Mechanisms Shaping Intra-species Interactions in Streptococcus pneumoniae

Detalhes bibliográficos
Autor(a) principal: Baía, Inês Sofia Lima
Data de Publicação: 2022
Tipo de documento: Dissertação
Idioma: eng
Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
Texto Completo: http://hdl.handle.net/10362/131998
Resumo: Streptococcus pneumoniae, pneumococcus, is a frequent asymptomatic colonizer of the human upper respiratory tract. Although infrequently, pneumococci can reach other body sites and cause diseases. These are associated with high morbidity and mortality worldwide. Co-colonization by more than one pneumococcal strain occurs and interactions can be established between strains. These interactions may be important to shape the pneumococcal population and how it evolves. Although the mechanisms underlying these interactions are poorly studied, they are important to understand vaccine impact and colonization dynamics. A previous study from the group identified interactions occurring between pneumococcal strains grown in an in vitro biofilm model. Two strong interactions were identified: i) an amensalism interaction in which strain 1990-19F inhibited strain 7031-NT’s growth; and ii) a commensalism interaction in which strain 5435-NT benefited when in co-culture with strain 7632-15A. The aim of this thesis was to identify the molecular mechanisms underlying these specific interactions. Experiments were conducted using variants of each strain labelled with green or red fluorescent protein (GFP or RFP, respectively) which enabled cell counts and strain discrimination by flow cytometry in experiments using mixed cultures. The amensalism interaction was also detectable in planktonic cultures. The interaction was shown to be mediated by a secreted protein and possibly other factors. Competence-induced fratricide (a competitive mechanism mediated by protein effectors), medium acidification and differences in the growth patterns of strains did not seem to play a determinant role in the interaction. The commensalism interaction was inconsistent in planktonic growth and, in a transwell experiment, this interaction was shown to be cell contact dependent. The 15A capsule itself was suggested to be important for the beneficial effect observed on the NT strain. In conclusion, this work provided new insights into the factors that may mediate amensalism and commensalism interactions between pneumococcal strains.
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spelling Mechanisms Shaping Intra-species Interactions in Streptococcus pneumoniaeStreptococcus pneumoniaeintra-species interactionsbiofilmsamensalismcommensalismbacteriocinsDomínio/Área Científica::Engenharia e Tecnologia::Outras Engenharias e TecnologiasStreptococcus pneumoniae, pneumococcus, is a frequent asymptomatic colonizer of the human upper respiratory tract. Although infrequently, pneumococci can reach other body sites and cause diseases. These are associated with high morbidity and mortality worldwide. Co-colonization by more than one pneumococcal strain occurs and interactions can be established between strains. These interactions may be important to shape the pneumococcal population and how it evolves. Although the mechanisms underlying these interactions are poorly studied, they are important to understand vaccine impact and colonization dynamics. A previous study from the group identified interactions occurring between pneumococcal strains grown in an in vitro biofilm model. Two strong interactions were identified: i) an amensalism interaction in which strain 1990-19F inhibited strain 7031-NT’s growth; and ii) a commensalism interaction in which strain 5435-NT benefited when in co-culture with strain 7632-15A. The aim of this thesis was to identify the molecular mechanisms underlying these specific interactions. Experiments were conducted using variants of each strain labelled with green or red fluorescent protein (GFP or RFP, respectively) which enabled cell counts and strain discrimination by flow cytometry in experiments using mixed cultures. The amensalism interaction was also detectable in planktonic cultures. The interaction was shown to be mediated by a secreted protein and possibly other factors. Competence-induced fratricide (a competitive mechanism mediated by protein effectors), medium acidification and differences in the growth patterns of strains did not seem to play a determinant role in the interaction. The commensalism interaction was inconsistent in planktonic growth and, in a transwell experiment, this interaction was shown to be cell contact dependent. The 15A capsule itself was suggested to be important for the beneficial effect observed on the NT strain. In conclusion, this work provided new insights into the factors that may mediate amensalism and commensalism interactions between pneumococcal strains.Streptococcus pneumoniae (pneumococo) coloniza assintomaticamente o trato respiratório superior humano. Em algumas circunstâncias o pneumococo causa infeções que estão associadas a uma elevada morbilidade e mortalidade a nível mundial. A colonização simultânea por mais de uma estirpe de pneumococos é frequente, criando oportunidade para estas interagirem. Estas interações parecem ser importantes para a evolução da população pneumocócica. Assim, o estudo das interações que ocorrem é importante para compreender a dinâmica de colonização e avaliar o impacto das vacinas. Um estudo realizado previamente pelo grupo, identificou interações entre estirpes de pneumococos num modelo in vitro de biofilme. Nesse estudo identificaram-se duas