Inactivation Data Research Articles

Thermal inactivation of non-proteolytic Clostridium botulinum spores in chilled plant-based foods

To support innovation in the dynamic plant-based foods sector, a study was conducted to set fit for purpose and risk-based alternative thermal processing conditions to the usually used safe harbour heat treatment of 90 °C - 10 min that ensures food safety of extended shelf life refrigerated products. To do so, inactivation data were collected to determine the thermal resistance properties (D- and z-values) of non-proteolytic Clostridium botulinum Type E spores (reference strain NCTC 8266) then build and validate specific inactivation models for two plant-based products: Konjac-based Seafood Analogue (Product 1) and Canola-based Eggs Analogue (Product 2) with desired sensorial properties not compatible with the application of the classical heat treatment of 90 °C - 10 min.D-values were determined at five temperatures ranging from 78 °C to 86 °C for each matrix to build the model and additional data were collected at 75 °C and 80 °C to validate the model. Dref80°C-values were estimated at 1.90 and 0.80 min and z-values at 6.12 °C and 6.84 °C respectively for products 1 and 2. Simulations were performed to estimate the time required to reach the required Performance Objective (PO) of 6 log reductions to ensure food safety at different temperatures for each product. The targeted PO could be reached in both products with lower temperatures and shorter times compared to the safe harbor. Results showed that the inactivation of non-proteolytic C. botulinum Type E spores was faster in product 2 compared to product 1. The developed and validated models, together with the proposed methodology can be used to support food industries in collecting specific data to justify setting fit for purpose heat treatments ensuring food safety with regards to the control of non-proteolytic C. botulinum type E spores without compromising on the desired sensorial properties such as those required for some plant-based products.

Inactivation kinetics for surrogates of common foodborne pathogens during food residue drying.

Postconsumer household food residues can act as useful substrates for other industries, but transporting high-moisture material corresponds to high fuel use and associated greenhouse gas production. Drying food residues at the household level reduces transportation weight, increases stability, and preserves the nutritional quality of recovered material. Mitigating foodborne microbiological safety risks is crucial to encourage the development of novel methods to rapidly dry and stabilize food residues. The objective of this study was to improve the prediction of bacterial pathogen inactivation under various thermal and drying processes in a synthetic mixture of residual food material (RFM). The log reduction rate was measured forEscherichia coli, Enterococcus faecium, and Listeria innocua (surrogates of common foodborne pathogens) in RFM under different moisture contents (12% and 25% by fresh weight) and temperatures (50, 55, and 60°C). Inactivation data were used to determine D- and z-values and to fit a multiple regression model to predict log(D-values) in response to temperature and moisture content. Across conditions, D-values were measured to be 5.1-120, 4.6-123, and 32-545min for E. coli, L. innocua, and E. faecium, respectively. Temperature sensitivities were significantly higher in lower moisture RFM for E. coli and L. innocua. Applying E. coli inactivation models during RFM at 55°C yielded inactivation rates that aligned with experimental values after 5min (0.1vs. 0-0.1logs), 30min (2.1vs. 1.3-2.3logs), and 90min (7.2vs. 7.1-8.9logs). These results can inform the design of RFM drying and stabilization processes to promote pathogen inactivation and safety in downstream applications of dried material.

Estimation of the UV susceptibility of aerosolized SARS-CoV-2 to 254 nm irradiation using CFD-based room disinfection simulations

The recent COVID-19 pandemic has raised interest in efficient air disinfection solutions. The application of germicidal ultraviolet (GUV) irradiation is an excellent contender to prevent airborne transmission of COVID-19, as well as other existing and future infectious airborne diseases. While GUV has already been proven effective in inactivating SARS-CoV-2, quantitative data on UV susceptibility and dose requirements, needed to predict and optimize the performance of GUV solutions, is still limited. In this study, the UV susceptibility of aerosolized SARS-CoV-2 to 254 nm ultraviolet (UV) irradiation is investigated. This is done by employing 3D computational fluid dynamics based simulations of SARS-CoV-2 inactivation in a test chamber equipped with an upper-room UV-C luminaire and comparing the results to previously published measurements performed in the same test chamber. The UV susceptibility found in this study is (0.6 ± 0.2) m2/J, which is equivalent to a D90 dose between 3 and 6 J/m2. These values are in the same range as previous estimations based on other corona viruses and inactivation data reported in literature.

