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      Resazurin-based 96-well plate microdilution method for the determination of minimum inhibitory concentration of biosurfactants

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          Abstract

          Objectives

          To develop and validate a microdilution method for measuring the minimum inhibitory concentration (MIC) of biosurfactants.

          Results

          A standardized microdilution method including resazurin dye has been developed for measuring the MIC of biosurfactants and its validity was established through the replication of tetracycline and gentamicin MIC determination with standard bacterial strains.

          Conclusion

          This new method allows the generation of accurate MIC measurements, whilst overcoming critical issues related to colour and solubility which may interfere with growth measurements for many types of biosurfactant extracts.

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          Most cited references11

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          Antimicrobial susceptibility testing: a review of general principles and contemporary practices.

          James H. Jorgensen, Mary Jane Ferraro (2009)
          An important task of the clinical microbiology laboratory is the performance of antimicrobial susceptibility testing of significant bacterial isolates. The goals of testing are to detect possible drug resistance in common pathogens and to assure susceptibility to drugs of choice for particular infections. The most widely used testing methods include broth microdilution or rapid automated instrument methods that use commercially marketed materials and devices. Manual methods that provide flexibility and possible cost savings include the disk diffusion and gradient diffusion methods. Each method has strengths and weaknesses, including organisms that may be accurately tested by the method. Some methods provide quantitative results (eg, minimum inhibitory concentration), and all provide qualitative assessments using the categories susceptible, intermediate, or resistant. In general, current testing methods provide accurate detection of common antimicrobial resistance mechanisms. However, newer or emerging mechanisms of resistance require constant vigilance regarding the ability of each test method to accurately detect resistance.
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            Investigation of the Alamar Blue (resazurin) fluorescent dye for the assessment of mammalian cell cytotoxicity.

            I Wilson, T Orton, F Pognan … (2000)
            We show here the identity of Alamar Blue as resazurin. The 'resazurin reduction test' has been used for about 50 years to monitor bacterial and yeast contamination of milk, and also for assessing semen quality. Resazurin (blue and nonfluorescent) is reduced to resorufin (pink and highly fluorescent) which is further reduced to hydroresorufin (uncoloured and nonfluorescent). It is still not known how this reduction occurs, intracellularly via enzyme activity or in the medium as a chemical reaction, although the reduced fluorescent form of Alamar Blue was found in the cytoplasm and of living cells nucleus of dead cells. Recently, the dye has gained popularity as a very simple and versatile way of measuring cell proliferation and cytotoxicity. This dye presents numerous advantages over other cytotoxicity or proliferation tests but we observed several drawbacks to the routine use of Alamar Blue. Tests with several toxicants in different cell lines and rat primary hepatocytes have shown accumulation of the fluorescent product of Alamar Blue in the medium which could lead to an overestimation of cell population. Also, the extensive reduction of Alamar Blue by metabolically active cells led to a final nonfluorescent product, and hence an underestimation of cellular activity.
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              Biosurfactants: potential applications in medicine.

              José Teixeira, Ibrahim M. Banat, Ligia Rodrigues … (2006)
              The use and potential commercial application of biosurfactants in the medical field has increased during the past decade. Their antibacterial, antifungal and antiviral activities make them relevant molecules for applications in combating many diseases and as therapeutic agents. In addition, their role as anti-adhesive agents against several pathogens indicates their utility as suitable anti-adhesive coating agents for medical insertional materials leading to a reduction in a large number of hospital infections without the use of synthetic drugs and chemicals. This review looks at medicinal and therapeutic perspectives on biosurfactant applications.
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                Author and article information

                Contributors
                Ibrahim M. Banat:
                ORCID: http://orcid.org/0000-0001-8964-2840
                im.banat@ulster.ac.uk , http://www.ulster.ac.uk
                Journal
                Journal ID (nlm-ta): Biotechnol Lett
                Journal ID (iso-abbrev): Biotechnol. Lett
                Title: Biotechnology Letters
                Publisher: Springer Netherlands (Dordrecht )
                ISSN (Print): 0141-5492
                ISSN (Electronic): 1573-6776
                Publication date (Electronic): 11 March 2016
                Publication date PMC-release: 11 March 2016
                Publication date (Print): 2016
                Volume: 38
                Pages: 1015-1019
                Affiliations
                School of Biomedical Sciences, Pharmaceutical Science and Practice Research Group, Ulster University, Cromore Road, Coleraine, County Londonderry BT52 1SA Northern Ireland, UK
                Author information
                http://orcid.org/0000-0001-8964-2840
                Article
                Publisher ID: 2079
                DOI: 10.1007/s10529-016-2079-2
                PMC ID: 4853446
                PubMed ID: 26969604
                SO-VID: 47ac301c-2627-4248-9a01-7a242e1badb1
                Copyright © © The Author(s) 2016
                License:

                Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License ( http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.

                History
                Date received : 15 January 2016
                Date accepted : 25 February 2016
                Funding
                Funded by: EU FP7
                Award ID: project 289219
                Award Recipient :
                Categories
                Subject: Original Research Paper
                Custom metadata
                issue-copyright-statement © Springer Science+Business Media Dordrecht 2016

                ScienceOpen disciplines: Biotechnology
                Keywords: antibiotics,biosurfactant,minimum inhibitory concentration,resazurin
                Data availability:
                ScienceOpen disciplines: Biotechnology
                Keywords: antibiotics, biosurfactant, minimum inhibitory concentration, resazurin

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