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Comparison of methods to assess biofilm disinfection and recovery by drinking water-isolated bacteria

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Resumo:Drinking water (DW) distribution systems are known to harbour biofilms even in the presence of disinfectants. DW biofilms are constituted by microbial communities adapted to low nutrient concentrations and high chlorine levels. Biofilm formation and resistance to disinfection have been recognized as important factors that contribute to the survival and persistence of microbial contamination in DW. The purpose of this work was the comparison of diverse methods to assess the disinfection of biofilms formed by six DW-isolated opportunistic bacteria (Acinetobacter calcoaceticus, Burkholderia cepacia, Methylobacterium sp., Mycobacterium mucogenicum, Sphingomonas capsulata and Staphylococcus sp.) by sodium hypochlorite (SHC). Single and multi-species biofilms (composed of combinations of 6 and 5 bacteria) were developed in 96- wells microtiter plates for 3 days, afterwards, were exposed to several independent SHC concentrations (0.1, 0.5, 1 and 10 mg/L) during 1 h. The potential of biofilms to recover was assessed 24 h after disinfection. The disinfection efficacy and recovery were assessed in terms of variation in: biofilm mass (crystal violet staining); metabolic activity (XTT staining); cultivability (CFUs) and viability (Live/Dead staining). The results indicated that biomass removal increased with increasing SHC concentration, but total biofilm mass removal was not achieved. The effects of SHC on the biofilm activity, cultivability and viability were also concentration dependent. Total biofilm inactivation was achieved only for A. calcoaceticus biofilms and for multi-species biofilms without A. calcoaceticus, when exposed to high SHC concentrations. Almost all multispecies biofilms were more resistant to removal and inactivation than the single biofilms. Methylobacterium sp. and A. calcoaceticus formed the most resistant and the most susceptible biofilms, respectively. On the other hand, biofilm combination with the six DW bacteria was the most resistant to SHC and combination without A. calcoaceticus was the least resistant, for all concentration tested. The several methods used to assess of biofilm activity (metabolic activity, cultivability and viability) provided comparable results. However the viability results provide the worst case scenario in terms of biofilm control analysis (higher number of viable cells for all the SHC concentrations tested). The recovery results demonstrated that only biofilms without A.calcoaceticus were not able to recover their biomass from the SHC treatments. Also, those biofilms had a decreased ability to recover their metabolic activity, cultivability and viability. Conversely, multi-species biofilms without Staphylococcus sp. had the highest ability to recover from disinfection. Biofilm mass and activity recovery were not correlated for all the biofilms tested. However, the data of biofilm recovery in terms of metabolic activity, cultivability and viability also provided comparable results.
Autores principais:Simões, Lúcia C.
Outros Autores:Simões, M.; Vieira, M. J.
Assunto:Biofilm disinfection Drinking water bacteria Methods, recovery Resistance Sodium hypochlorite
Ano:2012
País:Portugal
Tipo de documento:outro
Tipo de acesso:acesso aberto
Instituição associada:Universidade do Minho
Idioma:inglês
Origem:RepositóriUM - Universidade do Minho
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author Simões, Lúcia C.
author2 Simões, M.
Vieira, M. J.
author2_role author
author
author_facet Simões, Lúcia C.
Simões, M.
Vieira, M. J.
author_role author
contributor_name_str_mv Universidade do Minho
country_str PT
creators_json_txt [{\"Person.name\":\"Simões, Lúcia C.\"},{\"Person.name\":\"Simões, M.\"},{\"Person.name\":\"Vieira, M. J.\"}]
datacite.contributors.contributor.contributorName.fl_str_mv Universidade do Minho
datacite.creators.creator.creatorName.fl_str_mv Simões, Lúcia C.
Simões, M.
Vieira, M. J.
datacite.date.Accepted.fl_str_mv 2012-01-01T00:00:00Z
datacite.date.available.fl_str_mv 2013-10-14T11:17:43Z
datacite.date.embargoed.fl_str_mv 2013-10-14T11:17:43Z
datacite.rights.fl_str_mv http://purl.org/coar/access_right/c_abf2
datacite.subjects.subject.fl_str_mv Biofilm disinfection
Drinking water bacteria
Methods, recovery
Resistance
Sodium hypochlorite
datacite.titles.title.fl_str_mv Comparison of methods to assess biofilm disinfection and recovery by drinking water-isolated bacteria
dc.contributor.none.fl_str_mv Universidade do Minho
dc.creator.none.fl_str_mv Simões, Lúcia C.
Simões, M.
