Phenotypic resistance in photodynamic inactivation unravelled at the single bacterium level

Autores
Martinez, Sol Romina; Palacios, Yohana Belén; Heredia, Daniel Alejandro; Agazzi, Maximiliano Luis; Durantini, Andres Matías
Año de publicación
2019
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Herein we report a simple fluorescence microscopy methodology that, jointly with four photosensitizers (PSs) and a cell viability marker, allows monitoring of phenotypic bacterial resistance to photodynamic inactivation (PDI) treatments. The PSs, composed of BODIPY dyes, were selected according to their ability to interact with the cell wall and the photoinactivating mechanism involved (type I or type II). In a first approach, the phenotypic heterogeneity allowing bacteria to persist during PDI treatment was evaluated in methicillin-resistant Staphylococcus aureus (MRSA) and Escherichia coli as Gram-positive and Gram-negative models, respectively. By means of propidium iodide (PI), we monitored with spatiotemporal resolution cell viability at the single bacterium level. All the PSs were effective at inactivating pathogens; however, the cationic nonhalogenated PS (compound 1) surpassed the others and was capable of photoinactivating E. coli even under optimal growth conditions. Compound 1 was further tested on two other Gram-negative strains, Pseudomonas aeruginosa and Klebsiella pneumoniae, with outstanding results. All bacterial strains used here are well-known ESKAPE pathogens, which are the leading cause of nosocomial infections worldwide. Thorough data analysis of individual cell survival times revealed clear phenotypic variation expressed in the cell wall that affected PI permeation and thus its intercalation with DNA. For the same bacterial sample, death times may vary from seconds to hours. In addition, the PI incorporation time is also a parameter governed by the phenotypic characteristics of the microbes. Finally, we demonstrate that the results gathered for the bacteria provide direct and unique experimental evidence that supports the time-kill curve profiles.
Fil: Martinez, Sol Romina. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Departamento de Química; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; Argentina
Fil: Palacios, Yohana Belén. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Departamento de Química; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; Argentina
Fil: Heredia, Daniel Alejandro. Universidad Nacional de Río Cuarto. Instituto para el Desarrollo Agroindustrial y de la Salud. - Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto para el Desarrollo Agroindustrial y de la Salud; Argentina. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Departamento de Química; Argentina
Fil: Agazzi, Maximiliano Luis. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Departamento de Química; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; Argentina
Fil: Durantini, Andres Matías. Universidad Nacional de Río Cuarto. Instituto para el Desarrollo Agroindustrial y de la Salud. - Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto para el Desarrollo Agroindustrial y de la Salud; Argentina. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Departamento de Química; Argentina
Materia
ESKAPE PATHOGENS
FLUORESCENCE MICROSCOPY
PHENOTYPIC RESISTANCE
PHOTOACTIVE MOLECULES
PHOTODYNAMIC INACTIVATION
SINGLE CELL
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:11336/132326
Acceder
id CONICETDig_690eba868dcbe64d151f493e35e73171
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network_acronym_str CONICETDig
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network_name_str CONICET Digital (CONICET)
spelling Phenotypic resistance in photodynamic inactivation unravelled at the single bacterium levelMartinez, Sol RominaPalacios, Yohana BelénHeredia, Daniel AlejandroAgazzi, Maximiliano LuisDurantini, Andres MatíasESKAPE PATHOGENSFLUORESCENCE MICROSCOPYPHENOTYPIC RESISTANCEPHOTOACTIVE MOLECULESPHOTODYNAMIC INACTIVATIONSINGLE CELLhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Herein we report a simple fluorescence microscopy methodology that, jointly with four photosensitizers (PSs) and a cell viability marker, allows monitoring of phenotypic bacterial resistance to photodynamic inactivation (PDI) treatments. The PSs, composed of BODIPY dyes, were selected according to their ability to interact with the cell wall and the photoinactivating mechanism involved (type I or type II). In a first approach, the phenotypic heterogeneity allowing bacteria to persist during PDI treatment was evaluated in methicillin-resistant Staphylococcus aureus (MRSA) and Escherichia coli as Gram-positive and Gram-negative models, respectively. By means of propidium iodide (PI), we monitored with spatiotemporal resolution cell viability at the single bacterium level. All the PSs were effective at inactivating pathogens; however, the cationic nonhalogenated PS (compound 1) surpassed the others and was capable of photoinactivating E. coli even under optimal growth conditions. Compound 1 was further tested on two other Gram-negative strains, Pseudomonas aeruginosa and Klebsiella pneumoniae, with outstanding results. All bacterial strains used here are well-known ESKAPE pathogens, which are the leading cause of nosocomial infections worldwide. Thorough data analysis of individual cell survival times revealed clear phenotypic variation expressed in the cell wall that affected PI permeation and thus its intercalation with DNA. For the same bacterial sample, death times may vary from seconds to hours. In addition, the PI incorporation time is also a parameter governed by the phenotypic characteristics of the microbes. Finally, we demonstrate that the results gathered for the bacteria provide direct and unique experimental evidence that supports the time-kill curve profiles.Fil: Martinez, Sol Romina. