Shrimp are major seafood in the aquaculture industry, particularly in Southeast Asia. However, Vibrio -related infections frequently occur in shrimp farms. Antibiotics are widely used in shrimp farming both preventative and therapeutic purposes in the fight against bacterial growth. However, overuse of antibiotics in aquatic environment has resulted in the establishment of antimicrobial resistance (AMR). Therefore, the study aims to provide a comprehensive database on the distribution of Vibrio spp. as well virulence genes and antimicrobial resistance patterns in Malaysian shrimp farms. Briefly, a total of 225 presumptive Vibrio isolates were isolated from seven farms cultured shrimp which were subjected to molecular detection using pyrH gene analysis. Eventually, 13 different Vibrio spp. were successfully isolated and characterized. The isolates were then analyzed with virulence genes resulting in eight of the eleven virulence genes were detected, including pirA , pirB , tlh , flaC , toxR , chiA , luxR , and hlyA . Other virulence genes, such as tdh , trh , and ctxA , were not detected in any isolates. From the virulence gene assessment, three Vibrio isolates were chosen for the pathogenicity test based on highest number present in each of the species. Meanwhile, the majority of the species tested were resistant to penicillin G (100%), but susceptible to norfloxacin (96%). According to the multiple antibiotic resistance (MAR) index, 84% of Vibrio spp. exhibited a MAR index value of more than 0.2. As a result, the findings suggested that a large number of Vibrio spp. were resistant to several antibiotics and had been exposed to antibiotics on a regular basis at the farms. In conclusion, ongoing monitoring of antibiotic-resistant microorganisms as well as understanding the genetic basis of resistance are essential to implement a strategy for minimizing AMR strain yet to improve shrimp farming management in Malaysia.