c Polychromatic UV irradiation is a common method of pathogen inactivation in the water treatment industry. To improve its disinfection efficacy, more information on the mechanisms of UV inactivation on microorganisms at wavelengths throughout the germicidal UV spectrum, particularly at below 240 nm, is necessary. This work examined UV inactivation of bacteriophage MS2, a common surrogate for enteric pathogens, as a function of wavelength. The bacteriophage was exposed to monochromatic UV irradiation from a tunable laser at wavelengths of between 210 nm and 290 nm. To evaluate the mechanisms of UV inactivation throughout this wavelength range, RT-qPCR (reverse transcription-quantitative PCR) was performed to measure genomic damage for comparison with genomic damage at 253.7 nm. The results indicate that the rates of RNA damage closely mirror the loss of viral infectivity across the germicidal UV spectrum. This demonstrates that genomic damage is the dominant cause of MS2 inactivation from exposure to germicidal UV irradiation. These findings contrast those for adenovirus, for which MS2 is used as a viral surrogate for validating polychromatic UV reactors. UV irradiation is a common method of disinfection in the water treatment industry. UV light induces damage to the genomes of bacteria, protozoa, and viruses, breaking bonds and forming photodimeric lesions in nucleic acids, DNA, and RNA (1, 2). These lesions prevent both transcription and replication and ultimately lead to inactivation of the microorganisms (3, 4). Direct UV damage to nucleic acids occurs at the wavelengths absorbed by DNA and RNA, in the germicidal UV region between 200 and 300 nm (5, 6). In this wavelength range, however, UV light also damages other cellular and viral components, causing, for example, photochemical reactions in proteins and enzymes (7,8). For this reason, UV sources that emit polychromatic light, across the germicidal UV spectrum, are considered more effective at inactivating certain pathogens than sources that emit monochromatic light at 253.7 nm (9-12). As polychromatic sources become more common, more research is being undertaken to understand the mechanisms of inactivation occurring in pathogens exposed to polychromatic UV irradiation.Male-specific (MS2) coliphage is a single-stranded RNA virus. It infects strains of Escherichia coli that produce F ϩ pili, which serve as viral receptors. The virion consists of a short singlestranded RNA genome (3,569 bases) surrounded by an icosahedral protein capsid, 27 nm in diameter (13). MS2 is commonly used in the water treatment industry as a surrogate for enteroviruses because of its similar size, shape, and genome composition (9,14). It serves as a biodosimeter for UV disinfection studies (15) and for UV reactor validation in North America (14, 16). For reactor validation, MS2 is also used as a surrogate for Cryptosporidium and adenovirus, despite the differences in UV sensitivity and spectral sensitivity between these microorganisms. Recent interest has grown regarding microbial ...

Adenovirus is regarded as the most resistant pathogen to ultraviolet (UV) disinfection due to its demonstrated resistance to monochromatic, low-pressure (LP) UV irradiation at 254 nm. This resistance has resulted in high UV dose requirements for all viruses in regulations set by the United States Environmental Protection Agency. Polychromatic, medium-pressure (MP) UV irradiation has been shown to be much more effective than 254 nm, although the mechanisms of polychromatic UV inactivation are not completely understood. This research analyzes the wavelength-specific effects of UV light on adenovirus type 2 by analyzing in parallel the reduction in viral infectivity and damage to the viral genome. A tunable laser from the National Institute of Standards and Technology was used to isolate single UV wavelengths. Cell culture infectivity and PCR were employed to quantify the adenoviral inactivation rates using narrow bands of irradiation (<1 nm) at 10 nm intervals between 210 and 290 nm. The inactivation rate corresponding to adenoviral genome damage matched the inactivation rate of adenovirus infectivity at 253.7 nm, 270 nm, 280 nm, and 290 nm, suggesting that damage to the viral DNA was primarily responsible for loss of infectivity at those wavelengths. At 260 nm, more damage to the nucleic acid was observed than reduction in viral infectivity. At 240 nm and below, the reduction of viral infectivity was significantly greater than the reduction of DNA amplification, suggesting that UV damage to a viral component other than DNA contributed to the loss of infectivity at those wavelengths. Inactivation rates were used to develop a detailed spectral sensitivity or action spectrum of adenovirus 2. This research has significant implications for the water treatment industry with regard to polychromatic inactivation of viruses and the development of novel wavelength-specific UV disinfection technologies.

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Even extremely low dosages of ultraviolet light can be highly effective for inactivating Cryptosporidium oocysts.Recent studies have shown that Cryptosporidium parvum oocysts demonstrate high susceptibility to low dosages of medium‐pressure ultraviolet (UV) light. These investigations have raised several questions, which include determination of minimum medium‐pressure UV dosages necessary to inactivate C. parvum oocysts, elucidation of differences (if any) between medium‐ and low‐pressure UV light for inactivating C. parvum oocysts, and evaluation of medium‐pressure UV effectiveness in inactivating oocysts suspended in poorer quality water. To compare low‐ and medium‐pressure UV, the authors exposed oocysts suspended in deionized water to UV delivered by either medium‐ or low‐pressure UV lamps at bench scale using a collimated beam apparatus. The applied UV dosages ranged from 3 to 33 mJ/cm2, and oocyst inactivation was assessed using the neonatal mouse infectivity assay. At 3 mJ/cm2, medium‐pressure UV showed a 3.4‐log inactivation of oocysts, and low‐pressure UV showed a 3.0‐log inactivation, demonstrating medium‐ and low‐pressure UV did not differ significantly in inactivating C. parvum oocysts.

In vivo studies indicate the infectivity of treated Cryptosporidium parvum oocysts more reliably than do in vitro assays. Inactivation of Cryptosporidium parvum oocysts in finished drinking water by medium‐pressure ultraviolet (UV) light was investigated at bench scale using a collimated beam apparatus and at demonstration scale using a UV reactor. Oocyst viability was assessed in vitro (using 4',6‐diamidino‐2‐phenylindole with propidium iodide and maximized in vitro excystation) and in vivo (using neonatal mouse infectivity assays). In vivo bench‐scale studies showed > 4‐log inactivation at UV dosages as low as 41 mJ cm–2, although in vitro surrogate assays showed little or no inactivation at this or higher UV dosages. The in vitro assays, which indicate oocyst viability, grossly overestimated the UV dosages required to prevent oocyst infection in susceptible hosts. Results of demonstration studies, carried out under the Environmental Technology Verification program of the National Sanitation Foundation and the US Environmental Protection Agency, agreed with the bench‐scale results and showed that a UV dosage as low as 19 mJ cm–2 provided 3.9‐log inactivation of Cryptosporidium oocysts.