Isolation and characterization of pathogenic Escherichia coli bacteriophages from chicken and beef offal

Abstract

Objective: This study was conducted to isolate and characterize lytic bacteriophages for pathogenic Escherichia coli from chicken and beef offal, and analyze their capability as biocontrol for several foodborne pathogens. Methods done in this research are bacteriophage isolation, purification, titer determination, application, determination of host range and minimum multiplicity of infection (miMOI), and bacteriophage morphology. Results: Six bacteriophages successfully isolated from chicken and beef offal using EPEC and EHEC as host strain. Bacteriophage titers observed between 10 9 and 10 10 PFU mL −1. CS EPEC and BL EHEC bacteriophage showed high efficiency in reduction of EPEC or EHEC contamination in meat about 99.20% and 99.04%. The lowest miMOI was 0.01 showed by CS EPEC bacteriophage. CI EPEC and BL EPEC bacteriophage suspected as Myoviridae family based on its micrograph from Transmission Electron Microscopy (TEM). Refers to their activity, bacteriophages isolated in this study have a great potential to be used as biocontrol against several foodborne pathogens.

Isolation and Identification of Escherichia coli O157:H7 Lytic Bacteriophage from Environment Sewage

“…Bacteriophages could affect bacterial lysis because they are able to recognize receptors on the surface of the host bacteria, so that bacteriophages are able to transfer their genetic material into the host cell and replicate in the host cell causing lysis [ 33 ]. From Table 3 , it can be seen that the diameter size was around 0.7 to 3.5 mm.…”

“…Bacteriophage quantification was measured by counting the amount of plaque formed, and the concentration obtained was of 5 × 10 3 to 10 × 10 8 PFU/mL. The number of bacteriophages previously detected was >10 8 PFU/mL of Bacillus cereus bacteriophage [ 33 ], 10 15 PFU/mL of Klebsiella pneumoniae bacteriophage [ 43 ], 5 × 10 12 PFU/mL of E. coli bacteriophage [ 44 ], 1.2 × 10 16 PFU/mL [ 45 ], and 5 × 10 6 PFU/mL for T4 bacteriophage [ 46 ], and 1.3 × 10 11 PFU/mL [ 23 ], 2.1 × 10 10 PFU/mL [ 34 ], and 2.62 × 10 10 PFU/mL of E. coli bacteriophage [ 33 ]. Marti et al conducted a study with Salmonella spp.…”

“…Bao et al found the bacteriophage Siphoviridae with a tail length of 103.57 nm and a head diameter of 57.14 nm and Myoviridae bacteriophage with a head diameter of 74.3 nm and a tail length of 114.2 nm from chicken waste in chicken farms in China [ 48 ], while Lukman et al also found EPEC bacteriophages with a head diameter of 67-70 nm and a tail length of 83-90 nm as Myoviridae from chickens and beef offal from traditional markets in Tangerang, Indonesia [ 33 ]. To ensure bacteriophage classification, more study, such as bacteriophage genome sequencing, is required.…”

“…In addition, the ability of lysis was also influenced by the multiplicity of infection (MOI), which is the ratio of bacteriophages to the number of target bacteria [ 53 ]. Bacterial growth will increase along with the decrease in MOI [ 33 ], but MOI cannot completely inhibit cell growth [ 55 ].…”

“…Chen et al found bacteriophages which lysed Pasteurella multocida A, and P capsular type was not able to lyse strains with D or F capsular type or other Gram-negative bacteria, including E. coli, Salmonella spp., and Bordetella bronchiseptica [ 59 ]. Lukman et al also found a bacteriophage that can lyse EPEC and EHEC [ 33 ]. Further research on bacteriophage host specificity on another pathogenic E. coli is required based on these traits.…”