A new diatom growth inhibition assay using the XTT colorimetric method

Comp Biochem Physiol C Toxicol Pharmacol. 2016 Jul-Aug:185-186:13-19. doi: 10.1016/j.cbpc.2016.02.004. Epub 2016 Mar 3.

Abstract

Marine biofouling, which leads to significant operational stress and economic damage on marine infrastructures, is a major problem in marine related industries. Currently, the most common way to avoid marine biofouling involves the use of biocidal products in surface coatings. However, the need for environmentally friendly antibiofouling compounds has increased rapidly with the recent global prohibition of harmful antifoulants, such as tributyltin (TBT). In particular, periphytic diatoms have been shown to contribute significantly to biofilms, which play an important role in biofouling. Therefore, inhibiting the proliferation of fouling diatoms is a very important step in the prevention of marine biofouling. In this study, we developed a new, rapid, accurate, and convenient growth inhibition assay using the XTT colorimetric method to prevent the growth of the fouling periphytic diatom, Nitzschia amabilis Hidek. Suzuki (replaced synonym, Nitzschia laevis Hustedt). The feasibility of this method was verified by determining the growth inhibition activities of two standard photosynthetic inhibitors, DCMU and CuSO4. However, neither inhibitor had any cytotoxic activities at the range of concentrations tested. Moreover, this method was applied by screening and purification of herbicidic but non-cytotoxic compounds from cyanobacteria extracts. Our results demonstrate the utility of this newly established growth inhibition assay for the identification of marine anti-biofouling compounds.

Keywords: Biofouling; Cyanobacterium; Cytotoxicity; Growth inhibition; Nitzschia amabilis; Periphytic diatom; XTT.

Publication types

  • Evaluation Study

MeSH terms

  • Animals
  • Biofilms / drug effects*
  • Biofilms / growth & development
  • Biofouling / prevention & control*
  • Biological Assay / methods*
  • Cell Line, Tumor
  • Cell Proliferation / drug effects
  • Colorimetry*
  • Copper Sulfate / pharmacology*
  • Copper Sulfate / toxicity
  • Diatoms / drug effects*
  • Diatoms / growth & development
  • Disinfectants / pharmacology*
  • Disinfectants / toxicity
  • Diuron / pharmacology*
  • Diuron / toxicity
  • Dose-Response Relationship, Drug
  • Feasibility Studies
  • Indicators and Reagents / chemistry*
  • Mice
  • Tetrazolium Salts / chemistry
  • Time Factors

Substances

  • Disinfectants
  • Indicators and Reagents
  • Tetrazolium Salts
  • 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-5-((phenylamino)carbonyl)-2H-tetrazolium hydroxide
  • Diuron
  • Copper Sulfate