Polyhydroxyalkanoate granules quantification in mixed microbial cultures using image analysis: Sudan Black B versus Nile Blue A staining

Daniela P. Mesquita, A. Luis Amaral, Cristiano Leal, Adrian Oehmen, Maria A. M. Reis, Eugenio C. Ferreira

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

Polyhydroxyalkanoates (PHA) can be produced and intracellularly accumulated as inclusions by mixed microbial cultures (MMC) for bioplastic production and in enhanced biological phosphorus removal (EBPR) systems. Classical methods for PHA quantification use a digestion step prior to chromatography analysis, rendering them labor intensive and time-consuming. The present work investigates the use of two quantitative image analysis (QIA) procedures specifically developed for PHA inclusions identification and quantification. MMC obtained from an EBPR system were visualized by bright-field and fluorescence microscopy for PHA inclusions detection, upon Sudan Black B (SBB) and Nile Blue A (NBA) staining, respectively. The captured color images were processed by QIA techniques and the image analysis data were further treated using multivariate statistical analysis. Partial least squares (PLS) regression coefficients of 0.90 and 0.86 were obtained between QIA parameters and PHA concentrations using SBB and NBA, respectively. It was found that both staining procedures might be seen as alternative methodologies to classical PHA determination. (C) 2015 Elsevier B.V. All rights reserved.

Original languageEnglish
Pages (from-to)8-15
Number of pages8
JournalAnalytica Chimica Acta
Volume865
DOIs
Publication statusPublished - 20 Mar 2015

Keywords

  • Image analysis
  • Enhanced biological phosphorus removal (EBPR) systems
  • Mixed microbial cultures (MMC)
  • Polyhydroxyalkanoates (PHA)
  • Sudan Black B (SBB)
  • Nile Blue A (NBA)
  • BIOLOGICAL PHOSPHORUS REMOVAL
  • PARTIAL LEAST-SQUARES
  • POLYPHOSPHATE ACCUMULATING ORGANISMS
  • WATER TREATMENT-PLANT
  • ACTIVATED-SLUDGE
  • PHA PRODUCTION
  • STORAGE POLYMERS
  • MICROORGANISMS
  • COMPETITION
  • PROPIONATE

Cite this