Amoebicidal effect of chlorine dioxide gas against pathogenic Naegleria fowleri and Acanthamoeba polyphaga.

The pathogenic free-living amoebae, Naegleria fowleri and Acanthamoeba polyphaga, are found in freshwater, soil, and unchlorinated or minimally chlorinated swimming pools. N. fowleri and A. polyphaga are becoming problematic as water leisure activities and drinking water are sources of infection. Chlorine dioxide (ClO2) gas is a potent disinfectant that is relatively harmless to humans at the concentration used for disinfection. In this study, we examined the amoebicidal effects of ClO2 gas on N. fowleri and A. polyphaga. These amoebae were exposed to ClO2 gas from a ready-to-use product (0.36 ppmv/h) for 12, 24, 36, and 48 h. Microscopic examination showed that the viability of N. fowleri and A. polyphaga was effectively inhibited by treatment with ClO2 gas in a time-dependent manner. The growth of N. fowleri and A. polyphaga exposed to ClO2 gas for 36 h was completely inhibited. In both cases, the mRNA levels of their respective actin genes were significantly reduced following treatment with ClO2 gas. ClO2 gas has an amoebicidal effect on N. fowleri and A. polyphaga. Therefore, ClO2 gas has been proposed as an effective agent for the prevention and control of pathogenic free-living amoeba contamination.

Antimicrobial efficiency of chlorine dioxide and its potential use as anti-SARS-CoV-2 agent: mechanisms of action and interactions with gut microbiota.

Chlorine dioxide (ClO2) is a disinfectant gas with strong antifungal, antibacterial, and antiviral activities. Applied on hard, non-porous surfaces as an aqueous solution or gas, the ClO2 exerts antimicrobial activity through its interaction and destabilization of cell membrane proteins, as well as through DNA/RNA oxidation, triggering cell death. As for viruses, the ClO2 promotes protein denaturalization mechanisms, preventing the union between the human cells and the viral envelope. Currently, ClO2 has been pointed out as a potential anti-SARS-CoV-2 clinical treatment for use in humans with the ability to oxidize the cysteine residues in the spike protein of SARS-CoV-2, inhibiting the subsequent binding with the Angiotensin-converting enzyme type 2 receptor, located in the alveolar cells. Orally administered ClO2 reaches the gut tract and exacerbates the symptoms of COVID-19, generating a dysbiosis with gut inflammation and diarrhea as side effects, and once absorbed, produces toxic effects including methemoglobinemia and hemoglobinuria, which can trigger respiratory diseases. These effects are dose-dependent and may not be entirely consistent between individuals since the gut microbiota composition is highly heterogeneous. However, to support the use of ClO2 as an anti-SARS-CoV-2 agent, further studies focused on its effectiveness and safety both in healthy and immunocompromised individuals, are needed.

Phthalate acid esters promoted the enrichment of chlorine dioxide-resistant bacteria and their functions related to human diseases in rural polyvinyl chloride distribution pipes.

Polyvinyl chloride (PVC) pipes are widely used as drinking water distribution pipes in rural areas of China. However, whether phthalate acid esters (PAEs) released from PVC pipes will affect tap water quality is still unknown. The influence of released PAEs on the water quality was analysed in this study, especially after ClO2 disinfection. The results indicated that ClO2 disinfection could control the growth of total coliforms and heterotrophic bacteria (HPC). However, when the ClO2 residual decreased to below 0.10 mg/L, HPC and opportunistic pathogens, including Mycobacterium avium and Pseudomonas aeruginosa, increased significantly. In addition, after ClO2 disinfection, PAEs concentrations increased from 10.6-22.2 µg/L to 21.2-58.8 µg/L in different sampling cites. Linear discriminant analysis (LDA) effect size (LEfSe) and statistical analysis of metagenomic profiles (Stamp) showed that ClO2 disinfection induced the enrichment of Pseudomonas, Bradyrhizobium, and Mycobacterium and functions related to human diseases, such as pathogenic Escherichia coli infection, shigellosis, Staphylococcus aureus infection, and Vibrio cholerae infection. The released PAEs not only promoted the growth of these ClO2-resistant bacterial genera but also enhanced their functions related to human diseases. Moreover, these PAEs also induced the enrichment of other bacterial genera, such as Blastomonas, Dechloromonas, and Kocuria, and their functions, such as chronic myeloid leukaemia, African trypanosomiasis, leishmaniasis, hepatitis C and human T-cell leukaemia virus 1 infection. The released PAEs enhanced the microbial risk of the drinking water. These results are meaningful for guaranteeing water quality in rural areas of China.

