Application of a novel lytic phage to control enterotoxigenic Escherichia coli in dairy food matrices

Abstract

A novel Escherichia coli phage designated as EC BD was isolated from cattle dung samples. Transmission electron microscopy demonstrated that the morphology of phage EC BD belongs to the family Myoviridae. The efficiency of plating (EOP) and scanning electron microscopy revealed the strong lytic activity of phage EC BD with a large burst size and a short latent period. Whole genome data analysis suggested that phage EC BD was inclined towards being completely lytic and revealed the absence of toxins, virulence and antibiotic resistance genes. Phylogenomic analysis of phage EC BD receptor binding proteins (RBPs) showed 74–100 % similarity with sixteen Enterobacter phages, representing their broad host range. The phage genome contains 262 ORFs, of which 107 displayed a unique pattern and additionally, the presence of a tRNA gene directed their fast replication and high stability. Comparative genome analysis suggested phage EC BD as a novel member of the genus Duplodnaviria and family Myoviridae. The efficiency of phage EC BD was determined in dairy food matrices (milk, cheese and paneer) artificially contaminated with enterotoxigenic E. coli strains ETEC H10407, ETEC K 12S and ETEC PB 176 with a significant reduction of 4.8, 6.0 and 5.3 log CFU/mL in milk and a substantial 4.9, 5.8 and 4.6 log CFU/mL reduction in cheese samples after 6 days at low storage temperature (4 °C); furthermore, within the similar conditions, paneer samples showed 4, 5.1 and 3.5 log CFU/mL reduction. EC BD phage treatment represents the complete eradication of three ETEC strains in liquid and semisolid dairy food matrices. This study suggested that phage EC BD might have potential as a biocontrol approach against ETEC foodborne infections.