The agar microdilution method – a new method for antimicrobial susceptibility testing for essential oils and plant extracts
Aims To develop a new agar microdilution technique suitable for the assessment of the antimicrobial activity of natural plant products such as essential oils or plant extracts as well as to evaluate the antimicrobial effect of several essential oils and plant extracts. Methods and Results The propos...
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| Veröffentlicht in: | Journal of applied microbiology 2016-11, Vol.121 (5), p.1291-1299 |
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| container_title | Journal of applied microbiology |
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| creator | Golus, J. Sawicki, R. Widelski, J. Ginalska, G. |
| description | Aims
To develop a new agar microdilution technique suitable for the assessment of the antimicrobial activity of natural plant products such as essential oils or plant extracts as well as to evaluate the antimicrobial effect of several essential oils and plant extracts.
Methods and Results
The proposed agar microdilution method was evolved on the basis of the CLSI agar dilution method, approved for conventional antimicrobials. However, this new method combines convenience and time/cost effectiveness typical for microtitre methods with the advantages of the agar dilution of hydrophobic or coloured substances. Different concentrations of the tested agents were added to Eppendorf tubes with molten Mueller‐Hinton agar, vortexed and dispensed into the 96‐well microplate in a small volume of 100 μl per well which allows for rapid, easy and economical preparation of samples as well as providing a uniform and stable dispersion without separation of the oil–water phases which occurs in methods with liquid medium. Next, the agar microdilution plates were inoculated with four reference bacterial strains. The results of our study demonstrated that the minimal inhibitory concentrations (MICs) were successfully determined using the agar microdilution method even with hydrophobic essential oils or strongly coloured plant extracts.
Conclusions
The new agar microdilution method avoids the problems associated with testing of water insoluble, oily or strongly coloured plant natural products. Moreover, it enables the reliable, cheap and easy MIC determination of such agents.
Significance and Impact of the Study
In the era of increasing antibiotic resistance high hopes are associated with new drugs of plant origin. However, the lack of standardized and reliable testing methods for assessing antibacterial activity of plant natural products causes impediment to research into this area. This study demonstrated that the agar microdilution method can be used successfully for testing oily and coloured substances. |
| doi_str_mv | 10.1111/jam.13253 |
| format | Article |
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To develop a new agar microdilution technique suitable for the assessment of the antimicrobial activity of natural plant products such as essential oils or plant extracts as well as to evaluate the antimicrobial effect of several essential oils and plant extracts.
Methods and Results
The proposed agar microdilution method was evolved on the basis of the CLSI agar dilution method, approved for conventional antimicrobials. However, this new method combines convenience and time/cost effectiveness typical for microtitre methods with the advantages of the agar dilution of hydrophobic or coloured substances. Different concentrations of the tested agents were added to Eppendorf tubes with molten Mueller‐Hinton agar, vortexed and dispensed into the 96‐well microplate in a small volume of 100 μl per well which allows for rapid, easy and economical preparation of samples as well as providing a uniform and stable dispersion without separation of the oil–water phases which occurs in methods with liquid medium. Next, the agar microdilution plates were inoculated with four reference bacterial strains. The results of our study demonstrated that the minimal inhibitory concentrations (MICs) were successfully determined using the agar microdilution method even with hydrophobic essential oils or strongly coloured plant extracts.
Conclusions
The new agar microdilution method avoids the problems associated with testing of water insoluble, oily or strongly coloured plant natural products. Moreover, it enables the reliable, cheap and easy MIC determination of such agents.
