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Metaproteomic analysis of an enriched anaerobic rumen consortium (ERAC)

ABSTRACT: Metaproteomic analysis of an enriched anaerobic rumen consortium (ERAC) using sugarcane bagasse and rumen as unique carbon and microbial sources

INSTRUMENT(S): LTQ Orbitrap Velos

ORGANISM(S): Metagenomes

TISSUE(S): Enrichment Culture

SUBMITTER: Geizecler Tomazetto

LAB HEAD: Geizecler Tomazetto

PROVIDER: PXD019219 | Pride | 2020-07-15

REPOSITORIES: pride

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Project description:AIMS: To explore and validate the utility of rumen endoscopy for collection of rumen papillae for gene expression measurement. METHODS: Four adult Coopworth ewes were fasted for either 4 or 24 hours. Animals were sedated, placed in a dorsally recumbent position at 45 degrees with the head upright, and an endoscope inserted via a tube inserted into the mouth. Biopsies of rumen papillae were taken from the ventral surface of the rumen atrium under visual guidance. Two biopsies were collected from one of the animals that had been fasted for 4 hours, and three from one of the animals that had been fasted for 24 hours. Video of the rumen atrium and reticulum was also collected. The animals recovered uneventfully. Biopsies were subsequently used for extraction and sequencing of mRNA. RESULTS: The ventral surface of the rumen atrium was accessible after 4 hours off pasture, but a larger region was accessible after 24 hours of fasting. Sedation allowed access for endoscope use for around 5 to 10 minutes after which increased saliva flow was noted. Rumen papillae biopsies were easily collected, with samples from a variety of sites collected in the ∼10 minute time window. High quality RNA was obtained for stranded mRNA sequencing. Of the resulting reads, 69–70% mapped uniquely to version 3.1 of the ovine genome, and 48–49% to a known gene. The rumen mRNA profiles were consistent with a previously reported study. CONCLUSIONS: This method for obtaining rumenal tissue was found to be rapid and resulted in no apparent short or long term effects on the animal. High quality RNA was successfully extracted and amplified from the rumen papillae biopsies, indicating that this technique could be used for future gene expression studies. The use of rumen endoscopy could be extended to collection of a variety of rumen and reticulum anatomical measurements and deposition and retrieval of small sensors from the rumen. Rumen endoscopy offers an attractive and cost effective approach to repeated rumen biopsies compared with serial slaughter or use of cannulated animals.

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	06jun_Geize_1C.raw	Raw
	06jun_Geize_1D.raw	Raw
	06jun_Geize_2C.raw	Raw
	06jun_Geize_2D.raw	Raw
	06jun_Geize_3C.raw	Raw

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Metaproteomic analysis of an enriched anaerobic rumen consortium (ERAC)

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