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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

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          Abstract

          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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          Development and application of a resazurin-based biomass activity test for activated sludge plant management.

          Janet W. McGrath, B. McNicholl, Virginia Quinn (2006)
          A rapid, robust and cost-effective method of assaying the metabolic activity of the biomass of activated sludge plants would be a valuable process control tool in the wastewater treatment industry. We have developed and optimised a simple colorimetric test protocol, based on the redox dye resazurin, in which levels of reduction of the dye are proportional to cell biomass and respiration rate in both freshly sampled municipal sludges and in a surrogate activated sludge culture, Polytox. The method has been used to assess the impact of trade wastes on the activities of two municipal activated sludge populations of differing characteristics.
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            Screening seeds of Scottish plants for antibacterial activity.

            Yashodharan Kumarasamy, Philip Cox, Marcel Jaspars … (2002)
            Based on ethnopharmacological and taxonomic information, seeds of 21 Scottish plant species from 14 different families were obtained from authentic seed suppliers. Their n-hexane, dichloromethane and methanol extracts were assessed for antibacterial activity against 11 pathogenic bacterial species. Methanol extracts of 11 plant species showed significant antibacterial activity. Malva moschata and Prunus padus were active against five bacterial species, Reseda lutea against four, Centaurium erythraea and Crithmum maritimum against three, Calluna vulgaris against two, and Armeria maritima, Centaurea scabiosa, Daucus carota, Rosa canina and Stellaria holostea against one bacterial species. C. erythraea and P. padus were also active against methicillin resistant Staphylococcus aureus.
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              Comparison of cation-adjusted Mueller-Hinton broth with Iso-Sensitest broth for the NCCLS broth microdilution method.

              David I. Phillips, Mike H. Knight, Laura J. May-Collado … (2000)
              Comparison of MIC results obtained in different parts of the world is currently difficult because of variations in methods. In this study, cation-adjusted Mueller-Hinton broth, the NCCLS-recommended medium, was compared with Iso-Sensitest broth, which is widely used in Europe. Microbroth dilution testing, using the NCCLS procedure, was performed on 124 Gram-positive (staphylococci and enterococci) and Gram-negative (Enterobacteriaceae and Pseudomonas aeruginosa) isolates from the CDC reference set, with the only variable being the medium used. Twelve antimicrobial agents were tested: amoxycillin-clavulanic acid, ampicillin, ciprofloxacin, erythromycin, gentamicin, imipenem, levofloxacin, oxacillin, gemifloxacin, trimethoprim- sulphamethoxazole, tetracycline and vancomycin. Vancomycin, erythromycin and oxacillin were only evaluated for the Gram-positive organisms. Trimethoprim-sulphamethoxazole was only evaluated for a subset of Gram-negative organisms because of off-scale results. The 124 isolates were tested in one American and one UK laboratory with two batches of cation-adjusted Mueller-Hinton broth and two of Iso-Sensitest broth. A statistical evaluation of the data used a 24 fully specified factorial analysis to determine if there were significant differences in results owing to Gram reaction, site of testing and type and/or batch of broth. In addition, the cumulative results for each antimicrobial agent in each broth were plotted against the range of MIC dilutions tested. MICs of ciprofloxacin, levofloxacin, gemifloxacin, gentamicin and tetracycline were slightly higher (half a doubling dilution) with Iso-Sensitest broth than with Mueller-Hinton broth. MIC results for the other antimicrobial agents were equivalent. Essential and category agreement rates were comparable for all agents (88.4-100% and 88.2-99.0%, respectively).
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                Author and article information

                Journal
                Journal ID (nlm-ta): Methods
                Title: Methods (San Diego, Calif.)
                Publisher: Academic Press
                ISSN (Print): 1046-2023
                ISSN (Electronic): 1095-9130
                Publication date PMC-release: August 2007
                Publication date (Print): August 2007
                Volume: 42
                Issue: 4
                Pages: 321-324
                Affiliations
                [a ]School of Biomedical Sciences, University of Ulster at Coleraine, Cromore Road, Coleraine BT52 1SA, Co. Londonderry, Northern Ireland, UK
                [b ]Department of Chemistry, University of Aberdeen, Aberdeen AB24 3UE, Scotland, UK
                [c ]School of Pharmacy, The Robert Gordon University, Schoolhill, Aberdeen AB10 1FR, Scotland, UK
                Author notes
                [* ]Corresponding author. Fax: +44 28 7032 4965. s.sarker@ 123456ulster.ac.uk
                Article
                Publisher ID: YMETH2134
                DOI: 10.1016/j.ymeth.2007.01.006
                PMC ID: 1895922
                PubMed ID: 17560319
                SO-VID: 8d7e9201-1c38-47e7-934c-a4ef0217e84e
                Copyright © .
                License:

                This document may be redistributed and reused, subject to certain conditions.

                History
                Date accepted : 12 January 2007
                Categories
                Subject: Article

                ScienceOpen disciplines: Life sciences
                Keywords: resazurin,antibacterial assay,plant extract,serial dilution
                Data availability:
                ScienceOpen disciplines: Life sciences
                Keywords: resazurin, antibacterial assay, plant extract, serial dilution

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