Research > Microbe > Bacteria > Staphylococcus


Common Generic Names
  • Electrolytically Generated Hypochlorous Acid (HOCl)
  • Neutral Electrolyzed Water (NEW)
  • Electrolyzed Oxidizing Water (EOW)
  • Electro-chemically Activated Water (ECA)
  • Super-oxidized water (SOW)

Results: 33 published articles


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Microbe(s): Staphylococcus aureus, Pseudomonas aeruginosa


Many antiseptics have been used to treat wounds. To compare the microbicidal efficacy of ClHO (Clortech) with other antiseptics used on wounds, healthy skin and mucous membranes. The microbicidal efficacy of 13 antiseptic products on eight micro-organisms (three Gram-positive three Gram-negative two yeasts) inoculated on organic germ-carriers was studied. In addition, the loss of efficacy against Staphylococcus aureus and Pseudomonas aeruginosa with biofilm was assessed with the six best-performing products. Chlorhexidine (1) had the highest microbicidal effect at 1 min. At 5 min, 500 and 1500 mg/L ClHO showed similar, or better, activity than the other antiseptics studied. The ClHO concentration of 300 mg/L achieved this same efficacy at 10 min. The product that lost the most efficacy due to biofilm was 1 chlorhexidine, while 1 PVP-I and ClHO at either 300 or 500 mg/L were moderately affected by biofilm. The most effective in the presence of biofilm was ClHO at 1500 mg/L. ClHO at mediumlow concentrations (300 or 500 mg/L) is a good antiseptic that can be used on wounds and mucous membranes for 510 min. Lower concentrations of ClHO, as well as of the other antiseptics studied, were less effective or more altered by the biofilm. ClHO at a concentration of 1500 mg/L is very effective in the presence or absence of biofilm that can be used on healthy skin for 5 min.


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Microbe(s): Acinetobacter baumannii, Staphylococcus aureus, Pseudomonas aeruginosa


Biofilm formation causes prolonged wound infections due to the dense biofilm structure, differential gene regulation to combat stress, and production of extracellular polymeric substances. Acinetobacter baumannii, Staphylococcus aureus, and Pseudomonas aeruginosa are three difficult-to-treat biofilm-forming bacteria frequently found in wound infections. This work describes a novel wound dressing in the form of an electrochemical scaffold (e-scaffold) that generates controlled, low concentrations of hypochlorous acid (HOCl) suitable for killing biofilm communities without substantially damaging host tissue. Production of HOCl near the e-scaffold surface was verified by measuring its concentration using needle-type microelectrodes. E-scaffolds producing 17, 10 and 7mM HOCl completely eradicated S. aureus, A. baumannii, and P. aeruginosa biofilms after 3hours, 2hours, and 1hour, respectively. Cytotoxicity and histopathological assessment showed no discernible harm to host tissues when e-scaffolds were applied to explant biofilms. The described strategy may provide a novel antibiotic-free strategy for treating persistent biofilm-associated infections, such as wound infections.


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Microbe(s): Staphylococcus aureus, Pseudomonas aeruginosa


Sodium hypochlorite (NaClO, SHC)/hypochlorous acid (HClO, HCA) wound irrigation solutions have experienced a renaissance in the prevention and treatment of low-level wound infections. They are attributed with lower cytotoxicity and have therefore gained increasing attention in daily clinical practice. To determine the cytotoxicity and antimicrobial efficacy of six NaClO/HClO wound irrigation solutions. For cytotoxicity evaluation (based on DIN EN 10993-5), human keratinocytes (HaCaT) and human skin fibroblasts (BJ) were used. Staphylococcus aureus and Pseudomonas aeruginosa were used for antimicrobial efficacy evaluation (based on DIN EN 13727). Solutions were evaluated after 1, 5 and 15min of exposure. Additionally, physicochemical properties (pH and oxidationreduction potential values) were investigated. Efficacy and cytotoxicity varied significantly between solutions. Generally, increasing antimicrobial activity was associated with decreasing cell viability. Furthermore, a concentration- and time-dependent impact on pathogens and cells was observed: cytotoxic and antimicrobial activity increased with rising NaClO/HClO solution concentrations and extended exposure times. Based on these in vitro evaluations, the following ranking (lowest to highest microbicidal effect and cytotoxic impact) was found: Microdacyn60 (SHC/HCA-M)


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Microbe(s): Staphylococcus aureus, Pseudomonas aeruginosa


In-vitro and in-vivo studies have supported antimicrobial, anti-inflammatory, and other biologic properties of hypochlorous acid (HOCl), which has led to its in the treatment of skin wounds, pruritus, diabetic ulcers, and some inflammatory skin disorders. Research has also shown that the physiochemical properties of HOCl after application to skin are highly dependent on both pH and formulation stability. In this review, the authors discuss a core HOCI formulation that is stable for up to two years, noncytotoxic, and pH-neutralized to augment therapeutic activity, skin tolerability, and stability. The authors summarize relevant study outcomes and potential modes of action related to this core HOCI formulation, as well as describe its ready-to-vehicles that are approved and available for topical application.