Vieira, M. J.
dc.date.Accepted.fl_str_mv 2012-01-01T00:00:00Z
dc.date.available.fl_str_mv 2013-10-14T11:17:43Z
dc.date.embargoed.fl_str_mv 2013-10-14T11:17:43Z
dc.format.none.fl_str_mv application/pdf
dc.identifier.none.fl_str_mv https://hdl.handle.net/1822/25684
dc.language.none.fl_str_mv eng
dc.publisher.none.fl_str_mv International Conference on Antimicrobial Research (ICAR 2012)
dc.rights.none.fl_str_mv http://purl.org/coar/access_right/c_abf2
dc.subject.none.fl_str_mv Biofilm disinfection
Drinking water bacteria
Methods, recovery
Resistance
Sodium hypochlorite
dc.title.fl_str_mv Comparison of methods to assess biofilm disinfection and recovery by drinking water-isolated bacteria
dc.type.none.fl_str_mv http://purl.org/coar/resource_type/c_1843
description Drinking water (DW) distribution systems are known to harbour biofilms even in the presence of disinfectants. DW biofilms are constituted by microbial communities adapted to low nutrient concentrations and high chlorine levels. Biofilm formation and resistance to disinfection have been recognized as important factors that contribute to the survival and persistence of microbial contamination in DW. The purpose of this work was the comparison of diverse methods to assess the disinfection of biofilms formed by six DW-isolated opportunistic bacteria (Acinetobacter calcoaceticus, Burkholderia cepacia, Methylobacterium sp., Mycobacterium mucogenicum, Sphingomonas capsulata and Staphylococcus sp.) by sodium hypochlorite (SHC). Single and multi-species biofilms (composed of combinations of 6 and 5 bacteria) were developed in 96- wells microtiter plates for 3 days, afterwards, were exposed to several independent SHC concentrations (0.1, 0.5, 1 and 10 mg/L) during 1 h. The potential of biofilms to recover was assessed 24 h after disinfection. The disinfection efficacy and recovery were assessed in terms of variation in: biofilm mass (crystal violet staining); metabolic activity (XTT staining); cultivability (CFUs) and viability (Live/Dead staining). The results indicated that biomass removal increased with increasing SHC concentration, but total biofilm mass removal was not achieved. The effects of SHC on the biofilm activity, cultivability and viability were also concentration dependent. Total biofilm inactivation was achieved only for A. calcoaceticus biofilms and for multi-species biofilms without A. calcoaceticus, when exposed to high SHC concentrations. Almost all multispecies biofilms were more resistant to removal and inactivation than the single biofilms. Methylobacterium sp. and A. calcoaceticus formed the most resistant and the most susceptible biofilms, respectively. On the other hand, biofilm combination with the six DW bacteria was the most resistant to SHC and combination without A. calcoaceticus was the least resistant, for all concentration tested. The several methods used to assess of biofilm activity (metabolic activity, cultivability and viability) provided comparable results. However the viability results provide the worst case scenario in terms of biofilm control analysis (higher number of viable cells for all the SHC concentrations tested). The recovery results demonstrated that only biofilms without A.calcoaceticus were not able to recover their biomass from the SHC treatments. Also, those biofilms had a decreased ability to recover their metabolic activity, cultivability and viability. Conversely, multi-species biofilms without Staphylococcus sp. had the highest ability to recover from disinfection. Biofilm mass and activity recovery were not correlated for all the biofilms tested. However, the data of biofilm recovery in terms of metabolic activity, cultivability and viability also provided comparable results.
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eu_rights_str_mv openAccess
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fulltext.url.fl_str_mv https://prod-dspace.uminho.pt/bitstreams/8f544ecc-d1bb-4188-ac8f-13a89df23f2f/download
id rum_0e7e4a6c4c7fd32a7ec8a3eb376f30d2
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instacron_str repositorium
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instname_str Universidade do Minho
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oai_identifier_str oai:repositorium.uminho.pt:1822/25684
organization_str_mv urn:organizationAcronym:repositorium
person_str_mv Simões, Lúcia C.
Simões, M.
