title: Combination Inhibition Activity of Nisin and Ethanol on the Growth Inhibition of Pathogenic Gram Negative Bacteria and Their Application as Disinfectant Solution.
abstract: Nisin and ethanol have been used as antimicrobial agents in food industry. However, nisin alone could not inhibit the growth of gram-negative bacteria, except in combination with a chelating agent, EDTA, or organic acid. This research aimed to study the survival of Escherichia coli O157: H7, Salmonella Typhimurium TISTR 292 and Salmonella Enteritidis DMST 17368 after treatment with nisin at 100, 200, 300, 500, 800, or 1000 IU/mL and ethanol at 70%, 50%, 30%, 20%, or 10% (v/v) alone and in combination. None of all nisin concentrations could reduce the growth of target strains. While 20% ethanol (v/v) having no negative effect on human health, could slightly reduce the growth of target strains. However, the combination of nisin at 500, 800 or 1000 IU/mL and 20% ethanol displayed significant growth reduction at 15 min were below 1 log CFU/mL. Thus, the minimum inhibitory concentration of nisin and ethanol was 500 IU/mL and 20% (v/v), respectively. The release of fatty acid, genetic materials and scanning electron microscope suggested that nisin-ethanol treated cells have altered permeability causing bacterial growth inhibition. Comparison treatment of combined solution and commercial chloride based sanitizer were done for all target strains on stainless steel surface. Survivals of three target strains were below 1 log CFU/mL. The result suggested that combined solution of nisin and ethanol may be a beneficial sanitizer for food industry to inhibit the growth of E. coli O157:H7 and Salmonella sp.
  
  
  CONCLUSIONS
  The combination of nisin and ethanol, first report, could inhibit growth of Gram negative bacteria under the mild condition and short time. This combination solution could be used as a sanitizer solution for food industry and reduce the risk of consumer directly contacting on strong chemical agents.
authors.affiliation: Specialized Research Unit of Probiotics and Prebiotics for Health, Dept. of Biotechnology, Faculty of Agro-Industry, Kasetsart Univ, Bangkok, 10900, Thailand.
authors.forename: Komsan
authors.initials: K
authors.lastname: Phongphakdee
authors.affiliation: Specialized Research Unit of Probiotics and Prebiotics for Health, Dept. of Biotechnology, Faculty of Agro-Industry, Kasetsart Univ, Bangkok, 10900, Thailand.
authors.forename: Sunee
authors.initials: S
authors.lastname: Nitisinprasert
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D004202:Disinfectants / Q000494:pharmacology*; D019453:Escherichia coli O157 / Q000254:growth & development; D000431:Ethanol / Q000494:pharmacology*; D006090:Gram-Negative Bacteria / Q000187:drug effects* / Q000254:growth & development; D008826:Microbial Sensitivity Tests; D009561:Nisin / Q000494:pharmacology*; D012477:Salmonella enteritidis / Q000254:growth & development; D012486:Salmonella typhimurium / Q000187:drug effects
chemical_list: D000900:Anti-Bacterial Agents; D004202:Disinfectants; D009561:Nisin; D000431:Ethanol
keywords: Gram negative bacteria; Nisin; combination; ethanol; inhibition activity