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Microbe(s): Staphylococcus aureus, coagulase-negative staphylococci (CNS), Pseudomonas aeruginosa


The purpose of this study was to determine whether a commercial formulation of hypochlorous acid hygiene solution (0.01), Avenova, can destroy existing biofilms formed by ocular clinical bacterial isolates, including blepharitis isolates of Staphylococcus aureus and coagulase-negative staphylococci (CNS), and a keratitis isolate of Pseudomonas aeruginosa. Biofilms grown in bacterial growth media on disposable contact lens cases were challenged with hypochlorous acid hygiene solution. At various time points, surviving bacteria were quantified by serial dilution and colony counts. S. aureus biofilms formed on glass were challenged using a hypochlorous acid hygiene solution, and imaged using vital staining and confocal laser scanning microscopy. Bactericidal activity (3 Log10 99.9) was observed for all tested bacterial species after a 30-minute exposure. S. aureus biofilms had a bactericidal level of killing by 10 minutes (p<0.01), S. capitis by 5 minutes (p<0.001), S. epidermidis by 30 minutes (p<0.001), and P. aeruginosa by 10 minutes (p<0.01). Confocal microscopy and crystal violet staining analysis of bacterial biofilms treated with hypochlorous acid solution both demonstrated that biofilm bacteria were readily killed, but biofilm structure was largely maintained. Hypochlorous acid (0.01) hygiene solution was able to achieve bactericidal levels of killing of bacteria in biofilms, but did not disrupt biofilm structures. Susceptibility of tested staphylococcal blepharitis isolates varied by species, with S. capitis being the most susceptible and S. epidermidis being the least susceptible.


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Microbe(s): Propionibacterium acnes, Corynebacterium, Staphylococcus aureus, Staphylococcus epidermidis


Purpose To examine the magnitude of bacterial reduction on the surface of the periocular skin 20 minutes after application of a saline hygiene solution containing 0.01 pure hypochlorous acid (HOCl). Methods Microbiological specimens were collected immediately prior to applying the hygiene solution and again 20 minutes later. Total microbial colonies were counted and each unique colony morphology was processed to identify the bacterial species and to determine the susceptibility profile to 15 out altering the diversity of bacterial species remaining on the skin under the lower eyelid.


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Microbe(s): Staphylococcus aureus


Slightly acidic electrolyzed water (SAEW), considered as a broad-spectrum and high-performance bactericide are increasingly applied in the food industry. However, its disinfection mechanism has not been completely elucidated. This study aims to examine the disinfection efficacy and mechanism of SAEW on Staphylococcus aureus, compared with that of sodium hypochlorite (NaClO) and hydrochloric acid (HCl). SAEW treatment significantly reduced S. aureus by 5.8 log CFU/mL in 1 min, while 3.26 and 2.73 log reductions were obtained with NaClO and HCl treatments, respectively. A series of biological changes including intracellular potassium leakage, TTC-dehydrogenase relative activity and bacterial ultrastructure destruction were studied following disinfection treatment of S. aureus. The results showed that SAEW decreased the relative activity of TTC-dehydrogenase by 65.84%. Comparing intracellular potassium leakage, the SAEW treatment caused a greater percent of protein leakage (108.34%) than the NaClO (18.75%) or HCl (0.84%) treatments. These results demonstrated the potent impact SAEW had on the permeability of cell membranes. In addition, the ranking of partly agglutinated cellular inclusion formation was HCl > SAEW > NaClO. It appeared that HCl, along with its low pH value, are responsible for most of the cytoplasmic disruptions. Overall, this study demonstrated that the disinfection mechanism of SAEW was disrupting the permeability of cell membrane and the cytoplasmic ultrastructures in S. aureus cells.


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Microbe(s): Staphylococcus aureus, Escherichia coli


The article focuses on investigation of the effects of usage of acidic electrolyzed water (AEW) with different sodium chloride concentration (0.001, 0.01, and 0.1) for the preparation of carrageenan and gelatine hydrosols and hydrogels. To determine physiochemical properties of hydrosols, the pH, oxidation-reduction potential (ORP), available chloride concentration (ACC) and rheological parameters such us gelation and flow temperatures were measured. The samples were also 0.1 w/v). These results suggest that hydrogels and hydrosols incorporated with AEW may be used for food preservation.


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Microbe(s): Escherichia coli, Salmonella enteritidis, Staphylococcus aureus


The objective of this study was to evaluate the effectiveness of slightly acidic electrolyzed water (SAEW) in reducing pathogens on pure cultures and on cotton fabric surfaces in the presence of organic matter and estimate its efficacy in comparison with povidone iodine solution for reducing pathogenic microorganisms on internal surfaces of layer houses. Pure cultures of E.coli, S.enteritidis, and S.aureus and cotton fabric surfaces inoculated with these strains were treated with SAEW in the presence of bovine serum albumin (BSA). In the absence of BSA, complete inactivation of all strains in pure cultures and on cotton fabric surfaces was observed after 2.5 and 5 min treatment with SAEW at 40 mg/L of available chlorine concentration (ACC), respectively. The bactericidal efciency of SAEW increased with increasing ACC, but decreased with increasing BSA concentration. Then, the surfaces of the layer houses were sprayed with SAEW at 60, 80, and 100 mg/L of ACC and povidone iodine using the automated disinfection system at a rate of 110 mL/m2, respectively. Samples from the floor, wall, feed trough, and egg conveyor belt surfaces were collected with sterile cotton swabs before and after spraying disinfection. Compared to tap water, SAEW and povidone iodine significantly reduced microbial populations on each surface of the layer houses. SAEW with 80 or 100 mg/L of ACC showed significantly higher efficacy than povidone iodine for total aerobic bacteria, staphylococci, coliforms, or yeasts and moulds on the floor and feed trough surfaces (P < 0.05). SAEW was more effective than povidone iodine at reducing total aerobic bacteria, coliforms, and yeasts and moulds on the wall surface. Additionally, SAEW had similar bactericidal activity with povidone iodine on the surface of the egg conveyor belt. Results suggest that SAEW exerts a higher or equivalent bactericidal efficiency for the surfaces compared to povidone iodine, and it may be used as an effective alternative for reducing microbial contamination on surfaces in layer houses.


