Evaluating UV-C Sensitivity of Calonectria pseudonaviculata in Model Buffer Solution Using UV-C Light-Emitting-Diode System.

Calonectria pseudonaviculata, responsible for boxwood blight, produces sticky conidia that pose a contamination risk in boxwood production, cross-contamination tools and equipment and other resources. This study evaluated UV-C LED irradiation (263-287 nm) as a disinfection method by examining its effectiveness in inactivating conidia and determining the UV-C sensitivity. Conidial suspensions were exposed to quantifiable UV-C doses under dynamic stirring condition. Average volumetric intensity was quantified by accounting for UV gradients and UV dose was calculated as a product of average fluence rate (mW/cm2) and exposure time (s). UV-C irradiation effectively inactivated the tested pathogen following log-linear + shoulder kinetics as identified by parameters of goodness of model fit (i.e. high R2 and low RMSE values). The model predicted the UV sensitivity of C. pseudonaviculata conidia as 46.6 mJ/cm2/log. A total of 2.04 log reductions of the population could be obtained by an exposure of 60 mJ.cm-2 of UV-C dose. The calculated D10 was 13.53 ± 0.98 mJ.cm-2 (R2 = 0.97, RMSE = 0.14), Kmax = 0.17 ± 0.01, and shoulder length (Sl) = 33.06 ± 1.81 mJ.cm-2. These findings indicate that UV-C irradiation could be a viable option for disinfecting tools, equipment, and possibly propagation cuttings in nurseries.

Inactivation of B. cereus spores in whole milk and almond milk by serpentine path coiled tube UV-C system: Numerical simulation of flow field, lipid peroxidation and volatiles analysis.

A novel continuous thin-film (1.59 mm) serpentine path coiled tube (SPCT) UV system operating at 254 nm wavelength was designed and compared with flow field distribution of whole milk with helical path coiled tube (HPCT) UV system using computational fluid dynamics. The results revealed efficient velocity magnitude distribution at serpentine bend geometric locations of the SPCT UV system. Further in this study, we evaluatedBacillus cereusSpores inactivation in whole milk (WM) and almond milk (AM) using the developed SPCT UV system. Experimental data showed that > 4 log reduction of spores was achieved after six and ten passes of WM and AM at a flow rate of 70 and 162 mL/min, respectively. The bio-dosimetry method was used to verify the delivered reduction equivalent fluence (REF) and reported as 33 ± 0.73 and 36.5 ± 1.9 mJ/cm2. We noticed no significant effect on lipid oxidation and volatiles profile (p > 0.05) up to delivered REF of 60 mJ/cm2. This study demonstrated that high levels of inactivation ofB. cereusspores could be feasible with minimal impact on product quality by UV-C processing of dairy and non-dairy opaque scattering fluids.

Modeling the UV-C Inactivation Kinetics and Determination of Fluence Required for Incremental Inactivation of Cronobacter spp.

ABSTRACT: A study was undertaken to model the UV-C inactivation kinetics and determine the fluences required for the incremental inactivation of several strains of Cronobacter spp. suspended in clear phosphate-buffered saline (PBS). In total, 13 strains of Cronobacter spp. were individually suspended in PBS and treated with UV-C doses of 0, 2, 4, 6, 8, and 10 mJ cm-2 with a collimated beam device emitting UV-C at 253.7 nm. The log reduction from each treatment was identified using the plate count method and plotted against the UV-C dose and then curve fitted using several mathematical models. The UV-C dose required for incremental inactivation of each isolate was determined using both linear and nonlinear regression. For the 13 strains tested, a UV-C dose of 10 mJ cm-2 inactivated between 3.66 ± 0.101 and 5.04 ± 0.465 log CFU mL-1. The survival behavior of all strains was best fitted to the Weibull+tail model, with correlation coefficients between 97.17 and 99.71%, and was used to determine the fluences required for incremental inactivation. The UV-C fluences needed to inactivate 1 log (D10-value) of Cronobacter spp. in buffer were between 3.53 and 5.50 mJ cm-2, whereas a fluence greater than 6.57 mJ cm-2 was required to achieve a 4-log inactivation. A clear understanding of the UV-C dose-response of several strains of Cronobacter spp. lays the foundation to design effective UV-based disinfection systems.

