skip to main content


Title: Evaluation of parameters governing dark and photo-repair in UVC-irradiated Escherichia coli
After decades of UV disinfection practice and numerous studies on the potential for pathogens to undergo dark or photo-repair after UV exposure, recent advances in UV light emitting diode (LED) technologies prompt renewed attention to bacterial reactivation and regrowth processes after UV exposure. The aspect of photorepair conditions warrants particular attention, because even studies on conventional mercury vapor lamps have not sufficiently characterized these parameters. Wastewater encounters a wide range of environmental conditions upon discharge ( e.g. , solar irradiation and dissolved organics) which may affect repair processes and ultimately lead to overestimations of pathogen removal. Escherichia coli was used here to investigate the impacts of changing reactivation conditions after UV 254 and UV 278 irradiation. UV 254 and UV 278 doses of 13.75 ± 0.4 mJ cm −2 and 28.3 ± 0.8 mJ cm −2 were required to induce a 3.0 log inactivation of E. coli , respectively. Specifically, photoreactivation conditions were varied across dissolved organic matter (DOM) content and photoreactivation wavelengths and intensities. Photoreactivation achieved higher log recoveries than dark repair, ranging from 0.8 to 1.8 log differences, but a secondary disinfection effect occurred under UVA irradiation. During photoreactivation, humic acid inhibited the initial repair of UV 278 -dosed E. coli , but culture media enhanced recovery for both dosage wavelengths. Photoreactivation profiles under UV 395 , UV 365 , and visible light depended on both fluence and time, with more regrowth observed upon exposure to visible light and the least under 365 nm. The susceptibility of E. coli to UVA was increased by prior exposure to UVC.  more » « less
Award ID(s):
2046660 1952409
NSF-PAR ID:
10321852
Author(s) / Creator(s):
; ; ;
Date Published:
Journal Name:
Environmental Science: Water Research & Technology
Volume:
8
Issue:
2
ISSN:
2053-1400
Format(s):
Medium: X
Sponsoring Org:
National Science Foundation
More Like this
  1. Abstract The ongoing COVID-19 global pandemic has necessitated evaluating various disinfection technologies for reducing viral transmission in public settings. Ultraviolet (UV) radiation can inactivate pathogens and viruses but more insight is needed into the performance of different UV wavelengths and their applications. We observed greater than a 3-log reduction of SARS-CoV-2 infectivity with a dose of 12.5 mJ/cm 2 of 254 nm UV light when the viruses were suspended in PBS, while a dose of 25 mJ/cm 2 was necessary to achieve a similar reduction when they were in an EMEM culture medium containing 2%(v/v) FBS, highlighting the critical effect of media in which the virus is suspended, given that SARS-CoV-2 is always aerosolized when airborne or deposited on a surface. It was found that SARS-CoV-2 susceptibility (a measure of the effectiveness of the UV light) in a buffer such as PBS was 4.4-fold greater than that in a cell culture medium. Furthermore, we discovered the attenuation of UVC disinfection by amino acids, vitamins, and niacinamide, highlighting the importance of determining UVC dosages under a condition close to aerosols that wrap the viruses. We developed a disinfection model to determine the effect of the environment on UVC effectiveness with three different wavelengths, 222 nm, 254 nm, and 265 nm. An inverse correlation between the liquid absorbance and the viral susceptibility was observed. We found that 222 nm light was most effective at reducing viral infectivity in low absorbing liquids such as PBS, whereas 265 nm light was most effective in high absorbing liquids such as cell culture medium. Viral susceptibility was further decreased in N95 masks with 222 nm light being the most effective. The safety of 222 nm was also studied. We detected changes to the mechanical properties of the stratum corneum of human skins when the 222 nm accumulative exposure exceeded 50 J/cm 2 .The findings highlight the need to evaluate each UV for a given application, as well as limiting the dose to the lowest dose necessary to avoid unnecessary exposure to the public. 
    more » « less
  2. null (Ed.)
