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The Portable Chemical Sterilizer (PCS), D-FENS, and D-FEND ALL: novel chlorine dioxide decontamination technologies for the military
Abstract
There is a stated Army need for a field-portable, non-steam sterilizer technology that can be used by Forward Surgical Teams, Dental Companies, Veterinary Service Support Detachments, Combat Support Hospitals, and Area Medical Laboratories to sterilize surgical instruments and to sterilize pathological specimens prior to disposal in operating rooms, emergency treatment areas, and intensive care units. The following ensemble of novel, 'clean and green' chlorine dioxide technologies are versatile and flexible to adapt to meet a number of critical military needs for decontamination(6,15). Specifically, the Portable Chemical Sterilizer (PCS) was invented to meet urgent battlefield needs and close critical capability gaps for energy-independence, lightweight portability, rapid mobility, and rugged durability in high intensity forward deployments(3). As a revolutionary technological breakthrough in surgical sterilization technology, the PCS is a Modern Field Autoclave that relies on on-site, point-of-use, at-will generation of chlorine dioxide instead of steam. Two (2) PCS units sterilize 4 surgical trays in 1 hr, which is the equivalent throughput of one large steam autoclave (nicknamed "Bertha" in deployments because of its cumbersome size, bulky dimensions, and weight). However, the PCS operates using 100% less electricity (0 vs. 9 kW) and 98% less water (10 vs. 640 oz.), significantly reduces weight by 95% (20 vs. 450 lbs, a 4-man lift) and cube by 96% (2.1 vs. 60.2 ft(3)), and virtually eliminates the difficult challenges in forward deployments of repairs and maintaining reliable operation, lifting and transporting, and electrical power required for steam autoclaves.
... Utilizing techniques of power electronics, certain improvements to reduce high power requirements for the decontamination process are being explored. Very notable is the down-scaling of sterilizer size as a means of reducing power density during usage [26]. Medium-scale and bed-side sterilizers have been designed following this concept. ...
A major constraint in hospitals and clinics in the interior villages of low resource countries is the access to stable power supply. Decontamination of reusable metal-based surgical tools is an energy-intensive process, the power required for this procedure may not be accessible in many health centres in low resource countries. Hence, decontamination device with low-energy requirements could immensely benefit village clinical settings. The developed sterilizer utilizes a Fresnel lens in a multi-baffle multi-pass chamber to amplify radiation intensity. An intelligent scheme of air passage was achieved using Fuzzy logic control to ensure control of pressure and temperature regime thermal transport within the chamber. The Fuzzy logic controller program was designed on Matlab's fuzzy logic toolbox and simulated with Simulink to evaluate its accuracy. The performance evaluation of the device showed that at an ambient temperature of 27℃ and a solar radiation intensity of 1362 W/m2, the sterilizer was able to sterilize at a temperature of 169.69℃ which is within the range for efficient dry heat sterilization to take place (140℃-170℃). This work has demonstrated that fuzzy logic controlled dry air sterilizer could achieve temperature ~ 150℃ within the heater chamber which may be suitable for sterilizing used surgical equipment.
... It is possible that these inconsistencies could have resulted from differences in the method used to inoculate the produce (Rodgers et al ., 2004). In separate experiments, fresh-cut leaves (25 g) of iceberg lettuce or romaine were inoculated to eight logs by immersion in an aqueous suspensions of E. coli O157:H7 for 5 min, dried in a salad spinner, then exposed for 2 min to 20-200 ppm aqueous chlorine dioxide, which was effective in reducing E. coli O157:H7 populations on iceberg lettuce by up to 1.25 log and approximately 1 log on romaine lettuce (Keskinen et al ., 2009) The " D isinfectantsprayer for F oods and EN vironmentally-friendly S anitation" (D-FENS) is an aqueous chlorine dioxide technology invented at NSRDEC and intended for use in military deployments (Setlow et al ., 2009;Doona et al ., 2014). D-FENS is a convenient spray-and-wipe sanitizing device that generates aqueous chlorine dioxide on-site. ...
