Rumen fermentation and methanogenesis are vital metabolic processes in cattle and are carried out by microbial populations that are affected by dietary factors such as secondary metabolites, nutritional composition and degradability. The aim of this study was to monitor populations of total bacteria, total methanogens and Butirivibrio fibrisolvens in the rumen of Lucerne heifers fed on diets typical of intensive silvopastoral systems (ISS) or of a traditional (control) system. Rumen contents (RC) were collected orally from eight heifers consuming 100% Cynodon plectostachyus (control) and 76% C. plectostachyus + 24% Leucaena leucocephala (ISS) following a crossover design and DNA was extracted and quantified by quantitative PCR. Populations [Log10 (ng g-1 RC)] were 5.6 and 5.8 for total bacteria (p = 0.5343), 3.6 and 3.5 for B. fibrisolvens (p = 0.4742) and 5.0 and 5.3 for total methanogens (p = 0.2661) respectively in control and ISS diets. However, when measured in a separate parallel study, enteric methane emissions (g kg-1 of fermented dry matter) were significantly reduced with the inclusion of L. leucocephala. This fact indicated the importance of investigating the structure, function and interactions of populations beyond quantitative analysis to determine how diet affects rumen microbial populations and their function.

The inclusion of Tithonia diversifolia in pasture-based diets is a promising alternative to increase bovine productivity, due to its chemical composition and wide adaptation, but there are few in vivo studies to determine its effect on methane yield and animal production in grazing systems. The objective of this study was to determine the effects of the T. diversifolia inclusion in a basal diet of Brachiaria humidicola on methane (CH4) emissions by enteric fermentation, and on milk yield and quality in dual-purpose cows. The polytunnel technique was used for the determination of methane yield and two diets were evaluated (Diet 1: Brachiaria humidicola 100 %; Diet 2: T. diversifolia 15 % + B. humidicola 85 % dry matter basis) in the moderate rainy and rainy seasons using a cross-over experimental design; milk production was measured by daily milk weighing, and milk quality was determined using a LACTOSCAN analyzer. The inclusion of T. diversifolia did not increase the dry matter intake (P = 0.369), but increased the intake of crude protein and minerals, and reduced fiber intake, resulting in the increased yield of milk and its components in the moderate rainy season (P = 0.012). The inclusion of T. diversifolia reduced the absolute CH4 emissions (P = 0.016), Ym and emission intensity (per unit of fat, protein and kilogram fat and protein corrected milk yields) both in the moderate rainy and rainy seasons (P <0.05). We conclude that the inclusion of T. diversifolia in the forage feed base in the humid tropics such as the Amazon piedmont can be used as a tool to both mitigate enteric CH4 emissions and to increase animal productivity and hence reduce emissions intensity, and thus reduce pressure on the agricultural frontier in critical areas such as the Amazon.

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