    To better understand the elimination of transforming activity of antibiotic resistance genes (ARGs), this study investigated the deactivation of transforming activity of an ARG (in Escherichia coli as a host) and ARG degradation (according to quantitative PCR [qPCR] with different amplicon sizes) during UV (254 nm) and UV/H 2 O 2 treatments of plasmid pUC19 containing an ampicillin resistance gene ( amp R ). The required UV fluence for each log 10 reduction of the transforming activity during UV treatment was ∼37 mJ cm −2 for both extra- and intra-cellular pUC19 (the latter within E. coli ). The resulting fluence-based rate constant ( k ) of ∼6.2 × 10 −2 cm 2 mJ −1 was comparable to the k determined previously for transforming activity loss of plasmids using host cells capable of DNA repair, but much lower (∼10-fold) than that for DNA repair-deficient cells. The k value for pUC19 transforming activity loss was similarly much lower than the k calculated for cyclobutane-pyrimidine dimer (CPD) formation in the entire plasmid. These results indicate the significant role of CPD repair in the host cells. The degradation rate constants ( k ) of amp R measured by qPCR increased with increasing target amplicon size (192–851 bp) and were close to the k calculated for the CPD formation in the given amplicons. Further analysis of the degradation kinetics of plasmid-encoded genes from this study and from the literature revealed that qPCR detected most UV-induced DNA damage. In the extracellular plasmid, DNA damage mechanisms other than CPD formation ( e.g. , base oxidation) were detectable by qPCR and gel electrophoresis, especially during UV/H 2 O 2 treatment. Nevertheless, the enhanced DNA damage for the extracellular plasmids did not result in faster elimination of the transforming activity. Our results indicate that calculated CPD formation rates and qPCR analyses are useful for predicting and/or measuring the rate of DNA damage and predicting the efficiency of transforming activity elimination for plasmid-encoded ARGs during UV-based water disinfection and oxidation processes. 
    more » « less
  3. This study proposes a novel disinfection process by sequential application of peracetic acid (PAA) and ultra-violet light (UV), on the basis of elucidation of disinfection mechanisms under UV/PAA. Results show that hydroxyl radicals, generated by UV-activated PAA, contribute to the enhanced inactivation of Escherichia coli under UV/PAA compared to PAA alone or UV alone. Furthermore, the location of hydroxyl radical generation is a critical factor. Unlike UV/H2O2, which generates hydroxyl radicals mainly in the bulk solution, the hydroxyl radicals under UV/PAA are produced close to or inside E. coli cells, due to PAA diffusion. Therefore, hydroxyl radicals exert significantly stronger disinfection power under UV/PAA than under UV/H2O2 conditions. Pre-exposing E. coli to PAA in the dark followed by application of UV (i.e., a PAA-UV/PAA process) promotes diffusion of PAA to the cells and achieves excellent disinfection efficiency while saving more than half of the energy cost associated with UV compared to simultaneous application of UV and PAA. The effectiveness of this new disinfection strategy has been demonstrated not only in lab water but also in wastewater matrices. 
    more » « less
  4. null (Ed.)
    The photolysis of hypochlorous acid (HOCl) and hypochlorite (OCl − ) produces a suite of reactive oxidants, including hydroxyl radical (˙OH), chlorine radical (Cl˙), and ozone (O 3 ). Therefore, the addition of light to chlorine disinfection units could effectively convert existing drinking water treatment systems into advanced oxidation processes. This review critically examines existing studies on chlorine photolysis as a water treatment process. After describing the fundamental chemistry of chlorine photolysis, we evaluate the ability of chlorine photolysis to transform model probe compounds, target organic contaminants, and chlorine-resistant microorganisms. The efficacy of chlorine photolysis to produce reactive oxidants is dependent on solution and irradiation conditions ( e.g. , pH and irradiation wavelengths). For example, lower pH values result in higher steady-state concentrations of ˙OH and Cl˙, resulting in enhanced contaminant removal. We also present the current state of knowledge on the alteration of dissolved organic matter and subsequent formation of disinfection by-products (DBPs) during chlorine photolysis. Although the relative yields of DBPs during chlorine photolysis are also dependent on solution conditions ( e.g. , higher organic DBP yields at low pH values), there is conflicting evidence on whether chlorine photolysis increases or decreases DBP production compared to thermal reactions between chlorine and dissolved organic matter in the dark. We conclude the review by identifying knowledge gaps in the current body of literature. 
    more » « less
  5. The main objective of this study was to investigate the effectiveness of ultraviolet light (UV-C) emitting diodes for the decontamination of stainless steel food contact surfaces. Listeria monocytogenes (ATCC 19115), Escherichia coli (ATCC 25922), and Salmonella enterica serovar Typhimurium (ATCC 700720) were chosen as challenge microorganisms. Target microorganisms were subjected to UV-C dosages of 0, 2, 4, 6, and 8 mJ cm −2 at an average fluence of 0.163 mW/cm 2 using a near-collimated beam operating at 279 nm wavelength. Escherichia coli showed lower sensitivity to UV-C light compared to Salmonella Typhimurium and followed first-order kinetics. Escherichia coli and Salmonella Typhimurium were reduced by more than 3-log 10 cycles at the maximum UV dosage of 12 mJ cm −2 . In contrast, Listeria monocytogenes followed the Weibull model with an apparent shoulder in the initial doses. A maximum reduction of 4.4-log 10 was achieved at the highest exposure level. This study showed that UV-C LED devices represent an excellent alternative for the inactivation of foodborne microorganisms in droplets. Results clearly demonstrate that UV-C LED devices can serve as an additional sanitation method to routine cleaning practices, which are commonly utilized in the food industry. 
    more » « less