Evaluation of antimicrobial efficacies of chlorine dioxide gas released into the air towards pathogens present on the surfaces of inanimate objects.

The efficacy of ClO2 gas, as surface disinfectant at around 1,000 ppb against avian orthoavulaviruses type 1 (AOAV-1), infectious bronchitis virus (IBV), Escherichia coli (EC), and Salmonella Enteritidis (SE) was evaluated at the required level (≥99.9% reduction) on various surfaces. Exposing the surfaces to ClO2 gas for 1 hr reduced AOAV-1, except for rayon sheets which required 3 hr. However, 1 hr of exposure did not effectively reduced IBV titer. In the case of EC, glass plates and plastic carriers needed 1 hr of exposure, while rayon sheets required 2 hr. SE on rayon sheets required 1 hr exposure, but on the other tested surfaces showed inadequate reduction. Overall, ClO2 gas is an effective disinfectant for poultry farms.

Viral Disinfection of Porous Fomites Utilizing a Bacteriophage Model and Chlorine Dioxide Gas.

The pursuit of disinfecting porous materials or fomites to inactivate viral agents has special challenges. To address these challenges, a highly portable chlorine dioxide (ClO2) gas generation system was used to ascertain the ability of a gaseous preparation to inactivate a viral agent, the MS2 bacteriophage, when associated with potentially porous fomites of cloth, paper towel, and wood. The MS2 bacteriophage is increasingly used as a model to identify means of inactivating infectious viral agents of significance to humans. Studies showed that MS2 bacteriophage can be applied to and subsequently recovered from potential porous fomites such as cloth, paper towel, and wood. Paired with viral plaque assays, this provided a means for assessing the ability of gaseous ClO2 to inactivate bacteriophage associated with the porous materials. Notable results include 100% inactivation of 6 log bacteriophage after overnight exposure to 20 parts per million(ppm) ClO2. Reducing exposure time to 90 minutes and gas ppm to lower concentrations proved to remain effective in bacteriophage elimination in association with porous materials. Stepwise reduction in gas concentration from 76 ppm to 5 ppm consistently resulted in greater than 99.99% to 100% reduction of recoverable bacteriophage. This model suggests the potential of ClO2 gas deployment systems for use in the inactivation of viral agents associated with porous potential fomites. The ClO2 gas could prove especially helpful in disinfecting enclosed areas containing viral contaminated surfaces, rather than manually spraying and wiping them.

The effectiveness of chlorine dioxide gas in portable personal disinfectants to inhibit bacterial growth.

Disinfectants, especially air disinfectants, are necessary to prevent the potential spread of pathogens (bacteria and viruses) in the pandemic era and minimize the spread of pathogens. Some of the commercial disinfectant products that are often used generally contain chlorine dioxide (ClO2) gas. This study tested the effectiveness of two different commercial disinfectants, a liquid stick disinfectant and a powder disinfection card, to carry out the disinfection of pathogenic bacteria in the environment. These two disinfectants were used as a medium for releasing chlorine dioxide gas which has a much stronger bactericidal effect. In the form of liquid stick, ClO2 is more effective in the disinfection process rather than in the form of powder. The effectiveness of the liquid disinfectant in inhibiting the growth of pathogenic bacteria is influenced by the temperature and the area of the open space covered. Considering that the release from both disinfectants used is very small (0.002 ppmv/h), it takes a small area to ensure that the disinfection process runs effectively.

Could polyhexanide and chlorine dioxide be used as an alternative to chlorhexidine? A systematic review

