The greenhouse gases (GHGs) derived from agriculture include carbon dioxide, nitrous oxide, and methane (CH4). Of these GHGs, CH4, in particular, constitutes a major component of the GHG emitted by the agricultural sector. Along with environmental concerns, CH4 emission also leads to losses in gross energy intake with economic implications. While ruminants are considered the main source of CH4 from agriculture, nonruminant animals also contribute substantially, and the CH4 emission intensity of nonruminants remains comparable to that of ruminants. Means of mitigating CH4 emissions from enteric fermentation have therefore been sought. Methane is produced by methanogens-archaeal microorganisms that inhabit the digestive tracts of animals and participate in fermentation processes. As the diversity of methanogen communities is thought to be responsible for the differences in CH4 production among nonruminant animals, it is necessary to investigate the archaeal composition of specific animal species. Methanogens play an important role in energy metabolism and adipose tissue deposition in animals. Higher abundances of methanogens, along with their higher diversity, have been reported to contribute to lean phenotype in pigs. In particular, a greater abundance of Methanosphaera spp. and early dominance of Methanobrevibacter smithii have been reported to correlate with lower body fat formation in pigs. Besides the contribution of methanogens to the metabolic phenotype of their hosts, CH4 release reduces the productivity that could be achieved through other hydrogen (H2) disposal pathways. Enhanced participation of acetogenesis in H2 disposal, leading to acetate formation, could be a more favorable direction for animal production and the environment. Better knowledge and understanding of the archaeal communities of the gastrointestinal tract (GIT), including their metabolism and interactions with other microorganisms, would thus allow the development of new strategies for inhibiting methanogens and shifting toward acetogenesis. There are a variety of approaches to inhibiting methanogens and mitigating methanogenesis in ruminants, which can find an application for nonruminants, such as nutritional changes through supplementation with biologically active compounds and management changes. We summarize the available reports and provide a comprehensive review of methanogens living in the GIT of various nonruminants, such as swine, horses, donkeys, rabbits, and poultry. This review will help in a better understanding of the populations and diversity of methanogens and the implications of their presence in nonruminant animals.

The efficacy of methane (CH₄) suppression using medium‐chain fatty acids (MCFA) remains inconclusive, despite a number of studies on this topic are available. We thus carried out a meta‐analysis to integrate the published data on different concentrations and types of MCFA such as lauric acid and myristic acid, which investigated ruminal methanogenesis and fermentation in in vitro and in vivo experiments. In vitro MCFA sources were classified either as pure MCFA (lauric acid, myristic acid and their combinations) or as natural MCFA‐rich oils (canola oil enriched with lauric acids, coconut oil, krabok oil and palm kernel oil). The MCFA sources used in the in vivo studies were coconut oil, lauric acid, myristic acid and the combination of lauric and myristic acids. A total of 41 studies (20 in vitro and 21 in vivo studies) were compiled in our database, which included the data on CH₄emission, digestibility, ruminal fermentation products and microbial populations. The results showed that the amount of CH₄production per unit of digested organic matter decreased linearly under in vitro conditions (p< .01) and tended to decrease quadratically under in vivo conditions (p< .07) with increasing doses of MCFA. Populations of protozoa (p< .01) in both in vitro and in vivo responded negatively in a linear manner, whereasArchaeapopulation diminished quadratically (p= .04) only in the in vitro conditions with increasing doses of MCFA. Increasing dietary MCFA concentrations also reduced the fibre digestibility linearly (p< .05) in both in vitro and in vivo conditions. CH₄production for different sources of MCFA decreased in following order: coconut oil > lauric acid > myristic acid > mixed lauric and myristic acids > palm kernel oil > canola oil enriched with lauric acids > krabok oil. It can be concluded that the effect of MCFA on ruminal methanogenesis depends on the amount and type of MCFA.

The study was conducted to evaluate the effects of partial replacement of soybean meal (SBM) by 15% raw or fermented rapeseed cake (RRC or FRC) to broilers’ diets on growth performance, nutrient utilization, methane emission, and breast muscle fatty acid (FA) composition. A total of 420 one-day-old female Ros 308 broilers were used in three independent experiments (300 birds in the first experiment and 60 in the second and third experiments). In each trial, three treatments were set up: a control group (without rapeseed), and diets replaced soybean meal with 15% addition of RRC or FRC. Birds fed the FRC diet experienced no effect (p > 0.05) on performance or nutrients utilization. Methane emission and total methanogen population in the ceca was decreased (p < 0.05) with the FRC diet. The concentrations of n-3 and n-6 FAs in the breast tissue of fourteen-day-old birds were not affected (p > 0.05) by FRC. However, the n-6/n-3 ratio in the breast muscle of 28-day-old birds was reduced (p < 0.001). In conclusion, the replacement of SBM by FRC in the broiler diets did not show any unfavorable effects on performance or nutrient utilization. Furthermore, the breast meat FA profile was improved, methanogen counts significantly decreased, and methane emission was limited.