Fresh and fresh-cut produce are often consumed raw and are an important part of a healthy, balanced diet. In recent years, the consumption of fresh produce has been associated with outbreaks of Escherichia coli O157:H7, hepatitis A virus (HAV), and human norovirus (NoV), for example. Methods for improving fresh and fresh-cut produce safety would have tremendous benefit both for consumers, in terms of health and safety, and for the industry. This chapter addresses the development of combinations of chemical sanitizers (chlorine, ozone, chlorine dioxide) and nonthermal processing technologies that have potential commercial applications, such as the use of ultrasound to dislodge pathogens from fresh produce surfaces, and the use of energy-independent ClO2 technologies developed for forward military deployments to disinfect produce or food contact and handling surfaces.
... The PCS produces gaseous ClO 2 and proceeds where no commercial device existed previously, with a 100% reduction in power usage, 98% reduction in water, 95% reduction in weight, and 96% reduction in cubic footprint compared to conventional steam autoclaves. The PCS used the NCC to sterilize live cultures of Geobacillus stearothermophilus spores in aqueous suspensions (recovered on Antibiotic Assay Medium with 1% soluble starch -see Feeherry et al., 1987;Doona et al., 2014) ...
Recently, global public health organizations such as Doctors without Borders (MSF), the World Health Organization (WHO), Public Health Canada, National Institutes of Health (NIH), and the U.S. government developed and deployed Field Decontamination Kits (FDKs), a novel, lightweight, compact, reusable decontamination technology to sterilize Ebola-contaminated medical devices at remote clinical sites lacking infra-structure in crisis-stricken regions of West Africa (medical waste materials are placed in bags and burned). The basis for effectuating sterilization with FDKs is chlorine dioxide (ClO2) produced from a patented invention developed by researchers at the US Army Natick Soldier RD&E Center (NSRDEC) and commercialized as a dry mixed-chemical for bacterial spore decontamination. In fact, the NSRDEC research scientists developed an ensemble of ClO2 technologies designed for different applications in decontaminating fresh produce; food contact and handling surfaces; personal protective equipment; textiles used in clothing, uniforms, tents, and shelters; graywater recycling; airplanes; surgical instruments; and hard surfaces in latrines, laundries, and deployable medical facilities. These examples demonstrate the far-reaching impact, adaptability, and versatility of these innovative technologies. We present herein the unique attributes of NSRDEC’s novel decontamination technologies and a Case Study of the development of FDKs that were deployed in West Africa by international public health organizations to sterilize Ebola-contaminated medical equipment. FDKs use bacterial spores as indicators of sterility. We review the properties and structures of spores and the mechanisms of bacterial spore inactivation by ClO2. We also review mechanisms of bacterial spore inactivation by novel, emerging, and established non-thermal technologies for food preservation, such as high pressure processing, irradiation, cold plasma, and chemical sanitizers, using an array of Bacillus subtilis mutants to probe mechanisms of spore germination and inactivation. We employ techniques of high-resolution atomic force microscopy and phase contrast microscopy to examine the effects of γ-irradiation on bacterial spores of Bacillus anthracis, Bacillus thuringiensis, and Bacillus atrophaeus spp. and of ClO2 on B. subtilis spores, and present in detail assays using spore bio-indicators to ensure sterility when decontaminating with ClO2.
Active food packaging concepts exhibiting antimicrobial activity were developed using a novel chemical system in conjunction with polymers (a film made of polylactic acid, PLA, or a sachet made of superabsorbent hydrogel) with applications in the food industry for ensuring microbial safety, extending shelf-life, and reducing waste of perishable foods such as fresh fruits, vegetables, and berries. One system produces chlorine dioxide in-packaging from precursors adhered to the surface of PLA biopolymeric films that can be used in food packaging. The second is a smart food packaging system designed with chemical precursors incorporated in an insertable sachet made of a superabsorbent hydrogel polymer and is called the Compartment of Defense (CoD). The CoD packaging system is characterized by the humidity-activated, time-released controlled and sustained production of low concentrations of gaseous ClO2 in-container and can be used to inactivate pathogens and prevent mold growth and extend the safe shelf-life while reducing economic losses of perishable foods that respire in-packaging (fresh berries, fruits, and vegetables). These next-generation chlorine dioxide technologies for microbial decontamination can be adapted for food industry applications (e.g., treating fresh produce, sanitizing food processing/handling surfaces, and cleaning-in-place methods for production lines) and other microbial decontamination applications (e.g., preventing mold on porous surfaces, disinfecting hard surfaces, self-decontaminating garments, and inactivating bacterial spores of Clostridiodes difficile and Bacillus anthracis).