Modeling and validation of delivered fluence of a continuous Dean flow pilot scale UV system: monitoring fluence by biodosimetry approach.

The inactivation of pathogenic microorganisms in water and high transmittance liquid foods has been studied extensively. The efficiency of the process is relatively low for treating opaque liquid foods using traditional UV systems. This study evaluated the ability of UV-C light to inactivate foodborne pathogens in a simulated opaque fluid (6.5 to 17 cm-1) at commercial relevant flow rates (31.70, 63.40, 95.10 gph) using a pilot-scale Dean Flow UV system. In this study, a mathematical model for the prediction of delivered fluence was developed by the biodosimetry method. The results revealed that increased Reduction equivalent fluence (REF) rates were observed with increased flow rates due to additional turbulence. The experimental and calculated REF were well correlated with the UV-C absorption coefficient range of 6.5 to 17 cm-1 indicating efficient mixing in the reactor. REF scaled up linearly at experimental conditions as an inverse function of flow rate and absorption coefficient, and a linear mathematical model (R2 > 0.99, p < 0.05) to predict delivered REF was developed. The model was tested and validated against independent experiments using Salmonella Typhimurium and Bacillus cereus endospores. The predicted and experimental REF values were in close agreement (p > 0.05). It is demonstrated that the developed model can predict the REF, thus microbial inactivation of microbial suspensions in simulated fluid with the absorption coefficient of 6.5-17 cm-1 and flow rates of 31.70-95.10 gph. The pilot system will be field-tested against microorganisms in highly absorbing and scattering fluids.

Phycobilins as Potent Food Bioactive Broad-Spectrum Inhibitors Against Proteases of SARS-CoV-2 and Other Coronaviruses: A Preliminary Study.

In the 21st century, we have witnessed three coronavirus outbreaks: SARS in 2003, MERS in 2012, and the ongoing pandemic coronavirus disease 2019 (COVID-19). The search for efficient vaccines and development and repurposing of therapeutic drugs are the major approaches in the COVID-19 pandemic research area. There are concerns about the evolution of mutant strains (e.g., VUI - 202012/01, a mutant coronavirus in the United Kingdom), which can potentially reduce the impact of the current vaccine and therapeutic drug development trials. One promising approach to counter the mutant strains is the "development of effective broad-spectrum antiviral drugs" against coronaviruses. This study scientifically investigates potent food bioactive broad-spectrum antiviral compounds by targeting main protease (Mpro) and papain-like protease (PLpro) proteases of coronaviruses (CoVs) using in silico and in vitro approaches. The results reveal that phycocyanobilin (PCB) shows potential inhibitor activity against both proteases. PCB had the best binding affinity to Mpro and PLpro with IC50 values of 71 and 62 µm, respectively. Also, in silico studies with Mpro and PLpro enzymes of other human and animal CoVs indicate broad-spectrum inhibitor activity of the PCB. As with PCB, other phycobilins, such as phycourobilin (PUB), phycoerythrobilin (PEB), and phycoviolobilin (PVB) show similar binding affinity to SARS-CoV-2 Mpro and PLpro.

Genomic Modeling as an Approach to Identify Surrogates for Use in Experimental Validation of SARS-CoV-2 and HuNoV Inactivation by UV-C Treatment.

Severe Acute Respiratory Syndrome coronavirus-2 (SARS-CoV-2) is responsible for the COVID-19 pandemic that continues to pose significant public health concerns. While research to deliver vaccines and antivirals are being pursued, various effective technologies to control its environmental spread are also being targeted. Ultraviolet light (UV-C) technologies are effective against a broad spectrum of microorganisms when used even on large surface areas. In this study, we developed a pyrimidine dinucleotide frequency based genomic model to predict the sensitivity of select enveloped and non-enveloped viruses to UV-C treatments in order to identify potential SARS-CoV-2 and human norovirus surrogates. The results revealed that this model was best fitted using linear regression with r 2 = 0.90. The predicted UV-C sensitivity (D 90 - dose for 90% inactivation) for SARS-CoV-2 and MERS-CoV was found to be 21.5 and 28 J/m2, respectively (with an estimated 18 J/m2 obtained from published experimental data for SARS-CoV-1), suggesting that coronaviruses are highly sensitive to UV-C light compared to other ssRNA viruses used in this modeling study. Murine hepatitis virus (MHV) A59 strain with a D 90 of 21 J/m2 close to that of SARS-CoV-2 was identified as a suitable surrogate to validate SARS-CoV-2 inactivation by UV-C treatment. Furthermore, the non-enveloped human noroviruses (HuNoVs), had predicted D 90 values of 69.1, 89, and 77.6 J/m2 for genogroups GI, GII, and GIV, respectively. Murine norovirus (MNV-1) of GV with a D 90 = 100 J/m2 was identified as a potential conservative surrogate for UV-C inactivation of these HuNoVs. This study provides useful insights for the identification of potential non-pathogenic (to humans) surrogates to understand inactivation kinetics and their use in experimental validation of UV-C disinfection systems. This approach can be used to narrow the number of surrogates used in testing UV-C inactivation of other human and animal ssRNA viral pathogens for experimental validation that can save cost, labor and time.

