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1.
Appl Environ Microbiol ; 82(4): 1035-1039, 2016 02 15.
Article in English | MEDLINE | ID: mdl-26637595

ABSTRACT

There is still great interest in controlling bacterial endospores. The use of chemical disinfectants and, notably, oxidizing agents to sterilize medical devices is increasing. With this in mind, hydrogen peroxide (H2O2) and peracetic acid (PAA) have been used in combination, but until now there has been no explanation for the observed increase in sporicidal activity. This study provides information on the mechanism of synergistic interaction of PAA and H2O2 against bacterial spores. We performed investigations of the efficacies of different combinations, including pretreatments with the two oxidizers, against wild-type spores and a range of spore mutants deficient in the spore coat or small acid-soluble spore proteins. The concentrations of the two biocides were also measured in the reaction vessels, enabling the assessment of any shift from H2O2 to PAA formation. This study confirmed the synergistic activity of the combination of H2O2 and PAA. However, we observed that the sporicidal activity of the combination is largely due to PAA and not H2O2. Furthermore, we observed that the synergistic combination was based on H2O2 compromising the spore coat, which was the main spore resistance factor, likely allowing better penetration of PAA and resulting in the increased sporicidal activity.


Subject(s)
Anti-Bacterial Agents/pharmacology , Drug Synergism , Hydrogen Peroxide/pharmacology , Microbial Viability/drug effects , Peracetic Acid/pharmacology , Spores, Bacterial/drug effects , Spores, Bacterial/physiology
2.
J Antimicrob Chemother ; 70(3): 773-9, 2015 Mar.
Article in English | MEDLINE | ID: mdl-25428922

ABSTRACT

OBJECTIVES: To elucidate the mechanisms of spore resistance to and killing by the oxidizing microbicide peracetic acid (PAA). METHODS: Mutants of Bacillus subtilis lacking specific spore structures were used to identify resistance properties in spores and to understand the mechanism of action of PAA. We also assessed the effect of PAA treatment on a number of spore properties including heat tolerance, membrane integrity and germination. RESULTS: The spore coat is essential for spore PAA resistance as spores with defective coats were greatly sensitized to PAA treatment. Small acid-soluble spore proteins apparently provide no protection against PAA. Defects in spore germination, specifically in germination via the GerB and GerK but not the GerA germination receptors, as well as leakage of internal components suggest that PAA is active at the spore inner membrane. It is therefore likely that the inner membrane is the major site of PAA's sporicidal activity. CONCLUSIONS: PAA treatment targets the spore membrane, with some of its activity directed specifically against the GerB and GerK germination receptors.


Subject(s)
Anti-Infective Agents/pharmacology , Bacillus subtilis/drug effects , Microbial Viability/drug effects , Peracetic Acid/pharmacology , Spores, Bacterial/drug effects , Bacillus subtilis/genetics , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Membrane Proteins/genetics , Membrane Proteins/metabolism , Membranes/drug effects , Oxidants/pharmacology , Spores, Bacterial/genetics
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