E.coli represents a major cause of mortality and morbidity worldwide. This study was carried out to investigate distribution of virulence genes and pathotypes of E.coli isolated from farm environment, diarrhoea and healthy cattle. The recovered isolates were studied for hemolysis, biofilm production and presence of virulence marker genes viz., ST, cnf-2 by polymerase chain reaction .Out of 188 samples, 112 (59.57%) bacterial isolates were biochemically identified as E. coli which comprises of soil (11), feed and fodder (12), water sample (15 from drinking and 1 from tap water), 11 from diarrhoeic cases and 36 from healthy cattle, drainage (16), silage (1), manure (9). Out of 112 isolates 74(66.07%) were found to cause lysis on sheep erythrocytes, whereas 87(77.67%) were found to be biofilm producer on congo red. Singleplex PCR revealed the presence of ST gene in 60 (53.57%) and cnf -2 in 48(42.85%) isolates. Eleven strains (9.82 %) possessed both genes.