Catalytic Stability of S-1-(4-Hydroxyphenyl)-Ethanol Dehydrogenase from Aromatoleum aromaticum.

Derived from the denitrifying bacterium Aromatoleum aromaticum EbN1 (Azoarcus sp.), the enzyme S-1-(4-hydroxyphenyl)-ethanol dehydrogenase (S-HPED) belongs to the short-chain dehydrogenase/reductase family. Using research techniques like UV-Vis spectroscopy, dynamic light scattering, thermal-shift assay and HPLC, we investigated the catalytic and structural stability of S-HPED over a wide temperature range and within the pH range of 5.5 to 9.0 under storage and reaction conditions. The relationship between aggregation and inactivation of the enzyme in various pH environments was also examined and interpreted. At pH 9.0, where the enzyme exhibited no aggregation, we characterized thermally induced enzyme inactivation. Through isothermal and multitemperature analysis of inactivation data, we identified and confirmed the first-order inactivation mechanism under these pH conditions and determined the kinetic parameters of the inactivation process. Additionally, we report the positive impact of glucose as an enzyme stabilizer, which slows down the dynamics of S-HPED inactivation over a wide range of pH and temperature and limits enzyme aggregation. Besides characterizing the stability of S-HPED, the enzyme's catalytic activity and high stereospecificity for 10 prochiral carbonyl compounds were positively verified, thus expanding the spectrum of substrates reduced by S-HPED. Our research contributes to advancing knowledge about the biocatalytic potential of this catalyst.

Demonstration of Inappropriate Validation Method for a Cracker Baking Process Using Predictive Modeling

Validation of baking processes for the inactivation of Salmonella is complicated by the combined effects of product heating and drying. The goal of this study was to quantitatively evaluate a previously disseminated approach to validating baking processes utilizing a predictive model developed using only isothermal and single-moisture inactivation data for the initially formulated dough. A simple cracker dough was formulated using flour inoculated with a five-strain cocktail of Salmonella. Side-by-side isothermal and baking experiments were performed to estimate Salmonella inactivation kinetics and to quantify survivors in a dynamic environment, respectively. Isothermal, single-moisture inactivation experiments were performed with cracker dough (water activity, aw = 0.956 ± 0.002; moisture content = 0.50 ± 0.01 dry basis) at three temperatures (56, 60, or 63°C) with ≥6 time intervals. Baking experiments were performed in a convection oven at 177°C with samples pulled every 30 s up to 360 s, with an endpoint product aw (25°C) of 0.45. The Salmonella isothermal, single-moisture inactivation kinetics in cracker dough resulted in D60°C and z−values of 4.6 min and 4.9°C, respectively; this model was then integrated over the dynamic product temperature profiles from the baking experiments. In the baking experiments, an average of 5-log reductions of Salmonella was achieved by 150 s of treatment; however, >100-log reductions were predicted by the dough-based models at that time point. This fail-dangerous overestimation of Salmonella lethality in crackers explicitly demonstrated that single-level moisture-based prediction models are inappropriate for describing inactivation in a process with both dynamic temperature and moisture, and that model-based validations must incorporate moisture/aw. Furthermore, end-users should exercise caution when utilizing unvalidated models to validate preventive control processes.