It was hypothesised that Nannochloropsis limnetica due to the specific chemical composition and fatty acids profile, may positively affect rumen fermentation. To confirm this hypothesis the batch culture experiment was conducted to evaluate N. limnetica supplemented at 0, 2, 4 and 6% of the substrate dry matter (DM) on both fermentation and fatty acid proportion in the ruminal culture. It was found that microalgae N. limnetica contain (g · kg−1 DM): crude protein 238, Ca 48.7, Na 31.8 and unsaturated fatty acids (51.7 g · 100 g−1 fatty acids). Moreover, leucine and lysine were the most abundant essential amino acids in the analysed microalgae. The total bacteria count was negatively affected if N. limnetica algae were supplemented at more than 4%. So, the research hypothesis that microalgae N. limnetica may affect rumen fermentation was confirmed, mainly by increasing propionic acid concentration without changes in the total volatile fatty acids concentration. However, a high dose (6%) decreased rumen bacteria count. Further research under commercial farm conditions should be conducted to confirm the usefulness of freshwater microalgae N. limnetica as a feed additive for ruminants.

Background: The use of industrial by-products rich in bioactive compounds as animal feeds can reduce greenhouse gas production. Paulownia leaves silage (PLS) was supplemented to dairy cows' diet and evaluated in vitro (Exp. 1; Rusitec) and in vivo (Exp. 2, cannulated lactating dairy cows and Exp. 3, non-cannulated lactating dairy cows). The study investigated the PLS effect on ruminal fermentation, microbial populations, methane production and concentration, dry matter intake (DMI), and fatty acid (FA) proportions in ruminal fluid and milk. Results: Several variables of the ruminal fluid were changed in response to the inclusion of PLS. In Exp. 1, the pH increased linearly and quadratically, whereas ammonia and total volatile fatty acid (VFA) concentrations increased linearly and cubically. A linear, quadratic, and cubical decrease in methane concentration was observed with increasing dose of the PLS. Exp. 2 revealed an increase in ruminal pH and ammonia concentrations, but no changes in total VFA concentration. Inclusion of PLS increased ruminal propionate (at 3 h and 6 h after feeding), isovalerate, and valerate concentrations. Addition of PLS also affected several populations of the analyzed microorganisms. The abundances of protozoa and bacteria were increased, whereas the abundance of archaea were decreased by PLS. Methane production decreased by 11% and 14% in PLS-fed cows compared to the control in Exp. 2 and 3, respectively. Exp. 3 revealed a reduction in the milk protein and lactose yield in the PLS-fed cows, but no effect on DMI and energy corrected milk yield. Also, the PLS diet affected the ruminal biohydrogenation process with an increased proportions of C18:3 cis-9 cis-12 cis-15, conjugated linoleic acid, C18:1 trans-11 FA, polyunsaturated fatty acids (PUFA), and reduced n6/n3 ratio and saturated fatty acids (SFA) proportion in milk. The relative transcript abundances of the 5 of 6 analyzed genes regulating FA metabolism increased. Conclusions: The dietary PLS replacing the alfalfa silage at 60 g/kg diet can reduce the methane emission and improve milk quality with greater proportions of PUFA, including conjugated linoleic acid, and C18:1 trans-11 along with reduction of SFA. Graphical Graphical abstract of the experimental roadmap

Active plant metabolites (APM) are recognized as modifiers of ruminal microbial fermentation including methanogenesis and biohydrogenation of fatty acids (FA). Coleus amboinicus Lour. leaves (CAL) are rich in several APM, which could serve as ruminal fermentation modulators. A phytochemical analysis showed that CAL contain phenolic acids (10.4 mg·g^-1 dry matter (DM); high in rosmarinic acid), flavonoids (2.6 mg·g^-1 DM), diterpenes (2 mg·g^-1 DM), and linolenic acid (35.4 g (100 g) ^-1 FA). The present study aimed to investigate the effect of CAL on ruminal methanogenesis and biohydrogenation as well as basic fermentation characteristics and microbial populations. The in vitro experiment was carried out using Hohenheim gas test system with 40 ml of buffered ruminal fluid incubated for 24 h at 39 °C in anaerobic conditions. Approximately 400 mg (DM basis) of total mixed ration (TMR) was used as a control substrate and the CAL substrate was used at doses of 10, 20, 40, and 80 mg DM replacing equal amounts of TMR. Addition of CAL decreased methane production up to 30% linearly as the amount of CAL increased (P < 0.05). In vitro dry matter digestibility and ammonia tended to increase with increasing doses of CAL. Concentration of total volatile fatty acids was not affected by the CAL although there appeared to be a minor positive linear trend; however, acetate, butyrate and isobutyrate proportion increased quadratically (P < 0.001). CAL tended to linearly increase α-linolenic acid and conjugated linoleic acid as well as increased stearic acid concentration in buffered ruminal fluid. CAL particularly increased total protozoa and bacterial populations during fermentation, but inhibited methanogens. It is concluded that the CAL leaves may be promishing to be used as a feed additive to decrease methanogenesis well as biohydrogenation of FA in the rumen.