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Master's Dissertation
DOI
https://doi.org/10.11606/D.87.2011.tde-27092011-095344
Document
Author
Full name
Priscila Ikeda Ushimaru
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2011
Supervisor
Committee
Pellizari, Vivian Helena (President)
Garboggini, Fabiana Fantinatti
Matte, Maria Helena
Title in Portuguese
Aplicação de metodologias de isolamento de bactérias ainda 'não-cultivadas' em ecossistemas marinhos.
Keywords in Portuguese
Bactérias (isolamento e purificação)
Ecossistemas marinhos
Fenótipos (análise)
Microbiologia ambiental
Sequenciamento genético
Abstract in Portuguese
Aplicou-se duas metodologias para isolamento de bactérias ainda "não-cultivadas" em amostras de água do mar de Ubatuba (SP, Brasil) e da Baía do Almirantado (Antártica). Pela adaptação do método de cultivo de alto desempenho (HTC), foi possível isolar 4 culturas bacterianas, nas quais duas podem ser consideradas ainda não-cultivadas e foram identificadas através das análises filogenéticas do gene rRNA 16S. Ao aplicar o método de inóculo em meio de cultura diluído 1/10, nas amostras de água do mar antártico, 81 isolados bacterianos foram identificados (gene rRNA 16S), sendo que um deles, é candidato a um isolado "não-cultivado". Outras abordagens fenotípicas e genômicas serão necessárias para definir a caracterização taxonômica destas bactérias "não-cultivadas" obtidas. A presença dos genes alk foi detectada em 11 isolados bacterianos antárticos, destes, um representante apresentou similaridade com uma sequência de gene alkM, recém-descrita em estudo prévio, em um dos clones de bibliotecas metagenômicas de sedimentos, na mesma região de amostragem.
Title in English
Applications of methods for isolating uncultured bacteria in marine environments.
Keywords in English
Bacteria (isolation and purification)
Environmental microbiology
Genetic sequencing
Marine ecosystems
Phenotypes (analysis)
Abstract in English
Two different methods were applied for culturing marine bacteria in order to isolate uncultured representatives from Ubatuba seawater (SP, Brazil) and from Admiralty Bay (Antarctica). The adapted high-throughput culturing (HTC) procedures allowed to obtain four isolates. Two of them are uncultured bacteria candidates from coastal seawater studied and they were identified by phylogenetic analysis for 16S rRNA genes. The use of traditional plating on 1/10 dilution of agar media for the Antarctica seawater samples, allowed to isolate 81 cultures that were identified (16S rRNA gene), and one of these isolates is most likely to be an uncultured micro-organism. For taxonomic purposes, several others phenotypic and genomic methods must be applied for the further characterization of these uncultured bacteria. The alk genes were detected in eleven bacterial isolates from Antarctic. One of them, showed similarity to the sequence of an alkM gene, described in previous work in environmental clones libraries od sediments, from the same sampling area.
 
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Publishing Date
2011-09-29
 
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