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Microbe(s): Methicillin-resistant Staphylococcus aureus, MRSA


OBJECTIVE: Biofilms represent a key challenge in the treatment of chronic wounds, as they are among the main reasons for delays in chronic wound healing. This in vitro study was aimed at evaluating the activity of a new acid-oxidizing solution (AOS) on biofilm formation. Acid-oxidizing solution contains free chlorine species with stabilized hypochlorous acid in high concentration (> 95) and is RP2). Different approaches were used to assess the prevention and eradication of methicillin-resistant Staphyloccocus aureus biofilm by the study products. Xylitol and chlorhexidine were used as positive controls. The activity of the study products on the biofilm structure was evaluated analyzing the ultrastructural modification by scanning electron microscopy, while skin compatibility was assessed on noncolonized tissues measuring the metabolic activity of the cells. RESULTS: In all experiments, AOS showed to be active on the biofilm matrix, modifying its structure and allowing bacterial release from the matrix. In all experiments, no cytotoxicity was observed in the tissues treated with the product suggesting a good compatibility of AOS with skin tissues. Reference product 1 affected the biofilm, suggesting a disruption effect RP2 was slightly less active than AOS in modifying the biofilm structure. CONCLUSION: Treatment with AOS affects biofilm by modifying its structure and therefore facilitating local bacteria accessibility to bactericidal agents, with consequent potential clinical benefits in the treatment of chronic wounds.


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Microbe(s): Acinetobacter baumannii, Escherichia coli, Enterococcus faecalis, Klebsiella pneumoniae, Pseudomonas aeruginosa, Staphylococcus aureus, Bacillus subtilis, Myroides spp, MRSA, VRE


The aim of this study was to investigate the in-vitro antimicrobial activity of usage and normal concentrations of electrolyzed water in hospital. In our study, the effects of different concentrations of electrolyzed water on two gram positive, four gram negative standard strains and clinical isolates of four gram negative, two gram positive, one spore-forming bacillus and Myroides spp strains that lead to hospital infections were researched. The effects of different concentrations and different contact times of Envirolyte electrolyzed water on cited strains were researched through method of qualitative suspension tests. Petri dishes fo bacteria have been incubated at 37 C 48 hours. Bactericidal disinfectant was interpreted to be effective at the end of the period due to the lack of growth. Solutions to which disinfectant were not added were prepared with an eye to control reproduction and controlcultures were made by using neutralizing agents. 1/1, 1/2, and 1/10 concentrations of Envirolyte electrolyzed water were found to be effective on the bacteria that lead to hospital infections used during all test times. As a conclusion, based upon the results we acquired, it was observed that Envirolyte electrolyzed water of 100 concentration would be convenient to be used for disinfection when diluted to a usage concentration of 1/10.


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Microbe(s): Escherichia coli O157:H7, Listeria monocytogenes, Staphylococcus aureus, and Salmonella Typhimurium


This study evaluated the efficacy of the individual treatments (slightly acidic electrolyzed water [SAcEW] or fumaric acid [FA]) and their combination to reduce Escherichia coli O157:H7, Listeria monocytogenes, Staphylococcus aureus, and Salmonella Typhimurium in fresh pork as well as to study the shelf life and sensory quality (color, odor, and texture) of pork during storage at 4 and 10 C. The inoculated pork samples (10 g) were dipped for 3 min in each treatment (tap water [TW], SAcEW, strong acidic electrolyzed water [StAEW], 0.5% FA, or SAcEW + 0.5% FA) with or without mild heat (40 C). Decontamination of fresh pork with SAcEW +0.5% FA at 40 C for 3 min showed greater bactericidal effect compared to other treatments, which significantly (P < 0.05) reduced E. coli O157:H7, L. monocytogenes, S. aureus, and S. Typhimurium by 2.59, 2.69, 2.38, and 2.99 log CFU/g, respectively. This combined treatment significantly (P < 0.05) yielded in a longer lag time of naturally occurring bacteria (TBC) on pork stored at 4 C. This combined treatment also prolonged the shelf life of pork up to 6 days and 4 5 days when stored at 4 C and 10 C, respectively, compared to those of the untreated pork. The results suggest that the combined treatment of SAcEW + 0.5% FA has potential as a novel method to enhance the microbial safety and quality of fresh pork.


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Microbe(s): Staphylococcus aureus, Bacillus cereus, Escherichia coli, Aspergillus fumigatus


Application of slightly acidic electrolyzed water (SAEW) in combination with ultrasound for decontamination of kashk was investigated. SAEW had a pH of 5.3-5.5, an oxidation reduction potential of 545-600 mV, and an available chlorine concentration of 20-22 mg/L. Kashk is a dairy product with a unique aroma and a high nutritive value produced in Iran. A 2/1 SAEW/kashk ratio showed 1.42, 1.13, 1.24, and 1.37 log CFU/mL microbial reductions in Staphylococcus aureus, Bacillus cereus, Escherichia coli, and Aspergillus fumigatus, respectively, at room temperature. A combination of SAEW treatment with ultrasound (SAEWultrasound) resulted in 1.87, 1.67, 1.71, and 1.91 log CFU/mL reductions in S. aureus, B. cereus, E. coli, and A. fumigatus, respectively. The developed hurdle approach can be a useful tool for sanitization of kashk and similar products. Application of SAEWultrasound in dairy microbial decontamination is first reported herein.


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Microbe(s): Acinetobacter baumannii, Escherichia coli, Enterococcus faecalis, Klebsiella pneumoniae, Pseudomonas aeruginosa, Staphylococcus aureus, MRSE, VRE Bacillus subtilis, Myroides spp.


