Direct and culture-enriched 16S rRNA sequencing of cecal content of healthy horses and horses with typhlocolitis.

Autor: Zakia LS; Department of Clinical Studies, Ontario Veterinary College, University of Guelph, Guelph, Ontario, Canada., Gomez DE; Department of Clinical Studies, Ontario Veterinary College, University of Guelph, Guelph, Ontario, Canada., Caddey BB; Department of Production Animal Health, Faculty of Veterinary Medicine, University of Calgary, Calgary, Alberta, Canada., Boerlin P; Department of Pathobiology, Ontario Veterinary College, University of Guelph, Guelph, Ontario, Canada., Surette MG; Department of Biochemistry and Biomedical Sciences, McMaster University, Hamilton, Ontario, Canada., Arroyo LG; Department of Clinical Studies, Ontario Veterinary College, University of Guelph, Guelph, Ontario, Canada.
Jazyk: angličtina
Zdroj: PloS one [PLoS One] 2023 Apr 13; Vol. 18 (4), pp. e0284193. Date of Electronic Publication: 2023 Apr 13 (Print Publication: 2023).
DOI: 10.1371/journal.pone.0284193
Abstrakt: Next generation sequencing has demonstrated that alpha diversity of the fecal microbiota is significantly altered in horses with typhlocolitis. The objective of this study was to evaluate the bacterial composition of the cecum content of horses with and without typhlocolitis through direct and culture-enriched 16S gene sequencing of six healthy horses and six horses with acute typhlocolitis; a case-control study design. Cecal content was collected after euthanasia. An aliquot was used for direct 16S gene sequencing. Another was serially diluted with brain heart infusion (BHI) and plated onto five different agar media. All culture medias, except for MacConkey, were incubated anaerobically. Bacterial colonies were harvested in bulk and used for DNA extraction, 16S PCR amplification, and sequenced using the Illumina MiSeq platform. Predominant phyla in healthy and diseased horses were Firmicutes, followed by Bacteroidetes in all cultured medias, except for MacConkey agar, in which Proteobacteria was the dominant phylum. Greater bacterial richness was identified in sequenced cecal contents as compared to cultured plates (P < 0.05). Culture-enriched molecular profiling combined with 16S rRNA gene sequencing offer an alternative method for the study of the gut microbiota of horses. For direct cecum content 16S gene amplification, the alpha diversity indices were lower in diarrheic horses compared to healthy horses (P < 0.05). A higher relative abundance of Fusobacteriota was found in 2/6 samples from diarrheic horses. The role of Fusobacteriota in equine colitis deserves investigation.
Competing Interests: The authors have declared that no competing interests exist.
(Copyright: © 2023 Zakia et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)
Databáze: MEDLINE
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