Ruminants are major source of methane emissions to the atmosphere, which negatively impacts global warming. A proven and effective means of reducing methane emissions are antibiotic feed additives, the use of which has been banned in the European Community, so we are looking for and alternative, which includes extracts of certain plants. In this research we used anaerobic batch bioreactor system to simulate the conditions in the rumen of sheep, and thus studied and compared in vitro the influence of extracts of garlic, hops, and chesnut tannin on the production of gases (total, CO2, CH4 and H2) and short-chain fatty acids (SCFA). Using molecular methods (TRFLP) and bioinformatic analyses, we studied the influence on these compounds on the composition of the bacterial and archaeal part of the rumen microbiome. The effect of the plant extracts was also compared with the effect of the antibiotic monensin (MON). We found that the addition of selected plant extracts had a statistically significant effect on total gas production as well as methane production in most cases. The greatest effect was obtained by the addition of garlic oil and beta acids from hops, which were even more effective than the addition of MON at higher concentrations. The addition of MON, garlic oil, and beta acids resulted in the production of hydrogen instead of methane, probably due to the inhibition of methanogenesis. Analysis of SCFA showed, that the addition of selected plant extracts led to a decrease in the production of these fatty acids, with the exception of diallyl disulfide (DADS), which increased their production. DADS did not change the ratio of acetic to propionic acid compared with the positive control, where as the other active ingredients decreased it (with the exception of alpha acids from hops). When analysing the profiles of the bacterial and archaeal parts of the microbiome, we found that the addition of each active ingredient had a specific effect on the profile of the microbiomes, with clear differences between them. The analysis of the bacterial part of the microbiomes showed a great similarity between the microbiomes of the samples with added plant extracts and the positive control samples, while the addition of monensin strongly changed that part of microbiomes. A different distribution is characteristic for the achaeal part of the microbiomes, which was most affected by the addition of DADS.
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