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Sodium polyphosphate and polyethylenimine enhance the antimicrobial activities of plant essential oils

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      Plant extracts have been used for millennia for treatment of disease, with much recent interest focusing on the antimicrobial activities of plant essential oils (EOs). Although EOs are active against common microbial pathogens, their effective use as topical, environmental, or food antimicrobials will require EO-based formulations with enhanced antimicrobial activities. In this study, two polyionic compounds, sodium polyphosphate (polyP, a polyanion) and polyethylenimine (PEI, a polycation), were evaluated for their abilities to enhance the antimicrobial activities of six EOs against the human pathogens Escherichia coli O157:H7, Salmonella enterica subsp. enterica ser. Minnesota, Pseudomonas aeruginosa, Listeria monocytogenes, Staphylococcus aureus, and Candida albicans. EOs tested were cinnamon, clove, regular and redistilled oregano, and two types of thyme oil. EOs were examined via disk diffusion and broth microdilution, either alone or in the presence of subinhibitory levels of polyP or PEI. Both polyP and PEI were found to be effective enhancers of EO activity against all strains examined, and calculation of fractional inhibitory indices for select EO/organism pairings demonstrated that true synergy was possible with this enhancement approach. Experiments with a deep-rough strain of S. Minnesota probed the role of the outer membrane in both intrinsic resistance to EOs and enhancement by polyions. The use of polyP and PEI for boosting the antimicrobial activities of EOs may eventually facilitate the development of more effective EO-based antimicrobial treatments for use in applications such as wound treatment, surface disinfection, or as generally recognized as safe antimicrobials for use in foods or on food contact surfaces.

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      Essential oils: their antibacterial properties and potential applications in foods--a review.

      In vitro studies have demonstrated antibacterial activity of essential oils (EOs) against Listeria monocytogenes, Salmonella typhimurium, Escherichia coli O157:H7, Shigella dysenteria, Bacillus cereus and Staphylococcus aureus at levels between 0.2 and 10 microl ml(-1). Gram-negative organisms are slightly less susceptible than gram-positive bacteria. A number of EO components has been identified as effective antibacterials, e.g. carvacrol, thymol, eugenol, perillaldehyde, cinnamaldehyde and cinnamic acid, having minimum inhibitory concentrations (MICs) of 0.05-5 microl ml(-1) in vitro. A higher concentration is needed to achieve the same effect in foods. Studies with fresh meat, meat products, fish, milk, dairy products, vegetables, fruit and cooked rice have shown that the concentration needed to achieve a significant antibacterial effect is around 0.5-20 microl g(-1) in foods and about 0.1-10 microl ml(-1) in solutions for washing fruit and vegetables. EOs comprise a large number of components and it is likely that their mode of action involves several targets in the bacterial cell. The hydrophobicity of EOs enables them to partition in the lipids of the cell membrane and mitochondria, rendering them permeable and leading to leakage of cell contents. Physical conditions that improve the action of EOs are low pH, low temperature and low oxygen levels. Synergism has been observed between carvacrol and its precursor p-cymene and between cinnamaldehyde and eugenol. Synergy between EO components and mild preservation methods has also been observed. Some EO components are legally registered flavourings in the EU and the USA. Undesirable organoleptic effects can be limited by careful selection of EOs according to the type of food.
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        The phenolic hydroxyl group of carvacrol is essential for action against the food-borne pathogen Bacillus cereus.

        The natural antimicrobial compound carvacrol shows a high preference for hydrophobic phases. The partition coefficients of carvacrol in both octanol-water and liposome-buffer phases were determined (3.64 and 3.26, respectively). Addition of carvacrol to a liposomal suspension resulted in an expansion of the liposomal membrane. Maximum expansion was observed after the addition of 0.50 micromol of carvacrol/mg of L-alpha-phosphatidylethanolamine. Cymene, a biological precursor of carvacrol which lacks a hydroxyl group, was found to have a higher preference for liposomal membranes, thereby causing more expansion. The effect of cymene on the membrane potential was less pronounced than the effect of carvacrol. The pH gradient and ATP pools were not affected by cymene. Measurement of the antimicrobial activities of compounds similar to carvacrol (e.g., thymol, cymene, menthol, and carvacrol methyl ester) showed that the hydroxyl group of this compound and the presence of a system of delocalized electrons are important for the antimicrobial activity of carvacrol. Based on this study, we hypothesize that carvacrol destabilizes the cytoplasmic membrane and, in addition, acts as a proton exchanger, thereby reducing the pH gradient across the cytoplasmic membrane. The resulting collapse of the proton motive force and depletion of the ATP pool eventually lead to cell death.
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          Agents that increase the permeability of the outer membrane.

          The outer membrane of gram-negative bacteria provides the cell with an effective permeability barrier against external noxious agents, including antibiotics, but is itself a target for antibacterial agents such as polycations and chelators. Both groups of agents weaken the molecular interactions of the lipopolysaccharide constituent of the outer membrane. Various polycations are able, at least under certain conditions, to bind to the anionic sites of lipopolysaccharide. Many of these disorganize and cross the outer membrane and render it permeable to drugs which permeate the intact membrane very poorly. These polycations include polymyxins and their derivatives, protamine, polymers of basic amino acids, compound 48/80, insect cecropins, reptilian magainins, various cationic leukocyte peptides (defensins, bactenecins, bactericidal/permeability-increasing protein, and others), aminoglycosides, and many more. However, the cationic character is not the sole determinant required for the permeabilizing activity, and therefore some of the agents are much more effective permeabilizers than others. They are useful tools in studies in which the poor permeability of the outer membrane poses problems. Some of them undoubtedly have a role as natural antibiotic substances, and they or their derivatives might have some potential as pharmaceutical agents in antibacterial therapy as well. Also, chelators (such as EDTA, nitrilotriacetic acid, and sodium hexametaphosphate), which disintegrate the outer membrane by removing Mg2+ and Ca2+, are effective and valuable permeabilizers.

            Author and article information

            Rapid Microbial Detection and Control Laboratory, Department of Food Science and Human Nutrition, Iowa State University, Ames, IA, USA
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            [* ]Corresponding author’s e-mail address: byron@
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            ScienceOpen Research
            29 December 2016
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            © 2017 Wright and Brehm-Stecher

            This work has been published open access under Creative Commons Attribution License CC BY 4.0 , which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Conditions, terms of use and publishing policy can be found at .

            Figures: 7, Tables: 3, References: 41, Pages: 13
            Original article

            General agriculture, Microbiology & Virology

            Natural antimicrobials


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