Evaluation of Escherichia coli viability by flow cytometry: A method for determining bacterial responses to antibiotic exposure
Background In this study, we check for the presence of specific resistance genes by polymerase chain reaction (PCR) and then we used flow cytometry (FCM) to evaluate antibiotic‐induced effects in different strains of Escherichia coli (E. coli). Methods The presence of resistance genes was investigat...
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| Veröffentlicht in: | Cytometry. Part B, Clinical cytometry Clinical cytometry, 2015-05, Vol.88 (3), p.149-153 |
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| creator | Boi, Paola Manti, Anita Pianetti, Anna Sabatini, Luigia Sisti, Davide Rocchi, Marco Bruno Bruscolini, Francesca Galluzzi, Luca Papa, Stefano |
| description | Background
In this study, we check for the presence of specific resistance genes by polymerase chain reaction (PCR) and then we used flow cytometry (FCM) to evaluate antibiotic‐induced effects in different strains of Escherichia coli (E. coli).
Methods
The presence of resistance genes was investigated by PCR in 10 strains of E. coli isolated from Foglia River. Bacterial responses to different antibiotics were also tested with FCM techniques by evaluating both the degree of decrease in viability and the light scatter changes in all of the strains.
Results
PCR revealed that only one strain exhibits the presence of one resistance gene. Despite this, analyses of strains using FCM evidenced the presence of viable subpopulations after antibiotic treatment. Furthermore, analyses of scatter signals revealed profound changes in the Forward Scatter and Side Scatter of the bacterial populations as a consequence of antibiotic exposure, confirming the viability and membrane potential data. The riverine strains were in general less sensitive to antibiotics than the reference strain (ATCC 25922).
Conclusions
Antibiotic resistance is a widespread phenomena. The multiparametric approach based on FCM used in this study, providing results about different aspects (cell viability, membrane potential, light scatter changes), may overcome the limitation of PCR and could represent an adequate method for the evaluation of bacteria responses to antibiotic exposure. © 2014 International Clinical Cytometry Society © 2014 International Clinical Cytometry Society |
| doi_str_mv | 10.1002/cyto.b.21214 |
| format | Article |
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In this study, we check for the presence of specific resistance genes by polymerase chain reaction (PCR) and then we used flow cytometry (FCM) to evaluate antibiotic‐induced effects in different strains of Escherichia coli (E. coli).
Methods
The presence of resistance genes was investigated by PCR in 10 strains of E. coli isolated from Foglia River. Bacterial responses to different antibiotics were also tested with FCM techniques by evaluating both the degree of decrease in viability and the light scatter changes in all of the strains.
Results
PCR revealed that only one strain exhibits the presence of one resistance gene. Despite this, analyses of strains using FCM evidenced the presence of viable subpopulations after antibiotic treatment. Furthermore, analyses of scatter signals revealed profound changes in the Forward Scatter and Side Scatter of the bacterial populations as a consequence of antibiotic exposure, confirming the viability and membrane potential data. The riverine strains were in general less sensitive to antibiotics than the reference strain (ATCC 25922).
Conclusions
Antibiotic resistance is a widespread phenomena. The multiparametric approach based on FCM used in this study, providing results about different aspects (cell viability, membrane potential, light scatter changes), may overcome the limitation of PCR and could represent an adequate method for the evaluation of bacteria responses to antibiotic exposure. © 2014 International Clinical Cytometry Society © 2014 International Clinical Cytometry Society</description><identifier>ISSN: 1552-4949</identifier><identifier>EISSN: 1552-4957</identifier><identifier>DOI: 10.1002/cyto.b.21214</identifier><identifier>PMID: 25532721</identifier><language>eng</language><publisher>United States: Wiley Subscription Services, Inc</publisher><subject>Anti-Bacterial Agents - pharmacology ; antibiotic effects ; Carbocyanines ; Cell Membrane - drug effects ; Drug Resistance, Multiple, Bacterial ; Escherichia coli ; Escherichia coli - classification ; Escherichia coli - drug effects ; Escherichia coli - genetics ; Escherichia coli - isolation & purification ; Flow Cytometry ; Fluorescent Dyes ; Humans ; Membrane Potentials - drug effects ; Microbial Sensitivity Tests ; Microbial Viability - drug effects ; Polymerase Chain Reaction ; Rivers - microbiology ; Water Microbiology</subject><ispartof>Cytometry. Part B, Clinical cytometry, 2015-05, Vol.88 (3), p.149-153</ispartof><rights>2014 International Clinical Cytometry Society</rights><rights>2014 International Clinical Cytometry Society.</rights><rights>2015 International Clinical Cytometry Society</rights><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c4304-36569ad0deee36e0293f88ae4d69db34f1b29448148efbcb60c8f53ae6c6d9393</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://onlinelibrary.wiley.com/doi/pdf/10.1002%2Fcyto.b.21214$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1002%2Fcyto.b.21214$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>314,780,784,1417,1433,27924,27925,45574,45575,46409,46833</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/25532721$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Boi, Paola</creatorcontrib><creatorcontrib>Manti, Anita</creatorcontrib><creatorcontrib>Pianetti, Anna</creatorcontrib><creatorcontrib>Sabatini, Luigia</creatorcontrib><creatorcontrib>Sisti, Davide</creatorcontrib><creatorcontrib>Rocchi, Marco Bruno</creatorcontrib><creatorcontrib>Bruscolini, Francesca</creatorcontrib><creatorcontrib>Galluzzi, Luca</creatorcontrib><creatorcontrib>Papa, Stefano</creatorcontrib><title>Evaluation of Escherichia coli viability by flow cytometry: A method for determining bacterial responses to antibiotic exposure</title><title>Cytometry. Part B, Clinical cytometry</title><addtitle>Cytometry B Clin Cytom</addtitle><description>Background
In this study, we check for the presence of specific resistance genes by polymerase chain reaction (PCR) and then we used flow cytometry (FCM) to evaluate antibiotic‐induced effects in different strains of Escherichia coli (E. coli).