Evaluating the impact of pulsed light treatment on microbial and enzyme inactivation and quality attributes in fresh‐cut watermelon

AbstractThis study investigated the impact of pulsed light (PL) applied at a fluence of 4–12 J cm−2 (complying with U.S. Food and Drug Administration standards) on fresh‐cut watermelon, focusing on its efficacy in inactivating various food‐borne pathogenic gram‐positive (Escherichia coli and Staphylococcus aureus) and gram‐negative (Listeria monocytogenes and Salmonella) bacteria. The inactivation data were used to fit the log‐linear and Weibull kinetic models. Additionally, the impact of PL on physicochemical properties, lycopene content, total phenolic content, antioxidant activity, and pectin methylesterase (PME) residual activity (RA) was studied. The results demonstrated that the inactivation rate increases with the increase in fluence. Salmonella was most sensitive to PL, resulting in a reduction of 2.55 ± 0.08 log CFU g−1 and a higher kmax, whereas S. aureus was most resistant, with a reduction of 1.87 ± 0.24 log CFU g−1 and a lower kmax. The Weibull model was the best to predict the inactivation kinetics based on standard statistics and information theory criteria. For all assayed food‐borne pathogens, the inactivation curve showed non‐linearity with upward concavity except for L. monocytogenes, which followed a concave downward curve (β >1). Results showed that PL was effective in the retention of color, firmness, pH levels, total soluble solids, and titratable acidity, and also had a positive effect on enhancing the levels of essential components such as lycopene, total phenolic compounds, and antioxidant activity. Additionally, the PME RA significantly decreased with an increase in fluence due to structural modification. Therefore, PL can be explored to enhance fresh‐cut watermelon's safety and quality attributes.Practical ApplicationsThe implementation of this research holds significant potential for practical benefits in the fresh‐cut fruit industry. The findings could revolutionize processing practices, ensuring enhanced microbial safety, enzyme inactivation, and an extended shelf‐life of fresh‐cut watermelon. The study strongly contributes to preserving the desirable texture, color, and overall quality of the product, enhancing the retention of lycopene and other valuable antioxidants by standardizing the pulsed light treatment parameters while adhering to U.S. Food and Drug Administration standards for food application. Moreover, this research will scientifically contribute to addressing the critical challenges of foodborne outbreaks in the fresh‐cut fruit sector while providing consumers with a healthy, convenient food choice.

Unraveling the Biophysical Mechanisms of How Antiviral Detergents Disrupt Supported Lipid Membranes: Toward Replacing Triton X-100.

Triton X-100 (TX-100) is a membrane-disrupting detergent that is widely used to inactivate membrane-enveloped viral pathogens, yet is being phased out due to environmental safety concerns. Intense efforts are underway to discover regulatory acceptable detergents to replace TX-100, but there is scarce mechanistic understanding about how these other detergents disrupt phospholipid membranes and hence which ones are suitable to replace TX-100 from a biophysical interaction perspective. Herein, using the quartz crystal microbalance-dissipation (QCM-D) and electrochemical impedance spectroscopy (EIS) techniques in combination with supported lipid membrane platforms, we characterized the membrane-disruptive properties of a panel of TX-100 replacement candidates with varying antiviral activities and identified two distinct classes of membrane-interacting detergents with different critical micelle concentration (CMC) dependencies and biophysical mechanisms. While all tested detergents formed micelles, only a subset of the detergents caused CMC-dependent membrane solubilization similarly to that of TX-100, whereas other detergents adsorbed irreversibly to lipid membrane interfaces in a CMC-independent manner. We compared these biophysical results to virus inactivation data, which led us to identify that certain membrane-interaction profiles contribute to greater antiviral activity and such insights can help with the discovery and validation of antiviral detergents to replace TX-100.

Modeling the influence of propionic acid concentration and pH on the kinetics of Salmonella Typhimurium

Salmonella Typhimurium is a foodborne pathogen often found in the poultry production chain. Antibiotics have been used to reduce S. Typhimurium contamination in poultry aviaries and improve chicken growth. However, antibiotics were banned in several countries. Alternatively, organic acids, such as propionic acid (PA), can control pathogens. This study determined the PA minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC), and mathematically modeled S. Typhimurium growth/inactivation kinetics under the influence of PA at different pH values (4.5, 5.5, and 6.5) which are within the pH range of the chicken gastrointestinal tract. The PA MIC against S. Typhimurium was pH-dependent, resulting in 5.0, 3.5 and 9.0 mM undissociated PA at pH 4.5, 5.5, and 6.5, respectively. The Baranyi and Roberts and the Weibull model fit growth and inactivation data well, respectively. Secondary models were proposed. The validated model predicted 3-log reduction of S. Typhimurium in 3 h at 68.2 mM of undissociated PA and pH 4.5. The models presented a good capacity to describe the kinetics of S. Typhimurium subjected to PA, representing a useful tool to predict PA antibacterial action depending on the pH.

