Isolation and Characterization of EPEC Phage from Domestic Waste in Indonesia

Abstract

Aims: Enteropathogenic Escherichia coli (EPEC) K1.1 is one of pathogenic bacteria-causing diarrheal disease. This bacterium was resistant to tetracycline and ampicillin. The use of lytic phage can be a good solution to reduce antibiotic resistance of pathogenic bacteria. This study was to isolate and characterize phage that can lyse EPEC K1.1 cells. Methodology and results: Lytic phage was isolated from domestic waste at Darmaga, Bogor, West Java, Indonesia. It uses double layer plaque technique. The kind of phages was determined by the plaque morphology, the structure, the host range, and the protein characteristics of phages as well as storage stability of phage in different buffer conditions. Lytic phage only infects the EPEC K1.1. It does not infect the other bacteria in this research. Electron microscope observation showed that phage FBd3 has icosahedral head with 57.14 nm in diameter and short-non contractile tail with 18.2 nm in length. It was grouped into family Phodoviridae. Lytic phage had good storage stability in Ringers buffer at low temperature (4 °C). It showed that viability of phage decreased by 28.04% after 9 days of storage. Phage FBd3 has six different proteins with various molecular weight: 33 kDa, 42 kDa, 49 kDa, 57 kDa, 77 kDa and 106.7 kDa. This study indicates that EPEC K1.1 can be infected by more than one type of lytic phages. Conclusion, significance and impact of study: Lytic phage FBd3 reduces the population of PEC K1.1 effectively. It can be used as a bio-control agent to EPEC that contaminates food and water. Lytic phage found in this study potentially prevent bacterial contamination in food and beverage processing for diarrheal disease prevention.