ABSTRACT BACKGROUND: Maintenance of oral microbiota balance is the simplest way to prevent infectious oral diseases, through controlling dental biofilm. Combined use of mouthwash and mechanical removal has been shown to be a very effective way for this. OBJECTIVES: To identify clinical studies comparing the antimicrobial effect and possible adverse effects and/or side effects of chlorhexidine-based mouthwashes with those of mouthwashes containing chlorine dioxide and/or polyhexanide, for controlling oral microbiota. DESIGN AND SETTING: Systematic review designed by the stomatology sector of postgraduation in applied dental sciences of Bauru Dentistry School, University of São Paulo, Brazil. METHODS: A systematic review was conducted using online databases (PubMed, Embase, Web of Science and Science Direct) up to April 8, 2020. The search was conducted using the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. RESULTS: The studies included comprised eight articles published between 2001 and 2017. A total of 295 young adults, adults and elderly people were evaluated (males 44.75% and females 55.25%). Three articles compared polyhexanide with chlorhexidine and five articles compared chlorine dioxide with chlorhexidine. No studies comparing all three mouthwashes were found. The concentrations of the study solutions were quite varied, and all rinses had an antimicrobial effect. In four studies, it was stated that no side effects or adverse effects had been found. Three studies did not address these results and only one study addressed side effects and/or adverse effects. CONCLUSION: Mouthwashes containing chlorine dioxide and polyhexanide are viable alternatives to chlorhexidine, since they reduce oral biofilm and have little or no reported side or adverse effects.

pH-Regulating Nanoplatform for the "Double Channel Chase " of Tumor Cells by the Synergistic Cascade between Chlorine Treatment and Methionine-Depletion Starvation Therapy.

During rapid proliferation and metabolism, tumor cells show a high dependence on methionine. The deficiency of methionine exhibits significant inhibition on tumor growth, which provides a potential therapeutic target in tumor therapy. Herein, ClO2-loaded nanoparticles (fluvastatin sodium&metformin&bupivacaine&ClO2@CaSiO3@MnO2-arginine-glycine-aspatic acid (RGD) (MFBC@CMR) NPs) were prepared for synergistic chlorine treatment and methionine-depletion starvation therapy. After outer layer MnO2 was degraded in the high glutathione (GSH) tumor microenvironment (TME), MFBC@CMR NPs released metformin (Me) to target the mitochondria, thus interfering with the tricarboxylic acid (TCA) cycle and promoting the production of lactate. In addition, released fluvastatin sodium (Flu) by the NPs acted on monocarboxylic acid transporter 4 (MCT4) in the cell membrane to inhibit lactate leakage and induce a decrease of intracellular pH, further prompting the NPs to release chlorine dioxide (ClO2), which then oxidized methionine, inhibited tumor growth, and produced large numbers of Cl- in the cytoplasm. Cl- could enter mitochondria through the voltage-dependent anion channel (VDAC) channel, which was opened by bupivacaine (Bup). The disruption of Cl- homeostasis promotes mitochondrial damage and membrane potential decline, leading to the release of cytochrome C (Cyt-C) and apoptosis inducing factor (AIF) and further inducing cell apoptosis. To sum up, the pH-regulating and ClO2-loaded MFBC@CMR nanoplatform can achieve cascade chlorine treatment and methionine-depletion starvation therapy toward tumor cells, which is of great significance for improving the clinical tumor treatment effect.

Antimicrobial activity of ClO2 gas against Salmonella Enteritidis on almonds.

Nuts, including almonds, are occasionally contaminated with Salmonella spp. In this study, we used chlorine dioxide (ClO2) gas to inactivate S. enterica subsp. Enterica serovar Enteritidis on almonds. Almonds inoculated with a single strain of S. Enteritidis (8.95 log cfu/mL) were exposed to ClO2 gas generated from 1.0 or 1.5 mL ClO2 solution in a sealed container at 50 or 60 °C (43% relative humidity) for up to 10 h. The concentration of ClO2 gas peaked at 354-510 and 750-786 ppm within 0.5 h upon deposition of 1.0 and 1.5 mL of aqueous ClO2, respectively, and gradually decreased thereafter. Population of S. Enteritidis on almonds treated at 50 °C decreased to 1.70-2.32 log cfu/sample within 1 h of exposure to ClO2 gas and decreased to below the detection limit (1.7 log cfu/sample) at all ClO2 concentrations after 8 h. At 60 °C, the microbial population fell below the detection limit within 1 h, regardless of the volume of ClO2 solution supplied. Microbial survival on almonds treated with ClO2 gas and stored at 12 or 25 °C was observed for up to 8 weeks and the organism was not recovered from the almonds treated for 10 h and stored at 12 °C for 2-8 weeks. The lightness (L value) and redness (a value) of almonds treated for 10 h were not changed by ClO2 gas treatment, but yellowness (b value) increased. Results showed that Salmonella on almonds was successfully inactivated by ClO2 gas treatment and the microbial survival did not occur during storage.

Efficacy of Chlorine Dioxide Gas Against Hepatitis A Virus on Blueberries, Blackberries, Raspberries, and Strawberries.

