Quantitative assessment of individual populations within polymicrobial biofilms

• Type: Article in International Scientific Journal
• Date: 2018

Authors

Susana Patrícia Lopes (Author) Other; Nuno Filipe Azevedo (Author) FEUP; Maria Olívia Pereira (Author) Other

Journal

Title: Scientific Reports

Serial No. 1 Vol. 8 No. 9494

Pages: 1-13

ISSN: 2045-2322

Publisher: Springer Nature

Indexing

ISI Web of Knowledge - 4 Citations

Scopus - 29 Citations

Associated Projects

LEPABE - Laboratório de Engenharia de Processos, Ambiente, Biotecnologia e Energia

LEPABE-2-ECO-INNOVATION - LEPABE-2-ECO-INNOVATION

Coded-FISH - Desenvolvimento de um novo método de RING FISH codificado por cores: uma ferramenta inovadora para a deteção de patogénios resistentes a antibióticos

Other information

Authenticus ID: P-00R-A8M

DOI: 10.1038/s41598-018-27497-9

Abstract (EN): Selecting appropriate tools providing reliable quantitative measures of individual populations in biofilms is critical as we now recognize their true polymicrobial and heterogeneous nature. Here, plate count, quantitative real-time polymerase chain reaction (q-PCR) and peptide nucleic acid probe-fluorescence in situ hybridization (PNA-FISH) were employed to quantitate cystic fibrosis multispecies biofilms. Growth of Pseudomonas aeruginosa, lnquilinus limosus and Dolosigranulum pigrum was assessed in dual- and triple-species consortia under oxygen and antibiotic stress. Quantification methods, that were previously optimized and validated in planktonic consortia, were not always in agreement when applied in multispecies biofilms. Discrepancies in culture and molecular outcomes were observed, particularly for triple-species consortia and antibiotic-stressed biofilms. Some differences were observed, such as the higher bacterial counts obtained by q-PCR and/or PNA-FISH (<= 4 log(10) cells/cm(2)) compared to culture. But the discrepancies between PNA-FISH and q-PCR data (eg D. pigrum limited assessment by q-PCR) demonstrate the effect of biofilm heterogeneity in method's reliability. As the heterogeneity in biofilms is a reflection of a myriad of variables, tailoring an accurate picture of communities' changes is crucial. This work demonstrates that at least two, but preferentially three, quantification techniques are required to obtain reliable measures and take comprehensive analysis of polymicrobial biofilm-associated infections.

Language: English

Type (Professor's evaluation): Scientific

No. of pages: 13