International guidelines stipulate that autoclavation is necessary to sterilize surgical equipment. World Health Organization (WHO) guidelines for decontamination of medical devices require four levels of decontamination: cleaning, low- and high-level disinfection, as well as sterilization. Following disasters, there is a substantial need for wound care surgery. This requires prompt availability of a significant volume of instruments that are adequately decontaminated. Ideally, they should be sterilized using an autoclave, but due to the resource-limited field context, this may be impossible. The aim of this study was therefore to identify whether there are portable and less resource-demanding techniques to decontaminate surgical instruments for safe wound care surgery in disasters. A scoping review was chosen, and searches were performed in three scientific databases, grey literature, and included data from organizations and journals. Articles were scanned for decontamination techniques feasible for use in the resource-scarce disaster setting given that: they achieved at least high-level disinfected instruments, were portable, and did not require electricity. A total of 401 articles were reviewed, yielding 13 articles for inclusion. The study identified three techniques: pressure cooking, boiling, and liquid chemical immersion, all achieving either sterilized or high-level disinfected instruments. It was concluded that besides autoclaves, there are less resource-demanding decontamination techniques available for safe wound surgery in disasters. This study provides systematic information to guide optimal standard setting for sterilization of surgical material in resource-limited disaster settings.
High pressure processing (HPP) is an emerging non-thermal technology that is growing exponentially in use worldwide for the pasteurization of commercial foodstuffs. At combinations of elevated pressures and temperatures, HPP inactivates bacterial spores, but HPP has not yet been implemented commercially for food sterilization. Studies of the mechanisms of bacterial spore inactivation by HPP using primarily spores of Bacillus species have shown that spore germination precedes inactivation, with the release of dipicolinic acid from the spore core as the rate-determining step. Investigations probing spore resistance to and germination by HPP using Bacillus subtilis, a number of selected B. subtilis mutants, Bacillus amyloliquefaciens, and Clostridium difficile spores have compiled a wealth of detailed mechanistic information, while also accumulating abundant germination kinetics data that has not previously been analyzed by predictive models. Presently, we devise a “quasi-chemical” model for bacterial spore germination dynamics by HPP. This quasi-chemical germination model (QCGM) hypothesizes a three-step mechanism and derives a set of ordinary differential equations to model the observed germination dynamics. The results with this model are viewed in the context of historical studies of spore activation, germination, and inactivation, with an eye toward potentially integrating differential equation models for germination and inactivation into a single, comprehensive model for spore dynamics by HPP. With the increasing use of high hydrostatic pressure to investigate mechanisms of bacterial spore resistance and physiology, the QCGM results help promote the efficient control of bacterial spores, whether for the inactivation of Clostridium botulinum spores in low-acid foods or aerosolized Bacillus anthracis spores on textiles used in protective clothing, tents, or shelters.
Inactivation of Escherichia coli O157:H7, Salmonella typhimurium, and Listeria monocytogenes in iceberg lettuce by aqueous chlorine dioxide (ClO(2)) treatment was evaluated. Iceberg lettuce samples were inoculated with approximately 7 log CFU/g of E. coli O157:H7, S. typhimurium, and L. monocytogenes. Iceberg lettuce samples were then treated with 0, 5, 10, or 50 ppm ClO(2) solution and stored at 4 degrees C. Aqueous ClO(2) treatment significantly decreased the populations of pathogenic bacteria on shredded lettuce (P < 0.05). In particular, 50 ppm ClO(2) treatment reduced E. coli O157:H7, S. typhimurium, and L. monocytogenes by 1.44, 1.95, and 1.20 log CFU/g, respectively. The D(10)-values of E. coli O157:H7, S. typhimurium, and L. monocytogenes in shredded lettuce were 11, 26, and 42 ppm, respectively. The effect of aqueous ClO(2) treatment on the growth of pathogenic bacteria during storage was evaluated, and a decrease in the population size of these pathogenic bacteria was observed. Additionally, aqueous ClO(2) treatment did not affect the color of lettuce during storage. These results suggest that aqueous ClO(2) treatment can be used to improve the microbial safety of shredded lettuce during storage.
