Vibrio anguillarum causes severe economic losses in fish and shellfish species and poses public health concerns. This study aimed to provide some information about the prevalence and virulence determinants of V. anguillarum isolated from the farmed Oreochromisniloticus fish fingerlings nearby Qarun Lake, El-Fayoum Governorate, Egypt. PCRtargeting the rpoA (RNA polymerase alpha subunit)gene was done forconfirmation of Vibriospp. Moreover, V. anguillarum isolates were biochemically identified and confirmed by 16S rRNA gene sequence.V. anguillarum isolates produced caseinase, gelatinase, lipase, lecithinase, haemolysins, and possessed biofilm formation. The virulence-related genes;empA, vah1, vah3 and vah4, flaA,angM, and angRwere detected in all tested isolates. Therefore, proper identification of V.anguillarumis crucial to better understand the ecology and distribution patterns of such pathogen within Egyptian aquaculture. Moreover, the water quality in these farms near Qarun Lake was less supportive for larval rearing activities of O. niloticus fish.