Inactivation Kinetics of Pathogenic and Nonpathogenic Bacteria Upon In Vitro Treatment With Cold Atmospheric Pressure Plasma (CAPP).

In the present study, selected pathogenic (Salmonella Typhimurium, Escherichia coli, Pseudomonas aeruginosa, Listeria monocytogenes, Bacillus cereus, and Staphylococcus aureus) and nonpathogenic (Pseudomonas fragi, Pseudomonas fluorescens, Brochothrix thermosphacta, Bacillus subtilis, Lactiplantibacillus plantarum, and Leuconostoc mesenteroides) bacteria were subjected in vitro in cold atmospheric pressure plasma (CAPP) treatment for up to 15 min and the changes in the surviving microbial population were determined. Plasma treatments were carried out by a plasma jet device, operating with argon (Ar) as carrier gas under constant flow (4.0 L/min) at a frequency of 1 MHz and an electrical voltage of 2-6 kV. Microbial inactivation data were modelled using linear and nonlinear (Geeraerd, Weibull) models, through which the corresponding kinetic parameters were calculated. After 15 min of exposure to plasma radiation, the total reduction in the bacterial populations was 2.12 log10 CFU mL-1 for P. fragi, 1.77 log10 CFU mL-1 for P. fluorescens, 2.30 log10 CFU mL-1 for B. thermosphacta, 1.58 log10 CFU mL-1 for B. subtilis, 1.31 log10 CFU mL-1 for L. plantarum, 3.80 log10 CFU mL-1 for L. mesenteroides (highest reduction observed), 1.12 log10 CFU mL-1 for S. Typhimurium, 1.18 log10 CFU mL-1 for E. coli, 1.43 log10 CFU mL-1 for L. monocytogenes, 1.32 log10 CFU mL-1 for B. cereus, 0.88 log10 CFU mL-1 for S. aureus, and 0.73 log10 CFU mL-1 for P. aeruginosa. The results showed a higher reduction in the population of nonpathogenic microorganisms compared to pathogens. The relatively small decrease in the inactivation of bacteria indicates that parameter optimization is necessary to be considered to improve the efficacy of the treatment.

Dynamic Thermal Treatments in Green Coconut Water Induce Dynamic Stress Adaptation of Listeria innocua That Increases Its Thermal Resistance.

The global coconut water market is projected to grow in the upcoming years, attributed to its numerous health benefits. However, due to its susceptibility to microbial contamination and the limitations of non-thermal decontamination methods, thermal treatments remain the primary approach to ensure the shelf-life stability and the microbiological safety of the product. In this study, the thermal inactivation of Listeria innocua, a Listeria monocytogenes surrogate, was evaluated in coconut water and in tryptone soy broth (TSB) under both isothermal (50-60 °C) and dynamic conditions (from 30 to 60 °C, with temperature increases of 0.5, 1 and 5 °C/min). Mathematical models were used to analyse the inactivation data. The Geeraerd model effectively described the thermal inactivation of L. innocua in both TSB and coconut water under isothermal conditions, with close agreement between experimental data and model fits. Parameter estimates and analysis revealed that acidified TSB is a suitable surrogate medium for studying the thermal inactivation of L. innocua in coconut water, despite minor differences observed in the shoulder length of inactivation curves, likely attributed to the media composition. The models fitted to the data obtained at isothermal conditions fail to predict L. innocua responses under dynamic conditions. This is attributed to the stress acclimation phenomenon that takes place under dynamic conditions, where bacterial cells adapt to initial sub-lethal treatment stages, leading to increased thermal resistance. Fitting the Bigelow model directly to dynamic data with fixed z-values reveals a three-fold increase in D-values with lower heating rates, supporting the role of stress acclimation. The findings of this study aid in designing pasteurization treatments targeting L. innocua in coconut water and enable the establishment of safe, mild heat treatments for refrigerated, high-quality coconut water.

