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

Analytica Chimica Acta
Daniela P MesquitaEugénio C Ferreira

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.

References

Jul 23, 1999·International Journal of Biological Macromolecules·R C Fuller
Jun 19, 2002·Journal of Microbiological Methods·Luísa S SerafimMaria A M Reis
Sep 19, 2003·Bioprocess and Biosystems Engineering·M A M ReisM C M Van Loosdrecht
Apr 17, 2007·Water Research·Adrian OehmenMaria A M Reis
Aug 25, 2009·Colloids and Surfaces. B, Biointerfaces·Sureshbabu Ram Kumar PandianSangiliyandi Gurunathan

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Citations

Mar 10, 2016·International Journal of Systematic and Evolutionary Microbiology·Yue SuMin Wu
Feb 22, 2017·Environmental Microbiology·Timothy J WilliamsRicardo Cavicchioli
Dec 3, 2016·International Journal of Systematic and Evolutionary Microbiology·Yue SuMin Wu
Mar 3, 2020·International Journal of Systematic and Evolutionary Microbiology·Lu LuFang Peng
May 14, 2017·Environmental Science and Pollution Research International·António L AmaralEugénio C Ferreira

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