Optimization of an automatic counting system for the quantification of Staphylococcus epidermidis cells in biofilms
Autor(a) principal: | |
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Data de Publicação: | 2014 |
Outros Autores: | , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
Texto Completo: | http://hdl.handle.net/1822/32219 |
Resumo: | Biofilm formation is recognized as the main virulence factor in a variety of chronic infections. In vitro evaluation of biofilm formation is often achieved by quantification of viable or total cells. However, these methods depend on biofilm disruption, which is often achieved by vortexing or sonication. In this study, we investigated the effects of sonication on the elimination of Staphylococcus epidermidis cell clusters from biofilms grown over time, and quantification was performed by three distinct analytical techniques. Even when a higher number of sonication cycles was used, some stable cell clusters remained in the samples obtained from 48- and 72-h-old biofilms, interfering with the quantification of sessile bacteria by plate counting. On the other hand, the fluorescence microscopy automatic counting system allowed proper quantification of biofilm samples that had undergone any of the described sonication cycles, suggesting that this is a more accurate method for assessing the cell concentration in S. epidermidis biofilms, especially in mature biofilms. |
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Optimization of an automatic counting system for the quantification of Staphylococcus epidermidis cells in biofilmsBiofilmStaphylococcus epidermidisCell clustersSonicationAutomatic cell countingScience & TechnologyBiofilm formation is recognized as the main virulence factor in a variety of chronic infections. In vitro evaluation of biofilm formation is often achieved by quantification of viable or total cells. However, these methods depend on biofilm disruption, which is often achieved by vortexing or sonication. In this study, we investigated the effects of sonication on the elimination of Staphylococcus epidermidis cell clusters from biofilms grown over time, and quantification was performed by three distinct analytical techniques. Even when a higher number of sonication cycles was used, some stable cell clusters remained in the samples obtained from 48- and 72-h-old biofilms, interfering with the quantification of sessile bacteria by plate counting. On the other hand, the fluorescence microscopy automatic counting system allowed proper quantification of biofilm samples that had undergone any of the described sonication cycles, suggesting that this is a more accurate method for assessing the cell concentration in S. epidermidis biofilms, especially in mature biofilms.This work was funded by the Fundacao para a Ciencia e a Tecnologia (FCT) and the COMPETE grants PTDC/BIA-MIC/113450/2009 and FCOMP-01-0124-FEDER-014309.John Wiley and SonsUniversidade do MinhoFreitas, Ana IsabelVasconcelos, CarlosVilanova, ManuelCerca, Nuno20142014-01-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/1822/32219engFreitas, Ana Isabel; Vasconcelos, Carlos; Vilanova, Manuel; Cerca, Nuno, Optimization of an automatic counting system for the quantification of Staphylococcus epidermidis cells in biofilms. Journal of Basic Microbiology, 54(7), 750-757, 20140233-111X1521-402810.1002/jobm.20120060323686681info: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:RCAAP2023-07-21T12:47:53Zoai:repositorium.sdum.uminho.pt:1822/32219Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-19T19:46:01.326922Repositório 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 |
Optimization of an automatic counting system for the quantification of Staphylococcus epidermidis cells in biofilms |
title |
Optimization of an automatic counting system for the quantification of Staphylococcus epidermidis cells in biofilms |
spellingShingle |
Optimization of an automatic counting system for the quantification of Staphylococcus epidermidis cells in biofilms Freitas, Ana Isabel Biofilm Staphylococcus epidermidis Cell clusters Sonication Automatic cell counting Science & Technology |
title_short |
Optimization of an automatic counting system for the quantification of Staphylococcus epidermidis cells in biofilms |
title_full |
Optimization of an automatic counting system for the quantification of Staphylococcus epidermidis cells in biofilms |
title_fullStr |
Optimization of an automatic counting system for the quantification of Staphylococcus epidermidis cells in biofilms |
title_full_unstemmed |
Optimization of an automatic counting system for the quantification of Staphylococcus epidermidis cells in biofilms |
title_sort |
Optimization of an automatic counting system for the quantification of Staphylococcus epidermidis cells in biofilms |
author |
Freitas, Ana Isabel |
author_facet |
Freitas, Ana Isabel Vasconcelos, Carlos Vilanova, Manuel Cerca, Nuno |
author_role |
author |
author2 |
Vasconcelos, Carlos Vilanova, Manuel Cerca, Nuno |
author2_role |
author author author |
dc.contributor.none.fl_str_mv |
Universidade do Minho |
dc.contributor.author.fl_str_mv |
Freitas, Ana Isabel Vasconcelos, Carlos Vilanova, Manuel Cerca, Nuno |
dc.subject.por.fl_str_mv |
Biofilm Staphylococcus epidermidis Cell clusters Sonication Automatic cell counting Science & Technology |
topic |
Biofilm Staphylococcus epidermidis Cell clusters Sonication Automatic cell counting Science & Technology |
description |
Biofilm formation is recognized as the main virulence factor in a variety of chronic infections. In vitro evaluation of biofilm formation is often achieved by quantification of viable or total cells. However, these methods depend on biofilm disruption, which is often achieved by vortexing or sonication. In this study, we investigated the effects of sonication on the elimination of Staphylococcus epidermidis cell clusters from biofilms grown over time, and quantification was performed by three distinct analytical techniques. Even when a higher number of sonication cycles was used, some stable cell clusters remained in the samples obtained from 48- and 72-h-old biofilms, interfering with the quantification of sessile bacteria by plate counting. On the other hand, the fluorescence microscopy automatic counting system allowed proper quantification of biofilm samples that had undergone any of the described sonication cycles, suggesting that this is a more accurate method for assessing the cell concentration in S. epidermidis biofilms, especially in mature biofilms. |
publishDate |
2014 |
dc.date.none.fl_str_mv |
2014 2014-01-01T00:00:00Z |
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 |
http://hdl.handle.net/1822/32219 |
url |
http://hdl.handle.net/1822/32219 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
Freitas, Ana Isabel; Vasconcelos, Carlos; Vilanova, Manuel; Cerca, Nuno, Optimization of an automatic counting system for the quantification of Staphylococcus epidermidis cells in biofilms. Journal of Basic Microbiology, 54(7), 750-757, 2014 0233-111X 1521-4028 10.1002/jobm.201200603 23686681 |
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 |
John Wiley and Sons |
publisher.none.fl_str_mv |
John Wiley and Sons |
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 |
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Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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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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1799133026662219776 |