Seeking a means of sanitizing berries, the effectiveness of steady state levels of gaseous chlorine dioxide (ClO2) against hepatitis A virus (HAV) on laboratory-contaminated berries was determined. The generated ClO2 was maintained with 1 or 2 mg/l air inside a 269-l glove box to treat 50 g batches of blueberries, raspberries, and blackberries, and 100 g batches of strawberries that were immersion coated with HAV. Normalized data for ClO2 (ppm-h/g product) is reported as a function of ClO2 concentration, treatment time, and weight of treated product. Treatments of ClO2 ranging from 1.00 to 6.27 ppm-h/g berry were evaluated. When compared to untreated HAV-contaminated berries, log reductions of HAV were > 2.1 for all berry types and conditions tested indicating the gaseous ClO2 was effective. The average log reduction with strawberries, raspberries, blueberries and blackberries treated with 1.00 ppm-h/g, the lowest ClO2 treatment tested, were 2.44, 2.49, 3.23, and 3.45, respectively. The highest treatment of 6.27 ppm-h/g was applied at two different gas concentrations of 1 mg/l and 2 mg/l. Average log reductions for blueberries and strawberries treated with 6.27 ppm-h/g were 4.34 and 4.42, and 4.03 and 3.51, applied at 1 mg/l and 2 mg/l, respectively. For blackberries and raspberries 3.20 and 3.24, and 3.23 and 3.97 log reductions were observed for 6.27 ppm-h/g treatments applied at 1 mg/l and 2 mg/l, respectively. Results indicate that HAV contamination of berries can be substantially reduced by gaseous ClO2 and offer industry a waterless means of sanitizing berries against HAV.

Efficacy of novel aqueous photo-chlorine dioxide against a human norovirus surrogate, bacteriophage MS2 and Clostridium difficile endospores, in suspension, on stainless steel and under greenhouse conditions.

AIMS: The efficacy of a novel photochemical method for generating chlorine dioxide (photoClO2 ) was evaluated against human noroviruses (HuNoV) surrogate, bacteriophage MS2, and Clostridium difficile endospores. METHODS AND RESULTS: Chlorine dioxide was generated by mixing 1% sodium chlorite with 10 parts-per-million (ppm) Eosin Y and irradiating with a photo-activator-excitable light. PhotoClO2 efficacy was assessed against bacteriophage MS2 and C. difficile endospores in suspension, on hard surfaces and greenhouse conditions under soiled and unsoiled conditions. The estimated effective photoClO2 produced and consumed was 20·39 ± 0·16 ppm at a rate of 8·16 ppm per min in a 1% sodium chlorite solution. In suspension, MS2 phage was reduced by 3·35 and >5·10 log10 PFU per ml in 120 and 90 min, with and without soil, respectively. At the same time, when dried on stainless steel surface, MS2 phage was reduced by >4·53 log10 PFU per carrier in 30 min under both conditions. On the other hand, C. difficile endospores in suspension were reduced by 2·26 and 3·65 log10 CFU per ml in 120 min with and without soiling, respectively. However, on stainless steel surface, maximal reductions of the C. difficile endospores were 0·8 and 1·5 log10 CFU per carrier with and without soiling, respectively, and a maximal reduction of 2·97 log10 CFU per carrier under greenhouse conditions at 24 h. CONCLUSIONS: Overall, photoClO2 showed promise as a technology to control HuNoV contamination on environmental surfaces but requires further optimization and testing against C. difficile endospores. SIGNIFICANCE AND IMPACT OF THE STUDY: Results from this investigation will serve as a model for how to generate and quantify photoClO2 and how to appropriately evaluate this new class of disinfectants against environmentally resilient pathogens: viruses and bacterial endospores.

Extension of shelf life of semi-dry longan pulp with gaseous chlorine dioxide generating film.

