Radioresistance development of DNA repair deficient Escherichia coli DH5α in ground beef subjected to electron beam at sub-lethal doses

Abstract

Purpose: Electron beam (e-beam) efficiently and non-thermally inactivates microorganisms in food by lethal DNA changes (direct effects) and free radicals from water radiolysis (in-direct effects). Non-pathogenic Escherichia coli DH5α (α substrain of DH5 described by Hanahan , ‘DH’ stands for Douglas Hanahan) is a microorganism that lacks DNA repair capability, resulting in high radiosensitivity. Studying microbial inactivation of E. coli DH5α repeatedly subjected to sub-lethal e-beam in ground beef may enhance understanding of microbial radioresistance. The objective of this study was to determine if repetitive processing with e-beam at sub-lethal doses increases D-value (e-beam dose required to inactivate one log of microbial population) of E. coli DH5α in ground beef.

Materials and methods: Survivors from the highest e-beam dose were isolated and incubated in ground beef for the next cycle of e-beam processing. Five cycles were conducted. To acclimatise E. coli DH5α, first two cycles used low doses. D-values were determined following the third cycle.

Results: D-values increased (p < 0.05) significantly with each cycle. Thus, E. coli DH5α has a capability to develop greater radioresistance under these experimental conditions. Following the third cycle D-values were 0.32 ± 0.006 and 0.32 ± 0.002 kGy for survivors enumerated on non-selective and selective media, respectively; the fourth cycle 0.39 ± 0.007 and 0.40 ± 0.019 kGy; and the fifth cycle 0.46 ± 0.006 and 0.46 ± 0.020 kGy. D-values on non-selective and selective media were similar (p > 0.05) indicating absence of cell recovery in E. coli DH5α.

Conclusions: E. coli DH5α increases radioresistance to e-beam as a result of repetitive exposure to sub-lethal doses despite its DNA repair deficiency.

Keywords:

• Escherichia coli
• electron beam
• food irradiation
• microbial inactivation
• inactivation kinetics

Acknowledgements

This research was funded by the USDA Hatch program (project no. WVA00429). Special thanks to Sterigenics International for allowing us to use e-beam facility. Our appreciation extends to Carl A. Zinn and Richard Vallejo of Sterigenics for invaluable technical expertise with e-beam and Aunchalee Aussanasuwamakul for assistance with statistical analysis.

Declaration of interest:

The authors report no conflicts of interest. The authors alone are responsible for the content and writing of the paper.