Super-oxidized water is one of the broad spectrum disinfectants, which was introduced recently. There are many researches to find reliable chemicals which are effective, inexpensive, easy to obtain and use, and effective for disinfection of microorganisms leading hospital infections. Antimicrobial activity of super-oxidized water is promising. The aim of this study was to investigate the in-vitro antimicrobial activity of different concentrations of Medilox super-oxidized water that is approved by the Food and Drug Administration (FDA) as high level disinfectant. Material and methods In this study, super-oxidized water obtained from Medilox Soosan E & C, Korea device, which had been already installed in our hospital, was used. Antimicrobial activities of different concentrations of super-oxidized water (1/1, 1/2, 1/5, 1/10, 1/20, 1/50, 1/100) at different exposure times (1, 2, 5, 10, 30 min) against six ATCC strains, eight antibiotic resistant bacteria, yeasts and molds were evaluated using qualitative suspension test. Dey-Engley Neutralizing Broth Sigma-Aldrich, USA was used as neutralizing agent. Results Medilox was found to be effective against all standard strains (Acinetobacter baumannii 19606, Escherichia coli 25922, Enterococcus faecalis 29212, Klebsiella pneumoniae 254988, Pseudomonas aeruginosa 27853, Staphylococcus aureus 29213), all clinical isolates (Acinetobacter baumannii, Escherichia coli, vancomycin-resistant Enterococcus faecium, Klebsiella pneumoniae, Pseudomonas aeruginosa, methicillin-resistant Staphylococcus aureus, Bacillus subtilis, Myroides spp.), and all yeastsat 1/1 dilution in 1 minute. It was found to be effective on Aspergillus flavus at 1/1 dilution in 2 minutes and on certain molds in 5 minutes. Conclusion Medilox super-oxidized water is a broad spectrum, on-site producible disinfectant, which is effective on bacteria and fungi and can be used for the control of nosocomial infection.


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Microbe(s): Salmonella enteritidis, Escherichia coli O157:H7 and Staphylococcus aureus


The efficacy of slightly acidic electrolyzed water (SAEW) to inactivate foodborne pathogens and indigenous microbiota on shell eggs was evaluated and compared to chlorine dioxide (CD), acidic electrolyzed water (AEW) and NaClO solution. The eggs were artificially inoculated with S. enteritidis, E. coli O157:H7 and S. aureus and sprayed or immersed with SAEW, alkaline electrolyzed water (AlEW) followed by SAEW (AlEWSAEW), CD, AEW and NaClO solution, respectively. The effect of SAEW on the natural microbiota of shell eggs was also determined. Spraying shell eggs with SAEW, CD and NaClO solution at an ACC of 60 mg/L had no significant bactericidal difference for foodborne pathogens and indigenous microbiota on shell eggs, and the difference of disinfection effect between SAEW and AEW was not significant, whereas the bactericidal activity of SAEW for E. coli O157:H7, S. aureus, total aerobic bacteria and moulds and yeasts was significantly higher than that of CD and NaClO solution at ACCs of 80 or 100 mg/L. SAEW was found to be more effective when used in conjunction with AlEW, and higher reductions were obtained with the immersion treatment. Results indicate that the disinfectant efficiency of SAEW is equivalent to or higher than that of chlorine dioxide and NaClO solution and therefore SAEW shows the potential to be used for sanitization of egg shells as an environmentally friendly disinfection agent.


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Microbe(s): Total Microbial Count, methicillin-susceptible Staphylococcus aureus (MSSA), methicillin-resistant Staphylococcus aureus (MRSA)


This study aimed to monitor the microbiological effect of cleaning near-patient sites over a 48-hour period with a novel disinfectant, electrolyzed water. One ward dedicated to acute care of the elderly population in a district general hospital in Scotland. Lockers, left and right cotsides, and overbed tables in 30 bed spaces were screened for aerobic colony count (ACC), methicillin-susceptible Staphylococcus aureus (MSSA), and methicillin-resistant S. aureus (MRSA) before cleaning with electrolyzed water. Sites were rescreened at varying intervals from 1 to 48 hours after cleaning. Microbial growth was quantified as colony-forming units (CFUs) per square centimeter and presence or absence of MSSA and MRSA at each site. The study was repeated 3 times at monthly intervals. There was an early and significant reduction in average ACC (360 sampled sites) from a before-cleaning level of 4.3 to 1.65 CFU/cm2 at 1 hour after disinfectant cleaning (P <.0001). Average counts then increased to 3.53 CFU/cm2 at 24 hours and 3.68 CFU/cm2 at 48 hours. Total MSSA/MRSA (34 isolates) decreased by 71% at 4 hours after cleaning but then increased to 155% (53 isolates) of precleaning levels at 24 hours. Cleaning with electrolyzed water reduced ACC and staphylococci on surfaces beside patients. ACC remained below precleaning levels at 48 hours, but MSSA/MRSA counts exceeded original levels at 24 hours after cleaning. Although disinfectant cleaning quickly reduces bioburden, additional investigation is required to clarify the reasons for rebound contamination of pathogens at near-patient sites.