A packaging system using gaseous chlorine dioxide generating film (CDGF) in a sealed container was developed to extend the shelf life of semi-dry longan pulp (moisture content 38.8 wt%; aw0.8). The antimicrobial properties, formation of chloroxyanion residues and effects of CDGF on the quality of semi-dry longan pulp were investigated. CDGF was triggered by the moisture vapor from semi-dry longan pulp in the sealed container and released gaseous ClO2 into the headspace of the container. The antifungal test showed that CDGF significantly inactivated artificially inoculated molds in semi-dry longan pulp and achieved reductions of over 3 log CFU/g after 28 days storage at room temperature (25 °C). CDGF reduced total aerobic bacterial populations by over 6.4 log CFU/g and maintained these population levels at around 2.0 log CFU/g throughout the 180-day storage period at room temperature. The residual concentrations of chloride, chlorate and perchlorate in longan pulp increased and then decreased during the 180-day storage. Residual chloride levels were maintained at 1.5 mg/g after Day 120 and residual chlorate and perchlorate levels were not detected after Day 120 and Day 180, respectively, in CDGF-treated samples. CDGF treatments reduced total polyphenol content but didn't have any significant impact on the levels of polysaccharides in samples. There were no significant differences between CDGF-treated and control samples in color changes during storage. The content of 5-hydroymethylfurfural (5-HMF) in both samples increased during storage, suggesting that the Maillard reaction occurred. This study demonstrated an effective approach to develop a new antimicrobial packaging system for semi-dry longan pulp.

Disinfection performance of chlorine dioxide gas at ultra-low concentrations and the decay rules under different environmental factors.

Gaseous chlorine dioxide (ClO2) is one of the most promising air disinfectants. In this study, an ultra-low concentration of ClO2 gas (< 1.2 mg/m3) was generated in an office at various levels of humidity and illuminance to investigate the decay law. The disinfection efficiency and metal corrosiveness of ultra-low concentrations of ClO2 gas were also studied using an experimental chamber. At 48% and 75% humidity, the decay rate constants of ClO2 gas were 0.0034 min-1 and 0.0036 min-1, respectively. The rate of decline of the ClO2 concentration increased as the humidity of the environment increased. The decay rate constant of ClO2 gas at an illuminance of 76 lux and 3429 lux was 0.0034 min-1 and 0.00427 min-1, respectively; hence, the decay rate increased with increased illumination. At a humidity of 72% and illuminance of 2112 lux, the decay rate constant reached 0.00880 min-1. The effects of humidity and illuminance on the attenuation of the ClO2 concentration were strongly synergistic. When the gas concentration was maintained below 0.9 mg/m3, the disinfection rate of ClO2 on bacteria (P. aeruginosa, V. mimicus and S. aureus) exceeded 99.9%; thus, ClO2 gas exhibited a high disinfection efficiency. In addition, there was no corrosion to various metals by ClO2 under the same conditions. Consequently, gaseous ClO2 at ultra-low concentrations has a high sterilization efficiency and is non-corrosive to metals. IMPLICATIONS: Humidity and illuminance can influence decay laws of extremely low concentration ClO2 gas. The gaseous ClO2 at ultra-low concentrations has a high sterilisation efficiency and is non-corrosive to metals.

Evaluation of Chlorine Dioxide Gas against Four Salmonella enterica Serovars Artificially Contaminated on Whole Blueberries.

ABSTRACT: Fresh produce, such as blueberries, continues to be a source of foodborne illness in the United States. Despite new practices and intervention technologies, blueberries and other produce are contaminated with foodborne pathogens, such as Salmonella. The aim of this study was to evaluate the efficacy of chlorine dioxide gas (CDG) against Salmonella enterica serovars Newport, Stanley, Muenchen, and Anatum on artificially contaminated whole fresh blueberries. Blueberries were dip inoculated into a 400-mL bath containing a Salmonella serovar cocktail of either ca. 6 or 9 log CFU/mL. Samples were dried for either 2 or 24 h before treatment with 1.5 or 3 mg of CDG/L of air to a final treatment of 3.55 to 6 ppm/h/g of blueberry. Salmonella cells were recovered by stomaching CDG-treated and nontreated control samples with 0.1% peptone and enumerated on xylose lysine Tergitol 4 agar. CDG treatments achieved up to a 5.63-log CFU/g reduction of the cocktail using 5.5 ppm/h/g, whereas the lowest treatment, 4 ppm/h/g (1.5 mg of CDG/L), was still capable of a 4.45-log CFU/g reduction. Incubation time significantly (P < 0.001) affected CDG efficacy against both inoculation concentrations. Additionally, all serovars responded similarly to CDG treatment when tested individually (P > 0.0691). Finally, the availability of a water reservoir during treatments did not have a significant effect (P = 0.9818) on CDG efficacy in this study. Our results demonstrate that CDG can be an efficacious treatment option for whole blueberry decontamination.

The ability of two chlorine dioxide chemistries to inactivate human papillomavirus-contaminated endocavitary ultrasound probes and nasendoscopes.