Aqueous spore suspensions of Bacillus stearothermophilus ATCC 12980 were heated at different temperatures for various time intervals in a resistometer, spread plated on antibiotic assay medium supplemented with 0.1% soluble starch without (AAMS) or with (AAMS-S) 0.9% NaCl, and incubated at 55 degrees C unless otherwise indicated. Uninjured spores formed colonies on AAMS and AAMS-S; injured spores formed colonies only on AAMS. Values of D, the decimal reduction time (time required at a given temperature for destruction of 90% of the cells), when survivors were recovered on AAMS were 62.04, 18.00, 8.00, 3.33, and 1.05 min at 112.8, 115.6, 118.3, 121.1, and 123.9 degrees C, respectively. Recovery on AAMS-S resulted in reduced decimal reduction time. The computed z value (the temperature change which will alter the D value by a factor of 10) for spores recovered on AAMS was 8.3 degrees C; for spores recovered on AAMS-S, it was 7.6 degrees C. The rates of inactivation and injury were similar. Injury (judged by salt sensitivity) was a linear function of the heating temperature. At a heating temperature of less than or equal to 118.3 degrees C, spore injury was indicated by the curvilinear portion of the survival curve (judged by salt sensitivity), showing that injury occurred early in the thermal treatment as well as during logarithmic inactivation (reduced decimal reduction time). Heat-injured spores showed an increased sensitivity not only to 0.9% NaCl but also to other postprocessing environmental factors such as incubation temperatures, a pH of 6.6 for the medium, and anaerobiosis during incubation.
Introduction Bacterial Spore Resistance to and Inactivation by Chemical Agents Novel Chlorine Dioxide Technologies for Eliminating Microbial Hazards from Fresh Produce and Food - Handling Environments Modified Atmosphere Packaging (MAP) Acknowledgments References
The reduction of chlorite ion by the hydrogen ascorbate ion in a neutral solution safely produces chlorine dioxide. The decrease in absorbance at 268 nm with the presence of dimethyl sulfoxide (DMSO) allows measurement of the ascorbate disappearance in the reaction with excess chlorite. The measured rate constant at 25 ± 0.02°C, 3.67 × 10−4 M DMSO, ionic strength 0.51 M (NaClO4), and in the presence of 3.32 × 10−9 M ethylenediaminetetraacetic acid is 13.81 ± 1.30 M−1 s−1. Rate constant measurements over the range 15–35°C gave an Arrhenius activation energy of 75.51 ± 4.53 kJ mol−1. This result is the first reported determination of the kinetics of this reaction and is consistent with either electron- or oxygen-transfer mechanisms. Anomalously, reduction of chlorite results in its oxidation, because intermediate hypochlorite oxidizes chlorite.
The slow reaction between peroxodisulfate and formate is significantly accelerated by ascorbate at room temperature. The products of this induced oxidation, CO2 and oxalate (C2O2–4), were analyzed by several methods and the kinetics of this reaction were measured. The overall mechanism involves free radical species. Ascorbate reacts with peroxodisulfate to initiate production of the sulfate radical ion (SO•–4), which reacts with formate to produce carbon dioxide radical ion (CO•–2) and sulfate. The carbon dioxide radical reacts with peroxodisulfate to form CO2 or self-combines to form oxalate. Competition occurring between these two processes determines the overall fate of the carbon dioxide radical species. As pH decreases, protonation of the carbon dioxide radical ion tends to favor production of CO2.
The kinetics of Staphylococcus aureus (strains A, B, D) growth in bread crumb were determined as a function of water activity (Aw) from 0.836 to 0.909 at pH 5.2 to 5.5 and at 35°c. Adding glycerol to the dough or equilibrating the bread over saturated salt solutions adjusted the Aw of the bread. Growth kinetics data, plotted as enumerated colony counts versus incubation time, were fitted using the logistic function to determine maximum growth rates. Similar maximum growth rates resulted, irrespective of the method used to adjust Aw. Extrapolation of growth rate-Aw results predicts the Aw corresponding to a zero growth rate.