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Departamento de Química; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; ArgentinaFil: Palacios, Yohana Belén. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Departamento de Química; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; ArgentinaFil: Heredia, Daniel Alejandro. Universidad Nacional de Río Cuarto. Instituto para el Desarrollo Agroindustrial y de la Salud. - Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto para el Desarrollo Agroindustrial y de la Salud; Argentina. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Departamento de Química; ArgentinaFil: Agazzi, Maximiliano Luis. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Departamento de Química; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; ArgentinaFil: Durantini, Andres Matías. Universidad Nacional de Río Cuarto. Instituto para el Desarrollo Agroindustrial y de la Salud. - Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto para el Desarrollo Agroindustrial y de la Salud; Argentina. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Departamento de Química; ArgentinaAmerican Chemical Society2019-07-09info: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/11336/132326Martinez, Sol Romina; Palacios, Yohana Belén; Heredia, Daniel Alejandro; Agazzi, Maximiliano Luis; Durantini, Andres Matías; Phenotypic resistance in photodynamic inactivation unravelled at the single bacterium level; American Chemical Society; ACS Infectious Diseases; 5; 9; 9-7-2019; 1624-16332373-8227CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/http://pubs.acs.org/doi/10.1021/acsinfecdis.9b00185info:eu-repo/semantics/altIdentifier/doi/10.1021/acsinfecdis.9b00185info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-10-15T14:23:32Zoai:ri.conicet.gov.ar:11336/132326instacron: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:34982025-10-15 14:23:33.114CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Phenotypic resistance in photodynamic inactivation unravelled at the single bacterium level
title Phenotypic resistance in photodynamic inactivation unravelled at the single bacterium level
spellingShingle Phenotypic resistance in photodynamic inactivation unravelled at the single bacterium level
Martinez, Sol Romina
ESKAPE PATHOGENS
FLUORESCENCE MICROSCOPY
PHENOTYPIC RESISTANCE
PHOTOACTIVE MOLECULES
PHOTODYNAMIC INACTIVATION
SINGLE CELL
title_short Phenotypic resistance in photodynamic inactivation unravelled at the single bacterium level
title_full Phenotypic resistance in photodynamic inactivation unravelled at the single bacterium level
title_fullStr Phenotypic resistance in photodynamic inactivation unravelled at the single bacterium level
title_full_unstemmed Phenotypic resistance in photodynamic inactivation unravelled at the single bacterium level
title_sort Phenotypic resistance in photodynamic inactivation unravelled at the single bacterium level
dc.creator.none.fl_str_mv Martinez, Sol Romina
Palacios, Yohana Belén
Heredia, Daniel Alejandro
Agazzi, Maximiliano Luis
Durantini, Andres Matías
author Martinez, Sol Romina
author_facet Martinez, Sol Romina
Palacios, Yohana Belén
Heredia, Daniel Alejandro
Agazzi, Maximiliano Luis
Durantini, Andres Matías
author_role author
author2 Palacios, Yohana Belén
Heredia, Daniel Alejandro
Agazzi, Maximiliano Luis
Durantini, Andres Matías
author2_role author
author
author
author
dc.subject.none.fl_str_mv ESKAPE PATHOGENS
FLUORESCENCE MICROSCOPY
PHENOTYPIC RESISTANCE
PHOTOACTIVE MOLECULES
PHOTODYNAMIC INACTIVATION
SINGLE CELL
topic ESKAPE PATHOGENS
FLUORESCENCE MICROSCOPY
PHENOTYPIC RESISTANCE
PHOTOACTIVE MOLECULES
PHOTODYNAMIC INACTIVATION
SINGLE CELL
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Herein we report a simple fluorescence microscopy methodology that, jointly with four photosensitizers (PSs) and a cell viability marker, allows monitoring of phenotypic bacterial resistance to photodynamic inactivation (PDI) treatments. The PSs, composed of BODIPY dyes, were selected according to their ability to interact with the cell wall and the photoinactivating mechanism involved (type I or type II). In a first approach, the phenotypic heterogeneity allowing bacteria to persist during PDI treatment was evaluated in methicillin-resistant Staphylococcus aureus (MRSA) and Escherichia coli as Gram-positive and Gram-negative models, respectively. By means of propidium iodide (PI), we monitored with spatiotemporal resolution cell viability at the single bacterium level. All the PSs were effective at inactivating pathogens; however, the cationic nonhalogenated PS (compound 1) surpassed the others and was capable of photoinactivating E. coli even under optimal growth conditions. Compound 1 was further tested on two other Gram-negative strains, Pseudomonas aeruginosa and Klebsiella pneumoniae, with outstanding results. All bacterial strains used here are well-known ESKAPE pathogens, which are the leading cause of nosocomial infections worldwide. Thorough data analysis of individual cell survival times revealed clear phenotypic variation expressed in the cell wall that affected PI permeation and thus its intercalation with DNA. For the same bacterial sample, death times may vary from seconds to hours. In addition, the PI incorporation time is also a parameter governed by the phenotypic characteristics of the microbes. Finally, we demonstrate that the results gathered for the bacteria provide direct and unique experimental evidence that supports the time-kill curve profiles.
