Evaluation of detachment methods for the enumeration ofBacteroides fragilis in sediments via propidium monoazide quantitative PCR, in comparison with Enterococcus faecalis and Esch...

Autores
Kim, M.; Gutiérrez Cacciabue, Dolores; Schriewer, Alex; Rajal, Verónica Beatriz; Wuertz, Stefan
Año de publicación
2014
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Aims The aim was to develop an optimized detachment method for separating Bacteroidales from sediments to allow enumeration via PMA-qPCR. The effectiveness of four different detachment treatments in removing Bacteroides fragilis was compared as a function of time as well as in relation to Enterococcus faecalis and Escherichia coli as detected by cultivation and qPCR. Methods and Results Cells were inoculated into four sediments from sea water (SW) and freshwater (FW) beaches. Sediment samples were taken on days 1 and 7 and subjected to four different treatments for separation of micro-organisms. On day 1, the detachment treatments performed equally well in removing intact Bact. fragilis cells. In contrast, 7 days later the detachment treatment with Tween 80 and handshaking (TH) resulted in up to eightfold higher 16S rRNA gene concentrations of intact and total Bact. fragilis cells compared to other detachment treatments. Total Ent. faecalis cells based on the 23S rRNA gene were also preferentially recovered by treatment TH. Cultivable Ent. faecalis or E. coli numbers detached from sediments were similar for all methods in most sediments tested. Conclusions Handshaking and 1% Tween 80/NaOH (pH 7·0) eluant was the most efficient technique to recover intact as well as total Bact. fragilis cells in sediment samples with different salinities and after prolonged sediment cell contact time. Significance and Impact of the Study The optimized detachment method enables the application of PMA-qPCR to sediment samples to detect the presence of Bacteroidales cells and their DNA in future microbial source tracking studies.
Fil: Kim, M.. University of California. Department of Civil and Environmental Engineering; Estados Unidos
Fil: Gutiérrez Cacciabue, Dolores. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Salta. Instituto de Investigación Para la Industria Química (i); Argentina
Fil: Schriewer, Alex. University of California. Department of Civil and Environmental Engineering; Estados Unidos
Fil: Rajal, Verónica Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Salta. Instituto de Investigación Para la Industria Química (i); Argentina
Fil: Wuertz, Stefan. University of California. Department of Civil and Environmental Engineering; Estados Unidos
Materia
Bacteroidales
Enterococcus
Qpcr-Pma
Sediments
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/4744
Acceder
id CONICETDig_97c93dc107309d2d921d88389780f950
oai_identifier_str oai:ri.conicet.gov.ar:11336/4744
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Evaluation of detachment methods for the enumeration ofBacteroides fragilis in sediments via propidium monoazide quantitative PCR, in comparison with Enterococcus faecalis and Escherichia coliKim, M.Gutiérrez Cacciabue, DoloresSchriewer, AlexRajal, Verónica BeatrizWuertz, StefanBacteroidalesEnterococcusQpcr-PmaSedimentshttps://purl.org/becyt/ford/2.8https://purl.org/becyt/ford/2Aims The aim was to develop an optimized detachment method for separating Bacteroidales from sediments to allow enumeration via PMA-qPCR. The effectiveness of four different detachment treatments in removing Bacteroides fragilis was compared as a function of time as well as in relation to Enterococcus faecalis and Escherichia coli as detected by cultivation and qPCR. Methods and Results Cells were inoculated into four sediments from sea water (SW) and freshwater (FW) beaches. Sediment samples were taken on days 1 and 7 and subjected to four different treatments for separation of micro-organisms. On day 1, the detachment treatments performed equally well in removing intact Bact. fragilis cells. In contrast, 7 days later the detachment treatment with Tween 80 and handshaking (TH) resulted in up to eightfold higher 16S rRNA gene concentrations of intact and total Bact. fragilis cells compared to other detachment treatments. Total Ent. faecalis cells based on the 23S rRNA gene were also preferentially recovered by treatment TH. Cultivable Ent. faecalis or E. coli numbers detached from sediments were similar for all methods in most sediments tested. Conclusions Handshaking and 1% Tween 80/NaOH (pH 7·0) eluant was the most efficient technique to recover intact as well as total Bact. fragilis cells in sediment samples with different salinities and after prolonged sediment cell contact time. Significance and Impact of the Study The optimized detachment method enables the application of PMA-qPCR to sediment samples to detect the presence of Bacteroidales cells and their DNA in future microbial source tracking studies.Fil: Kim, M.. University of California. Department of Civil and Environmental Engineering; Estados UnidosFil: Gutiérrez Cacciabue, Dolores. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Salta. Instituto de Investigación Para la Industria Química (i); ArgentinaFil: Schriewer, Alex. University of California. Department of Civil and Environmental Engineering; Estados UnidosFil: Rajal, Verónica Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Salta. Instituto de Investigación Para la Industria Química (i); ArgentinaFil: Wuertz, Stefan. University of California. Department of Civil and Environmental Engineering; Estados UnidosWiley2014-08info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/4744Kim, M.; Gutiérrez Cacciabue, Dolores; Schriewer, Alex; Rajal, Verónica Beatriz; Wuertz, Stefan; Evaluation of detachment methods for the enumeration ofBacteroides fragilis in sediments via propidium monoazide quantitative PCR, in comparison with Enterococcus faecalis and Escherichia coli; Wiley; Journal of Applied Microbiology; 117; 5; 8-2014; 1513-15221364-5072enginfo:eu-repo/semantics/altIdentifier/url/http://onlinelibrary.wiley.com/doi/10.1111/jam.12630/abstractinfo:eu-repo/semantics/altIdentifier/doi/10.1111/jam.12630info:eu-repo/semantics/altIdentifier/issn/1364-5072info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T09:37:59Zoai:ri.conicet.gov.ar:11336/4744instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-09-29 09:38:00.114CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Evaluation of detachment methods for the enumeration ofBacteroides fragilis in sediments via propidium monoazide quantitative PCR, in comparison with Enterococcus faecalis and Escherichia coli
title Evaluation of detachment methods for the enumeration ofBacteroides fragilis in sediments via propidium monoazide quantitative PCR, in comparison with Enterococcus faecalis and Escherichia coli
spellingShingle Evaluation of detachment methods for the enumeration ofBacteroides fragilis in sediments via propidium monoazide quantitative PCR, in comparison with Enterococcus faecalis and Escherichia coli
Kim, M.
Bacteroidales
Enterococcus
Qpcr-Pma
Sediments
title_short Evaluation of detachment methods for the enumeration ofBacteroides fragilis in sediments via propidium monoazide quantitative PCR, in comparison with Enterococcus faecalis and Escherichia coli
title_full Evaluation of detachment methods for the enumeration ofBacteroides fragilis in sediments via propidium monoazide quantitative PCR, in comparison with Enterococcus faecalis and Escherichia coli
title_fullStr Evaluation of detachment methods for the enumeration ofBacteroides fragilis in sediments via propidium monoazide quantitative PCR, in comparison with Enterococcus faecalis and Escherichia coli
title_full_unstemmed Evaluation of detachment methods for the enumeration ofBacteroides fragilis in sediments via propidium monoazide quantitative PCR, in comparison with Enterococcus faecalis and Escherichia coli
title_sort Evaluation of detachment methods for the enumeration ofBacteroides fragilis in sediments via propidium monoazide quantitative PCR, in comparison with Enterococcus faecalis and Escherichia coli
dc.creator.none.fl_str_mv Kim, M.
Gutiérrez Cacciabue, Dolores
Schriewer, Alex
Rajal, Verónica Beatriz
Wuertz, Stefan
author Kim, M.
author_facet Kim, M.
Gutiérrez Cacciabue, Dolores
Schriewer, Alex
Rajal, Verónica Beatriz
Wuertz, Stefan
author_role author
author2 Gutiérrez Cacciabue, Dolores
Schriewer, Alex
Rajal, Verónica Beatriz
Wuertz, Stefan
author2_role author
author
author
author
dc.subject.none.fl_str_mv Bacteroidales
Enterococcus
Qpcr-Pma
Sediments
topic Bacteroidales
Enterococcus
Qpcr-Pma
Sediments
purl_subject.fl_str_mv https://purl.org/becyt/ford/2.8
https://purl.org/becyt/ford/2
dc.description.none.fl_txt_mv Aims The aim was to develop an optimized detachment method for separating Bacteroidales from sediments to allow enumeration via PMA-qPCR. The effectiveness of four different detachment treatments in removing Bacteroides fragilis was compared as a function of time as well as in relation to Enterococcus faecalis and Escherichia coli as detected by cultivation and qPCR. Methods and Results Cells were inoculated into four sediments from sea water (SW) and freshwater (FW) beaches. Sediment samples were taken on days 1 and 7 and subjected to four different treatments for separation of micro-organisms. On day 1, the detachment treatments performed equally well in removing intact Bact. fragilis cells. In contrast, 7 days later the detachment treatment with Tween 80 and handshaking (TH) resulted in up to eightfold higher 16S rRNA gene concentrations of intact and total Bact. fragilis cells compared to other detachment treatments. Total Ent. faecalis cells based on the 23S rRNA gene were also preferentially recovered by treatment TH. Cultivable Ent. faecalis or E. coli numbers detached from sediments were similar for all methods in most sediments tested. Conclusions Handshaking and 1% Tween 80/NaOH (pH 7·0) eluant was the most efficient technique to recover intact as well as total Bact. fragilis cells in sediment samples with different salinities and after prolonged sediment cell contact time. Significance and Impact of the Study The optimized detachment method enables the application of PMA-qPCR to sediment samples to detect the presence of Bacteroidales cells and their DNA in future microbial source tracking studies.