The effect of emulsifier type and oil fraction on Salmonella Typhimurium growth and thermal inactivation in oil-in-water emulsion.

High water activity oil-in-water emulsions can promote survival and growth of Salmonella Typhimurium. Nevertheless, the precise effect of emulsifier type and oil content on bacterial growth and inactivation is not fully understood. Here, emulsions were prepared using different emulsifiers (Tween 20, Tween 80, and Triton X-100) and different oil fractions (20%, 40%, and 60% (v/v)). TSB (control), emulsifier solutions, and emulsions were inoculated with S. Typhimurium. Bacterial growth rate was measured at 7, 22, and 37°C, whereas thermal inactivation was performed at 55°C. Growth and inactivation data was fitted into Logistic and Weibull models, respectively. At an incubation temperature of 37°C, the presence of high amount of oil (60%) in Tween 20 and Triton X stabilized emulsions extended the lag phase (5.83±2.20 and 9.43±1.07h, respectively, compared to 2.28±1.54h for TSB, p<0.05), whereas individual emulsifiers had no effect on growth behavior compared to TSB. This effect was also prevalent but attenuated at 22°C, whereas no growth was observed at 7°C. In thermal inactivation, we observed protective effect in Tween 80 and Triton X-100 solutions, where time required for five-log reduction was 1914.70±706.35min and 795.34±420.09min, respectively, compared to 203.89±10.18min for TSB (p<0.05). Interestingly, the presence of high amount of oil did not offer protective effect during thermal inactivation. We hypothesize that oleic acid in Tween 80 and lower hydrophobicity value of Triton X-100 help maintain membrane integrity and improve the resistance of bacteria to heat inactivation.

Lethality Validation for Human Pathogenic Salmonella enterica on Chicken Feathers and Blood during Simulated Commercial Low-Temperature Dry Rendering.

Poultry rendering is the process of upcycling inedible poultry carcass materials into useful animal food/feed components as well as other valuable commercial products. Microbiological safety validation is nonetheless critical to ensuring the prevention of food safety hazard(s) transmission. This study determined the death kinetics of the thermotolerant Salmonella enterica serovar Senftenberg isolate 775W in chicken feathers and blood in low-temperature dry rendering (i.e., no direct contact with heating medium) to validate pathogen inactivation in commercial processing. Chicken feathers and blood were inoculated with Salmonella Senftenberg 775W and heated to 60, 70, or 80 °C for up to 60, 20, and 5 min, respectively. Three identically completed replicates (N = 3) for each product were conducted. Pathogen inactivation data were fitted to a non-linear model, providing for the detection and characterization of shoulder, log-linear death, and tailing components in death curves. The analysis showed a >7-log10 reduction in Salmonella was achieved across all processing temperatures, with t7D values (time for 7.0 log-cycle lethality) ranging from 21.68, 7.30, and 4.26 min for feathers and 18.38, 5.03, and 2.79 min in blood at 60, 70, and 80 °C, respectively. Study findings validate that low-temperature processing conditions can inactivate Salmonella in poultry-rendered offal.

Art of the Kill: Designing and Testing Viral Inactivation Procedures for Highly Pathogenic Negative Sense RNA Viruses.