Effect of UV-C irradiation on the inactivation kinetics of oxidative enzymes, essential amino acids and sensory properties of coconut water.

The impact of ultraviolet light (UV-C) irradiation on oxidative enzymes [Polyphenol oxidase (PPO) and Peroxidase (POD)], free essential amino acids and sensory profile of coconut water were investigated. PPO and POD activities were lost to 94 and 93%, respectively of its original value at fluence level of 400 mJ/cm2. Inactivation kinetics of both enzymes were fitted to nonlinear Weibull model with an increase in UV dosage with a high coefficient of determination (R2 > 0.97) and low root mean square error (RMSE < 0.06). No significant change was observed in all essential amino acids (p > 0.05) after UV-C treatment up to maximum delivered fluence of 400 mJ/cm2. Sensory attributes of coconut water up to a treated UV-C fluence level of 200 mJ/cm2 were well retained in terms of chosen descriptors (p > 0.05). This study allow to further investigate the development of UV-C light technology for inhibition of spoilage enzymes and prolonged shelf-life of low acid beverages.

Performance of a UV-A LED system for degradation of aflatoxins B1 and M1 in pure water: kinetics and cytotoxicity study.

The efficacy of a UV-A light emitting diode system (LED) to reduce the concentrations of aflatoxin B1, aflatoxin M1 (AFB1, AFM1) in pure water was studied. This work investigates and reveals the kinetics and main mechanism(s) responsible for the destruction of aflatoxins in pure water and assesses the cytotoxicity in liver hepatocellular cells. Irradiation experiments were conducted using an LED system operating at 365 nm (monochromatic wave-length). Known concentrations of aflatoxins were spiked in water and irradiated at UV-A doses ranging from 0 to 1,200 mJ/cm2. The concentration of AFB1 and AFM1 was determined by HPLC with fluorescence detection. LC-MS/MS product ion scans were used to identify and semi-quantify degraded products of AFB1 and AFM1. It was observed that UV-A irradiation significantly reduced aflatoxins in pure water. In comparison to control, at dose of 1,200 mJ/cm2 UV-A irradiation reduced AFB1 and AFM1 concentrations by 70 ± 0.27 and 84 ± 1.95%, respectively. We hypothesize that the formation of reactive species initiated by UV-A light may have caused photolysis of AFB1 and AFM1 molecules in water. In cell culture studies, our results demonstrated that the increase of UV-A dosage decreased the aflatoxins-induced cytotoxicity in HepG2 cells, and no significant aflatoxin-induced cytotoxicity was observed at UV-A dose of 1,200 mJ/cm2. Further results from this study will be used to compare aflatoxins detoxification kinetics and mechanisms involved in liquid foods such as milk and vegetable oils.

Ultraviolet inactivation of bacteria and model viruses in coconut water using a collimated beam system.

