title: Metagenomic Analysis of the Rumen Microbiome of Steers with Wheat-Induced Frothy Bloat.
abstract: Frothy bloat is a serious metabolic disorder that affects stocker cattle grazing hard red winter wheat forage in the Southern Great Plains causing reduced performance, morbidity, and mortality. We hypothesize that a microbial dysbiosis develops in the rumen microbiome of stocker cattle when grazing on high quality winter wheat pasture that predisposes them to frothy bloat risk. In this study, rumen contents were harvested from six cannulated steers grazing hard red winter wheat (three with bloat score "2" and three with bloat score "0"), extracted for genomic DNA and subjected to 16S rDNA and shotgun sequencing on 454/Roche platform. Approximately 1.5 million reads were sequenced, assembled and assigned for phylogenetic and functional annotations. Bacteria predominated up to 84% of the sequences while archaea contributed to nearly 5% of the sequences. The abundance of archaea was higher in bloated animals (P < 0.05) and dominated by Methanobrevibacter. Predominant bacterial phyla were Firmicutes (65%), Actinobacteria (13%), Bacteroidetes (10%), and Proteobacteria (6%) across all samples. Genera from Firmicutes such as Clostridium, Eubacterium, and Butyrivibrio increased (P < 0.05) while Prevotella from Bacteroidetes decreased in bloated samples. Co-occurrence analysis revealed syntrophic associations between bacteria and archaea in non-bloated samples, however; such interactions faded in bloated samples. Functional annotations of assembled reads to Subsystems database revealed the abundance of several metabolic pathways, with carbohydrate and protein metabolism well represented. Assignment of contigs to CaZy database revealed a greater diversity of Glycosyl Hydrolases dominated by oligosaccharide breaking enzymes (>70%) in non-bloated samples. However, the abundance and diversity of CaZymes were greatly reduced in bloated samples indicating the disruption of carbohydrate metabolism. We conclude that mild to moderate frothy bloat results from tradeoffs both within and between microbial domains due to greater competition for substrates that are of limited availability as a result of biofilm formation.
authors.affiliation: Department of Clinical Studies, School of Veterinary Medicine, University of Pennsylvania Kennett Square, PA, USA.
authors.forename: D W
authors.initials: DW
authors.lastname: Pitta
authors.affiliation: Texas A&M AgriLife Research Vernon, TX, USA.
authors.forename: W E
authors.initials: WE
authors.lastname: Pinchak
authors.affiliation: Department of Clinical Studies, School of Veterinary Medicine, University of Pennsylvania Kennett Square, PA, USA.
authors.forename: N
authors.initials: N
authors.lastname: Indugu
authors.affiliation: Department of Clinical Studies, School of Veterinary Medicine, University of Pennsylvania Kennett Square, PA, USA.
authors.forename: B
authors.initials: B
authors.lastname: Vecchiarelli
authors.affiliation: Department of Microbiology, Perelman School of Medicine, University of Pennsylvania Kennett Square, PA, USA.
authors.forename: R
authors.initials: R
authors.lastname: Sinha
authors.affiliation: Texas A&M AgriLife Research Vernon, TX, USA.
authors.forename: J D
authors.initials: JD
authors.lastname: Fulford
keywords: frothy bloat; metagenomics; rumen microbiome; steers; wheat forage