Evaluation of DNA damage reversal during medium-pressure UV disinfection

Abstract

Ultraviolet (UV) disinfection relies on the principal that DNA exposure to UV irradiation leads to the formation of cytotoxic lesions resulting in the inactivation of microorganisms. Cyclobutane pyrimdine dimers (CPDs) account for the majority of DNA lesions upon UV exposure. Past research has demonstrated reversal of CPDs in extracted DNA formed at high UV-C wavelength irradiation (280 nm) upon subsequent irradiation at lower UVC wavelengths (230-240 nm). Medium-pressure (MP) UV lamps produce a polychromatic emission giving rise to the possibility that cellular DNA in a target pathogen may undergo simultaneous damage and repair when exposed to multiple wavelengths during the disinfection process, decreasing the efficiency of MP UV lamp disinfection. Culture techniques and a quantitative polymerase chain reaction (qPCR) assay were used to examine cell viability and DNA damage reversal. qPCR results indicated direct photoreversal of UV-induced DNA damage through sequential irradiations of 280 nm followed by 228 nm in Escherichia coli DNA. However, significant photoreversal was only observed after high initial doses and secondary doses of UV light. The doses where significant photoreversal took place were more than 10 times higher than those typically used in UV disinfection. Despite evidence of CPD photoreversal, bacterial growth assays showed no indication that sequential-wavelength irradiations result in higher survival rates than single-wavelength irradiations.

Keywords: Cyclobutane pyrimidine dimer; DNA damage; Medium-pressure UV; Photoreversal; Ultraviolet light.