This study investigated the effect of ultraviolet-C irradiation on the inactivation of microorganisms in coconut water, a highly opaque liquid food (1.01 ± 0.018 absorption coefficient). Ultraviolet-C inactivation kinetics of two bacteriophages (MS2, T1UV) and three surrogate bacteria (Escherichia coli, Salmonella Typhimurium, Listeria monocytogenes) in 0.1% (w/v) peptone and coconut water were investigated. Ultraviolet-C irradiation at 254 nm was applied to stirred samples, using a collimated beam device. A series of known ultraviolet-C doses (0-40 mJ cm-2) were applied for ultraviolet-C treatment except for MS2 where higher doses were delivered (100 mJ cm-2). Inactivation levels of all organisms were proportional to ultraviolet-C dose. At the highest dose of 40 mJ cm-2, three surrogates of pathogenic bacteria were inactivated by more than 5-log10 (p < 0.05) in 0.1% (w/v) peptone and coconut water. Results showed that ultraviolet-C irradiation effectively inactivated bacteriophage and surrogate bacteria in highly opaque coconut water. The log reduction kinetics of microorganisms followed log-linear and exponential models with higher R2 (>0.95) and low root mean square error values. The D10 values of 3, 5.48, and 4.58 mJ cm-2 were obtained from the inactivation of E. coli, S. Typhimurium, and L. monocytogenes, respectively. Models for predicting log reduction as a function of ultraviolet-C irradiation dose were found to be significant (p < 0.05). Fluid optics were the key controlling parameters for efficient microbial inactivation. Therefore, the ultraviolet-C dose must be calculated not only from the incident ultraviolet-C intensity but must also consider the attenuation in the samples. The results from this study imply that adequate log reduction of vegetative cells and model viruses is achievable in coconut water and suggested significant potential for ultraviolet-C treatment of other liquid foods.

Inactivation of Bacillus and Clostridium Spores in Coconut Water by Ultraviolet Light.

Bacterial spores are generally more resistant than vegetative bacteria to ultraviolet (UV) inactivation. The UV sensitivity of these spores must be known for implementing UV disinfection of low acid liquid foods. UV inactivation kinetics of bacterial spores in coconut water (CW) and distilled sterile water was studied. Populations of Bacillus cereus and Clostridium sporogenes dormant spores were reduced by more than 5.5 log10 at the UV-C photon fluence of 1142 µE·m-2 and 1919 µE·m-2 respectively. C. sporogenes spores showed higher UV-C resistance than B. cereus, with the photon fluence 300 µE·m-2 required for one log inactivation (D10) and 194 µE·m-2, respectively. No significant difference was observed in D10 values of spores suspended in the two fluid types (p > 0.05). The inactivation kinetics of microorganisms were described by log linear models with low root mean square error and high coefficient of determination (R2 > 0.98). This study clearly demonstrated that high levels of inactivation of bacterial spores can be achieved in CW. The baseline data generated from this study will be used to conduct spore inactivation studies in continuous flow UV systems. Further proliferation of the technology will include conducting extensive pilot studies.

UV-C Irradiation on the Quality of Green Tea: Effect on Catechins, Antioxidant Activity, and Cytotoxicity.

Short-wavelength ultraviolet (UV-C) irradiation is a nonthermal processing technique that is a possible alternative to the heat-pasteurization of tea beverages. This study investigated the effect of UV-C irradiation on the polyphenolic and total phenolic contents of a green tea beverage and analyzed cytotoxicity of irradiated green tea using a novel continuous flow-through UV system. UV-C fluence levels ranging from 0 to 240 mJ/cm2 were delivered to green tea, and polyphenols were chemically profiled. Continuous-flow UV-C irradiation of the green tea beverage at a fluence of 68 mJ/cm2  induced a minor reduction in the concentration of the most abundant catechin in green tea, (-)-epigallocatechin gallate (EGCG), from 145 to 131.1 µg/mL. The total phenolic content of the green tea beverage was 0.19 µg GAE/uL and remained constant at all UV fluence levels. The UV-treated green tea beverage showed no cytotoxic effects on normal intestinal cells with healthy colonic cells (CCD-18Co) maintained at 90% viability for the UV-treated green tea beverages and the control. The treated and nontreated green tea have comparable inhibitory effects on the survival of human colon cancer cells. Overall, these results demonstrate that the UV-C irradiation did not significantly deplete catechins or produce cytotoxic byproducts. PRACTICAL APPLICATION: Short wavelength ultraviolet (UV-C) irradiation is a nonthermal processing technique that is a possible alternative to the heat pasteurization of tea beverages. This study investigated the effect of UV-C irradiation on the antioxidant concentration of green tea and analyzed cytotoxicity of irradiated a green tea beverage using a novel continuous flow-through UV system. The results demonstrated that the UV-C irradiation did not significantly deplete catechins or produce cytotoxic byproducts.