interações fortes: i) uma interação de amensalismo em que a estirpe 1990-19F inibiu a estirpe 7031-NT; e ii) uma interação de comensalismo em que a estirpe 5435-NT beneficiou ao crescer com a estirpe 7632-15A. Esta tese teve como objetivo identificar os mecanismos moleculares que modelam as interações acima descritas. Nas experiências realizadas utilizaram-se variantes das estirpes marcadas com GFP ou RFP (green ou red fluorescente protein, respetivamente) que permitiram a contagem de células e a distinção entre estirpes por citometria de fluxo em experiências com co-culturas. A interação de amensalismo foi também detetada em culturas planctónicas. Identificou-se que a interação deverá ser mediada por uma proteína secretada, possivelmente em conjunto com outros fatores. Observou-se que o fratricídio induzido por competência (mediado por efetores proteicos), alterações de pH ou diferenças nos padrões de crescimento das duas estirpes não são determinantes para a interação. A interação de comensalismo não foi consistente em crescimento planctónico. Através de um ensaio transwell, identificou-se que esta deverá ser dependente de contacto celular. Observou-se que a cápsula 15A poderá ser importante para a interação. Em conclusão, este trabalho permitiu identificar potenciais mecanismos mediadores de interações de amensalismo e comensalismo entre estirpes de pneumococos.Sá-Leão, RaquelFilipe, SérgioRUNBaía, Inês Sofia Lima2022-01-172025-01-17T00:00:00Z2022-01-17T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfhttp://hdl.handle.net/10362/131998enginfo:eu-repo/semantics/embargoedAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2024-03-11T05:10:40Zoai:run.unl.pt:10362/131998Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-20T03:47:18.854535Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse
dc.title.none.fl_str_mv Mechanisms Shaping Intra-species Interactions in Streptococcus pneumoniae
title Mechanisms Shaping Intra-species Interactions in Streptococcus pneumoniae
spellingShingle Mechanisms Shaping Intra-species Interactions in Streptococcus pneumoniae
Baía, Inês Sofia Lima
Streptococcus pneumoniae
intra-species interactions
biofilms
amensalism
commensalism
bacteriocins
Domínio/Área Científica::Engenharia e Tecnologia::Outras Engenharias e Tecnologias
title_short Mechanisms Shaping Intra-species Interactions in Streptococcus pneumoniae
title_full Mechanisms Shaping Intra-species Interactions in Streptococcus pneumoniae
title_fullStr Mechanisms Shaping Intra-species Interactions in Streptococcus pneumoniae
title_full_unstemmed Mechanisms Shaping Intra-species Interactions in Streptococcus pneumoniae
title_sort Mechanisms Shaping Intra-species Interactions in Streptococcus pneumoniae
author Baía, Inês Sofia Lima
author_facet Baía, Inês Sofia Lima
author_role author
dc.contributor.none.fl_str_mv Sá-Leão, Raquel
Filipe, Sérgio
RUN
dc.contributor.author.fl_str_mv Baía, Inês Sofia Lima
dc.subject.por.fl_str_mv Streptococcus pneumoniae
intra-species interactions
biofilms
amensalism
commensalism
bacteriocins
Domínio/Área Científica::Engenharia e Tecnologia::Outras Engenharias e Tecnologias
topic Streptococcus pneumoniae
intra-species interactions
biofilms
amensalism
commensalism
bacteriocins
Domínio/Área Científica::Engenharia e Tecnologia::Outras Engenharias e Tecnologias
description Streptococcus pneumoniae, pneumococcus, is a frequent asymptomatic colonizer of the human upper respiratory tract. Although infrequently, pneumococci can reach other body sites and cause diseases. These are associated with high morbidity and mortality worldwide. Co-colonization by more than one pneumococcal strain occurs and interactions can be established between strains. These interactions may be important to shape the pneumococcal population and how it evolves. Although the mechanisms underlying these interactions are poorly studied, they are important to understand vaccine impact and colonization dynamics. A previous study from the group identified interactions occurring between pneumococcal strains grown in an in vitro biofilm model. Two strong interactions were identified: i) an amensalism interaction in which strain 1990-19F inhibited strain 7031-NT’s growth; and ii) a commensalism interaction in which strain 5435-NT benefited when in co-culture with strain 7632-15A. The aim of this thesis was to identify the molecular mechanisms underlying these specific interactions. Experiments were conducted using variants of each strain labelled with green or red fluorescent protein (GFP or RFP, respectively) which enabled cell counts and strain discrimination by flow cytometry in experiments using mixed cultures. The amensalism interaction was also detectable in planktonic cultures. The interaction was shown to be mediated by a secreted protein and possibly other factors. Competence-induced fratricide (a competitive mechanism mediated by protein effectors), medium acidification and differences in the growth patterns of strains did not seem to play a determinant role in the interaction. The commensalism interaction was inconsistent in planktonic growth and, in a transwell experiment, this interaction was shown to be cell contact dependent. The 15A capsule itself was suggested to be important for the beneficial effect observed on the NT strain. In conclusion, this work provided new insights into the factors that may mediate amensalism and commensalism interactions between pneumococcal strains.
publishDate 2022
dc.date.none.fl_str_mv 2022-01-17
2022-01-17T00:00:00Z
2025-01-17T00:00:00Z
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