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Microbe(s): Salmonella typhimurium, Listeria monocytogenes, Staphylococcus aureus, Escherichia coli


Salmonella spp. may be found in the nest box of breeder chickens, cold egg-storage rooms at the farm, on the hatchery truck, or in the hatchery environment (5). These bacteria may then be spread to fertilized hatching eggs on the shell or, in some cases, may penetrate the shell and reside just beneath the surface of the eggshell.Research has demonstrated that contamination of raw poultry products with Salmonella spp. may be attributable to cross-contamination in the hatchery from Salmonella infected eggs or surfaces to uninfected baby chicks during the hatching process. Cox et al. (6 and 7) reported that broiler and breeder hatcheries were highly contaminated with Salmonella spp. Within the broiler hatchery, 71 percent of eggshell fragments, 80 percent of chick conveyor belts swabs, and 74 percent of pad samples placed under newly hatched chicks contained Salmonella spp. (6).Cason et al. (4) reported that, although fertile hatching eggs were contaminated with high levels of Salmonella typhimurium, they were still able to hatch. The authors stated that paratyphoid salmonellae do not caadverse health affects to the developing and hatching chick. During the hatching process, Salmonella spp. is readily spread throughout the hatching cabinet due to rapid air movement by circulation fans. When eggs were inoculated with a marker strain of Salmonella during hatching, greater than 80 percent of the chicks in the trays above and below the inoculated eggs were contaminated (4). In an earlier study, Cason et al. (3) demonstrated that salmonellae on the exterior of eggs or in eggshell membranes could be transmitted to baby chicks during pipping.Salmonella may persist in hatchery environments for long periods of time. When chick fluff contaminated with Salmonella was held for 4 years at room temperature, up to 1,000,000 Salmonella cells per gram could be recovered from these samples (12).Researchers have demonstrated a link between cross-contamination in the hatchery and contaminated carcasses during processing. Goren et al. (8) isolated salmonellae from three different commercial hatcheries in Europe and reported that the same serotypes found in the hatcheries could be found on processed broiler chicken carcass skin. Proper disinfection of the hatchery environment and fertile hatching eggs, therefore, is essential for reducing Salmonella on ready-to-cook carcasses.


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Microbe(s): Staphylococcus epidermidis


The purpose of this study was to investigate the mechanism by which a direct electrical current reduced the viability of Staphylococcus epidermidis biofilms in conjunction with ciprofloxacin at physiologic saline conditions meant to approximate those in an infected artificial joint. Biofilms grown in CDC biofilm reactors were exposed to current for 24 hours in 1/10th strength tryptic soy broth containing 9 g/L total NaCl. Dose-dependent log reductions up to 6.7 log10 CFU/cm2 were observed with the application of direct current at all four levels (0.7 to 1.8 mA/cm2) both in the presence and absence of ciprofloxacin. There were no significant differences in log reductions for wells with ciprofloxacin compared to those without at the same current levels. When current exposures were repeated without biofilm or organics in the medium, significant generation of free chlorine was measured. Free chlorine doses equivalent to the 24 hour endpoint concentration for each current level were shown to mimic killing achieved by current application. Current exposure (1.8 mA/cm2) in medium lacking chloride and amended with sulfate, nitrate, or phosphate as alternative electrolytes produced diminished kills of 3, 2, and 0 log reduction, respectively. Direct current also killed Pseudomonas aeruginosa biofilms when NaCl was present. Together these results indicate that electrolysis reactions generating hypochlorous acid from chloride are likely a main contributor to the efficacy of direct current application. A physiologically relevant NaCl concentration is thus a critical parameter in experimental design if direct current is to be investigated for in vivo medical applications.


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Microbe(s): Staphylococcus aureus


Staphylococcus aureus is a major pathogen. It can form biofilm on the surfaces of medical devices and food equipment, which makes it more difficult to eradicate. To develop a novel method to eradicate S. aureus biofilm, the effects of electrolyzed water on removing and killing S. aureus biofilm were investigated in this study. By using a biofilm biomass assay with safranin staining and visualization of biofilm architecture with scanning electron microscopy, it was shown that basic electrolyzed water (BEW) could effectively remove established biofilm. The pH of electrolyzed water affected removal efficacy. Using a biofilm viability assay with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide staining, acidic electrolyzed water (AEW) efficiently killed biofilm-imbedded S. aureus. The available chlorine in AEW may be a main contributing factor for bactericidal activity. Additionally, BEW had a removal efficacy for S. aureus biofilm equivalent to 2% NaOH, and AEW had a bactericidal capability for S. aureus in biofilm equivalent to 2% HCl. These data suggested that AEW and BEW could be applied as a bactericide and removing agent for S. aureus in biofilm, respectively.


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Microbe(s): Staphylococcus aureus


The objective of this study was to investigate the combined effect of temperature (1535C), pH (3-9), and dipping time (15 min) on the inactivation of Staphylococcus aureus in broth treated with low concentration electrolyzed water (LcEW). Reductions of 1.447.12 log CFU/mL were observed in different combinations of the 3 factors. Also, a quadratic equation for S. aureus inactivation kinetic was developed by multiple regression analysis using response surface methodology. The predicted values were shown to be significantly in good agreement with experimental values as a result of the level of significance was p<0.0001. Besides, the developed model was validated by fitting with literature data and the results showed that the predictions had a good agreement with the observed data with a satisfied determination of coefficient (R2=0.963).


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Microbe(s): Escherichia coli O157:H7, Staphylococcus aureus


The use of different available chlorine concentrations (ACCs) of slightly acidic electrolyzed water (SAEW; 0.5 to 30 mg/liter), different treatment times, and different temperatures for inactivating Escherichia coli O157:H7 and Staphylococcus aureus was evaluated. The morphology of both pathogens also was analyzed with transmission electron microscopy. A 3-min treatment with SAEW (pH 6.0 to 6.5) at ACCs of 2 mg/liter for E. coli O157:H7 and 8 mg/liter for S. aureus resulted in 100% inactivation of two cultures (7.92- to 8.75-log reduction) at 25 C. The bactericidal activity of SAEW was independent of the treatment time and temperature at a higher ACC (P > 0.05). E. coli O157:H7 was much more sensitive than S. aureus to SAEW. The morphological damage to E. coli O157:H7 cells by SAEW was significantly greater than that to S. aureus cells. At an ACC as high as 30 mg/liter, E. coli O157:H7 cells were damaged, but S. aureus cells retained their structure and no cell wall damage or shrinkage was observed. SAEW with a near neutral pH may be a promising disinfectant for inactivation of foodborne pathogens.