Fil: Martinez, Sol Romina. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Departamento de Química; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; Argentina
Fil: Palacios, Yohana Belén. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Departamento de Química; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; Argentina
Fil: Heredia, Daniel Alejandro. Universidad Nacional de Río Cuarto. Instituto para el Desarrollo Agroindustrial y de la Salud. - Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto para el Desarrollo Agroindustrial y de la Salud; Argentina. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Departamento de Química; Argentina
Fil: Agazzi, Maximiliano Luis. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Departamento de Química; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; Argentina
Fil: Durantini, Andres Matías. Universidad Nacional de Río Cuarto. Instituto para el Desarrollo Agroindustrial y de la Salud. - Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto para el Desarrollo Agroindustrial y de la Salud; Argentina. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Departamento de Química; Argentina
description Herein we report a simple fluorescence microscopy methodology that, jointly with four photosensitizers (PSs) and a cell viability marker, allows monitoring of phenotypic bacterial resistance to photodynamic inactivation (PDI) treatments. The PSs, composed of BODIPY dyes, were selected according to their ability to interact with the cell wall and the photoinactivating mechanism involved (type I or type II). In a first approach, the phenotypic heterogeneity allowing bacteria to persist during PDI treatment was evaluated in methicillin-resistant Staphylococcus aureus (MRSA) and Escherichia coli as Gram-positive and Gram-negative models, respectively. By means of propidium iodide (PI), we monitored with spatiotemporal resolution cell viability at the single bacterium level. All the PSs were effective at inactivating pathogens; however, the cationic nonhalogenated PS (compound 1) surpassed the others and was capable of photoinactivating E. coli even under optimal growth conditions. Compound 1 was further tested on two other Gram-negative strains, Pseudomonas aeruginosa and Klebsiella pneumoniae, with outstanding results. All bacterial strains used here are well-known ESKAPE pathogens, which are the leading cause of nosocomial infections worldwide. Thorough data analysis of individual cell survival times revealed clear phenotypic variation expressed in the cell wall that affected PI permeation and thus its intercalation with DNA. For the same bacterial sample, death times may vary from seconds to hours. In addition, the PI incorporation time is also a parameter governed by the phenotypic characteristics of the microbes. Finally, we demonstrate that the results gathered for the bacteria provide direct and unique experimental evidence that supports the time-kill curve profiles.
publishDate 2019
dc.date.none.fl_str_mv 2019-07-09
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/11336/132326
Martinez, Sol Romina; Palacios, Yohana Belén; Heredia, Daniel Alejandro; Agazzi, Maximiliano Luis; Durantini, Andres Matías; Phenotypic resistance in photodynamic inactivation unravelled at the single bacterium level; American Chemical Society; ACS Infectious Diseases; 5; 9; 9-7-2019; 1624-1633
2373-8227
CONICET Digital
CONICET
url http://hdl.handle.net/11336/132326
identifier_str_mv Martinez, Sol Romina; Palacios, Yohana Belén; Heredia, Daniel Alejandro; Agazzi, Maximiliano Luis; Durantini, Andres Matías; Phenotypic resistance in photodynamic inactivation unravelled at the single bacterium level; American Chemical Society; ACS Infectious Diseases; 5; 9; 9-7-2019; 1624-1633
2373-8227
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/http://pubs.acs.org/doi/10.1021/acsinfecdis.9b00185
info:eu-repo/semantics/altIdentifier/doi/10.1021/acsinfecdis.9b00185
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 American Chemical Society
publisher.none.fl_str_mv American Chemical Society
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 13.051919

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