Assessment of saliva interference with UV-based disinfection technologies.

Worldwide shortages of personal protective equipment during COVID-19 pandemic has forced the implementation of methods for decontaminating face piece respirators such as N95 respirators. The use of UV irradiation to reduce bioburden of used respirators attracts attention, making proper testing protocols of uttermost importance. Currently artificial saliva is used but its comparison to human saliva from the UV disinfection perspective is lacking. Here we characterize UV spectra of human and artificial saliva, both fresh and after settling, to test for possible interference for UV-based disinfection. ASTM 2720 artificial saliva recipe (with either porcine or bovine mucin) showed many discrepancies from average (N = 18) human saliva, with different mucins demonstrating very different UV absorbance spectra, resulting in very different UV transmittance at different wavelength. Reducing porcine mucin concentration from 3 to 1.7 g/L brought UVA254 in the artificial saliva to that of average human saliva (although not for other wavelengths), allowing 254 nm disinfection experiments. Phosphate saline and modified artificial saliva were spiked with 8.6 log CFU/ml B. subtilis spores (ATCC 6633) and irradiated at dose of up to 100 mJ/cm2, resulting in 5.9 log inactivation for a saline suspension, and 2.8 and 1.1 log inactivation for ASTM-no mucin and ASTM-1.7 g/L porcine mucin 2 µL dried droplets, respectively. UVC irradiation of spores dried in human saliva resulted in 2.3 and 1.5 log inactivation, depending on the size of the droplets (2 vs 10 µL, respectively) dried on a glass surface. Our results suggest that in the presence of the current standard dried artificial saliva it is unlikely that UVC can achieve 6 log inactivation of B. subtilis spores using a realistic UV dose (e.g. less than 2 J/cm2) and the ATSM saliva recipe should be revised for UV decontamination studies.

A novel method for biosynthesis of different polymorphs of TiO2 nanoparticles as a protector for Bacillus thuringiensis from Ultra Violet.

Bacillus thuringiensis (Bt) were used for biosynthesis of amorphous TiO2 converted to distinct polymorphs (anatase, rutile, mix) under different temperature conditions. Characterizations of TiO2 nanoparticles were performed by using X-ray diffraction spectroscopy (XRD), Fourier-transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM) and, energy-dispersive X-ray spectroscopy (EDX) analysis. Stability of five formulations under ultraviolet (UV) radiation with spore viability and mortality test on Ephestia kuehniella Zeller larvae were investigated. TiO2(mix) showed the highest viabilities of 79.76% after exposure to ultraviolet (UVA385 nm), while viabilities of non-protected spores under these conditions were 41.32%. The mortality of TiO2(mix), TiO2(anatase), TiO2(rutile), TiO2(amorphous) and free spore formulations on second-instar larvae of Ephestia kuehniella were 73.76%, 71.24%, 57.12%, 51.32%, and 50.32%, respectively on the 10th day of the experiment. The obtained results suggest that TiO2(amorphous) does not increase Bt resistance, but both phases of TiO2 nanoparticles synthesized (anatase and rutile) through the Bacillus thuringiensis and phase mixture can increase the persistence of Bt to the UV light. Furthermore, the combination of both crystalline phases of TiO2(mix) has the highest performance in improving the Bt resistance.

Photocatalytic inactivation of bioaerosols in a fixed-bed reactor with TiO2-coated glass rings.

The photocatalytic inactivation of Bacillus subtilis spores in air was evaluated employing a fixed-bed reactor with TiO2-coated glass rings, under artificial UV-A radiation. Calculations of the radiation effectively absorbed inside the reactor were carried out by Monte Carlo simulations. The photocatalytic inactivation was assessed by analyzing the viability of the microorganisms retained by the coated glass rings inside the reactor at different irradiation periods. The initial concentration of the spores was reduced by almost 55% at the end of the experiment (12 h). Complementary assays were carried out employing Bacillus subtilis vegetative cells, obtaining a reduction of more than 96% under the same conditions. Two efficiency parameters were computed to assess the reactor performance: the photonic efficiency and the quantum efficiency of inactivation. Results of the efficiency parameters allow an objective comparison of the reactor performance under different experimental conditions and configurations.

Glycerol-based sterilization bioindicator system from Bacillus atrophaeus: development, performance evaluation, and cost analysis.

The development of new value-added applications for glycerol is of worldwide interest because of the environmental and economic problems that may be caused by an excess of glycerol generated from biodiesel production. A novel use of glycerol as a major substrate for production of a low-cost sterilization biological indicator system (BIS; spores on a carrier plus a recovery medium) was investigated. A sequential experimental design strategy was applied for product development and optimization. The proposed recovery medium enables germination and outgrowth of heat-damaged spores, promoting a D (160 °C) value of 6.6 ± 0.1 min. Bacillus atrophaeus spores production by solid-state fermentation reached a 2.3 ± 1.2 × 10(8) CFU/g dry matter. Sporulation kinetics results allowed this process to be restricted in 48 h. Germination kinetics demonstrated the visual identification of nonsterile BIS within 24 h. Performance evaluation of the proposed BIS against dry-heat and ethylene oxide sterilization showed compliance with the regulatory requirements. Cost breakdowns were from 41.8 (quality control) up to 72.8 % (feedstock). This is the first report on sterilization BIS production that uses glycerol as a sole carbon source, with significant cost reduction and the profitable use of a biodiesel byproduct.