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Microbe(s): Escherichia coli O157:H7, Listeria monocytogenes, Staphylococcus aureus, Salmonella Typhimurium


Strong acid electrolyzed water (SAEW) has a very limited application due to its low pH value (< 2.7) and corrosive characteristics. Thus, we developed new low concentration electrolyzed water (LcEW). The efficacy of LcEW under various treatment conditions for the inactivation of different foodborne pathogens in pure culture was evaluated and compared with SAEW. The efficiency of LcEW and SAEW for the inactivation of predominant foodborne pathogens (Escherichia coli O157:H7, Listeria monocytogenes, Staphylococcus aureus and Salmonella Typhimurium) with different dipping times (1, 3, 5, 7 and 10 min), pH values (2.5, 4.0, 5.0, 6.0 and 9.0) and temperatures (4, 15, 23, 35 and 50 C) were determined. Reductions of bacterial populations of 1.7 to 6.6 log10 CFU/mL in various treated conditions in cell suspensions were observed after treatment with LcEW and SAEW, compared to the untreated control. Dip washing (1 min at 35 C) of lettuce leaves in both electrolyzed water resulted in 2.5 to 4.0 log10 CFU/g compared to the unwashed control. Strong inactivation effects were observed in LcEW, and no significant difference (p > 0.05) was observed between LcEW and SAEW. The effective form of chlorine compounds in LcEW was almost exclusively hypochlorous acid (HOCl), which has strong antimicrobial activity and leaves no residuals due to the low concentration of residual chlorine. Thus, LcEW could be widely applied as a new sanitizer in the food industry.


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Microbe(s): Escherichia coli O157: H7, Staphylococcus aureus


The use of different available chlorine concentrations (ACCs) of slightly acidic electrolyzed water (SAEW; 0.5 to 30 mg/


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Microbe(s): Salmonella Enteritidis, Escherichia coli, Staphylococcus aureus, Listeria monocytogenes, and Bacillus cereus


The bactericidal effect of slightly acidic hypochlorous water (SAHW) on Salmonella Enteritidis, Escherichia coli, Staphylococcus aureus, Listeria monocytogenes, and Bacillus cereus, as well as some bacterial strains isolated from fresh lettuce was evaluated. Viable counts of all tested bacterial samples decreased immediately after treatment by SAHW. Most bacterial cells with the exception of B. cereus, and S. aureus were not culturable on TSA after treatment by 1 to 30 mg/L SAHW. Likewise, Pseudomonas sp., and Flavobacterium or Xanthomonas sp., Kurthia sp., Micrococcus sp., and Corynebacterium or Microbacterium sp. were not culturable on TSA after treatment by 30 mg/L SAHW. Viable counts of S. aureus, E. coli, Flavobacterium or Xanthomonas sp., and Pseudomonas sp. showed a 5 to 6 log cfu/mL reduction at day 0 and maintained a count of less than 1 log cfu/mL from day 1 to day 7 following treatment by 30 mg/L SAHW. Sodium hypochlorite (NaOCl, 0.5-1.0 mg/L) decreased the viable counts of S. Enteritidis to less than the lower limit of detection, 1 log cfu/mL, from day 1 to day 7 following treatment by 1 mg/L. NaOCl was not sufficient at 0.5-0.75 mg/L in reducing viable counts of S. Enteritidis because of a 2 to 5 log cfu/mL increase from day 2 to day 5 due to recovery from injury. Initial counts of S. Enteritidis after hydrogen


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Microbe(s): Escherichia coli, Staphylococcus aureus, Salmonella spp.


In the current study, the effectiveness of slightly acidic electrolyzed water (SAEW) on an in vitro inactivation of Escherichia coli (E. coli), Staphylococcus aureus (S. aureus) and Salmonella spp. was evaluated and compared with other sanitizers. SAEW (pH 5.6, 23 mg/l available chlorine concentration; ACC; and 940 mV oxidation reduction potential; ORP) was generated by electrolysis of dilute solution of HCl (2%) in a chamber of a non-membrane electrolytic cell. One milliliter of bacteria suspension (ca. 10-11 log10CFU/ml) was mixed with 9 ml of SAEW, strong acidic electrolyzed water (StAEW; ca. 50 mg/l ACC), sodium hypochlorite solution (NaOCl; ca.120 mg/l ACC) and distilled water (DW) as control and treated for 60 s. SAEW effectively reduced the population of E. coli, S. aureus and Salmonella spp. by 5.1, 4.8, and 5.2 log10CFU/ml. Although, ACC of SAEW was more than 5 times lower than that of NaOCl solution, they showed no significant bactericidal difference (p > 0.05). However, the bactericidal effect of StAEW was significantly higher (p < 0.05) than SAEW and NaOCl solution in all cases. When tested with each individual test solution, E. coli, S. aureus and Salmonella spp. reductions were not significantly different (p > 0.05). These findings indicate that SAEW with low available chlorine concentration can equally inactivate E. coli, S. aureus and Salmonella spp. as NaOCl solution and therefore SAEW shows a high potential of application in agriculture and food industry as an environmentally friendly disinfection agent.


