Evaluating methods for purifying cyanobacterial cultures by qPCR and high-throughput Illumina sequencing

dc.contributor.authorHeck, Karina
dc.contributor.authorMachineski, Gabriela Silva
dc.contributor.authorAlvarenga, Danillo Oliveira
dc.contributor.authorMarcal Vieira Vaz, Marcelo Gomes
dc.contributor.authorVarani, Alessandro de Mello [UNESP]
dc.contributor.authorFiore, Marli Fatima
dc.contributor.institutionUniversidade de São Paulo (USP)
dc.contributor.institutionUniversidade Estadual Paulista (Unesp)
dc.date.accessioned2018-11-27T21:34:10Z
dc.date.available2018-11-27T21:34:10Z
dc.date.issued2016-10-01
dc.description.abstractCyanobacteria are commonly found in association with other microorganisms, which constitutes a great challenge during the isolation of cyanobacterial strains. Although several methods have been published for obtaining axenic cyanobacterial cultures, their efficiency is usually evaluated by observing the growth of non-cyanobacteria in culture media. In order to verify whether uncultured bacteria should be a concern during cyanobacterial isolation, this work aimed to detect by molecular methods sequences from cyanobacteria and other bacteria present before and after a technique for obtaining axenic cultures from plating and exposure of Fischerella sp. CENA161 akinetes to the Extran detergent and sodium hypochlorite. Solutions containing 0.5, 1, and 2% sodium hypochlorite were able to remove contaminant bacterial CFUs from the culture. However, qPCR pointed that the quantity of sequences amplified with universal bacteria primers was higher than the number of cyanobacteria-specific sequences before and after treatments. The presence of uncultured bacteria in post-hypochlorite cultures was confirmed by high-throughput Illumina sequencing. These results suggest that culturing may overlook the presence of uncultured bacteria associated to cyanobacterial strains and is not sufficient for monitoring the success of cyanobacterial isolation by itself. Molecular methods such as qPCR could be employed as an additional measure for evaluating axenity in cyanobacterial strains. (C) 2016 Elsevier B.V. All rights reserved.en
dc.description.affiliationUniv Sao Paulo, Ctr Nucl Energy Agr, Sao Paulo, Brazil
dc.description.affiliationUniv Estadual Paulista, Fac Ciencias Agr & Vet, Sao Paulo, Brazil
dc.description.affiliationUnespUniv Estadual Paulista, Fac Ciencias Agr & Vet, Sao Paulo, Brazil
dc.description.sponsorshipFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
dc.description.sponsorshipCoordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
dc.description.sponsorshipConselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
dc.description.sponsorshipIdFAPESP: FAPESP 2013/09192-0
dc.description.sponsorshipIdFAPESP: 2013/20142-4
dc.description.sponsorshipIdFAPESP: 2011/08092-6
dc.description.sponsorshipIdFAPESP: 2010/18732-0
dc.description.sponsorshipIdCNPq: 310244/2015-3
dc.format.extent55-60
dc.identifierhttp://dx.doi.org/10.1016/j.mimet.2016.07.023
dc.identifier.citationJournal Of Microbiological Methods. Amsterdam: Elsevier Science Bv, v. 129, p. 55-60, 2016.
dc.identifier.doi10.1016/j.mimet.2016.07.023
dc.identifier.fileWOS000383941500009.pdf
dc.identifier.issn0167-7012
dc.identifier.urihttp://hdl.handle.net/11449/165322
dc.identifier.wosWOS:000383941500009
dc.language.isoeng
dc.publisherElsevier B.V.
dc.relation.ispartofJournal Of Microbiological Methods
dc.relation.ispartofsjr0,696
dc.rights.accessRightsAcesso aberto
dc.sourceWeb of Science
dc.subjectAkinetes
dc.subjectAxenic cultures
dc.subjectFischerella
dc.subjectMetagenomics
dc.subjectSymbiosis
dc.titleEvaluating methods for purifying cyanobacterial cultures by qPCR and high-throughput Illumina sequencingen
dc.typeArtigo
dcterms.licensehttp://www.elsevier.com/about/open-access/open-access-policies/article-posting-policy
dcterms.rightsHolderElsevier B.V.
unesp.author.lattes9429712259649346[5]
unesp.author.orcid0000-0002-8876-3269[5]

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