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Rumen function is generally suboptimal leading to losses in methane and nitrogen. Analysis of the rumen microbiome is thus important to understanding the underlying microbial activity under different feeding strategies. This study investigated the effect of forage conservation method and vitamin E supplementation on rumen function using a rumen simulation technique. Ryegrass (GRA) or ryegrass hay (HAY) was supplemented with 20% concentrate containing zero or 50 IU/d vitamin E, as α-tocopheryl acetate, according to a 2 × 2 factorial design. The forage conservation method did not substantially change the nutrient composition but had a profound impact on the structure and diversity of the rumen microbiome. HAY diets promoted a more complex bacterial community (+38 OTUs) dominated by  Firmicutes . This bacterial adaptation, together with increased rumen protozoa levels and methanogen diversity, was associated with greater fiber disappearance (+12%) in HAY diets, but also with greater rumen true N degradability (+7%) than GRA diets. HAY diets also had a higher metabolic H recovery and methane production (+35%) suggesting more efficient inter-species H transfer between bacteria, protozoa and methanogens. Contrarily, GRA diets promoted more simplified methanogen and bacterial communities, which were dominated by  Bacteroidetes  and  Lactobacillus , thus lactate formation may have acted as an alternative H sink in GRA diets. Moreover the structure of the bacterial community with GRA diets was highly correlated with N utilization, and GRA diets promoted greater bacterial growth and microbial protein synthesis (+16%), as well as a more efficient microbial protein synthesis (+22%). A dose-response experiment using batch cultures revealed that vitamin E supplementation increased rumen fermentation in terms of total VFA and gas production, with protozoal activity higher when supplying α-tocopheryl acetate vs. α-tocopherol. Moreover, α-tocopheryl acetate promoted a small increase in feed degradability (+8%), possibly as a result of its antioxidant properties which led to higher bacterial and protozoal levels. Vitamin E supplementation also modified the levels of some methanogen species indicating that they may be particularly sensitive to oxidative stresses. Our findings suggested that when possible, grass should be fed instead of grass hay, in order to improve rumen function and to decrease the environmental impact of livestock agriculture.

An Integrated Multi-Omics Approach Reveals the Effects of Supplementing Grass or Grass Hay with Vitamin E on the Rumen Microbiome and Its Function