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Microbe(s): Escherichia coli, Staphylococcus aureus


Suspension quantitative germicidal test showed that electrolyzed oxidizing water (EO water) was an efficient and rapid disinfectant. Disinfection rates towards E. coli (available chlorine concentration ACC: 12.40 mg/L) and Staphylococcus aureus (ACC: 37.30 mg/L) could reach 100% at 1 and 3 min, respectively. Disinfection mechanism of EO water was investigated at a molecular biological level by detecting a series of biochemical indices. The results showed that the dehydrogenase activities of E. coli and S. aureus decreased rapidly, respectively, at the rates of 45.9% and 32% in the 1st minute treatment with EO water. EO water also improved the bacterial membrane permeability, causing the rise of conductivities and the rapid leakages of intracellular DNA, K+, and proteins in 1 min. The leakages of DNA and K+ tended to slow down after about 1 min while those of proteins began to decrease a little after reaching the peak values. The sodium dodecyl sulfonate polyacrylamide gel electrophoresis (SDS-PAGE) showed that EO water destroyed intracellular proteins. The protein bands got fainter and even disappeared as the treatment proceeded. EO water s effects on the bacterial ultrastructures were also verified by the transmission electronic microscopy (TEM) photos. The disinfection mechanism of EO water was composed of several comprehensive factors including the destruction of bacterial protective barriers, the increase of membrane permeability, the leakage of cellular inclusions, and the activity decrease of some key enzymes.


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Microbe(s): Escherichia coli, Staphylococcus epidermidis


This study investigated residual bacteria and different food types left on tableware items after various washing and sanitization protocols. Escherichia coli K-12 and Staphylococcus epidermidis were inoculated into whole milk and soft cream cheese. The milk was used to contaminate regular drinking glasses and the cheese was used to contaminate plates and silverware. These tableware items were washed in manual (43 C) and mechanical (49 C) washers and sanitized with different sanitizers (24 C) for 5 s. Quaternary ammonium compound, sodium hypochlorite, peroxyacetic acid, neutral electrolyzed water (NEW), and a combination of citric acid with sodium dodecylbenzene sulfonate (acidic formulation) were used as the chemical sanitizers. Tap water was used as a control. Results showed that at least 5-log reductions in both bacterial numbers were achieved for all sanitizers in both types of washers, except for the control. With mechanical dishwashing, the NEW and acidic formulation treatments reduced bacterial populations by >6.9 and >6.0 log CFU per tableware item, respectively. With the manual operation, bacterial numbers were reduced by >5.4 and >6.0 log CFU per tableware item, respectively. This study revealed that NEW and the acidic formulation are as effective as the other chemical sanitizers for food contact surface sanitization in manual and mechanical ware washing.


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Microbe(s): Escherichia coli, Salmonella typhimurium, Staphylococcus aureus, Listeria monocytogenes, and Enterococcus faecalis


Food safety issues and increases in food borne illnesses have promulgated the development of new sanitation methods to eliminate pathogenic organisms on foods and surfaces in food service areas. Electrolyzed oxidizing water (EO water) shows promise as an environmentally friendly broad spectrum microbial decontamination agent. EO water is generated by the passage of a dilute salt solution (1% NaCl) through an electrochemical cell. This electrolytic process converts chloride ions and water molecules into chlorine oxidants (Cl2, HOCl/ClO-). At a near-neutral pH (pH 6.3-6.5), the predominant chemical species is the highly biocidal hypochlorous acid species (HOCl) with the oxidation reduction potential (ORP) of the solution ranging from 800 to 900 mV. The biocidal activity of near-neutral EO water was evaluated at 25 C using pure cultures of Escherichia coli, Salmonella typhimurium, Staphylococcus aureus, Listeria monocytogenes, and Enterococcus faecalis. Treatment of these organisms, in pure culture, with EO water at concentrations of 20, 50, 100, and 120 ppm total residual chlorine (TRC) and 10 min of contact time resulted in 100% inactivation of all five organisms (reduction of 6.1-6.7 log10 CFU/mL). Spray treatment of surfaces in food service areas with EO water containing 278-310 ppm TRC (pH 6.38) resulted in a 79-100% reduction of microbial growth. Dip (10 min) treatment of spinach at 100 and 120 ppm TRC resulted in a 4.0-5.0 log10 CFU/mL reduction of bacterial counts for all organisms tested. Dipping (10 min) of lettuce at 100 and 120 ppm TRC reduced bacterial counts of E. coli by 0.24-0.25 log10 CFU/mL and reduced all other organisms by 2.43-3.81 log10 CFU/mL.


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Microbe(s): Escherichia coli, Listeria monocytogenes, Pseudomonas aeruginosa, Staphylococcus aureus


This study evaluated the efficacy of neutral electrolyzed water (NEW; 64.1 mg/liter of active chlorine) to reduce populations of Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, and Listeria monocytogenes on plastic and wooden kitchen cutting boards. Its effectiveness was compared with that of a sodium hypochlorite solution (NaClO; 62.3 mg/liter of active chlorine). Inoculated portions of cutting boards were rinsed in either NEW or NaClO solutions, or deionized water (control). Plastic boards were rinsed for 1 min and wooden boards for 1 and 5 min. After each treatment, the surviving population of each strain was determined on the surface and in the soaking water. No significant difference (P 0.05) was found between the final populations of each strain with regard to the treatment solutions (NEW or NaClO). However, a significant difference (P 0.05) was revealed between surface materials after 1 min of washing. Whereas in plastic boards the initial bacterial populations were reduced by 5 log CFU/50 cm2, in wooden cutting boards they underwent a reduction of <3 log CFU/50 cm2. A 5-min exposure time yielded reductions of about 4 log CFU/50 cm2. The surviving populations of all bacteria in NEW and NaClO washing solutions were <1 log CFU/ml after soaking both surfaces. This study revealed that NEW treatment is an effective method for reducing microbial contamination on plastic and wooden cutting boards. NEW efficacy was comparable to that of NaClO, with the advantage of having a larger storage time.