Microbial inactivation and cytotoxicity evaluation of UV irradiated coconut water in a novel continuous flow spiral reactor.

A continuous-flow UV reactor operating at 254nm wave-length was used to investigate inactivation of microorganisms including bacteriophage in coconut water, a highly opaque liquid food. UV-C inactivation kinetics of two surrogate viruses (MS2, T1UV) and three bacteria (E. coli ATCC 25922, Salmonella Typhimurium ATCC 13311, Listeria monocytogenes ATCC 19115) in buffer and coconut water were investigated (D10 values ranging from 2.82 to 4.54mJ·cm-2). A series of known UV-C doses were delivered to the samples. Inactivation levels of all organisms were linearly proportional to UV-C dose (r2>0.97). At the highest dose of 30mJ·cm-2, the three pathogenic organisms were inactivated by >5 log10 (p<0.05). Results clearly demonstrated that UV-C irradiation effectively inactivated bacteriophage and pathogenic microbes in coconut water. The inactivation kinetics of microorganisms were best described by log linear model with a low root mean square error (RMSE) and high coefficient of determination (r2>0.97). Models for predicting log reduction as a function of UV-C irradiation dose were found to be significant (p<0.05) with low RMSE and high r2. The irradiated coconut water showed no cytotoxic effects on normal human intestinal cells and normal mouse liver cells. Overall, these results indicated that UV-C treatment did not generate cytotoxic compounds in the coconut water. This study clearly demonstrated that high levels of inactivation of pathogens can be achieved in coconut water, and suggested potential method for UV-C treatment of other liquid foods. INDUSTRIAL RELEVANCE: This research paper provides scientific evidence of the potential benefits of UV-C irradiation in inactivating bacterial and viral surrogates at commercially relevant doses of 0-120mJ·cm-2. The irradiated coconut water showed no cytotoxic effects on normal intestinal and healthy mice liver cells. UV-C irradiation is an attractive food preservation technology and offers opportunities for horticultural and food processing industries to meet the growing demand from consumers for healthier and safe food products. This study would provide technical support for commercialization of UV-C treatment of beverages.

Atmospheric Cold Plasma Inactivation of Salmonella and Escherichia coli on the Surface of Golden Delicious Apples.

The contamination of fruits with human pathogens is a reoccurring concern in the fresh produce industry. Atmospheric cold plasma (ACP) is a potential alternate to customary approaches for non-thermal decontamination of foods. In this study, the efficacy of a dielectric barrier discharge ACP system against Salmonella (Salmonella Typhimurium, ATCC 13311; Salmonella Choleraesuis, ATCC 10708) and Escherichia coli (ATCC 25922, ATCC 11775) was explored. For each bacteria, a two-strain mixture at 8 log10 CFU/ml was spot inoculated on the surface of Golden Delicious apples, air dried, and exposed to ACP at a fixed distance of 35 mm, input power of 200 W for 30, 60, 120, 180, and 240 s. Bacterial inactivation was achieved in all treatment times with highest reduction of 5.3 log10 CFU/cm2 for Salmonella and 5.5 log10 CFU/cm2 for E. coli. Our results showed that reductions were interrelated to exposure time and ranged from 1.3 to 5.3 and 0.6 to 5.5 log10 CFU/cm2 for Salmonella and E. coli, respectively. Salmonella and E. coli significantly decreased (>5.0 log) at 180 and 240 s as compared to 30, 60, and 120 s exposure. Microbial inactivation data was modeled by using Weibull distribution. These findings demonstrate the potential of ACP as a postharvest technology to effectively reduce pathogens on apples, with reference to Salmonella and E. coli.

Effect of UV irradiation on aflatoxin reduction: a cytotoxicity evaluation study using human hepatoma cell line.

