Changes in the Rumen Microbiota of Cows in Response to Dietary Supplementation with Nitrate, Linseed, and Saponin Alone or in Combination

Abstract

Dietary supplementation with linseed, saponins, and nitrate is a promising methane mitigation strategy in ruminant production. Here, we aimed to assess the effects of these additives on the rumen microbiota in order to understand underlying microbial mechanisms of methane abatement. Two 2-by-2 factorial design studies were conducted simultaneously, which also allowed us to make a broad-based assessment of microbial responses. Eight nonlactating cows were fed diets supplemented with linseed or saponin in order to decrease hydrogen production and nitrate to affect hydrogen consumption; also, combinations of linseed plus nitrate or saponin plus nitrate were used to explore the interaction between dietary treatments. Previous work assessed effects on methane and fermentation patterns. Rumen microbes were studied by sequencing 18S and 16S rRNA genes and ITS1 amplicons. Methanogen activity was monitored by following changes inmcrAtranscript abundance. Nitrate fed alone or in combination in both studies dramatically affected the composition and structure of rumen microbiota, although impacts were more evident in one of the studies. Linseed moderately modified only bacterial community structure. Indicator operational taxonomic unit (OTU) analysis revealed that both linseed and nitrate reduced the relative abundance of hydrogen-producingRuminococcaceae. Linseed increased the proportion of bacteria known to reduce succinate to propionate, whereas nitrate supplementation increased nitrate-reducing bacteria and decreased the metabolic activity of rumen methanogens. Saponins had no effect on the microbiota. Inconsistency found between the two studies with nitrate supplementation could be explained by changes in microbial ecosystem functioning rather than changes in microbial community structure.IMPORTANCEThis study aimed at identifying the microbial mechanisms of enteric methane mitigation when linseed, nitrate, and saponins were fed to nonlactating cows alone or in a combination. Hydrogen is a limiting factor in rumen methanogenesis. We hypothesized that linseed and saponins would affect hydrogen producers and nitrate would affect hydrogen consumption, leading to reduced methane production in the rumen. Contrary to what was predicted, both linseed and nitrate had a deleterious effect on hydrogen producers; linseed also redirected hydrogen consumption toward propionate production, whereas nitrate stimulated the growth of nitrate-reducing and, hence, hydrogen-consuming bacterial taxa. This novel knowledge of microbial mechanisms involved in rumen methanogenesis provides insights for the development and optimization of methane mitigation strategies.

Calcium salts of long-chain fatty acids from linseed oil decrease methane production by altering the rumen microbiome in vitro

“…Dai et al [79] demonstrated that Ruminococcus primarily synthesized putative cellulases and hemicellulases. It is well known that long chain fatty acids inhibit the growth of gram-positive bacteria [80], and supplementary linseed oil reduces Ruminococcaceae [11,81], which agrees with the results in the present study. Rikenellaceae may be associated with either primary or secondary degradation of structural carbohydrates [82].…”

Fecal and Ruminal Microbiome Components Associated With Methane Emission in Beef Cattle.

“…Also, PCoA analysis with the beta-diversity metric Unweighted Unifrac showed the existence of distinct clusters for treatment groups A and B for bacteria and archaea but not for protozoa. Altogether, these results indicate that feed is an important modulator of the microbiome, which agrees with previous studies in which the impact of different diets and feed components were evaluated [40,41] .…”

Fecal and Ruminal Microbiome Components Associated With Methane Emission in Beef Cattle.

“…Also, PCoA analysis with the beta-diversity metric Unweighted Unifrac showed the existence of distinct clusters for treatment groups A and B for bacteria and archaea but not for protozoa. Altogether, these results indicate that feed is an important modulator of the microbiome, which agrees with previous studies in which the impact of different diets and feed components were evaluated [40,41] .…”