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

Detalhes bibliográficos
Autor(a) principal: Mesquita, D. P.
Data de Publicação: 2015
Outros Autores: Amaral, A. L., Leal, C., Oehmen, Adrian, Reis, Maria A. M., Ferreira, Eugénio C.
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
Texto Completo: https://hdl.handle.net/1822/34593
Resumo: 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.
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spelling Polyhydroxyalkanoate granules quantification in mixed microbial cultures using image analysis: Sudan Black B versus Nile Blue A stainingImage analysisEnhanced biological phosphorus removal (EBPR) systemsMixed microbial cultures (MMC)Polyhydroxyalkanoates (PHA)Sudan Black B (SBB)Nile Blue A (NBA)Science & TechnologyPolyhydroxyalkanoates (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.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.ElsevierUniversidade do MinhoMesquita, D. P.Amaral, A. L.Leal, C.Oehmen, AdrianReis, Maria A. M.Ferreira, Eugénio C.2015-03-202015-03-20T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttps://hdl.handle.net/1822/34593engMesquita, D. P.; Amaral, A. L.; Leal, C.; Oehmen, A.; Reis, M. A.; Ferreira, E. C., Polyhydroxyalkanoate granules quantification in mixed microbial cultures using image analysis: Sudan Black B versus Nile Blue A staining. Analytica Chimica Acta, 865, 8-15, 20150003-26701873-432410.1016/j.aca.2015.01.01825732579http://www.elsevier.com/locate/issn/00032670info:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2023-07-21T12:31:08Zoai:repositorium.sdum.uminho.pt:1822/34593Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-19T19:26:22.453465Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse
dc.title.none.fl_str_mv Polyhydroxyalkanoate granules quantification in mixed microbial cultures using image analysis: Sudan Black B versus Nile Blue A staining
title Polyhydroxyalkanoate granules quantification in mixed microbial cultures using image analysis: Sudan Black B versus Nile Blue A staining
spellingShingle Polyhydroxyalkanoate granules quantification in mixed microbial cultures using image analysis: Sudan Black B versus Nile Blue A staining
Mesquita, D. P.
Image analysis
Enhanced biological phosphorus removal (EBPR) systems
Mixed microbial cultures (MMC)
Polyhydroxyalkanoates (PHA)
Sudan Black B (SBB)
Nile Blue A (NBA)
Science & Technology
title_short Polyhydroxyalkanoate granules quantification in mixed microbial cultures using image analysis: Sudan Black B versus Nile Blue A staining
title_full Polyhydroxyalkanoate granules quantification in mixed microbial cultures using image analysis: Sudan Black B versus Nile Blue A staining
title_fullStr Polyhydroxyalkanoate granules quantification in mixed microbial cultures using image analysis: Sudan Black B versus Nile Blue A staining
title_full_unstemmed Polyhydroxyalkanoate granules quantification in mixed microbial cultures using image analysis: Sudan Black B versus Nile Blue A staining
title_sort Polyhydroxyalkanoate granules quantification in mixed microbial cultures using image analysis: Sudan Black B versus Nile Blue A staining
author Mesquita, D. P.
author_facet Mesquita, D. P.
Amaral, A. L.
Leal, C.
Oehmen, Adrian
Reis, Maria A. M.
Ferreira, Eugénio C.
author_role author
author2 Amaral, A. L.
Leal, C.
Oehmen, Adrian
Reis, Maria A. M.
Ferreira, Eugénio C.
author2_role author
author
author
author
author
dc.contributor.none.fl_str_mv Universidade do Minho
dc.contributor.author.fl_str_mv Mesquita, D. P.
Amaral, A. L.
Leal, C.
Oehmen, Adrian
Reis, Maria A. M.
Ferreira, Eugénio C.
dc.subject.por.fl_str_mv Image analysis
Enhanced biological phosphorus removal (EBPR) systems
Mixed microbial cultures (MMC)
Polyhydroxyalkanoates (PHA)
Sudan Black B (SBB)
Nile Blue A (NBA)
Science & Technology
topic Image analysis
Enhanced biological phosphorus removal (EBPR) systems
Mixed microbial cultures (MMC)
Polyhydroxyalkanoates (PHA)
Sudan Black B (SBB)
Nile Blue A (NBA)
Science & Technology
description 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.
publishDate 2015
dc.date.none.fl_str_mv 2015-03-20
2015-03-20T00:00:00Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
format article
status_str publishedVersion
dc.identifier.uri.fl_str_mv https://hdl.handle.net/1822/34593
url https://hdl.handle.net/1822/34593
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv Mesquita, D. P.; Amaral, A. L.; Leal, C.; Oehmen, A.; Reis, M. A.; Ferreira, E. C., Polyhydroxyalkanoate granules quantification in mixed microbial cultures using image analysis: Sudan Black B versus Nile Blue A staining. Analytica Chimica Acta, 865, 8-15, 2015
0003-2670
1873-4324
10.1016/j.aca.2015.01.018
25732579
http://www.elsevier.com/locate/issn/00032670
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Elsevier
publisher.none.fl_str_mv Elsevier
dc.source.none.fl_str_mv reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
instacron:RCAAP
instname_str Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
instacron_str RCAAP
institution RCAAP
reponame_str Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
collection Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
repository.name.fl_str_mv Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
repository.mail.fl_str_mv
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