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The beneficiary bacteria isolated from healthy shrimp gut were found to be active against the growth of Vibrio pathogens and executed maximum zone of inhibition. The main objective of the research is to identify and screen probiotic isolates from healthy shrimp gut as biocontrol measures against Vibrio pathogenic species. The probiotics isolated from the shrimp gut were analyzed for different parameters including Gram staining, morphology, IMViC, acids, bile, NaCl, and antimicrobial activity against Vibrio pathogens. A total of 36 isolates were isolatedout of which 9 were screened as best microbes. By using MHA media swabbed with Vibrio pathogen species in the replica method, 3 isolates showed a maximum zone of inhibition against test pathogens. These test pathogen Vibrio cholera, Vibrio parahaemolyticus, Vibrio alginolyticus, Vibrio harveyi were collected and grown on TCBS media. The three isolates identified asLactobacillus rhamnosus,Lactobacillus paracasei,Pediococcus acidilacticiexhibited reproducible probiotic activity against test pathogen by well diffusion method. This study indicated that probiotic foods in shrimp farming will help them to fightagainst pathogenic organisms.