SARS-CoV-2 inactivation by ultraviolet radiation and visible light is dependent on wavelength and sample matrix.

Numerous studies have demonstrated that SARS-CoV-2 can be inactivated by ultraviolet (UV) radiation. However, there are few data available on the relative efficacy of different wavelengths of UV radiation and visible light, which complicates assessments of UV decontamination interventions. The present study evaluated the effects of monochromatic radiation at 16 wavelengths from 222 nm through 488 nm on SARS-CoV-2 in liquid aliquots and dried droplets of water and simulated saliva. The data were used to generate a set of action spectra which quantify the susceptibility of SARS-CoV-2 to genome damage and inactivation across the tested wavelengths. UVC wavelengths (≤280 nm) were most effective for inactivating SARS-CoV-2, although inactivation rates were dependent on sample type. Results from this study suggest that UV radiation can effectively inactivate SARS-CoV-2 in liquids and dried droplets, and provide a foundation for understanding the factors which affect the efficacy of different wavelengths in real-world settings.

National Institute of Standards and Technology transportable tunable ultraviolet laser irradiance facility for water pathogen inactivation.

A method of ultraviolet germicidal irradiation (UVGI) for water pathogen inactivation effectiveness using tunable, narrowband laser light is described. A transportable tunable UV (TTUV) laser system for providing a known irradiance (µW/cm2) or dose (mJ/cm2) suitable for irradiating water samples in Petri dishes over the wavelength range of 210 nm-300 nm was developed by the National Institute of Standards and Technology. The TTUV facility, consisting of a 1 kHz pulsed UV laser and light-tight enclosure containing the optics necessary to uniformly irradiate a water sample, was used in a microbiology laboratory to dose drinking water pathogens and surrogates as part of a Water Research Foundation study in the summer and fall of 2012. The approach demonstrated improved accuracy and simplified spectral analysis over conventional pathogen inactivation sources consisting of broadband UV sources and bandpass filters. In this work, the TTUV facility design and key components are described, including modifications in the field to provide the required irradiance levels. The irradiance and dose levels produced by the tunable UV laser during the project are also presented. The transportability of the TTUV system enabled it to be brought to a microbiology facility allowing the water samples (microbial suspensions) to be irradiated in a location with experienced staff and facilities for preparing, handling, analyzing, storing, and shipping the many samples studied. These results, published elsewhere, established that the tunable UV laser system provides unique UVGI capabilities for use with water pathogens and has applications for other pathogen experiments, for example, air-purification studies.

Calibration of night vision goggles: an SI-units-based gain measurement technique.

A gain measurement technique for the calibration of night vision goggles (NVG) is proposed and evaluated. This technique is based on the radiance measurements at the input and output of the NVG. In contrast to the old definition, which uses a non-International System of Units (SI) traceable luminance, the "equivalent luminance unit," the suggested technique utilizes the radiance quantities that are traceable to the SI units through National Institute of Standards and Technology (NIST) standards. Due to the implementation of the scaling coefficients originating from the NVG spectral responsivities, the same NVG gain is expected within both techniques. The suggested method was evaluated at the NIST night vision calibration facility and the experimental data were compared to the results obtained with a commercial NVG test set. The comparison of the radiometric quantities obtained using the two different methods indicated differences up to 15% due to different calibration conditions. However, at proper calibration, equal NVG gains within both the suggested and old gain definitions were measured for the goggles equipped with a filmless image tube. The NVG gain uncertainty analysis, including the effect of no-moon night sky radiation, was performed for goggle types A, B, and C.

Comparison of UV-Induced Inactivation and RNA Damage in MS2 Phage across the Germicidal UV Spectrum.

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.

Action spectra for validation of pathogen disinfection in medium-pressure ultraviolet (UV) systems.

Ultraviolet (UV) reactors used for disinfecting water and wastewater must be validated and monitored over time. The validation process requires understanding the photochemical properties of the pathogens of concern and the challenge microorganisms used to represent them. Specifically for polychromatic UV systems, the organisms' dose responses to UV light and their sensitivity across the UV spectrum must be known. This research measured the UV spectral sensitivity, called action spectra, of Cryptosporidium parvum, and MS2, T1UV, Q Beta, T7, and T7m Coliphages, as well as Bacillus pumilus spores. A tunable laser from the National Institute of Standards and Technology was used to isolate single UV wavelengths at 10 nm intervals between 210 and 290 nm. Above 240 nm, all bacteria and viruses tested exhibited a relative peak sensitivity between 260 and 270 nm. Of the coliphage, MS2 exhibited the highest relative sensitivity below 240 nm, relative to its sensitivity at 254 nm, followed by Q Beta, T1UV, T7m and T7 coliphage. B. pumilus spores were more sensitive to UV light at 220 nm than any of the coliphage. These spectra are required for calculating action spectra correction factors for medium pressure UV system validation, for matching appropriate challenge microorganisms to pathogens, and for improving UV dose monitoring. Additionally, understanding the dose response of these organisms at multiple wavelengths can improve polychromatic UV dose calculations and enable prediction of pathogen inactivation from wavelength-specific disinfection technologies such as UV light emitting diodes (LEDs).

Wavelength dependent UV inactivation and DNA damage of adenovirus as measured by cell culture infectivity and long range quantitative PCR.

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.

Comparison of absolute spectral irradiance responsivity measurement techniques using wavelength-tunable lasers.

Independent methods for measuring the absolute spectral irradiance responsivity of detectors have been compared between the calibration facilities at two national metrology institutes, the Helsinki University of Technology (TKK), Finland, and the National Institute of Standards and Technology (NIST). The emphasis is on the comparison of two different techniques for generating a uniform irradiance at a reference plane using wavelength-tunable lasers. At TKK's Laser Scanning Facility (LSF) the irradiance is generated by raster scanning a single collimated laser beam, while at the NIST facility for Spectral Irradiance and Radiance Responsivity Calibrations with Uniform Sources (SIRCUS), lasers are introduced into integrating spheres to generate a uniform irradiance at a reference plane. The laser-based irradiance responsivity results are compared to a traditional lamp-monochromator-based irradiance responsivity calibration obtained at the NIST Spectral Comparator Facility (SCF). A narrowband filter radiometer with a 24 nm bandwidth and an effective band-center wavelength of 801 nm was used as the artifact. The results of the comparison between the different facilities, reported for the first time in the near-infrared wavelength range, demonstrate agreement at the uncertainty level of less than 0.1%. This result has significant implications in radiation thermometry and in photometry as well as in radiometry.