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The composition of Escherichia coli in the neonatal gut has rarely been studied in developing countries. To gain insight into the composition of E. coli in the neonatal gut and to assess factors that could influence colonization by E. coli, analysis of the phylogenetic groups and virulence determinants of E. coli isolated from the guts of neonates in a tertiary care hospital was carried out. The distribution of the phylogroups of 124 E. coli isolates recovered showed that phylogroups A (23 %) and B1 (49 %) accounted for 72 % of the isolates. Isolates of the phylogenetic group B2 were rare (8 %). Virulence factors were also rare with the exception of aerobactin (iucC), which was detected in 45 % of the isolates and was significantly associated with phylogroup B1. Multinomial logistic regression established that colonization with phylogroup B1 was associated with a stay in the neonatal intensive care unit; phylogroup A was associated with a stay on the ward; and phylogroups B2 and D were associated with neonates delivered vaginally. Evaluation of the effect of different E. coli phylogroups, with and without identified virulence determinants, on the gut of neonatal mice showed histopathological changes in the mucosa. The severity of the changes could be correlated with the presence of virulence determinants, irrespective of the phylogroup.