Fil: Kim, M.. University of California. Department of Civil and Environmental Engineering; Estados Unidos
Fil: Gutiérrez Cacciabue, Dolores. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Salta. Instituto de Investigación Para la Industria Química (i); Argentina
Fil: Schriewer, Alex. University of California. Department of Civil and Environmental Engineering; Estados Unidos
Fil: Rajal, Verónica Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Salta. Instituto de Investigación Para la Industria Química (i); Argentina
Fil: Wuertz, Stefan. University of California. Department of Civil and Environmental Engineering; Estados Unidos
description Aims The aim was to develop an optimized detachment method for separating Bacteroidales from sediments to allow enumeration via PMA-qPCR. The effectiveness of four different detachment treatments in removing Bacteroides fragilis was compared as a function of time as well as in relation to Enterococcus faecalis and Escherichia coli as detected by cultivation and qPCR. Methods and Results Cells were inoculated into four sediments from sea water (SW) and freshwater (FW) beaches. Sediment samples were taken on days 1 and 7 and subjected to four different treatments for separation of micro-organisms. On day 1, the detachment treatments performed equally well in removing intact Bact. fragilis cells. In contrast, 7 days later the detachment treatment with Tween 80 and handshaking (TH) resulted in up to eightfold higher 16S rRNA gene concentrations of intact and total Bact. fragilis cells compared to other detachment treatments. Total Ent. faecalis cells based on the 23S rRNA gene were also preferentially recovered by treatment TH. Cultivable Ent. faecalis or E. coli numbers detached from sediments were similar for all methods in most sediments tested. Conclusions Handshaking and 1% Tween 80/NaOH (pH 7·0) eluant was the most efficient technique to recover intact as well as total Bact. fragilis cells in sediment samples with different salinities and after prolonged sediment cell contact time. Significance and Impact of the Study The optimized detachment method enables the application of PMA-qPCR to sediment samples to detect the presence of Bacteroidales cells and their DNA in future microbial source tracking studies.
publishDate 2014
dc.date.none.fl_str_mv 2014-08
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/4744
Kim, M.; Gutiérrez Cacciabue, Dolores; Schriewer, Alex; Rajal, Verónica Beatriz; Wuertz, Stefan; Evaluation of detachment methods for the enumeration ofBacteroides fragilis in sediments via propidium monoazide quantitative PCR, in comparison with Enterococcus faecalis and Escherichia coli; Wiley; Journal of Applied Microbiology; 117; 5; 8-2014; 1513-1522
1364-5072
url http://hdl.handle.net/11336/4744
identifier_str_mv Kim, M.; Gutiérrez Cacciabue, Dolores; Schriewer, Alex; Rajal, Verónica Beatriz; Wuertz, Stefan; Evaluation of detachment methods for the enumeration ofBacteroides fragilis in sediments via propidium monoazide quantitative PCR, in comparison with Enterococcus faecalis and Escherichia coli; Wiley; Journal of Applied Microbiology; 117; 5; 8-2014; 1513-1522
1364-5072
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/http://onlinelibrary.wiley.com/doi/10.1111/jam.12630/abstract
info:eu-repo/semantics/altIdentifier/doi/10.1111/jam.12630
info:eu-repo/semantics/altIdentifier/issn/1364-5072
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
dc.publisher.none.fl_str_mv Wiley
publisher.none.fl_str_mv Wiley
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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score 13.070432

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