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TitleAn in vitro fermentation model to study the impact of bacteriophages targeting shiga toxin-encoding escherichia coli on the colonic microbiota
Author(s)Pinto, Graça
Shetty, Sudarshan A.
Zoetendal, Erwin G.
Gonçalves, Raquel F. S.
Pinheiro, Ana Cristina
Almeida, Carina Manuela Fernandes
Azeredo, Joana
Smidt, Hauke
Applied microbiology
Next-generation sequencing
Issue date26-Sep-2022
PublisherNature Publishing Group
JournalNpj Biofilms and Microbiomes
CitationPinto, Graça; Shetty, Sudarshan; Zoetendal, Erwin G.; Gonçalves, Raquel F. S.; Pinheiro, Ana Cristina; Almeida, Carina; Azeredo, Joana; Smidt, Hauke, An in vitro fermentation model to study the impact of bacteriophages targeting Shiga toxin-encoding Escherichia coli on the colonic microbiota. npj Biofilms and Microbiomes, 8(74), 2022
Abstract(s)Lytic bacteriophages are considered safe for human consumption as biocontrol agents against foodborne pathogens, in particular in ready-to-eat foodstuffs. Phages could, however, evolve to infect different hosts when passing through the gastrointestinal tract (GIT). This underlines the importance of understanding the impact of phages towards colonic microbiota, particularly towards bacterial families usually found in the colon such as the Enterobacteriaceae. Here we propose in vitro batch fermentation as model for initial safety screening of lytic phages targeting Shiga toxin-producing Escherichia coli (STEC). As inoculum we used faecal material of three healthy donors. To assess phage safety, we monitored fermentation parameters, including short chain fatty acid production and gas production/intake by colonic microbiota. We performed shotgun metagenomic analysis to evaluate the outcome of phage interference with colonic microbiota composition and functional potential. During the 24h incubation, concentrations of phage and its host were also evaluated. We found the phage used in this study, named E. coli phage vB_EcoS_Ace (Ace), to be safe towards human colonic microbiota, independently of the donors faecal content used. This suggests that individuality of donor faecal microbiota did not interfere with phage effect on the fermentations. However, the model revealed that the attenuated STEC strain used as phage host perturbed the faecal microbiota as based on metagenomic analysis, with potential differences in metabolic output. We conclude that the in vitro batch fermentation model used in this study is a reliable safety screening for lytic phages intended to be used as biocontrol agents.
Publisher version
AccessOpen access
Appears in Collections:CEB - Publicações em Revistas/Séries Internacionais / Publications in International Journals/Series

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