Lactic acid bacteria (LAB) with prominent antimicrobial effects against pathogens have been reported in several milk-based and plant-based foods. Borhani is a popular beverage prepared from the ingredients of both dairy and plant origins and is believed to be highly beneficial for health. Herein, we report the pathogen-inhibitory activity of two borhani-associated lactic acid bacteria (LAB), Limosilactobacillus fermentum strain LAB-1 and Levilactobacillus brevis strain LAB-5. Their antimicrobial activity was primarily assessed using the cell free supernatant (CFS) by agar diffusion technique in which both strains showed strong antimicrobial effects against several pathogenic and spoilage microorganisms including Acinetobacter baumannii, Bacillus cereus, Bacillus subtilis, Escherichia coli, Klebsiella pneumonia, Pseudomonas aeruginosa, Salmonella abony, Salmonella typhi, Shigella flexneri, and Staphylococcus aureus. The L. fermentum strain with its ability to inhibit all the target pathogens appeared to be more effective with larger inhibition-zone formation as compared to the L. brevis strain which also successfully inhibited all pathogens but had relatively little effects on A. baumannii. The extent of their inhibitory effect was further assessed by co-culture inhibition assay in which growth of the test microbes was monitored for 24 hours in presence of the CFS. The CFS of both lactic acid bacteria could effectively inhibit growth of the pathogenic microbes for a significant period of time. While the L. fermentum strain could almost completely stop growth of all test organisms, the L. brevis strain was particularly effective against Shigella flexneri and the Salmonella species. Our study, therefore, suggests the presence of beneficial lactic acid bacteria in borhani which can be of important use as antimicrobial agents in functional foods and therapeutics to help acquire protection against drug resistant pathogens.