The study of highly pathogenic viruses handled under BSL-4 conditions and classified as Select Agents frequently involves the transfer of inactivated materials to lower containment levels for downstream analyses. Adhering to Select Agent and BSL-4 safety regulations requires validation or verification of the inactivation procedures, which comes with an array of challenges for each method. This includes the use of cytotoxic reagents for chemical inactivation and defining the precise inactivation parameters for physical inactivation. Here, we provide a workflow for various inactivation methods using Ebola, Nipah, and Lassa viruses as our examples. We choose three distinct inactivation methods (TRIzol/TRIzol LS, aldehyde fixation using different fixatives, and heat) to highlight the challenges of each method and provide possible solutions. We show that, whereas published chemical inactivation methods are highly reliable, the parameters for heat inactivation must be clearly defined to ensure complete inactivation. In addition to the inactivation data, we also provide examples and templates for the documentation required for approval and use of inactivation SOPs, including an inactivation report, the procedure sections of developed SOPs, and an electronic inactivation certificate that accompanies inactivated samples. The provided information can be used as a roadmap for similar studies at high and maximum containment laboratories.

Evaluation of blue light therapy in treating viral pulmonary infection with a viral surrogate and lung cells

RNA viruses can cause severe diseases, which necessitates finding effective treatment of viral infection. Anti-microbial blue light therapies, whose effectiveness has been proved in inhibiting bacteria and fungi, can also be an alternative. However, few studies have been devoted to antiviral blue light therapies so far. Efforts have been made in this work to especially investigate the suitability of blue light in treating viral infections in lungs. First, for bio-safety concerns, experimental methods using lentiviral vectors as the surrogates for hazardous RNA viruses were developed. The lung fibroblast cell line (MRC-5) was adopted as the host cells. Second, various experimental and dosimetric parameters were applied and compared, which showed that the 415 nm light at 27 J cm−2 provided the best therapeutic effect among the three candidates of 405, 415 and 450 nm, without compromising the viability of the host cells. Third, the intracellular reactive oxygen species (ROS) were measured, which proved that although the blue light cannot directly induce toxicity to the virus, the therapeutic effects of the blue light are actually attributed to the toxic ROS generated via the host cells. More interestingly, a strong correlation between the viral inactivation data and the light induced ROS levels in the host cells was discovered, independent of the wavelength. In conclusion, the effectiveness of the antiviral blue light therapy has been demonstrated in the early treatment of viral pulmonary infection.

Inactivation of Salmonella Typhimurium in fresh-cut lettuce during chlorine washing: Assessing the impacts of free chlorine concentrations and exposure times

The aim of this study was to evaluate, quantify and model the inactivation of Salmonella in fresh-cut lettuce during washings with chlorinated water at different free chlorine concentrations (FCC, 0–150 mg/L). Individual fresh-cut lettuce samples (4 cm2) were inoculated with a Salmonella culture (ca. 4 log CFU/cm2) and washed with 100-mL solutions with different FCC for different times (0–150 s). The surviving Salmonella cells recovered from samples were enumerated by plate count methodology. A fast decay on Salmonella counts was marked in the first 20 s of washing, followed by a slowing down on reductions. A maximum of 2.6 log-decrease was observed after 2.5-min washing regardless of FCC. The log-linear with tail primary model coupled with a linear secondary model was fitted to inactivation data obtained at FCC from 50 to 150 mg/L through global regression analysis, yielding a suitable model to describe Salmonella concentrations as a function of FCC and washing times (RMSE = 0.34, R2adj = 0.84). Simulations using the developed model showed inactivation rates varying from 0.17 log CFU/s at 50 mg/L to 0.86 log CFU/s at 150 mg/L. Disinfection models on lettuce are valuable tools for the validation of control measures in the fresh-cut produce industry and for quantitative risk assessments.

Effects of ambient temperature and humidity on viruses activity on different architectural coatings and kinetics study

The inactivation rates of non-encapsulated virus (MS2, φA039, enterovirus EV71) and enveloped virus (influenza A virus H3N2) on architectural coating with different porosity were measured at different temperatures and relative humidity (RH). To study the variation of virus activity on different architectural coating with temperature and humidity and provide reference for the control of indoor building surface virus. The inactivation rate of the virus on different surfaces was analyzed at 15 °C, 25 °C, 35 °C and 45 °C and at RHs of 30 %, 60 %, and 90 % for 12 h or 24 h. Overall, the architectural porous coating is adsorbent to viruses. Under low temperatures and high RH conditions (15 °C, 90 % RH), the viruses can survive longer on the surface of the architectural nonporous (few pores) coating. At low humidity, water can evaporate rapidly on the porous surface, which contributes to virus inactivation, especially for nonenveloped viruses. Architectural porous coating can absorb and retain more water, which reduces the impact of high temperature on the virus under high humidity conditions. The inactivation rate of H3N2 on porous surfaces decreased with the decrease of humidity. In addition, the first-order kinetic and nonlinear Weibull models are used to describe the virus inactivation data. The inactivation of the virus on the surface of the architectural coating and the goodness of fit of the virus inactivation rate is related to the porosity of the architectural coating, temperature, RH, and the type of virus.

