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      Chemical Composition, Antibacterial Properties and Mechanism of Action of Essential Oil from Clove Buds against Staphylococcus aureus

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          Abstract

          The essential oil of clove has a wide range of pharmacological and biological activities and is widely used in the medicine, fragrance and flavoring industries. In this work, 22 components of the essential oil obtained from clove buds were identified. Eugenol was the major component (76.23%). The essential oil exhibited strong antibacterial activity against Staphylococcus aureus ATCC 25923 with a minimum inhibitory concentration (MIC) of 0.625 mg/mL, and the antibacterial effects depended on its concentration and action time. Kill-time assays also confirmed the essential oil had a significant effect on the growth rate of surviving S. aureus. We hypothesized that the essential oil may interact with the cell wall and membrane first. On the one hand it destroys cell wall and membranes, next causing the losses of vital intracellular materials, which finally result in the bacterial death. Besides, essential oil penetrates to the cytoplasmic membrane or enters inside the cell after destruction of cell structure, and then inhibits the normal synthesis of DNA and proteins that are required for bacterial growth. These results suggested that the effects of the clove essential oil on the growth inhibition of S. aureus may be at the molecular level rather than only physical damage.

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          Microtitre plate-based antibacterial assay incorporating resazurin as an indicator of cell growth, and its application in the in vitro antibacterial screening of phytochemicals

          The resazurin assay utilising microtitre-plate, described by Drummond and Waigh in 2000, has been modified to achieve more accuracy in the determination of the minimum inhibitory concentration (MIC) values of natural products, including crude extracts, chromatographic fractions or purified compounds against various bacterial strains. This modified resazurin method is simple, sensitive, rapid, robust and reliable, and could be used successfully to assess antibacterial properties of natural products.
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            Antimicrobial herb and spice compounds in food

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              Eugenol (an essential oil of clove) acts as an antibacterial agent against Salmonella typhi by disrupting the cellular membrane.

              To evaluate the antibacterial activity of eugenol and its mechanism of bactericidal action against Salmonella typhi. The antibacterial activity was checked by disc-diffusion method, MIC, MBC, time course assay and pH sensitivity assay. The chemo-attractant property of eugenol was verified by chemotaxis assay. The mode of action of eugenol was determined by crystal violet assay, measurement of release of 260 nm absorbing material, SDS-PAGE, FT-IR spectroscopy, AFM and SEM. Treatment with eugenol at their MIC (0.0125%) and MBC (0.025%) reduced the viability and resulted in complete inhibition of the organism. Eugenol inactivated Salmonella typhi within 60 min exposure. The chemo-attractant property of eugenol combined with the observed high antibacterial activity at alkaline pH favors the fact that the compound can work more efficiently when given in vivo. Eugenol increased the permeability of the membrane, as evidenced by crystal violet assay. The measurement of release of 260 nm absorbing intracellular materials, SDS-PAGE, SEM and AFM analysis confirmed the disruptive action of eugenol on cytoplasmic membrane. The deformation of macromolecules in the membrane, upon treatment with eugenol was verified by FT-IR spectroscopy. The results suggest that the antibacterial activity of eugenol against Salmonella typhi is due to the interaction of eugenol on bacterial cell membrane. Copyright (c) 2010 Elsevier Ireland Ltd. All rights reserved.
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                Author and article information

                Contributors
                Role: Academic Editor
                Journal
                Molecules
                Molecules
                molecules
                Molecules
                MDPI
                1420-3049
                08 September 2016
                September 2016
                : 21
                : 9
                : 1194
                Affiliations
                [1 ]College of Food Sciences, Shanxi Normal University, Linfen 041004, China; sxsd456@ 123456sina.com
                [2 ]College of Life Sciences, Shanxi Normal University, Linfen 041004, China; liuyu1961119@ 123456163.com (T.L.); hqp72@ 123456163.com (Q.-P.H.)
                Author notes
                [* ]Correspondence: xujg@ 123456sxnu.edu.cn ; Tel.: +86-357-205-1247
                Article
                molecules-21-01194
                10.3390/molecules21091194
                6274078
                27617990
                7a277103-8863-49a9-8b92-9b0c5c376ce8
                © 2016 by the authors.

                Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC-BY) license ( http://creativecommons.org/licenses/by/4.0/).

                History
                : 03 August 2016
                : 05 September 2016
                Categories
                Article

                essential oil,staphylococcus aureus,alkaline phosphatase,membrane permeability,electron microscope,protein,dna

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