Use of MTT assay for determination of the biofilm formation capacity of microorganisms in metalworking fluids

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• 作者：El?bieta Anna Trafny (1)
  Rafa? Lewandowski (1)
  Irena Zawistowska-Marciniak (1)
  Ma?gorzata St?pińska (1)
  
• 关键词：MTT assay ; Metalworking fluids ; Biofilm ; Biocides
• 刊名：World Journal of Microbiology and Biotechnology
• 出版年：2013
• 出版时间：September 2013
• 年：2013
• 卷：29
• 期：9
• 页码：1635-1643
• 全文大小：381KB
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• 作者单位：El?bieta Anna Trafny (1)
  Rafa? Lewandowski (1)
  Irena Zawistowska-Marciniak (1)
  Ma?gorzata St?pińska (1)
  
  1. Department of Microbiology, Military Institute of Hygiene and Epidemiology, Kozielska 4, 01-163, Warsaw, Poland
  

文摘

Biofilm formation is a well-known problem in management of metalworking fluid systems. Due to persistence of microorganisms within biofilms, the reappearance of various species of bacteria, including nontuberculous mycobacteria is often observed after the use of biocides and/or cleaning of delivery systems and replacement of cooling fluid. The aim of this study was to determine the usefulness of the tetrazolium salt assay (MTT assay) for assessing the viability of bacteria in biofilms formed in vitro in fresh and used cutting oils, as well as their susceptibility to antimicrobial biocides. Biofilms were established in the microtiter plate format. The results showed that quantification of formazan, a product of the tetrazolium salt reduction by electron transport system could be used for determination of the propensity of bacteria to form biofilms in these complex media. The use of the assay allows also determination of antimicrobial activity of biocides against biofilms in fresh and used metalworking fluids. Biofilms produced by Gram-negative isolates recovered from field metalworking fluids as well as the wild bacterial communities differed in metabolic activity depending on the type of fresh coolants. The MTT assay has high-throughput potential and can be efficiently used for determination of biofilm-forming capacity of microorganisms from individual machines in metalworking industry. The use of the assay may also guide the selection of the most appropriate biocide to fight these microorganisms.