A dual-wavelength UV-C LED unit, emitting at peaks of 260 nm, 280 nm, and the combination of 260|280 nm together was evaluated for its inactivation efficacy and energy efficiency at disinfecting Escherichia coli, MS2 coliphage, human adenovirus type 2 (HAdV2), and Bacillus pumilus spores, compared to conventional low-pressure and medium-pressure UV mercury vapor lamps. The dual-wavelength unit was also used to measure potential synergistic effects of multiple wavelengths on bacterial and viral inactivation and DNA and RNA damage. All five UV sources demonstrated similar inactivation of E. coli. For MS2, the 260 nm LED was most effective. For HAdV2 and B. pumilus, the MP UV lamp was most effective. When measuring electrical energy per order of reduction, the LP UV lamp was most efficient for inactivating E. coli and MS2; the LP UV and MP UV mercury lamps were equally efficient for HAdV2 and B. pumilus spores. Among the UV-C LEDs, there was no statistical difference in electrical efficiency for inactivating MS2, HAdV2, and B. pumilus spores. The 260 nm and 260|280 nm LEDs had a statistical energy advantage for E. coli inactivation. For UV-C LEDs to match the electrical efficiency per order of log reduction of conventional LP UV sources, they must reach efficiencies of 25-39% or be improved on by smart reactor design. No dual wavelength synergies were detected for bacterial and viral inactivation nor for DNA and RNA damage.

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.

No abstract

Adenovirus, a waterborne pathogen responsible for causing bronchitis, pneumonia, and gastrointestinal infections, is highly resistant to UV disinfection and therefore drives the virus disinfection regulations set by the U.S. Environmental Protection Agency. Polychromatic UV irradiation has been shown to be more effective at inactivating adenovirus and other viruses than traditional monochromatic irradiation emitted at 254 nm; the enhanced efficacy has been attributed to UV-induced damage to viral proteins. This research shows UV-induced damage to adenoviral proteins across the germicidal UV spectrum at wavelength intervals between 200 and 300 nm. A deuterium lamp with bandpass filters and UV light-emitting diodes (UV LEDs) isolated wavelengths in approximate 10 nm intervals. Sodium dodecyl sulfate polyacrylamide gel electrophoresis and image densitometry were used to detect signatures for the hexon, penton, fiber, minor capsid, and core proteins. The greatest loss of protein signature, indicating damage to viral proteins, occurred below 240 nm. Hexon and penton proteins exposed to a dose of 28 mJ/cm emitted at 214 nm were approximately 4 times as sensitive and fiber proteins approximately 3 times as sensitive as those exposed to a dose of 50 mJ/cm emitted at 254 nm. At 220 nm, a dose of 38 mJ/cm reduced the hexon and penton protein quantities to approximately 33% and 31% of the original amounts, respectively. In contrast, a much higher dose of 400 mJ/cm emitted at 261 and 278 nm reduced the original protein quantity to between 66-89% and 80-93%, respectively. No significant damage was seen with a dose of 400 mJ/cm at 254 nm. This research directly correlates enhanced inactivation at low wavelengths with adenoviral protein damage at those wavelengths, adding fundamental insight into the mechanisms of inactivation of polychromatic germicidal UV irradiation for improving UV water disinfection.