Methods
The presence of resistance genes was investigated by PCR in 10 strains of E. coli isolated from Foglia River. Bacterial responses to different antibiotics were also tested with FCM techniques by evaluating both the degree of decrease in viability and the light scatter changes in all of the strains.
Results
PCR revealed that only one strain exhibits the presence of one resistance gene. Despite this, analyses of strains using FCM evidenced the presence of viable subpopulations after antibiotic treatment. Furthermore, analyses of scatter signals revealed profound changes in the Forward Scatter and Side Scatter of the bacterial populations as a consequence of antibiotic exposure, confirming the viability and membrane potential data. The riverine strains were in general less sensitive to antibiotics than the reference strain (ATCC 25922).
Conclusions
Antibiotic resistance is a widespread phenomena. The multiparametric approach based on FCM used in this study, providing results about different aspects (cell viability, membrane potential, light scatter changes), may overcome the limitation of PCR and could represent an adequate method for the evaluation of bacteria responses to antibiotic exposure. © 2014 International Clinical Cytometry Society © 2014 International Clinical Cytometry Society</description><subject>Anti-Bacterial Agents - pharmacology</subject><subject>antibiotic effects</subject><subject>Carbocyanines</subject><subject>Cell Membrane - drug effects</subject><subject>Drug Resistance, Multiple, Bacterial</subject><subject>Escherichia coli</subject><subject>Escherichia coli - classification</subject><subject>Escherichia coli - drug effects</subject><subject>Escherichia coli - genetics</subject><subject>Escherichia coli - isolation & purification</subject><subject>Flow Cytometry</subject><subject>Fluorescent Dyes</subject><subject>Humans</subject><subject>Membrane Potentials - drug effects</subject><subject>Microbial Sensitivity Tests</subject><subject>Microbial Viability - drug effects</subject><subject>Polymerase Chain Reaction</subject><subject>Rivers - microbiology</subject><subject>Water Microbiology</subject><issn>1552-4949</issn><issn>1552-4957</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2015</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqNkb1vFDEQxS0EIiHQUSNLNDR3-HvXdOF0fEiR0iRFKsv2znKOvOvD9iZsxb_OXhJSpEo1bzQ_vdHMQ-g9JWtKCPvs55rWbs0oo-IFOqZSspXQsnn5qIU-Qm9KuSaES6Ga1-iISclZw-gx-ru9sXGyNaQRpx5vi99BDn4XLPYpBnwTrAsx1Bm7Gfcx3eLDvgFqnr_gU7yIXepwnzLuoEIewhjGX9hZvzTBRpyh7NNYoOCasB1rcCHV4DH82acyZXiLXvU2Fnj3UE_Q5bftxebH6uz8-8_N6dnKC07EiiuptO1IBwBcAWGa921rQXRKd46LnjqmhWipaKF33ini215yC8qrTnPNT9Cne999Tr8nKNUMoXiI0Y6QpmKoahulFgv5DLSRbcMbSRb04xP0Ok15XA45UEK3VCuxUB8eqMkN0Jl9DoPNs_mfwgKIe-A2RJgf55SYQ8Lm8HHjzF3CZnN1cf71TvJ_uO6bdA</recordid><startdate>201505</startdate><enddate>201505</enddate><creator>Boi, Paola</creator><creator>Manti, Anita</creator><creator>Pianetti, Anna</creator><creator>Sabatini, Luigia</creator><creator>Sisti, Davide</creator><creator>Rocchi, Marco Bruno</creator><creator>Bruscolini, Francesca</creator><creator>Galluzzi, Luca</creator><creator>Papa, Stefano</creator><general>Wiley Subscription Services, Inc</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>7QO</scope><scope>7T5</scope><scope>8FD</scope><scope>FR3</scope><scope>H94</scope><scope>P64</scope><scope>7X8</scope><scope>7QL</scope><scope>C1K</scope></search><sort><creationdate>201505</creationdate><title>Evaluation of Escherichia coli viability by flow cytometry: A method for determining bacterial responses to antibiotic exposure</title><author>Boi, Paola ; Manti, Anita ; Pianetti, Anna ; Sabatini, Luigia ; Sisti, Davide ; Rocchi, Marco Bruno ; Bruscolini, Francesca ; Galluzzi, Luca ; Papa, Stefano</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c4304-36569ad0deee36e0293f88ae4d69db34f1b29448148efbcb60c8f53ae6c6d9393</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2015</creationdate><topic>Anti-Bacterial Agents - pharmacology</topic><topic>antibiotic effects</topic><topic>Carbocyanines</topic><topic>Cell Membrane - drug effects</topic><topic>Drug Resistance, Multiple, Bacterial</topic><topic>Escherichia coli</topic><topic>Escherichia coli - classification</topic><topic>Escherichia coli - drug effects</topic><topic>Escherichia coli - genetics</topic><topic>Escherichia coli - isolation & purification</topic><topic>Flow Cytometry</topic><topic>Fluorescent Dyes</topic><topic>Humans</topic><topic>Membrane Potentials - drug effects</topic><topic>Microbial Sensitivity Tests</topic><topic>Microbial Viability - drug effects</topic><topic>Polymerase Chain Reaction</topic><topic>Rivers - microbiology</topic><topic>Water Microbiology</topic><toplevel>online_resources</toplevel><creatorcontrib>Boi, Paola</creatorcontrib><creatorcontrib>Manti, Anita</creatorcontrib><creatorcontrib>Pianetti, Anna</creatorcontrib><creatorcontrib>Sabatini, Luigia</creatorcontrib><creatorcontrib>Sisti, Davide</creatorcontrib><creatorcontrib>Rocchi, Marco Bruno</creatorcontrib><creatorcontrib>Bruscolini, Francesca</creatorcontrib><creatorcontrib>Galluzzi, Luca</creatorcontrib><creatorcontrib>Papa, Stefano</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>Biotechnology Research Abstracts</collection><collection>Immunology Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>MEDLINE - Academic</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Environmental Sciences and Pollution Management</collection><jtitle>Cytometry. Part B, Clinical cytometry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Boi, Paola</au><au>Manti, Anita</au><au>Pianetti, Anna</au><au>Sabatini, Luigia</au><au>Sisti, Davide</au><au>Rocchi, Marco Bruno</au><au>Bruscolini, Francesca</au><au>Galluzzi, Luca</au><au>Papa, Stefano</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Evaluation of Escherichia coli viability by flow cytometry: A method for determining bacterial responses to antibiotic exposure</atitle><jtitle>Cytometry. Part B, Clinical cytometry</jtitle><addtitle>Cytometry B Clin Cytom</addtitle><date>2015-05</date><risdate>2015</risdate><volume>88</volume><issue>3</issue><spage>149</spage><epage>153</epage><pages>149-153</pages><issn>1552-4949</issn><eissn>1552-4957</eissn><abstract>Background
In this study, we check for the presence of specific resistance genes by polymerase chain reaction (PCR) and then we used flow cytometry (FCM) to evaluate antibiotic‐induced effects in different strains of Escherichia coli (E. coli).
Methods
The presence of resistance genes was investigated by PCR in 10 strains of E. coli isolated from Foglia River. Bacterial responses to different antibiotics were also tested with FCM techniques by evaluating both the degree of decrease in viability and the light scatter changes in all of the strains.
Results
PCR revealed that only one strain exhibits the presence of one resistance gene. Despite this, analyses of strains using FCM evidenced the presence of viable subpopulations after antibiotic treatment. Furthermore, analyses of scatter signals revealed profound changes in the Forward Scatter and Side Scatter of the bacterial populations as a consequence of antibiotic exposure, confirming the viability and membrane potential data. The riverine strains were in general less sensitive to antibiotics than the reference strain (ATCC 25922).
Conclusions
Antibiotic resistance is a widespread phenomena. The multiparametric approach based on FCM used in this study, providing results about different aspects (cell viability, membrane potential, light scatter changes), may overcome the limitation of PCR and could represent an adequate method for the evaluation of bacteria responses to antibiotic exposure. © 2014 International Clinical Cytometry Society © 2014 International Clinical Cytometry Society</abstract><cop>United States</cop><pub>Wiley Subscription Services, Inc</pub><pmid>25532721</pmid><doi>10.1002/cyto.b.21214</doi><tpages>5</tpages></addata></record> |
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| subjects | Anti-Bacterial Agents - pharmacology antibiotic effects Carbocyanines Cell Membrane - drug effects Drug Resistance, Multiple, Bacterial Escherichia coli Escherichia coli - classification Escherichia coli - drug effects Escherichia coli - genetics Escherichia coli - isolation & purification Flow Cytometry Fluorescent Dyes Humans Membrane Potentials - drug effects Microbial Sensitivity Tests Microbial Viability - drug effects Polymerase Chain Reaction Rivers - microbiology Water Microbiology |
| title | Evaluation of Escherichia coli viability by flow cytometry: A method for determining bacterial responses to antibiotic exposure |
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