Study of Pulsed Light-Induced Damage on Saccharomyces cerevisiae in Apple Juice by Flow Cytometry and Transmission Electron Microscopy

This work analyzed the pulsed light (PL) (0.0-71.6 J/cm2)-induced damage on Saccharomyces cerevisiae KE162 cells in peptone water (pH 3.5 or 5.6) and apple juice (pH 3.5) by applying flow cytometry (FCM) and transmission electronic microscopy. Cells were labeled with fluorescein diacetate (FDA) for...

Descripción completa

Guardado en:
Detalles Bibliográficos
Autores principales: Guerrero, Sandra, Alzamora, Stella Maris
Publicado: 2014
Materias:
Apple juice
Flow cytometry
Microscopy
Pulsed light
Saccharomyces cerevisiae
Acceso en línea:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_19355130_v7_n4_p1001_Ferrario
http://hdl.handle.net/20.500.12110/paper_19355130_v7_n4_p1001_Ferrario
Aporte de:
Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires
id paper:paper_19355130_v7_n4_p1001_Ferrario
record_format dspace
spelling paper:paper_19355130_v7_n4_p1001_Ferrario2023-06-08T16:32:03Z Study of Pulsed Light-Induced Damage on Saccharomyces cerevisiae in Apple Juice by Flow Cytometry and Transmission Electron Microscopy Guerrero, Sandra Alzamora, Stella Maris Apple juice Flow cytometry Microscopy Pulsed light Saccharomyces cerevisiae This work analyzed the pulsed light (PL) (0.0-71.6 J/cm2)-induced damage on Saccharomyces cerevisiae KE162 cells in peptone water (pH 3.5 or 5.6) and apple juice (pH 3.5) by applying flow cytometry (FCM) and transmission electronic microscopy. Cells were labeled with fluorescein diacetate (FDA) for detecting membrane integrity and esterase activity and with propidium iodide (PI) for monitoring membrane integrity. S. cerevisiae inactivation curves reached 6-7 log reductions (peptone water systems) and 3.9 log reductions (apple juice) after 60 s (71.6 J/cm2) of PL exposure. FCM revealed the same damage pattern (although at different doses) in all media: at low doses, there was an increase in population in PI+-FDA+ quadrant, while at high doses, most of the population was located at quadrant PI+-FDA-, indicating that PL provoked rupture of the cytoplasm membrane allowing PI to penetrate cells and there was progressive loss of esterase activity. Comparison of conventional culture technique with FCM revealed the occurrence of certain cell subpopulations in peptone water with pH 3.5 which were stressed and lost their ability to grow in agar but still showed metabolic activity. Transmission electron microphotographs of PL-treated cells clearly indicated that various cell structures other than plasma membranes were altered and/or destroyed in a different degree depending on exposure time and type of medium. © 2013 Springer Science+Business Media New York. Fil:Guerrero, S. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Alzamora, S.M. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. 2014 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_19355130_v7_n4_p1001_Ferrario http://hdl.handle.net/20.500.12110/paper_19355130_v7_n4_p1001_Ferrario
institution Universidad de Buenos Aires
institution_str I-28
repository_str R-134
collection Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA)
topic Apple juice
Flow cytometry
Microscopy
Pulsed light
Saccharomyces cerevisiae
spellingShingle Apple juice
Flow cytometry
Microscopy
Pulsed light
Saccharomyces cerevisiae
Guerrero, Sandra
Alzamora, Stella Maris
Study of Pulsed Light-Induced Damage on Saccharomyces cerevisiae in Apple Juice by Flow Cytometry and Transmission Electron Microscopy
topic_facet Apple juice
Flow cytometry
Microscopy
Pulsed light
Saccharomyces cerevisiae
description This work analyzed the pulsed light (PL) (0.0-71.6 J/cm2)-induced damage on Saccharomyces cerevisiae KE162 cells in peptone water (pH 3.5 or 5.6) and apple juice (pH 3.5) by applying flow cytometry (FCM) and transmission electronic microscopy. Cells were labeled with fluorescein diacetate (FDA) for detecting membrane integrity and esterase activity and with propidium iodide (PI) for monitoring membrane integrity. S. cerevisiae inactivation curves reached 6-7 log reductions (peptone water systems) and 3.9 log reductions (apple juice) after 60 s (71.6 J/cm2) of PL exposure. FCM revealed the same damage pattern (although at different doses) in all media: at low doses, there was an increase in population in PI+-FDA+ quadrant, while at high doses, most of the population was located at quadrant PI+-FDA-, indicating that PL provoked rupture of the cytoplasm membrane allowing PI to penetrate cells and there was progressive loss of esterase activity. Comparison of conventional culture technique with FCM revealed the occurrence of certain cell subpopulations in peptone water with pH 3.5 which were stressed and lost their ability to grow in agar but still showed metabolic activity. Transmission electron microphotographs of PL-treated cells clearly indicated that various cell structures other than plasma membranes were altered and/or destroyed in a different degree depending on exposure time and type of medium. © 2013 Springer Science+Business Media New York.
author Guerrero, Sandra
Alzamora, Stella Maris
author_facet Guerrero, Sandra
Alzamora, Stella Maris
author_sort Guerrero, Sandra
title Study of Pulsed Light-Induced Damage on Saccharomyces cerevisiae in Apple Juice by Flow Cytometry and Transmission Electron Microscopy
title_short Study of Pulsed Light-Induced Damage on Saccharomyces cerevisiae in Apple Juice by Flow Cytometry and Transmission Electron Microscopy
title_full Study of Pulsed Light-Induced Damage on Saccharomyces cerevisiae in Apple Juice by Flow Cytometry and Transmission Electron Microscopy
title_fullStr Study of Pulsed Light-Induced Damage on Saccharomyces cerevisiae in Apple Juice by Flow Cytometry and Transmission Electron Microscopy
title_full_unstemmed Study of Pulsed Light-Induced Damage on Saccharomyces cerevisiae in Apple Juice by Flow Cytometry and Transmission Electron Microscopy
title_sort study of pulsed light-induced damage on saccharomyces cerevisiae in apple juice by flow cytometry and transmission electron microscopy
publishDate 2014
url https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_19355130_v7_n4_p1001_Ferrario
http://hdl.handle.net/20.500.12110/paper_19355130_v7_n4_p1001_Ferrario
work_keys_str_mv AT guerrerosandra studyofpulsedlightinduceddamageonsaccharomycescerevisiaeinapplejuicebyflowcytometryandtransmissionelectronmicroscopy
AT alzamorastellamaris studyofpulsedlightinduceddamageonsaccharomycescerevisiaeinapplejuicebyflowcytometryandtransmissionelectronmicroscopy
_version_ 1768544201625042944

Ejemplares similares