Significance and Impact of the Study
In the era of increasing antibiotic resistance high hopes are associated with new drugs of plant origin. However, the lack of standardized and reliable testing methods for assessing antibacterial activity of plant natural products causes impediment to research into this area. This study demonstrated that the agar microdilution method can be used successfully for testing oily and coloured substances.</description><identifier>ISSN: 1364-5072</identifier><identifier>EISSN: 1365-2672</identifier><identifier>DOI: 10.1111/jam.13253</identifier><identifier>PMID: 27501239</identifier><identifier>CODEN: JAMIFK</identifier><language>eng</language><publisher>England: Oxford University Press</publisher><subject>Agar ; agar microdilution method ; Anti-Bacterial Agents - pharmacology ; antibacterial activity ; Antibiotics ; Drug resistance ; Drug Resistance, Bacterial ; essential oil ; Microbial Sensitivity Tests - methods ; Microbiology ; minimal inhibitory concentration ; Oils & fats ; Oils, Volatile - pharmacology ; plant extract ; Plant Extracts - pharmacology</subject><ispartof>Journal of applied microbiology, 2016-11, Vol.121 (5), p.1291-1299</ispartof><rights>2016 The Society for Applied Microbiology</rights><rights>2016 The Society for Applied Microbiology.</rights><rights>Copyright © 2016 The Society for Applied Microbiology</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c4213-2385e0330bc55415ea69da97ee0a8170f18d024b08b83851316c21150b43afc93</citedby><cites>FETCH-LOGICAL-c4213-2385e0330bc55415ea69da97ee0a8170f18d024b08b83851316c21150b43afc93</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://onlinelibrary.wiley.com/doi/pdf/10.1111%2Fjam.13253$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1111%2Fjam.13253$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>314,776,780,1411,27903,27904,45553,45554</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/27501239$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Golus, J.</creatorcontrib><creatorcontrib>Sawicki, R.</creatorcontrib><creatorcontrib>Widelski, J.</creatorcontrib><creatorcontrib>Ginalska, G.</creatorcontrib><title>The agar microdilution method – a new method for antimicrobial susceptibility testing for essential oils and plant extracts</title><title>Journal of applied microbiology</title><addtitle>J Appl Microbiol</addtitle><description>Aims
To develop a new agar microdilution technique suitable for the assessment of the antimicrobial activity of natural plant products such as essential oils or plant extracts as well as to evaluate the antimicrobial effect of several essential oils and plant extracts.
Methods and Results
The proposed agar microdilution method was evolved on the basis of the CLSI agar dilution method, approved for conventional antimicrobials. However, this new method combines convenience and time/cost effectiveness typical for microtitre methods with the advantages of the agar dilution of hydrophobic or coloured substances. Different concentrations of the tested agents were added to Eppendorf tubes with molten Mueller‐Hinton agar, vortexed and dispensed into the 96‐well microplate in a small volume of 100 μl per well which allows for rapid, easy and economical preparation of samples as well as providing a uniform and stable dispersion without separation of the oil–water phases which occurs in methods with liquid medium. Next, the agar microdilution plates were inoculated with four reference bacterial strains. The results of our study demonstrated that the minimal inhibitory concentrations (MICs) were successfully determined using the agar microdilution method even with hydrophobic essential oils or strongly coloured plant extracts.
Conclusions
The new agar microdilution method avoids the problems associated with testing of water insoluble, oily or strongly coloured plant natural products. Moreover, it enables the reliable, cheap and easy MIC determination of such agents.
Significance and Impact of the Study
In the era of increasing antibiotic resistance high hopes are associated with new drugs of plant origin. However, the lack of standardized and reliable testing methods for assessing antibacterial activity of plant natural products causes impediment to research into this area. This study demonstrated that the agar microdilution method can be used successfully for testing oily and coloured substances.</description><subject>Agar</subject><subject>agar microdilution method</subject><subject>Anti-Bacterial Agents - pharmacology</subject><subject>antibacterial activity</subject><subject>Antibiotics</subject><subject>Drug resistance</subject><subject>Drug Resistance, Bacterial</subject><subject>essential oil</subject><subject>Microbial Sensitivity Tests - methods</subject><subject>Microbiology</subject><subject>minimal inhibitory concentration</subject><subject>Oils & fats</subject><subject>Oils, Volatile - pharmacology</subject><subject>plant extract</subject><subject>Plant Extracts - pharmacology</subject><issn>1364-5072</issn><issn>1365-2672</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2016</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqN0ctKxDAUBuAgijOOLnwBCbjRRcdcmrazFPGK4kbXJW1PNUMvY5IyzkLwHXxDn8RjZ8aFIJhNQvjyc3IOIfucjTmuk6mux1wKJTfIkMtIBSKKxWZ_DgPFYjEgO85NGeOSqWibDESsGBdyMiRvD89A9ZO2tDa5bQtTdd60Da3BP7cF_Xz_oJo2MF9flK2luvGm15nRFXWdy2HmTWYq4xfUg_OmeeohOAdoEbWmcviuoLMKX1N49Vbn3u2SrVJXDvZW-4g8Xpw_nF0Ft_eX12ent0EeCi4DIRMFTEqW5UqFXIGOJoWexABMJzxmJU8KJsKMJVmClEse5YJzxbJQ6jKfyBE5WubObPvSYYVpbbDqCouBtnMpT2QsZSLD-D9UYfNUIpAe_qLTtrMNfuRbMYG9jhSq46XChjlnoUxn1tTaLlLO0u_xpTi-tB8f2oNVYpfVUPzI9bwQnCzB3FSw-DspvTm9W0Z-AQDDpE4</recordid><startdate>201611</startdate><enddate>201611</enddate><creator>Golus, J.