Effect of Chitosan and Naringin on Enteric Methane Emissions in Crossbred Heifers Fed Tropical Grass.

In order to meet consumer needs, the livestock industry is increasingly seeking natural feed additives with the ability to improve the efficiency of nutrient utilization, alternatives to antibiotics, and mitigate methane emissions in ruminants. Chitosan (CHI) is a polysaccharide with antimicrobial capability against protozoa and Gram-positive and -negative bacteria, fungi, and yeasts while naringin (NA) is a flavonoid with antimicrobial and antioxidant properties. First, an in vitro gas production experiment was performed adding 0, 1.5, 3.0 g/kg of CHI and NA under a completely randomized design. The substrate containing forage and concentrate in a 70:30 ratio on a dry matter (DM) basis. Compounds increased the concentration of propionic acid, and a significant reduction in methane production was observed with the inclusion of CHI at 1.5 g/kg in in vitro experiments (p < 0.001). In a dry matter rumen degradability study for 96 h, there were no differences in potential and effective degradability. In the in vivo study, six crossbred heifers fitted with rumen cannulas were assigned to a 6 × 6 Latin square design according to the following treatments: control (CTL), no additive; chitosan (CHI1, 1.5 g/kg DMI); (CHI2, 3.0 g/kg DMI); naringin (NA1, 1.5 g/kg DMI); (NA2, 3.0 g/kg DMI) and a mixture of CHI and NA (1.5 + 1.5 g/kg DMI) given directly through the rumen cannula. Additives did not affect rumen fermentation (p > 0.05), DM intake and digestibility of (p > 0.05), and enteric methane emissions (p > 0.05). CHI at a concentration of 1.5 g/kg DM in in vitro experiments had a positive effect on fermentation pattern increasing propionate and reduced methane production. In contrast, in the in vivo studies, there was not a positive effect on rumen fermentation, nor in enteric methane production in crossbred heifers fed a basal ration of tropical grass.

Role of Secondary Plant Metabolites on Enteric Methane Mitigation in Ruminants.

The rumen microbiome plays a fundamental role in all ruminant species, it is involved in health, nutrient utilization, detoxification, and methane emissions. Methane is a greenhouse gas which is eructated in large volumes by ruminants grazing extensive grasslands in the tropical regions of the world. Enteric methane is the largest contributor to the emissions of greenhouse gases originating from animal agriculture. A large variety of plants containing secondary metabolites [essential oils (terpenoids), tannins, saponins, and flavonoids] have been evaluated as cattle feedstuffs and changes in volatile fatty acid proportions and methane synthesis in the rumen have been assessed. Alterations to the rumen microbiome may lead to changes in diversity, composition, and structure of the methanogen community. Legumes containing condensed tannins such as Leucaena leucocephala have shown a good methane mitigating effect when fed at levels of up to 30-35% of ration dry matter in cattle as a result of the effect of condensed tannins on rumen bacteria and methanogens. It has been shown that saponins disrupt the membrane of rumen protozoa, thus decreasing the numbers of both protozoa and methanogenic archaea. Trials carried out with cattle housed in respiration chambers have demonstrated the enteric methane mitigation effect in cattle and sheep of tropical legumes such as Enterolobium cyclocarpum and Samanea saman which contain saponins. Essential oils are volatile constituents of terpenoid or non-terpenoid origin which impair energy metabolism of archaea and have shown reductions of up to 26% in enteric methane emissions in ruminants. There is emerging evidence showing the potential of flavonoids as methane mitigating compounds, but more work is required in vivo to confirm preliminary findings. From the information hereby presented, it is clear that plant secondary metabolites can be a rational approach to modulate the rumen microbiome and modify its function, some species of rumen microbes improve protein and fiber degradation and reduce feed energy loss as methane in ruminants fed tropical plant species.

Influence of energy supplementation on dietary nitrogen utilization and milk production in cows fed foliage of Leucaena leucocephala.

