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Diarrhoea continues to be one of the most common causes of morbidity and mortality among infants and children in developing countries. Escherichia coli is an emerging agent among pathogens that cause diarrhea. This research was carried out to study the prevalence of Escherichia coli in Ile-Ife, Osun State, Nigeria by using standard microbiological methods to analyze the diarrhoeic stool samples from children collected at three different Health Care Institutions Ile-Ife between the months of August 2013 to February 2014. Out of 100 stool samples collected, 22(22%) were positive for E. coli while 78(78%) were negative. showing that the prevalent of E. coli among the study population is 22%. 47 samples were from males with 8(17%) E. coli and 39(83%) negative while 53 samples were collected from females with 14(26.4%) E. coli and 39(73.6%) negative. 35 samples collected from patients who drink sachet water had 8(22.9%) E. coli, 32 samples collected from patients who drink well water, 7(21.9%) had E. coli, 7(24.1%) had E. coli out of 29 stool samples that were collected from patients that drink tap water. Patients < 6 months of age, had 7 (20%) E. coli, for age 6-11 months, 10(25%) had E. coli. Age 1-2 years had 3(15.7%) E. coli, ages 3-5 years had 2(33.3%) E. coli. Out of 58 watery stool samples collected 8(13.8%) had E. coli, 29 mucoid stool samples had, 8(27.6%) E. coli, 13 formed stool samples had 6(46.2%) E. coli, patients whose parents have no formal education had 2(50%) E. coli. 3 samples collected from patients whose parents have modern one level of education; E. coli was isolated from 1(33.3%) stool sample, patients whose parent have completed primary education had 8(44.4%) E. coli, patients whose parents have attained modern 3 level of education, E. coli was isolated in 3(15.8%), patients whose parents have completed SSCE had 4(14.8%), E. coli, patients whose parents have attained post-secondary school education had 4(13.8%) E. coli, patients whose parents’ income are <N10,000 in which 7(29.2%) had E. coli, parents’ income are>N10,000, 8(29.6%) had E. coli, patients whose parents’ have no stable source of income had 7(14.3%) E. coli. The isolates showed a resistance to Ampicillin (100%), Augmentin (95.5%), Cefuroxime (86.4%), Ceftazidine (72.7%), Ofloxacin (13.6%), Gentamycin (13.6%), Ciprofoxacin (13.6%) and Nitrofurantoin (4.5%). The isolates are sensitive to Nitrofurantoin (95.5%), Ofloxacin (86.4%), Gentamycin (72.7%), CPR (86.4%), Augmentin (4.5%), Cefuroxime (9%) and Ceftazidine (4.5%). The results of the study show that diarrhoegenic E. coli is still an important pathogen of health importance affecting children.