</creator><creator>Sawicki, R.</creator><creator>Widelski, J.</creator><creator>Ginalska, G.</creator><general>Oxford University Press</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QL</scope><scope>7QO</scope><scope>7T7</scope><scope>7TM</scope><scope>7U7</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>M7N</scope><scope>P64</scope><scope>RC3</scope><scope>7X8</scope></search><sort><creationdate>201611</creationdate><title>The agar microdilution method – a new method for antimicrobial susceptibility testing for essential oils and plant extracts</title><author>Golus, J. ; Sawicki, R. ; Widelski, J. ; Ginalska, G.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c4213-2385e0330bc55415ea69da97ee0a8170f18d024b08b83851316c21150b43afc93</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2016</creationdate><topic>Agar</topic><topic>agar microdilution method</topic><topic>Anti-Bacterial Agents - pharmacology</topic><topic>antibacterial activity</topic><topic>Antibiotics</topic><topic>Drug resistance</topic><topic>Drug Resistance, Bacterial</topic><topic>essential oil</topic><topic>Microbial Sensitivity Tests - methods</topic><topic>Microbiology</topic><topic>minimal inhibitory concentration</topic><topic>Oils & fats</topic><topic>Oils, Volatile - pharmacology</topic><topic>plant extract</topic><topic>Plant Extracts - pharmacology</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Golus, J.</creatorcontrib><creatorcontrib>Sawicki, R.</creatorcontrib><creatorcontrib>Widelski, J.</creatorcontrib><creatorcontrib>Ginalska, G.</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Biotechnology Research Abstracts</collection><collection>Industrial and Applied Microbiology Abstracts (Microbiology A)</collection><collection>Nucleic Acids Abstracts</collection><collection>Toxicology Abstracts</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Engineering Research Database</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Journal of applied microbiology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Golus, J.</au><au>Sawicki, R.</au><au>Widelski, J.</au><au>Ginalska, G.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>The agar microdilution method – a new method for antimicrobial susceptibility testing for essential oils and plant extracts</atitle><jtitle>Journal of applied microbiology</jtitle><addtitle>J Appl Microbiol</addtitle><date>2016-11</date><risdate>2016</risdate><volume>121</volume><issue>5</issue><spage>1291</spage><epage>1299</epage><pages>1291-1299</pages><issn>1364-5072</issn><eissn>1365-2672</eissn><coden>JAMIFK</coden><abstract>Aims
To develop a new agar microdilution technique suitable for the assessment of the antimicrobial activity of natural plant products such as essential oils or plant extracts as well as to evaluate the antimicrobial effect of several essential oils and plant extracts.
Methods and Results
The proposed agar microdilution method was evolved on the basis of the CLSI agar dilution method, approved for conventional antimicrobials. However, this new method combines convenience and time/cost effectiveness typical for microtitre methods with the advantages of the agar dilution of hydrophobic or coloured substances. Different concentrations of the tested agents were added to Eppendorf tubes with molten Mueller‐Hinton agar, vortexed and dispensed into the 96‐well microplate in a small volume of 100 μl per well which allows for rapid, easy and economical preparation of samples as well as providing a uniform and stable dispersion without separation of the oil–water phases which occurs in methods with liquid medium. Next, the agar microdilution plates were inoculated with four reference bacterial strains. The results of our study demonstrated that the minimal inhibitory concentrations (MICs) were successfully determined using the agar microdilution method even with hydrophobic essential oils or strongly coloured plant extracts.
Conclusions
The new agar microdilution method avoids the problems associated with testing of water insoluble, oily or strongly coloured plant natural products. Moreover, it enables the reliable, cheap and easy MIC determination of such agents.
Significance and Impact of the Study
In the era of increasing antibiotic resistance high hopes are associated with new drugs of plant origin. However, the lack of standardized and reliable testing methods for assessing antibacterial activity of plant natural products causes impediment to research into this area. This study demonstrated that the agar microdilution method can be used successfully for testing oily and coloured substances.</abstract><cop>England</cop><pub>Oxford University Press</pub><pmid>27501239</pmid><doi>10.1111/jam.13253</doi><tpages>9</tpages><oa>free_for_read</oa></addata></record> |
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| ispartof | Journal of applied microbiology, 2016-11, Vol.121 (5), p.1291-1299 |
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| source | MEDLINE; Wiley Online Library Journals Frontfile Complete; Oxford University Press Journals All Titles (1996-Current) |
| subjects | Agar agar microdilution method Anti-Bacterial Agents - pharmacology antibacterial activity Antibiotics Drug resistance Drug Resistance, Bacterial essential oil Microbial Sensitivity Tests - methods Microbiology minimal inhibitory concentration Oils & fats Oils, Volatile - pharmacology plant extract Plant Extracts - pharmacology |
| title | The agar microdilution method – a new method for antimicrobial susceptibility testing for essential oils and plant extracts |
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