A total of 100 random samples of fresh and marine fishes represented by Tilapia niloticus, Mugil cephalus, Sardine and Barboni (25 0f each) were collected from the different fish markets located in Gharbia governorate.  E. coli was identified by standard microbiological, biochemical tests, and further confirmed by multiplex PCR. The faecal coliform loads in fish samples were assessed. The bacterium was detected in 8(32%), 6(24%), 4(16%) and 3(12%) of the examined samples of Oreochromis niloticus, Mugil cephalus, Sardine and Barboni respectively. The accepted fish samples were 17 (68 %), 19 (76 %), 21 (86 %) and 22 (88 %) of the examined samples of O. niloticus, Mugil cephalus, Sardine and Barboni respectively while the unaccepted samples were 8 (32 %), 6 (24 %), 4 (16 %) and 3 (12 %) of the examined samples of O. niloticus, Mugilcephalus, Sardine and Barboni respectively according to Egyptian standards (E.S 3494/2005). the results cleared that, the main isolates include O26: H11; O44: H18, O86: H; O91: H21; O103: H2; O119: H6; O121: H7; O124: H; O128: H2; O146: H21 and it isolated mainly from O. niloticus, Mugil cephalus, then Sardine and Barboni. The work was aimed to study prevalence of Escherichia coli contamination in fish sold in different fish markets located in Gharbia governorate.