Use of UV-C radiation to disinfect non-critical patient care items: a laboratory assessment of the Nanoclave Cabinet.

BACKGROUND: The near-patient environment is often heavily contaminated, yet the decontamination of near-patient surfaces and equipment is often poor. The Nanoclave Cabinet produces large amounts of ultraviolet-C (UV-C) radiation (53 W/m2) and is designed to rapidly disinfect individual items of clinical equipment. Controlled laboratory studies were conducted to assess its ability to eradicate a range of potential pathogens including Clostridium difficile spores and Adenovirus from different types of surface. METHODS: Each test surface was inoculated with known levels of vegetative bacteria (10(6) cfu/cm(2)), C. difficile spores (10(2)-10(6) cfu/cm(2)) or Adenovirus (10(9) viral genomes), placed in the Nanoclave Cabinet and exposed for up to 6 minutes to the UV-C light source. Survival of bacterial contaminants was determined via conventional cultivation techniques. Degradation of viral DNA was determined via PCR. Results were compared to the number of colonies or level of DNA recovered from non-exposed control surfaces. Experiments were repeated to incorporate organic soils and to compare the efficacy of the Nanoclave Cabinet to that of antimicrobial wipes. RESULTS: After exposing 8 common non-critical patient care items to two 30-second UV-C irradiation cycles, bacterial numbers on 40 of 51 target sites were consistently reduced to below detectable levels (≥ 4.7 log10 reduction). Bacterial load was reduced but still persisted on other sites. Objects that proved difficult to disinfect using the Nanoclave Cabinet (e.g. blood pressure cuff) were also difficult to disinfect using antimicrobial wipes. The efficacy of the Nanoclave Cabinet was not affected by the presence of organic soils. Clostridium difficile spores were more resistant to UV-C irradiation than vegetative bacteria. However, two 60-second irradiation cycles were sufficient to reduce the number of surface-associated spores from 10(3) cfu/cm(2) to below detectable levels. A 3 log10 reduction in detectable Adenovirus DNA was achieved within 3 minutes; after 6 minutes, viral DNA was undetectable. CONCLUSION: The results of this study suggest that the Nanoclave Cabinet can provide rapid and effective disinfection of some patient-related equipment. However, laboratory studies do not necessarily replicate 'in-use' conditions and further tests are required to assess the usability, acceptability and relative performance of the Nanoclave Cabinet when used in situ.

Monte Carlo N-particle simulation of neutron-based sterilisation of anthrax contamination.

OBJECTIVE: To simulate the neutron-based sterilisation of anthrax contamination by Monte Carlo N-particle (MCNP) 4C code. METHODS: Neutrons are elementary particles that have no charge. They are 20 times more effective than electrons or γ-rays in killing anthrax spores on surfaces and inside closed containers. Neutrons emitted from a (252)Cf neutron source are in the 100 keV to 2 MeV energy range. A 2.5 MeV D-D neutron generator can create neutrons at up to 10(13) n s(-1) with current technology. All these enable an effective and low-cost method of killing anthrax spores. RESULTS: There is no effect on neutron energy deposition on the anthrax sample when using a reflector that is thicker than its saturation thickness. Among all three reflecting materials tested in the MCNP simulation, paraffin is the best because it has the thinnest saturation thickness and is easy to machine. The MCNP radiation dose and fluence simulation calculation also showed that the MCNP-simulated neutron fluence that is needed to kill the anthrax spores agrees with previous analytical estimations very well. CONCLUSION: The MCNP simulation indicates that a 10 min neutron irradiation from a 0.5 g (252)Cf neutron source or a 1 min neutron irradiation from a 2.5 MeV D-D neutron generator may kill all anthrax spores in a sample. This is a promising result because a 2.5 MeV D-D neutron generator output >10(13) n s(-1) should be attainable in the near future. This indicates that we could use a D-D neutron generator to sterilise anthrax contamination within several seconds.

The assessment of particle association and UV disinfection of wastewater using indigenous spore-forming bacteria.

Studies have shown that association between particles and coliform bacteria in wastewater influence the inactivation of these microorganisms by ultraviolet (UV) irradiation. This research investigated the potential use of indigenous aerobic spore-forming (ASF) bacteria for studying the particle - microorganism interaction and its effect on UV disinfection of protozoan pathogens, such as Giardia spp. and Cryptosporidium spp., present in effluents from full-scale municipal wastewater treatment plants. The effect of particle - ASF association was determined by homogenizing wastewater effluent samples before and after exposure to controlled UV doses delivered by a bench-scale collimated beam apparatus. Particle association between Bacillus subtilis spores added to wastewater and wastewater particles was also assessed. The results indicate that spores are not significantly associated with wastewater particulate matter and particle association does not significantly affect the inactivation of indigenous spores present in wastewater by UV radiation in this study.