Sexual transmission is the most common pathway for the spread of Human papillomavirus (HPV). However, the potential for iatrogenic HPV infections is also real. Even though cleared by the Food and Drug Administration and recommended by the World Federation for Ultrasound in Medicine and Biology, several disinfectants including glutaraldehyde and o-phthalaldehyde have shown a lack of efficacy for inactivating HPV. Other methods such as ultraviolet C and concentrated hydrogen peroxide have been shown highly effective at inactivating infectious HPV. In this study, two chlorine dioxide systems are also shown to be highly efficacious at inactivating HPV. An important difference in these present studies is that as opposed to testing in suspension or using a carrier, we dried the infectious virus directly onto endocavitary ultrasound probes and nasendoscopes, therefore, validating a more realistic system to demonstrate disinfectant efficacy.

Low-cost, Small-scale Decontamination of Laboratory Equipment by Using Chlorine Dioxide Gas.

A significant concern in laboratory animal medicine is contamination due to pathogen outbreaks and how to adequately decontaminate small equipment. Many factors play a role in the selection of the decontamination method including cost, efficacy, personnel time and safety. Chlorine dioxide (ClO2) gas is an effective method, but decontamination often requires a ClO2 gas generator with a specialized air-tight exposure chamber. Although this method works well for large-scale decon- tamination, the use of a gas generator may be impractical and too costly for smaller-scale decontamination. The goal of this study was to create and validate an effective, small-scale decontamination method that uses ClO2 gas and which is an affordable, efficient, safe, and reproducible. First, we identified a product that generates ClO2 gas after the combination of 2 dry reagents. To find an affordable exposure chamber, we evaluated the ability of 4 household totes with gasket-seal lid systems to retain ClO2 gas and relative humidity (RH). The efficacy of decontamination was validated by concurrently using 2 different biologic indicators (BI), Bacillus atrophaeus (B.a.) and Geobacillus stearothermophilus (G.s.). All household totes evaluated held sufficient gas and RH for a 15-h cycle, providing adequate contact time to inactivate both BI evaluated. Our results suggest that a total exposure dose of 71 ± 42 ppm-h of ClO2 gas over 15 h at 90% or greater RH is adequate to inactivate both B.a. and G.s. There was no statistical significance between the 2 BI as indicators for decontamination; 65 of 230 (28.3%) B.a. and 75 of 230 (32.6%) G.s spore strips were positive for growth (P = 0.36). In conclusion, we successfully combined a variety of low-cost materials to establish an effective, small-scale method to decontaminate laboratory equipment. Depending on the size of the tote and whether BI are used, the cost of our method is roughly 1% that of large-scale ClO2 gas generators used with specialized air-tight exposure chambers.

Photo-induced anti-inflammatory activities of chloro substituted subphthalocyanines on the mammalian macrophage in vitro.

In this study, a series of subphthalocyanines (SubPcs) derivatives were synthesized to generate unique immunomodulatory molecules that can be activated through photo-induction. Immunomodulatory agents have a great potential in medicine to manipulate the immune system according to our needs and prevent disease symptoms. Inflammation is one of these symptoms and macrophages play a crucial role in the generation of inflammatory responses. Being able to control the activity of these agents through photo-induction enables the fine tuning on their activities in a location specific and non-invasive manner with possibly minor side effects. Mammalian macrophages' pro-inflammatory activity was examined in the presence of our compounds as well as LPS as a danger mimic. These compounds exerted photo-induced anti-inflammatory activities on the macrophages. Number of Cl atoms was a defining factor in their photo-induced anti-inflammatory immunomodulatory efficiencies.

Effect of relative humidity on inactivation of foodborne pathogens using chlorine dioxide gas and its residues on tomatoes.