Impact of chlorine or peracetic acid on inactivation of Salmonella, Escherichia coli, and Listeria monocytogenes in agricultural water

Preharvest agricultural water has been recognized as one of the routes of contamination for foodborne pathogens during fruit and vegetable production. Several strategies have been proposed to reduce the risk of pathogens, including preharvest water chemigation, but literature is lacking with regards to microbiological inactivation of common bacterial foodborne pathogens associated with fresh produce contamination, Salmonella enterica, Shiga-toxigenic Escherichia coli (STEC), and Listeria monocytogenes, in surface irrigation water after exposure to chlorine and peracetic acid (PAA). Surface water supplied by a local irrigation district was collected over the summer of 2019. Water was autoclaved, divided into 100 mL samples, and inoculated with a cocktail of five Salmonella, STEC, or Listeria monocytogenes strains or a single strain non-pathogenic E. coli. Samples were then treated with 3, 5, or 7 ppm of free chlorine or PAA, and surviving populations were evaluated using a time-kill assay. A first-order kinetic model was used to fit the inactivation data and obtain the D-values. A secondary model was used to explain the changes due to the type of water, treatment, and microorganism. At a concentration of 3 ppm, the observed and predicted D-values of free chlorine treatments were higher than that of PAA treatments for ground and surface water. Results indicated that PAA was more effective inactivating bacteria than sodium hypochlorite at concentrations of 3 and 5 ppm for both water sources (surface and ground). However, at 7 ppm, the effectiveness of PAA and sodium hypochlorite showed no statistically significant difference for both surface and groundwater. Findings will provide information regarding efficacy of chemical sanitizers like chlorine and PAA for inactivation of Salmonella, Listeria, and STEC in surface water from which treatments can be derived. Ultimately benefitting growers in the selection of an appropriate method for in-field treatment of irrigation water if deemed necessary.

Pulsed light treatment of table grape juice: Influence of matrix pH on microbial and enzyme inactivation kinetics

The influence of pulsed light (PL) treatment and matrix pH was tested in the case of green table grape juice. Escherichia coli, Listeria monocytogenes, and Saccharomyces cerevisiae were inoculated into the table grape juices with matrix pH levels of 3.0, 3.5, and 4.0. Further, the juice was treated at 378–3186 J/cm2. The Weibull model better explained the microbial inactivation data. The shape parameters (β) from the Weibull model for E. coli, L. monocytogenes, and S. cerevisiae are 0.95 ± 0.02, 0.55 ± 0.04, and 0.40 ± 0.03, respectively. A lesser δ-value (scale parameter in J/cm2) was obtained for each microorganism at a lower pH. PL exposure resulted in a lack of cell visibility, cell wall, and cell membrane disruption in bacterial cells. The inactivation of polyphenol oxidase (PPO), peroxidase (POD), and pectin methyl esterase (PME) was studied at an elevated fluence level of 1152–3186 J/cm2. Enzymes in the juice were less sensitive to PL than microorganisms. The enzyme inactivation of >90% was achieved at 3186 J/cm2; thus, the juice was adjudged as the microbially safe and enzymatically stable pasteurized sample. The PL-induced enzyme inactivation followed nth-order kinetics with an inactivation order (n) of 1.1, 1.15, and 0.85 for PPO, POD, and PME, respectively. The enzyme inactivation rate was higher at a low pH. The PL-treated pasteurized juice at pH 3.5 showed the highest sensory acceptance (7.4 out of 9), with a minimal color change, a 7% loss in total phenolics, and a 12% loss in vitamin C and flavonoids.