In this proof-of-concept study, the efficacy of a medium-pressure UV (MPUV) lamp source to reduce the concentrations of aflatoxin B1, aflatoxin B2, and aflatoxin G1 (AFB1, AFB2, and AFG1) in pure water is investigated. Irradiation experiments were conducted using a collimated beam system operating between 200 to 360 nm. The optical absorbance of the solution and the irradiance of the lamp are considered in calculating the average fluence rate. Based on these factors, the UV dose was quantified as a product of average fluence rate and treatment time. Known concentrations of aflatoxins were spiked in water and irradiated at UV doses ranging from 0, 1.22, 2.44, 3.66, and 4.88 J cm-2. The concentration of aflatoxins was determined by HPLC with fluorescence detection. LC-MS/MS product ion scans were used to identify and semi-quantify degraded products of AFB1, AFB2, and AFG1. It was observed that UV irradiation significantly reduced aflatoxins in pure water (p < 0.05). Irradiation doses of 4.88 J cm-2 reduced concentrations 67.22% for AFG1, 29.77% for AFB2, and 98.25% for AFB1 (p < 0.05). Using this technique, an overall reduction of total aflatoxin content of ≈95% (p < 0.05) was achieved. We hypothesize that the formation of ˙OH radicals initiated by UV light may have caused photolysis of AFB1, AFB2, and AFG1 molecules. In cell culture studies, our results demonstrated that the increase of UV dosage decreased the aflatoxin-induced cytotoxicity in HepG2 cells. Therefore, our research finding suggests that UV irradiation can be used as an effective technique for the reduction of aflatoxins.

Treating cell culture media with UV irradiation against adventitious agents: minimal impact on CHO performance.

Sterility of cell culture media is an important concern in biotherapeutic processing. In large scale biotherapeutic production, a unit contamination of cell culture media can have costly effects. Ultraviolet (UV) irradiation is a sterilization method effective against bacteria and viruses while being non-thermal and non-adulterating in its mechanism of action. This makes UV irradiation attractive for use in sterilization of cell culture media. The objective of this study was to evaluate the effect of UV irradiation of cell culture media in terms of chemical composition and the ability to grow cell cultures in the treated media. The results showed that UV irradiation of commercial cell culture media at relevant disinfection doses impacted the chemical composition of the media with respect to several carboxylic acids, and to a minimal extent, amino acids. The cumulative effect of these changes, however, did not negatively influence the ability to culture Chinese Hamster Ovary cells, as evaluated by cell viability, growth rate, and protein titer measurements in simple batch growth compared with the same cells cultured in control media exposed to visible light.

The effect of delactosed whey permeate on phytochemical content of canned tomatoes.

The effect of delactosed whey permeate (DWP) treatment on antioxidant and phyto-chemical components of canned Irish plum tomatoes were investigated. Tomatoes were sterilised for 5 min (F(0)) at 120 °C and stored for 6 months. The DWP treatment retained significantly (p<0.05) higher levels of ascorbic acid and lycopene of tomatoes. The antioxidant activity of DWP treated tomatoes was higher (7%) than the control at the end of storage. The firmness in DWP-treated fruits was around 40% higher than that in control. All the parameters decreased significantly (p<0.05) during storage except lycopene and total phenols. Lycopene content showed no significant change and total phenols increased during storage. The changes in ascorbic acid, antioxidant activity and texture were fitted well to Weibull kinetic models with high coefficients of determination (R(2)) and low RMSE (root mean sum of squared error). The results clearly indicate that DWP enhanced the retention of antioxidant compounds in tomatoes during storage.

Optimization of ultrasound assisted extraction of antioxidant compounds from marjoram (Origanum majorana L.) using response surface methodology.

The present study optimized the ultrasound assisted extraction (UAE) conditions to maximize the antioxidant activity [Ferric ion Reducing Antioxidant Power (FRAP)], total phenol content (TP) and content of individual polyphenols of extracts from marjoram. Optimal conditions with regard to amplitude of sonication (24.4-61.0 µm) and extraction temperature (15-35 °C) and extraction time (5-15 min) were identified using response surface methodology (RSM). The results showed that the combined treatment conditions of 61 µm, 35 °C and 15 min were optimal for maximizing TP, FRAP, rosmarinic acid, luteolin-7-O-glucoside, apigenin-7-O-glucoside, caffeic acid, carnosic acid and carnosol values of the extracts. The predicted values from the developed quadratic polynomial equation were in close agreement with the actual experimental values with low average mean deviations (E%) ranging from 0.45% to 1.55%. The extraction yields of the optimal UAE were significantly (p < 0.05) higher than solid/liquid extracts. Predicted models were highly significant (p < 0.05) for all the parameters studied with high regression coefficients (R(2)) ranging from 0.58 to 0.989.