenorhabditis elegans. The protective effect of probiotics was evaluated in terms of increase in life span of the worm through colonization in the intestine. A total of 15 probiotic cultures were evaluated for their effect on mean life span, pharyngeal pumping, and normal reproduction behavior in the worms. The chemotactic behavior in terms of binary choice index was also evaluated. The adherence and colonization of the intestine of the worm by probiotics were monitored at different time intervals by enumerating the microbial population and fluorescent microscopic observations. The survival analysis-based Kaplan-Meier method indicated that the probiotic cultures increased the survival probability as compared to control strain E. coli OP50. There was no effect of feeding probiotics on physiological responses of the worm such as pharyngeal pumping and reproduction. The principal component analysis (PCA) of the results indicated Lactobacillus plantarum K90 and L. paracasei CD4 as potential probiotics with binary choice index of 0.8 as food preference of C. elegans. The strains exhibit higher adherence and colonization in the gut of worms and increased the life span by 5 days as compared to control E. coli OP50. In conclusion, feeding with probiotic cultures is effective in extending the lifespan of C. elegans; however, the colonization ability differs among the strains.