Background: A number of infections and deadly diseases caused by different bacteria, as Pseudomonas aeruginosa, is the main cause of nosocomial infections and other lethal infections among immunocompromised infants. Objective: The main purpose of this study was to isolate P. aeruginosa from infants’ sputum samples suffering from pulmonary infections and to assess their sensitivity against isolated Lactobacilli from different dairy products, meat and coconut water. Methodology: Sputum samples were collected from hospital. Isolation and screening of P. aeruginosa was done on different media and was confirmed by performing different biochemical tests. Simultaneously, Lactobacilli isolation from different samples was done. Molecular characterization of isolated pathogenic P. aeruginosa and Lactobacilli was done by ribotyping. Sequences were submitted at National Center for Biotechnology Information to obtain accession number of isolated strains. Antibacterial activity of Lactobacilli in comparison to other antibacterial agents was checked by agar well diffusion method. Results: Accession number of P. aeruginosa and Lactobacilli were obtained which confirm the isolation of desired strains. Results of antibacterial activity indicated that some antibiotics inhibited the growth of P. aeruginosa quickly but some strains of P. aeruginosa showed resistance against some generally used antibiotics. Similarly some strains of P. aeruginosa were sensitive to herbal extracts and exhibited zone of inhibition but some herbal extracts exhibited no antibacterial activity. But astonishingly, all strains of P. aeruginosa exhibited sensitivity against Lactobacilli. No doubt, Lactobacilli showed antibacterial activity slowly against pathogenic strains but it exhibited. Conclusion: From this study, it can be concluded that use of probiotics and herbal or plant based medicines to inhibit the growth of pathogen are beneficial than the use of antibiotics. As, antibiotics temporary effect the growth of pathogen rapidly but excessive use of antibiotics becomes the reason of pathogen’s resistance and also cause other side effects in patients.