A commercial fresh pack potato operation was used as a model to evaluate FIT fruit and vegetable wash effectiveness in reducing levels of microorganisms on potatoes and in flume water. Fresh potatoes were washed in flume water with or without FIT, or treated with a spray bar utilizing either FIT, 9 ppm chlorine dioxide (ClO2), or a water control. Both flume treatments were also evaluated for APC and Gram-negatives. There were no significant differences in reduction of these microorganisms on treated or control potatoes. However, levels of Gram-negative bacteria in FIT-amended flume water were reduced by 5.95 log CFU/g, and the APC was reduced by 1.43 log CFU/g. To validate plant trial findings, this test was repeated using solutions of sterile potato flume water from the fresh pack operation, containing a typical level of dissolved and suspended solids. Treatment solutions prepared with flume water or deionized water containing FIT, 9 ppm ClO2, or a water control were inoculated with E. coli O157:H7, Salmonella Typhimurium, or Pectobacterium carotovorum ssp. carotovorum. FIT and ClO2 prepared with deionized water reduced levels of microorganisms by >6.1 to 6.6 log CFU/g to below the detection limit. FIT prepared with flume water reduced levels of all organisms by >6.0 to 6.4 log CFU/g to below the detection limit, whereas ClO2 prepared from flume water reduced bacterial levels of all organisms by only 0.7 to 1.4 log CFU/g. Neither FIT nor ClO2 was particularly efficacious against E. coli O157:H7, S. Typhimurium, APC, yeasts, or molds on potato surfaces.
To determine the mechanisms of Bacillus subtilis spore killing by hypochlorite and chlorine dioxide, and its resistance against them.
Spores of B. subtilis treated with hypochlorite or chlorine dioxide did not accumulate damage to their DNA, as spores with or without the two major DNA protective alpha/beta-type small, acid soluble spore proteins exhibited similar sensitivity to these chemicals; these agents also did not cause spore mutagenesis and their efficacy in spore killing was not increased by the absence of a major DNA repair pathway. Spore killing by these two chemicals was greatly increased if spores were first chemically decoated or if spores carried a mutation in a gene encoding a protein essential for assembly of many spore coat proteins. Spores prepared at a higher temperature were also much more resistant to these agents. Neither hypochlorite nor chlorine dioxide treatment caused release of the spore core's large depot of dipicolinic acid (DPA), but hypochlorite- and chlorine dioxide-treated spores much more readily released DPA upon a subsequent normally sub-lethal heat treatment than did untreated spores. Hypochlorite-killed spores could not initiate the germination process with either nutrients or a 1 : 1 chelate of Ca2+-DPA, and these spores could not be recovered by lysozyme treatment. Chlorine dioxide-treated spores also did not germinate with Ca2+-DPA and could not be recovered by lysozyme treatment, but did germinate with nutrients. However, while germinated chlorine dioxide-killed spores released DPA and degraded their peptidoglycan cortex, they did not initiate metabolism and many of these germinated spores were dead as determined by a viability stain that discriminates live cells from dead ones on the basis of their permeability properties.
Hypochlorite and chlorine dioxide do not kill B. subtilis spores by DNA damage, and a major factor in spore resistance to these agents appears to be the spore coat. Spore killing by hypochlorite appears to render spores defective in germination, possibly because of severe damage to the spore's inner membrane. While chlorine dioxide-killed spores can undergo the initial steps in spore germination, these germinated spores can go no further in this process probably because of some type of membrane damage.
These results provide information on the mechanisms of the killing of bacterial spores by hypochlorite and chlorine dioxide.
Chlorine dioxide (ClO(2)) gas is a strong oxidizing and sanitizing agent that has a broad and high biocidal effectiveness and big penetration ability; its efficacy to prolong the shelf-life of a minimally processed (MP) vegetable, grated carrots (Daucus carota L.), was tested in this study. Carrots were sorted, their ends removed, hand peeled, cut, washed, spin dried and separated in 2 portions, one to be treated with ClO(2) gas and the other to remain untreated for comparisons. MP carrots were decontaminated in a cabinet at 91% relative humidity and 28 degrees C for up to 6 min, including 30 s of ClO(2) injection to the cabinet, then stored under equilibrium modified atmosphere (4.5% O(2), 8.9% CO(2), 86.6% N(2)) at 7 degrees C for shelf-life studies. ClO(2) concentration in the cabinet rose to 1.33 mg/l after 30 s of treatment, and then fell to nil before 6 min. The shelf-life study included: O(2) and CO(2) headspace concentrations, microbiological quality (mesophilic aerobic bacteria, psychrotrophs, lactic acid bacteria, and yeasts), sensory quality (odour, flavour, texture, overall visual quality, and white blushing), and pH. ClO(2) did not affect respiration rate of MP carrots significantly (alpha< or =0.05), and lowered the pH significantly (alpha< or =0.05). The applied packaging configuration kept O(2) headspace concentrations in treated samples in equilibrium and prevented CO(2) accumulation. After ClO(2) treatment, the decontamination levels (log CFU/g) achieved were 1.88, 1.71, 2.60, and 0.66 for mesophilic aerobic bacteria, psychrotrophs, and yeasts respectively. The initial sensory quality of MP carrots was not impaired significantly (alpha< or =0.05). A lag phase of at least 2 days was observed for mesophilic aerobic bacteria, psychrotrophs, and lactic acid bacteria in treated samples, while mesophilic aerobic bacteria and psychrotrophs increased parallelly. Odour was the only important attribute in sensory deterioration, but it reached an unacceptable score when samples were already rejected from the microbiological point of view. The shelf-life extension was limited to one day due to the restricted effect of the ClO(2) treatment on yeast counts. Nevertheless, ClO(2) seems to be a promising alternative to prolong the shelf-life of grated carrots.