Vieira, M. J.
publishDate 2012
publisher.none.fl_str_mv International Conference on Antimicrobial Research (ICAR 2012)
reponame_str RepositóriUM - Universidade do Minho
repository_id_str urn:repositoryAcronym:rum
service_str_mv urn:repositoryAcronym:rum
spelling engInternational Conference on Antimicrobial Research (ICAR 2012)porDrinking water (DW) distribution systems are known to harbour biofilms even in the presence of disinfectants. DW biofilms are constituted by microbial communities adapted to low nutrient concentrations and high chlorine levels. Biofilm formation and resistance to disinfection have been recognized as important factors that contribute to the survival and persistence of microbial contamination in DW. The purpose of this work was the comparison of diverse methods to assess the disinfection of biofilms formed by six DW-isolated opportunistic bacteria (Acinetobacter calcoaceticus, Burkholderia cepacia, Methylobacterium sp., Mycobacterium mucogenicum, Sphingomonas capsulata and Staphylococcus sp.) by sodium hypochlorite (SHC). Single and multi-species biofilms (composed of combinations of 6 and 5 bacteria) were developed in 96- wells microtiter plates for 3 days, afterwards, were exposed to several independent SHC concentrations (0.1, 0.5, 1 and 10 mg/L) during 1 h. The potential of biofilms to recover was assessed 24 h after disinfection. The disinfection efficacy and recovery were assessed in terms of variation in: biofilm mass (crystal violet staining); metabolic activity (XTT staining); cultivability (CFUs) and viability (Live/Dead staining). The results indicated that biomass removal increased with increasing SHC concentration, but total biofilm mass removal was not achieved. The effects of SHC on the biofilm activity, cultivability and viability were also concentration dependent. Total biofilm inactivation was achieved only for A. calcoaceticus biofilms and for multi-species biofilms without A. calcoaceticus, when exposed to high SHC concentrations. Almost all multispecies biofilms were more resistant to removal and inactivation than the single biofilms. Methylobacterium sp. and A. calcoaceticus formed the most resistant and the most susceptible biofilms, respectively. On the other hand, biofilm combination with the six DW bacteria was the most resistant to SHC and combination without A. calcoaceticus was the least resistant, for all concentration tested. The several methods used to assess of biofilm activity (metabolic activity, cultivability and viability) provided comparable results. However the viability results provide the worst case scenario in terms of biofilm control analysis (higher number of viable cells for all the SHC concentrations tested). The recovery results demonstrated that only biofilms without A.calcoaceticus were not able to recover their biomass from the SHC treatments. Also, those biofilms had a decreased ability to recover their metabolic activity, cultivability and viability. Conversely, multi-species biofilms without Staphylococcus sp. had the highest ability to recover from disinfection. Biofilm mass and activity recovery were not correlated for all the biofilms tested. However, the data of biofilm recovery in terms of metabolic activity, cultivability and viability also provided comparable results.application/pdfporComparison of methods to assess biofilm disinfection and recovery by drinking water-isolated bacteriaSimões, Lúcia C.Simões, M.Vieira, M. J.HostingInstitutionOrganizationalUniversidade do Minhoe-mailmailto:repositorium@usdb.uminho.ptrepositorium@usdb.uminho.pt2013-10-14T11:17:43Z20122012-01-01T00:00:00ZHandlehttps://hdl.handle.net/1822/25684http://purl.org/coar/access_right/c_abf2open accessBiofilm disinfectionDrinking water bacteriaMethods, recoveryResistanceSodium hypochlorite94166 bytesother research producthttp://purl.org/coar/resource_type/c_1843otherhttp://purl.org/coar/access_right/c_abf2application/pdffulltexthttps://prod-dspace.uminho.pt/bitstreams/8f544ecc-d1bb-4188-ac8f-13a89df23f2f/download
spellingShingle Comparison of methods to assess biofilm disinfection and recovery by drinking water-isolated bacteria
Simões, Lúcia C.
Biofilm disinfection
Drinking water bacteria
Methods, recovery
Resistance
Sodium hypochlorite
status SINGLETON
subject.fl_str_mv Biofilm disinfection
Drinking water bacteria
Methods, recovery
Resistance
Sodium hypochlorite
title Comparison of methods to assess biofilm disinfection and recovery by drinking water-isolated bacteria
title_full Comparison of methods to assess biofilm disinfection and recovery by drinking water-isolated bacteria
title_fullStr Comparison of methods to assess biofilm disinfection and recovery by drinking water-isolated bacteria
title_full_unstemmed Comparison of methods to assess biofilm disinfection and recovery by drinking water-isolated bacteria
title_short Comparison of methods to assess biofilm disinfection and recovery by drinking water-isolated bacteria
title_sort Comparison of methods to assess biofilm disinfection and recovery by drinking water-isolated bacteria
topic Biofilm disinfection
Drinking water bacteria
Methods, recovery
Resistance
Sodium hypochlorite
topic_facet Biofilm disinfection
Drinking water bacteria
Methods, recovery
Resistance
Sodium hypochlorite
url https://hdl.handle.net/1822/25684
visible 1