Abstract 1413: Chromosome arm aneuploidies as genetic vulnerabilities of triple-negative breast cancer

Abstract Triple-negative breast cancer (TNBC) is an aggressive subtype of breast cancer. It has no approved targeted therapies due to the lack of the expression of key biomarkers, namely Estrogen and Progesterone receptors, and HER2 amplification. As a result, TNBC has the worst prognosis compared to other breast cancer subtypes. We previously showed that chromosome 4p (chr4p) loss is recurrent, correlates with poor prognosis, is an early event in tumor evolution, and confers a proliferative advantage. Here, we propose to leverage chr4p loss as a genetic vulnerability of TNBC by identifying its genetic interactions (GIs) with genes whose inactivation leads to synthetic lethality i.e. loss of viability of chr4p copy loss cells, but not of the chr4p copy neutral cells, or suppression i.e., positive GIs. We first classified breast cancer cell lines with TNBC enriched molecular subtype called basal-like, as chr4p copy loss (9) and chr4p copy neutral (7), using copy number data from CCLE. Associated with chr4p copy loss, the differential expression analysis carried out using CCLE data revealed transcriptomic changes that distinctly cluster the chr4p copy loss and chr4p copy neutral cell lines. We then used the publicly available CRISPR-based genome-wide gene inactivation data from the DepMap project and quantified chr4p loss-associated GIs using multiple scoring methods such as drugZ and MAGeCK. In total, we identified ~200 negative and ~70 positive statistically significant GIs involving genes annotated to biological processes such as translation, metabolism, and others. We have prioritized a subset of them for subsequent experimental validation using CRISPR-Cas9-based gene editing in TNBC cancer cell line models. Alongside, using the publicly available drug sensitivity data obtained from the PRISM project, we identified compounds that impair cell growth in a chr4p loss-specific manner, the strongest of which targeted redox balance. Collectively, the set of GIs, the compound sensitivities, and the computational methods we have generated are unique resources for developing precision oncology therapeutic strategies for TNBC, as well as for other cancers harboring chr4p loss. Citation Format: Rohan Dandage, Michael Schwartz, Lynn Karam, Alain Pacis, Traver Hart, Guillaume Bourque, Morag Park, Elena Kuzmin. Chromosome arm aneuploidies as genetic vulnerabilities of triple-negative breast cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 1413.

Performance of UV-LED and UV-C treatments for the inactivation of Escherichia coli ATCC 25922 in food model solutions: Influence of optical and physical sample characteristics

Shortwave ultraviolet light (UV-C) and ultraviolet light-emitting diodes (UV-LEDs) are emergent technologies to inactivate pathogenic and spoilage microorganisms in food. However, the effectiveness of these technologies is influenced by the optical properties of the treated food. This work aimed to evaluate the effect of the optical properties of nine model food solutions on the efficacy of UV-C (at 255 nm) and UV-LEDs (at 255, 265 or 280 nm) to inactivate E. coli ATCC 25922. Model solutions were formulated with saccharose (SC), tartrazine (TT) and xanthan gum (XG), exposed from 0 to 50 min. The microbial population was reduced after 15 min of UV-C and UV-LED treatment by >6 log CFU/mL for water and TT, and by UV-C for SC, XG, TT + SC, and (XG + TT + SC) m solutions. The inactivation data were correlated using three different models. Colored compound (TT) showed 60% degradation by UV-C compared to 3% by UV-LED at 50 min. Industrial relevanceNon-thermal treatments such as those based on ultraviolet light, UV-C, and UV-LEDs could have high industrial relevance because of their simplicity of operation and reduced by-product formation, being a friendly alternative for food processing. Understanding the effect of different optical and physicochemical properties of liquid food to be treated by UV-based technologies is mandatory for the equipment's efficient design and operation. Furthermore, the proper selection of processing conditions, such as delivered dose and processing time, allows for obtaining safe and high-quality products.