Inactivation kinetics of inoculated Escherichia coli O157:H7, Listeria monocytogenes and Salmonella enterica on strawberries by chlorine dioxide gas at different concentrations (0.5, 1, 1.5, 3 and 5 mgl(-1)) for 10 min were studied. A cocktail of three strains of each targeted organism (100 microl) was spotted onto the surface of the strawberries (approximately 8-9 log ml(-1)) separately followed by air drying, and then treated with ClO(2) gas at 22 degrees C and 90-95% relative humidity. Approximately a 4.3-4.7 logCFU reduction per strawberry of all examined bacteria was achieved by treatment with 5 mgl(-1) ClO(2) for 10 min. The inactivation kinetics of E. coli O157:H7, L. monocytogenes and S. enterica were determined using first-order kinetic models to establish D-values and z-values. The D-values of E. coli, L. monocytogenes and S. enterica were 2.6+/-0.2, 2.3+/-0.2 and 2.7+/-0.7 min, respectively, at 5 mgl(-1) ClO(2). The z-values of E. coli, L. monocytogenes and S. enterica were 16.8+/-3.5, 15.8+/-3.5 and 23.3+/-3.3 mgl(-1), respectively. Furthermore, treatment with ClO(2) gas significantly (p < or = 0.05) reduced the initial microflora (mesophilic, psychrotrophic bacteria, yeasts and molds) on strawberries. Treatment with ClO(2) gas did not affect the color of strawberries and extended the shelf-life to 16 days compared to 8 days for the untreated control.
The purpose of this investigation was to study inactivation kinetics of inoculated Escherichia coli O157:H7 and Salmonella enterica on lettuce leaves by ClO(2) gas at different concentrations (0.5, 1.0, 1.5, 3.0, and 5.0 mg l(-1)) for 10 min and to determine the effect of ClO(2) gas on the quality and shelf life of lettuce during storage at 4 degrees C for 7 days. One hundred microliters of each targeted organism was separately spot-inoculated onto the surface (5 cm(2)) of lettuce (approximately 8-9 log CFU ml(-1)), air-dried, and treated with ClO(2) gas at 22 degrees C and 90-95% relative humidity for 10 min. Surviving bacterial populations on lettuce were determined using a membrane transferring method, which included a non-selective medium followed by a selective medium. The inactivation kinetics of E. coli O157:H7 and S. enterica was determined using first-order kinetics to establish D-values and z-values. The D-values of E. coli and S. enterica were 2.9+/-0.1 and 3.8+/-0.5 min, respectively, at 5.0 mg l(-1) ClO(2) gas. The z-values of E. coli and S. enterica were 16.2+/-2.4 and 21.4+/-0.5 mg l(-1), respectively. A 5 log CFU reduction (recommended by the United States Food and Drug Administration) for E. coli and S. enterica could be achieved with 5.0 mg l(-1) ClO(2) gas for 14.5 and 19.0 min, respectively. Treatment with ClO(2) gas significantly reduced inherent microflora on lettuce and microbial counts remained significantly (p<0.05) lower than the uninoculated control during storage at 4 degrees C for 7 days. However, treatment with ClO(2) gas had a significantly (p<0.05) negative impact on visual leaf quality. These results showed that treatment with ClO(2) gas significantly reduced selected pathogens and inherent microorganisms on lettuce; however, the processing conditions would likely need to be altered for consumer acceptance.