The effect of relative humidity (RH) on the antimicrobial efficacy of chlorine dioxide (ClO2 ) gas against foodborne pathogens on tomatoes was evaluated. Also, levels of ClO2 residues on tomatoes after exposure to ClO2 gas under different RH conditions were measured to determine the quantity of solubilized ClO2 gas on tomato surfaces. Escherichia coli O157:H7, Salmonella Typhimurium and Listeria monocytogenes were inoculated on tomatoes and exposed to ClO2 gas (5, 10, 20 and 30 ppmv) under different RH conditions (50, 70 and 90%). As ClO2 gas concentration and treatment time increased, significant differences (P < 0·05) were observed between inactivation levels under different RH conditions. Exposure to 30 ppmv of ClO2 gas (50% RH) for 20 min resulted in 1·22-1·52 log reductions of the three foodborne pathogens. Levels of the three foodborne pathogens were reduced to below the detection limit (0·48 log CFU per cm2 ) within 15 min when exposed to 30 ppmv of ClO2 gas at 70% RH and within 10 min at 90% RH. At a given ClO2 gas concentration, ClO2 residues on tomatoes significantly (P < 0·05) increased with increasing RH, and there were close correlations between log reductions of pathogens and ClO2 residues on tomatoes. SIGNIFICANCE AND IMPACT OF THE STUDY: This study reported on the correlation between the amount of ClO2 residues on produce surfaces and the level of inactivation of pathogens after ClO2 gas treatment. Variations in RH have great effect on the solubilization of ClO2 gas on tomato surfaces considering that ClO2 residues on tomatoes increased with increasing RH. Also, the amount of ClO2 residues on tomatoes is positively correlated with the level of inactivation of pathogens. The results of this study provide insights for predicting inactivation patterns of foodborne pathogens by ClO2 gas for practical application by the fresh produce industry.

Effect of temperature on chlorine dioxide inactivation of Escherichia coli O157:H7, Salmonella typhimurium, and Listeria monocytogenes on spinach, tomatoes, stainless steel, and glass surfaces.

The objective of this study was to evaluate how treatment temperature influences the solubility of ClO2 gas and the antimicrobial effect of ClO2 gas against Escherichia coli O157:H7, Salmonella Typhimurium, and Listeria monocytogenes on produce and food contact surfaces. Produce and food contact surfaces inoculated with a combined culture cocktail of three strains each of the three foodborne pathogens were processed in a treatment chamber with 20 ppmv ClO2 gas at 15 or 25 °C under the same conditions of absolute humidity (11.2-12.3 g/m3) for up to 30 min. As treatment time increased, ClO2 gas treatment at 15 °C caused significantly more (p < 0.05) inactivation of the three pathogens than treatment at 25 °C. ClO2 gas treatment at 25 °C for 30 min resulted in 1.15 to 1.54, 1.53 to 1.88, and 1.00 to 1.78 log reductions of the three pathogens on spinach leaves, tomatoes, and stainless steel No.4, respectively. ClO2 gas treatment at 15 °C for 30 min caused 2.53 to 2.88, 2.82 to 3.23, and 2.37 to 3.03 log reductions of the three pathogens on spinach leaves, tomatoes, and stainless steel No.4, respectively. Treatment with ClO2 gas at 25 °C for 20 min resulted in 1.88 to 2.31 log reductions of the three pathogens on glass while >5.91 to 6.82 log reductions of these pathogens occurred after 20 min when treated at 15 °C. Residual ClO2 levels after gas treatment at 15 °C were significantly (p < 0.05) higher than those at 25 °C. The results of this study can help the food processing industry establish optimum ClO2 gas treatment conditions for maximizing the antimicrobial efficacy of ClO2 gas.

Comparative stability and efficacy of selected chlorine-based biocides against Escherichia coli in planktonic and biofilm states.

Microbial contamination is an unavoidable problem in industrial processes. Sodium hypochlorite (SH) is the most common biocide used for industrial disinfection. However, in view of the current societal concerns on environmental and public health aspects, there is a trend to reduce the use of this biocide as it can lead to the formation of organochlorinated carcinogenic compounds. In this work the efficacy of SH was assessed against Escherichia coli in planktonic and biofilm states and compared with three alternative chlorine-based biocides: neutral electrolyzed oxidizing water (NEOW), chlorine dioxide (CD) and sodium dichloroisocyanurate (NaDCC). The planktonic tests revealed that SH had the fastest antimicrobial action, NaDCC exhibited the highest antimicrobial rate and NEOW caused the highest antimicrobial effects. Additionally, NEOW was the biocide that allowed the highest formation of reactive oxygen species (ROS). In biofilm control, NEOW and CD were the most efficient biocides causing 3.26 and 3.20 log CFU·cm-2 reduction, respectively. In terms of stability for chlorine depletion, NEOW had the longest decay time for chlorine loss (70days at 5°C) and the lowest chlorine loss rate (0.013ppm·min-1 at 5°C). CD and NaDCC had equivalent stability. The overall results demonstrated NEOW as a good alternative to SH due to its higher antimicrobial effects and lower chlorine depletion over time.