Antibiotic resistance among various pathogens has caused an alarming decrease in the effectiveness of available drugs. Biofilm mode of bacteria provide it strength to neglect the effects of antibiotics. Worldwide efforts are being made to overcome this problem. In this regard, lactic acid bacteria, specifically those of the genera Lactobacillus, are considered important because of their reported antagonistic potential. Present study was conducted to check the antimicrobial and antibiofilm activity of Lactobacillus brevis isolated from fecal matter of herbivores and carnivores, against three pathogenic bacteria i.e., Pseudomonas aeruginosa, Staphylococcus aureus and Escherichia coli. The strains An27 and An15 were isolated from horse and cow (herbivores), respectively, while Cat and G1 were isolated from cat and lion (carnivores), respectively. Antibacterial activity was accessed by agar well diffusion method and antibiofilm activity was accessed by crystal violet microtiter plate method. All the isolates showed significant antibacterial activity with zones of inhibition ranging from >10mm to ≤24mm. The inhibition of biofilm was shown by all the strains against all indicators, with a minimum inhibition of 52%, varying up to 89% in different strains. When the biofilm removing ability of strains was investigated against S. aureus, P. aeruginosa and E. coli, all the L. brevis strains showed removal of S. aureus biofilm to some extent, but the most significant activity was shown by Cat (strain of cat origin), i.e., 48% removal of S. aureus biofilm. But no removing ability against P. aeruginosa and E. coli biofilm was observed in any of the strains. The comparison of antibacterial activity of L. brevis from herbivores and carnivores has shown no significant difference in their activities. Similarly, no significant difference in the antibiofilm activity of L. brevis, isolated from herbivores and carnivores, was recorded.