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Microbe(s): Escherichia coli, Listeria monocytogenes, Pseudomonas aeruginosa, Staphylococcus aureus


Aim: To ascertain the efficacy of neutral electrolysed water (NEW) in reducing Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus and Listeria monocytogenes on glass and stainless steel surfaces. Its effectiveness for that purpose is compared with that of a sodium hypochlorite (NaClO) solution with similar pH, oxidation-reduction potential (ORP) and active chlorine content. Methods and Results: First, the bactericidal activity of NEW was evaluated over pure cultures (8-5 log CFU ml-1) of the abovementioned strains: all of them were reduced by more than 7 log CFU ml-1 within 5 min of exposure either to NEW (63 mg l-1 active chlorine) or to NaClO solution (62 mg l-1 active chlorine). Then, stainless steel and glass surfaces were inoculated with the same strains and rinsed for 1 min in either NEW, NaClO solution or deionized water (control). In the first two cases, the populations of all the strains decreased by more than 6 log CFU 50 cm-2. No significant difference (P 0 05) was found between the final populations of each strain with regard to the treatment solutions (NEW or NaClO solution) or to the type of surface. Conclusions: NEW was revealed to be as effective as NaClO at significantly reducing the presence of pathogenic and spoilage bacteria (in this study, E. coli, L. monocytogenes, P. aeruginosa and S. aureus) on stainless steel and glass surfaces. Significance and Impact of the Study: NEW has the advantage of being safer than NaClO and easier to handle. Hence, it represents an advantageous alternative for the disinfection of surfaces in the food industry.


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Microbe(s): Salmonella typhimurium, Staphylococcus aureus, and Listeria monocytogenes, Escherichia coli


Research was conducted to compare the effectiveness of electrolyzed oxidative (EO) water applied using an electrostatic spraying system (ESS) for killing populations of bacteria that are of concern to the poultry industry. Populations of pathogenic bacteria (Salmonella typhimurium, Staphylococcus aureus, and Listeria monocytogenes), and the indicator bacterium Escherichia coli were applied to eggs and allowed to attach for 1 h. EO water completely eliminated all Salmonella typhimurium on 3, 7, 1, and 8 out of 15 eggs in Repetitions (Rep) 1, 2, 3, and 4, respectively, even when very high inoculations were used. EO water completely eliminated all Staphylococcus aureus on 12, 11, 12, and 11 out of 15 eggs in Rep 1, 2, 3, and 4, respectively. EO water completely eliminated all Listeria monocytogenes on 8, 13, 12, and 14 out of 15 eggs in Reps 1, 2, 3, and 4, respectively. EO water completely eliminated all Escherichia coli on 9, 11, 15, and 11 out of 15 eggs in Reps 1, 2, 3, and 4, respectively. Even when very high concentrations of bacteria were inoculated onto eggs (many times higher than would be encountered in industrial situations), EO water was found to be effective when used in conjunction with electrostatic spraying for eliminating pathogenic and indicator populations of bacteria from hatching eggs.


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Microbe(s): Staphylococcus, Staphylococcal enterotoxin-A


Electrolyzed anodic NaCl solutions [EW(+)], prepared by the electrolysis of 0.1% NaCl, have been shown to instantly inactivate most pathogens that cause food-borne disease. Elimination of food-borne pathogens does not necessarily guarantee food safety because enterotoxins produced by pathogens may remain active. We have tested whether EW(+) can inactivate Staphylococcal enterotoxin A (SEA), one of the major enterotoxins responsible for food poisoning. Fixed quantities of SEA were mixed with increasing molar ratios of EW(+), and SEA was evaluated by reversed-phase passive latex agglutination (RPLA) test, immunoassay, native polyacrylamide gel electrophoresis (PAGE), and amino acid analysis after 30 min incubations. Exposure of 70 ng, or 2.6 pmol, of SEA in 25 L of PBS to a 10-fold volume of EW(+), or ca. 64.6 103-fold molar excess of HOCl in EW(+), caused a loss of immuno-reactivity between SEA and a specific anti-SEA antibody. Native PAGE indicated that EW(+) caused fragmentation of SEA, and amino acid analysis indicated a loss in amino acid content, in particular Met, Tyr, Ile, Asn, and Asp. Staphylococcal enterotoxin-A excreted into culture broth was also inactivated by exposure to an excess molar ratio of EW(+). Thus, EW(+) may be a useful management tool to ensure food hygiene by food processing industries.


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Microbe(s): Enterobacter aerogenes, Staphylococcus aureus


The effectiveness of electrolyzed (EO) water at killing Enterobacter aerogenes and Staphylococcus aureus in pure culture was evaluated. One milliliter (approximately 109 CFU/ml) of each bacterium was subjected to 9 ml of EO water or control water (EO water containing 10% neutralizing buffer) at room temperature for 30 s. Inactivation (reduction of >9 log10 CFU/ml) of both pathogens occurred within 30 s after exposure to EO water containing approximately 25 or 50 mg of residual chlorine per liter. The effectiveness of EO water in reducing E. aerogenes and S. aureus on different surfaces (glass, stainless steel, glazed ceramic tile, unglazed ceramic tile, and vitreous china) was also evaluated. After immersion of the tested surfaces in EO water for 5 min without agitation, populations of E. aerogenes and S. aureus were reduced by 2.2 to 2.4 log10 CFU/cm2 and by 1.7 to 1.9 log10 CFU/cm2, respectively, whereas washing with control water resulted in a reduction of only 0.1 to 0.3 log10 CFU/cm2. The washing of tested surfaces in EO water with agitation (50 rpm) reduced populations of viable cells on the tested surfaces to <1 CFU/cm2. For the control water treatment with agitation, the surviving numbers of both strains on the tested surfaces were approximately 3 log10 CFU/cm2. No viable cells of either strain were observed in the EO water after treatment, regardless of agitation. However, large populations of both pathogens were recovered from control wash solution after treatment.