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Establishment of a continuous model system to study helicobacter pylori survival in potable water biofilms

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Resumo:Close association of the pathogen Helicobacter pylori in drinking water biofilms has been suggested. Using a two-stage water model, the survival and development of the pathogen in potable water biofilms was monitored. Filter sterilized tap water was used as the growth medium and the inoculum consisted of a naturally occurring consortium of microorganisms. Biofilms were generated on removable stainless steel coupons that were placed in the second vessel. Novel technology peptide nucleic acid (PNA) molecular probes were used to detect and locate the pathogen in the biofilms. The PNA-labelled oligonucleotide probes were highly specific, and complementary to the helix 6 region of H. pylori 16S rRNA. The pathogen was tracked in the biofilms using epifluorescence microscopy and episcopic differential interference contrast microscopy. Results show that H. pylori can successfully incorporate within biofilms and its presence was detected for up to five days after inoculation. PNA probes provided an easy and quick way of performing fluorescence in situ hybridisation assays in heterogeneous biofilms.
Autores principais:Azevedo, N. F.
Outros Autores:Vieira, M. J.; Keevil, C. W.
Assunto:Biofilm Episcopic differential interference contrast microscopy Helicobacter pylori Peptide nucleic acids
Ano:2003
País:Portugal
Tipo de documento:comunicação em conferência
Tipo de acesso:acesso aberto
Instituição associada:Universidade do Minho
Idioma:inglês
Origem:RepositóriUM - Universidade do Minho
Descrição
Resumo:Close association of the pathogen Helicobacter pylori in drinking water biofilms has been suggested. Using a two-stage water model, the survival and development of the pathogen in potable water biofilms was monitored. Filter sterilized tap water was used as the growth medium and the inoculum consisted of a naturally occurring consortium of microorganisms. Biofilms were generated on removable stainless steel coupons that were placed in the second vessel. Novel technology peptide nucleic acid (PNA) molecular probes were used to detect and locate the pathogen in the biofilms. The PNA-labelled oligonucleotide probes were highly specific, and complementary to the helix 6 region of H. pylori 16S rRNA. The pathogen was tracked in the biofilms using epifluorescence microscopy and episcopic differential interference contrast microscopy. Results show that H. pylori can successfully incorporate within biofilms and its presence was detected for up to five days after inoculation. PNA probes provided an easy and quick way of performing fluorescence in situ hybridisation assays in heterogeneous biofilms.