The aim of the study was to assess the effect of four energy supplements (two highly fermentable; two starch-based carbohydrates) on blood urea nitrogen (BUN), urinary urea excretion, and milk yield, in dual-purpose cows fed foliage of Leucaena leucocephala (Leucaena). Five Holstein-Zebu cows with 450 kg body weight in their second third of lactation were used in a 5 × 5 Latin square design. Cows were fed (dry basis) a mixture of 45% Leucaena and 55% Pennisetum purpureum grass. Treatments were supplementation with (i) sugarcane molasses (Mo), (ii) sorghum grain (So), (iii) fresh citrus pulp (CitP) or (iv) rice polishing (RP), all of them incorporated into the diet at 25 MJ of ME/cow/day. There was a control group (Cont) without energy supplementation. The study comprised five periods of 20 days (15 days adaptation, 5 days measurements). Dry matter intake (kg/day) was lower (P < 0.05) for Cont (9.4) compared with Mo (12.1), So (12.0), CitP (11.9) and RP (11.9) but no difference was observed among energy supplements (P > 0.05). Milk yield (kg/day) was higher (P < 0.05) in cows supplemented with starch supplements (4.7 for So; 4.9 for RP) compared with Cont (3.3). Milk yield from highly fermentable supplements (Mo and CitP) did not differ (P > 0.05) from Cont or other treatments. Milk protein, fat and lactose were not different among treatments (P > 0.05). Blood urea nitrogen and urinary urea excretion were both reduced (P < 0.05) by energy supplementation. Urinary urea excretion was not different (P > 0.05) among cows fed different sources of energy. It is concluded that in dual-purpose cows fed Leucaena foliage, supplementation with sugarcane molasses, citrus pulp or rice polishing reduced blood urea nitrogen and urinary urea excretion. Milk yield was increased by sorghum and rice polishing whereas energy supplementation did not affect milk composition.

Effect of Dried Leaves of Leucaena leucocephala on Rumen Fermentation, Rumen Microbial Population, and Enteric Methane Production in Crossbred Heifers.

The effects of dietary inclusion of dried Leucaena leucocephala leaves (DLL) on nutrient digestibility, fermentation parameters, microbial rumen population, and production of enteric methane (CH4) in crossbred heifers were evaluated. Four heifers were used in a 4 × 4 Latin square design consisting of four periods and four levels of inclusion of DLL: 0%, 12%, 24%, and 36% of dry matter (DM) intake. Results showed that DM intake (DMI), organic matter intake, and gross energy intake (GEI) were similar (p > 0.05) among treatments. Apparent digestibility of organic matter, neutral detergent fiber, and energy decreased with increasing levels of DLL in the ration (p < 0.05). In contrast, digestible crude protein (CP) was higher (p < 0.05) in treatments with 12% and 24% DM of DLL. The inclusion of DLL did not affect (p > 0.05) rumen pH and total volatile fatty acids. Rumen microbial community was not affected (p > 0.05) by treatment. There was a linear reduction (p < 0.05) in CH4 emissions as the levels of DLL in the ration were increased. Results of this study suggest that an inclusion of 12% DM of ration as DLL enhances digestible CP and reduces daily production of enteric CH4 without adversely affecting DMI, rumen microbial population, and fermentation parameters.

Rumen function in vivo and in vitro in sheep fed Leucaena leucocephala.

The effect of Leucaena leucocephala inclusion in sheep diets upon rumen function was evaluated. Nine Pelibuey sheep, 32.6 ± 5.33 kg live weight (LW), fitted with rumen cannula were used. A complete randomized block design was employed. Two experimental periods of 60 days each, with 60-day intervals between them, were used. Experimental treatments were as follows (n = 6): T1 (control), 100 % Pennisetum purpureum grass; T2, 20 % L. leucocephala + 80 % P. purpureum; T3, 40 % L. leucocephala + 60 % P. purpureum. In situ rumen neutral detergent fiber (aNDF) and crude protein (CP) degradation, dry matter intake (DMI), volatile fatty acids (VFA) production, estimated methane (CH4) yield, rumen pH, ammonia nitrogen (N-NH3), and protozoa counts were measured. The aNDF in situ rumen degradation of P. purpureum and leucaena was higher (P < 0.05) in T2 and T3. Leucaena CP degradation was higher in T2 and T3 but for P. purpureum it was only significantly higher in T3. Leucaena aNDF and CP degradation rate (c) was 50 % higher (P < 0.05) in T2 and T3, but only higher in T3 for P. purpureum. Voluntary intake and rumen (N-NH3) was higher in T2 and T3 (P = 0.0001, P = 0.005, respectively). Molar VFA proportions were similar for all treatments (P > 0.05). Protozoa counts and in vitro gas production (48 h) were lower in T2 and T3 (P < 0.05, P < 0.0001). Estimated methane yield (mol CH4/day) was higher in sheep fed leucaena (P < 0.0001). However, CH4 yield relative to animal performance (mol CH4/g LW gain) was lower in T2 and T3 (P < 0.0001). In summary, these results indicate that including L. leucocephala in sheep diets did not modify rumen fermentation pattern (same VFA ratios) nor reduce the amount of CH4 per unit of DMI (mol CH4/g DMI). However, leucaena inclusion does increase rumen N-NH3, aNDF and CP digestibility, and voluntary intake.