Different concentrations (3, 30, 300, and 3000 mg/L of culture fluid) of garlic oil (GAR), diallyl sulfide (DAS), diallyl disulfide (DAD), allicin (ALL), and allyl mercaptan (ALM) were incubated for 24 h in diluted ruminal fluid with a 50:50 forage:concentrate diet (17.7% crude protein; 30.7% neutral detergent fiber) to evaluate their effects on rumen microbial fermentation. Garlic oil (30 and 300 mg/L), DAD (30 and 300 mg/L), and ALM (300 mg/L) resulted in lower molar proportion of acetate and higher proportions of propionate and butyrate. In contrast, at 300 mg/L, DAS only increased the proportion of butyrate, and ALL had no effects on volatile fatty acid proportions. In a dual-flow continuous culture of rumen fluid fed the same 50:50 forage:concentrate diet, addition of GAR (312 mg/L), DAD (31.2 and 312 mg/L), and ALM (31.2 and 312 mg/L) resulted in similar changes to those observed in batch culture, with the exception of the lack of effect of DAD on the proportion of propionate. In a third in vitro study, the potential of GAR (300 mg/L), DAD (300 mg/L), and ALM (300 mg/L) to decrease methane production was evaluated. Treatments GAR, DAD, and ALM resulted in a decrease in methane production of 73.6, 68.5, and 19.5%, respectively, compared with the control. These results confirm the ability of GAR, DAD, and ALM to decrease methane production, which may help to improve the efficiency of energy use in the rumen.

The growing demand for sustainable animal production is compelling researchers to explore the potential approaches to reduce emissions of greenhouse gases from livestock that are mainly produced by enteric fermentation. Some potential solutions, for instance, the use of chemical inhibitors to reduce methanogenesis, are not feasible in routine use due to their toxicity to ruminants, inhibition of efficient rumen function or other transitory effects. Strategies, such as use of plant secondary metabolites and dietary manipulations have emerged to reduce the methane emission, but these still require extensive research before these can be recommended and deployed in the livestock industry sector. Furthermore, immunization vaccines for methanogens and phages are also under investigation for mitigation of enteric methanogenesis. The increasing knowledge of methanogenic diversity in rumen, DNA sequencing technologies and bioinformatics have paved the way for chemogenomic strategies by targeting methane producers. Chemogenomics will help in finding target enzymes and proteins, which will further assist in the screening of natural as well chemical inhibitors. The construction of a methanogenic gene catalogue through these approaches is an attainable objective. This will lead to understand the microbiome function, its relation with the host and feeds, and therefore, will form the basis of practically viable and eco-friendly methane mitigation approaches, while improving the ruminant productivity.

Incubation trials were carried out with the rumen simulation technique (RUSITEC) to study the effects of four forms of N on the growth of ruminal micro-organisms and the fermentation variables when an all-fibre basal diet was incubated. The basal diet consisted of 10 g neutral-detergent fibre (NDF) from grass hay plus 2 g NDF from sugarbeet pulp. N forms were isolated soyabean protein, soyabean peptides, amino acids blended to profile soyabean protein and NH₃ as NH₄Cl. Half of the daily N supply was infused as NH₄Cl and the other half was infused as each of the four treatments described. Non-NH₃ N (NAN) forms increased NDF (P = 0·006), acid-detergent fibre (P = 0·003) and cellulose (P = 0·015) disappearance after 48 h incubation, CO₂ (P < 0·001), CH₄ (P = 0·002) and total volatile fatty acids production (P < 0·001), as well as the molar percentages of isobutyrate, isovalerate and valerate, which reflected the fermentation of amino acid C skeletons. NAN treatments also increased microbial N flow (P < 0·001) compared with NH₃, with peptides and protein supporting more (P = 0·036) than amino acids. The proportion of microbial N derived from NH₃ decreased successively (P < 0·05) with NH₃ > amino acids > peptides > protein treatments, indicating preferential uptake of peptides without passage through the NH₃ pool. Microbial efficiency (g microbial N/kg organic matter apparent disappearance) was greater (P = 0·002) for the NAN forms than for the NH₃ treatment, with peptides and protein treatments supporting higher (P = 0·009) values than amino acid treatment. These results indicate that N forms other than NH₃ are required for optimal fibre digestion and microbial growth.

The effects of disodium fumarate on microbial growth, CH₄production and fermentation of three diets differing in their forage content (800, 500 and 200 g/kg DM) by rumen micro-organismsin vitrowere studied using batch cultures. Rumen contents were collected from four Merino sheep. Disodium fumarate was added to the incubation bottles to achieve final concentrations of 0, 4 and 8 mm-fumarate, and¹⁵N was used as a microbial marker. Gas production was measured at regular intervals from 0 to 120 h of incubation. Fumarate did not affect (P>0·05) any of the measured gas production parameters. In 17 h incubations, the final pH and the production of acetate and propionate were increased linearly (P0·05) were observed for the other two diets. These results indicate that the effects of fumarate on rumen fermentation depend on the nature of the incubated substrate, the high-forage diet showing the greatest response.

The nutritive value of 26 agro-industrial by-products was assessed from their chemical composition, in vitro digestibility and rumen fermentation kinetics. By-products from sugar beet, grape, olive tree, almond, broccoli, lettuce, asparagus, green bean, artichoke, peas, broad beans, tomato, pepper, apple pomace and citrus were evaluated. Chemical composition, in vitro digestibility and fermentation kinetics varied largely across the by-products. Data were subjected to multivariate and principal component analyses (PCA). According to a multivariate cluster analysis chart, samples formed four distinctive groups (A–D). Less degradable by-products were olive tree leaves, pepper skins and grape seeds (group A); whereas the more degradable ones were sugar beet, orange, lemon and clementine pulps (group D). In the PCA plot, component 1 segregated samples of groups A and B from those of groups C and D. Considering the large variability among by-products, most of them can be regarded as potential ingredients in ruminant rations. Depending on the characteristic nutritive value of each by-product, these feedstuffs can provide alternative sources of energy (e.g., citrus pulps), protein (e.g., asparagus rinds), soluble fibre (e.g., sugar beet pulp) or less digestible roughage (e.g., grape seeds or pepper skin).