Assessment of Bacillus subtilis spores as a possible bioindicator for evaluation of the microbicidal efficacy of radiation processing of water.

Gamma and electron-beam irradiation of Bacillus subtilis spores suspended in different types of water was studied to evaluate the inactivation of the spores and assess their possible use as a bioindicator for radiation processing. We found that the inactivation proceeded endogenously, being dose-rate-dependent and affected by oxygen. The radiation resistance of the suspended spores was found to be rather high; therefore, B. subtilis spores used as a bioindicator for efficiency of water treatment by radiation under practical conditions might result in the spores being overly conservative surrogates for pathogenic microorganisms. Moreover, the doserate dependency impedes the use of the spores as a bioindicator. Thus, B. subtilis spores cannot be recommended as a bioindicator for evaluation of the microbicidal efficacy of ionizing radiation processing of water.

Experimental measurements of fluence distribution in a UV reactor using fluorescent microspheres.

One concern with current techniques of UV reactor validation is that they provide only a measure of the mean UV fluence. In this research, the actual fluence distribution of a UV reactor is measured through the use of photochemically active fluorescent microspheres. Experimental tests were performed in a pilot-scale monochromatic UV 254 nm reactor operated at two flow rates. Analysis of the fluorescence intensity decay was performed using collimated beam experiments for determination of decay rate kinetics. A stochastic hierarchal process involving Bayesian statistics, and the Markov chain Monte Carlo integration technique was used to correlate the microsphere fluorescence intensity distribution to the UV fluence distribution. The experimental UV fluence distribution was compared with the fluence distribution predicted using a computational fluid dynamics model. The results showed that the fluorescent microspheres measured a wider distribution of UV fluences with a flow rate of 3 gpm than with 7.5 gpm. The principal differences between the modeled and the measured distribution were in the low UV fluences where the microspheres predicted lower fluence levels than the model. The use of microspheres is demonstrated as a novel technique for measurement of the fluence distribution in UV reactors. This technique has both fundamental and practical implications for reactor evaluation and testing and could improve confidence in the future use of mathematical models for UV reactor characterization. It also serves as a complement to biodosimetry testing by providing greater insights regarding reactor behavior and validation.

High-performance, low-cost solar collectors for disinfection of contaminated water.

Although the germicidal action of sunlight has long been recognized, its potential for practical applications has to be researched more thoroughly. This paper summarizes the progress made toward a commercially practical collector for solar disinfection applications. Nontracking compound parabolic collectors (CPCs), developed originally for capturing solar photons for thermal energy applications, were examined as potential solar photoreactors. A field demonstration of solar disinfection treatment using commercially manufactured solar reactors was conducted. Field tests showed successful destruction of Escherichia coli and Enterococcus faecalis and have provided data for full-scale design of water treatment systems. From above observations, a throughput value of 50 L/m2 h for the low-cost CPC reactor tested was estimated. For a 190 m3/d (0.05 MGD) facility, the estimated total costs for disinfection using UV-A is U.S. $0.19/m3 ($0.70/1000 gal). The use of near-UV sunlight to disinfect water supplies seems promising in rural communities of developing countries where treated water is unavailable.

The track structures of ionizing particles and their application to radiation biophysics. I. A new analytical method for investigating two biophysical models.

A new approach to the interpretation of the effects of radiation on cells is described, in which sample particle tracks are constructed using a Monte Carlo computer program and the exposure of cellular targets to these tracks is simulated using a second program known as BIOPHYS. Data on the shapes and DNA contents of the cell nuclei are obtained from the literature. It is assumed that the sensitive material is DNA, and that the target is divided into cubes of approximately 2 nm (the diameter of the DNA helix) per side; the numbers of these cubes containing different numbers of ionizations are derived. Two different methods of analysing the output of BIOPHYS are described. In the first, it is assumed that lethality is caused by the occurrence of a number of ionizations equal to or greater than a certain threshold in one cube; in the second method, it is assumed that only two ionizations are required, in different parts of the cube, but that only some fraction of the cube is sensitive. These models have been applied to the interpretation of the variation of radiosensitivity with a linear energy transfer (LET) of spores of Bacillus subtilis exposed wet and dry, and good fits to the published experimental data were obtained using both models. Fits to experimental data for a range of other cell lines will be presented in a second paper.

Assessment of microwave sterilization of foods using intrinsic chemical markers.

The uniformity of microwave processing was investigated by measuring the formation of intrinsic chemical markers in disc-shaped and cylindrically-shaped whey protein gel model systems. These markers are formed as a result of thermally induced reactions of sugar and protein precursors. They were measured in samples placed in a pressurizable Teflon vessel and microwave heated to different peak temperatures using different power levels. Heating uniformity was mapped by sectioning the sample and analyzing for markers. The destruction of B. stearothermophilus spores in alginate beads was correlated with marker formation. The results show that the markers can be used to assess sterility and spatial time-temperature distributions in solid food samples.