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Abstract in this study, 61 tested flocks were examined from different sources of poultry ( grandparents and layers and broilers and broiler breeders ) of suspected E. coli septicaemia in Egypt in addition to 29 human samples of (urine and feces). Based on the obtained results, it was found that the prevalence of E.coli infection was 95.6 %. The 86 serotypes were distributed as 16 (18.16%) of serotypes O125:K70, 12 (13.95%) of serotypes O158: K-, 10 (11.62%) of serotypes O44:K74 and O114:K90 each, 9 (10.46%) of untypable isolates, 6 (6.97%) of serotypes O91: K-, 4 (4.48%) O111:K58, O78:K80 and O25:K11, 2 (11.62%) of serotypes O103: K- and O26:K60 and finally the lowest percentage 1 (1.16%) were detected in serotypes O86: K- and O127:K63 - Testing of the phenotypic properties of the 86 isolates it was found that all tested E. coli isolates were Congo red positive but with different intensities those with CR+++ constituted 46/86 (53.5%) and 40/86 (46.5%) were CR++. The isolates tested were positive in their motility test and haemolysis assay (100%), results showed that the two cell lines MDCK and Vero cells were infected with E. coli. Most of the cells were dead or indicated cytopathic changes. The results of Biofilm binding activity of 86 E.coli isolates indicated that 49 isolates showed weak results and 18 isolates as strong and 19 isolates as moderate isolates.The in vitro sensitivity of 86 E.coli isolates were conducted against 15 antimicrobial drugs, results revealed all sensitive strains for each of colistin while 100% resistance to oxytetracycline and streptomycin. Polymerase chain reaction of 10 of genes: The study showed the presence of uidA gene in all isolates tested (100%).Tetracycline gene tet (A) and Sulphonamide (sulI), (sulIl) and Trimethoprim (dhfrI) , (dhfrXIII) , (dhfrV) for the detection of antibiotic resistant genes associated with poultry. The result also revealed that the prevalence of resistance is tet (A) 62%, Sulphonamide (sulI ) 45% , (sulIl) 50% , (dhfrI) 55% , (dhfrXIII) 20% and (dhfrV) no amplification could be observed. The phylogenetic grouping result of 86 strains as determined by reference methods, were as follows: 17(20%) belonged to groupA, 7 (8%) belonged to groupB1, 36 (42%) belonged to groupD, and 26 (30%) belonged to groupB2. |