The current study was planned for the isolation and identification of Vibrio spp. from fish samples (tilapia,mugil, tuna, saurus, pagrus, and shrimp) retailed in Zagazig city, Sharkia Governorate, Egypt. In addition todetermine the antimicrobial susceptibility of the isolates. The PCR screening for virulence genes of V. parahaemolyticus and V. cholerae was also determined. The results in the present study revealed that the most contaminated samples with Vibrio spp. were pagrus (56%), followed by tilapia (52 %), then mugil (44 %), saurus (40 %), tuna (36 %), and shrimp (36 %). The most prevalent Vibrio spp. were V. parahaemolyticus (18.6%) followed by V. mimicus (11.3%), and V. alginolyticus (10%). V. cholerae was isolated in a percentage of 0.7%. Vibrio species in the current study were highly resistant to erythromycin (100%), ampicillin (75%), cephalothin (66.7%), sulphamethoxazole (66.7%), and amikacin (50%). PCR screening of virulence genes among various Vibrio spp. revealed that V. parahaemolyticus harbored tlh (100 %), tdh (80 %), and trh (80 %). While, V. cholerae was positive for rtxC, hylA, and ompU genes. As a result, the current data proves that Vibrio spp. contamination of fish, which displayed various degrees of antibiotic resistance. In addition, the isolated Vibrio spp. had virulent genes that could be dangerous to the consumer’s health.