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https://hdl.handle.net/1822/25955
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Campo DC | Valor | Idioma |
---|---|---|
dc.contributor.author | Cerca, Filipe | - |
dc.contributor.author | Trigo, Gabriela | - |
dc.contributor.author | Correia, Alexandra | - |
dc.contributor.author | Cerca, Nuno | - |
dc.contributor.author | Azeredo, Joana | - |
dc.contributor.author | Vilanova, Manuel | - |
dc.date.accessioned | 2013-11-04T16:06:37Z | - |
dc.date.available | 2013-11-04T16:06:37Z | - |
dc.date.issued | 2011 | - |
dc.identifier.issn | 0008-4166 | por |
dc.identifier.issn | 1480-3275 | por |
dc.identifier.uri | https://hdl.handle.net/1822/25955 | - |
dc.description.abstract | Staphylococcus epidermidis biofilms with different proportions of viable but nonculturable bacteria were used to show that SYBR green (SYBR) may be used as a probe to evaluate the bacterial physiological state using flow cytometry. Biofilms grown in excess glucose presented significantly higher proportions of dormant bacteria than biofilms grown in excess glucose with buffered pH conditions or with exponential-phase planktonic cultures. Bacteria obtained from biofilms with high or low proportions of viable but nonculturable cells were further cultured in broth medium and stained with SYBR at different time points. An association between bacterial growth and SYBR staining intensity was observed. In addition, bacteria presenting higher SYBR fluorescence intensity also stained more intensely with cyanoditolyl tetrazolium chloride, used as a probe to evaluate cellular metabolism. Accordingly, planktonic bacteria treated with rifampicin, an inhibitor of bacterial RNA transcription, presented lower SYBR and cyanoditolyl tetrazolium chloride staining intensity than nontreated bacteria. Overall, our results indicate that SYBR, in addition to being used as a component of LIVE/DEAD stain, may also be used as a probe to evaluate the physiological state of S. epidermidis cells. | por |
dc.description.abstract | Des biofilms formés de Staphylococcus epidermidis contenant différentes proportions de bactéries viables mais non cultivables (BVNC) ont été utilisés ici pour montrer que le SYBR green (SYBR) peut être employé comme sonde pour évaluer l’état physiologique bactérien par cytométrie en flux. Les biofilms cultivés en présence d’un excès de glucose présentaient des proportions significativement plus élevées de bactéries dormantes comparativement aux biofilms cultivés en présence d’un excès de glucose dans des conditions de pH tamponné, ou aux cultures planctoniques en phase exponentielle. Les bactéries obtenues des biofilms contenant des proportions élevées ou faibles de BVNC ont été cultivées dans un bouillon de culture et colorées au SYBR après différentes périodes de temps. Une association entre la croissance bactérienne et l’intensité de coloration au SYBR a été observée. De plus, les bactéries présentant une intensité de fluorescence au SYBR plus élevée étaient également colorées de façon plus intense au chlorure de cyanoditolyl tétrazolium (CTC), utilisé comme sonde pour évaluer le métabolisme cellulaire. Conséquemment, les bactéries planctoniques traitées à la rifampicine, un inhibiteur de la transcription bactérienne, présentaient une intensité de coloration au SYBR et au CTC plus faible que les bactéries non traitées. L’ensemble de nos résultats indique que le SYBR, en plus d’être utilisé comme composante du colorant vital LIVE/DEAD, peut aussi être employé comme sonde pour évaluer l’état physiologique de S. epidermidis. | por |
dc.description.sponsorship | The authors acknowledge the technical assistance of Encarnacao Rebelo. The work was funded by Fundacao para a Ciencia e a Tecnologia (FCT) and COMPETE grants PTDC/BIA-MIC/113450/2009 and FCOMP-01-0124-FEDER-014309. Filipe Cerca and Alexandra Correia were funded by FCT fellowship SFRH/BD/27638/2006 and SFRH/BD/31354/2006. | por |
dc.language.iso | eng | por |
dc.publisher | NRC Research Press (Canadian Science Publishing) | por |
dc.rights | restrictedAccess | por |
dc.subject | Staphylococcus epidermidis | por |
dc.subject | Biofilm | por |
dc.subject | Flow cytometry | por |
dc.subject | Viable but nonculturable bacteria | por |
dc.subject | VBNC | por |
dc.subject | SYBR green | por |
dc.subject | Cyanoditolyl tetrazolium chloride | por |
dc.subject | CTC | por |
dc.subject | LIVE/DEAD | por |
dc.subject | Cytométrie en flux | por |
dc.subject | Bactéries viables mais non cultivables | por |
dc.subject | BVNC | por |
dc.subject | Chlorure de cyanoditolyl tétrazolium | por |
dc.subject | Colorant vital LIVE/DEAD | por |
dc.title | SYBR green as a fluorescent probe to evaluate the biofilm physiological state of Staphylococcus epidermidis, using flow cytometry | por |
dc.type | article | eng |
dc.peerreviewed | yes | por |
sdum.publicationstatus | published | por |
oaire.citationStartPage | 850 | por |
oaire.citationEndPage | 856 | por |
oaire.citationIssue | 10 | por |
oaire.citationTitle | Canadian journal of microbiology | por |
oaire.citationVolume | 57 | por |
dc.identifier.doi | 10.1139/W11-078 | por |
dc.identifier.pmid | 21950962 | por |
dc.subject.wos | Science & Technology | por |
sdum.journal | Canadian journal of microbiology | por |
Aparece nas coleções: | CEB - Publicações em Revistas/Séries Internacionais / Publications in International Journals/Series |
Ficheiros deste registo:
Ficheiro | Descrição | Tamanho | Formato | |
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CJM(2011) FilipeCerca SYBR green as probe.pdf Acesso restrito! | 581,25 kB | Adobe PDF | Ver/Abrir |