TY - JOUR
T1 - Polyhydroxyalkanoate granules quantification in mixed microbial cultures using image analysis
T2 - Sudan Black B versus Nile Blue A staining
AU - Mesquita, Daniela P.
AU - Luis Amaral, A.
AU - Leal, Cristiano
AU - Oehmen, Adrian
AU - Reis, Maria A. M.
AU - Ferreira, Eugenio C.
N1 - Sem PDF.
The authors thank the Fundacao para a Ciencia e a Tecnologia (FCT) Strategic Project PEst-OE/EQB/LA0023/2013 and the Project "BioEnv - Biotechnology and Bioengineering for a sustainable world", REF. NORTE-07-0124-FEDER-000048, co-funded by the Programa Operacional Regional do Norte (ON.2 - O Novo Norte), QREN, FEDER. The authors also acknowledge the financial support to Daniela P. Mesquita through the postdoctoral grant (SFRH/BPD/82558/2011) and the project PTDC/EBB-EBI/103147/2008 provided by FCT.
PY - 2015/3/20
Y1 - 2015/3/20
N2 - 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.
AB - 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.
KW - Image analysis
KW - Enhanced biological phosphorus removal (EBPR) systems
KW - Mixed microbial cultures (MMC)
KW - Polyhydroxyalkanoates (PHA)
KW - Sudan Black B (SBB)
KW - Nile Blue A (NBA)
KW - BIOLOGICAL PHOSPHORUS REMOVAL
KW - PARTIAL LEAST-SQUARES
KW - POLYPHOSPHATE ACCUMULATING ORGANISMS
KW - WATER TREATMENT-PLANT
KW - ACTIVATED-SLUDGE
KW - PHA PRODUCTION
KW - STORAGE POLYMERS
KW - MICROORGANISMS
KW - COMPETITION
KW - PROPIONATE
U2 - 10.1016/j.aca.2015.01.018
DO - 10.1016/j.aca.2015.01.018
M3 - Article
C2 - 25732579
SN - 0003-2670
VL - 865
SP - 8
EP - 15
JO - Analytica Chimica Acta
JF - Analytica Chimica Acta
ER -