Sea turtles are air-breathing, marine reptiles. The past few years have witnessed a surge in research focusing on the causes of morbidity and mortality, along with intensified endeavors to conserve them. This research aimed to isolate and characterize vibrio alginolyticus from apparently healthy sea turtles, Antimicrobial sensitivity test, and Molecular identification. There were 60 red-eared turtle samples (30 buccal and 30 cloacal samples), and 18 green turtle samples (9 buccal and 9 cloacal samples). collected aseptically from the Red Sea, Pet stores, and Giza zoo and subjected to isolation and biochemical characterization of V. alginolyticu, V. alginolyticus are Gram-negative motile comma-shaped rodes that are facultatively anaerobic. Isolates of V. alginolyticus are divided into (9) positive isolates from Red-eared turtles, and (3) from Green turtles. All tested isolates were positive for catalase, oxidase, indole test, Methyl red, and citrate utilization and Negative for Vogus prosper. The V. alginolyticus strain exhibited major resistance to trimethoprim 5µg, ampicillin10µg, streptomycin 10µg based on the antibiotic sensitivity test, besides the intermediate sensitivity to Tobramycin 10µg, Naldixic acid 30µg and kanamycin (30µg). Furthermore, the bacterium demonstrated high sensitivity to Chloramphenicol 30µg, Tetracycline 30µg, and Ciprofloxacin 5µg. The molecular identification was achieved by utilizing species-specific primers to target the collagenase gene.