Redução da produção de metano por meio da modulação ruminal com suplementação de algas tropicais em capim-Napier

Short chain (C2-C6) fatty acids are produced in the colon through bacterial fermentation of mainly dietary fibre. Butyrate (C4) possesses antineoplastic effects on human colon carcinoma cells, and epidemiological studies indicate that high fibre diets may reduce the incidence of colorectal cancer. The role of dietary fibre during colorectal carcinogenesis might therefore be related to its fermentation to butyrate. Faecal concentrations of total short chain fatty acids and concentrations and ratios of the individual C2-C6 fatty acids did not differ between 16 healthy controls, 17 patients with colonic adenomas, and 17 patients with colonic cancer. Comparison of the molar production velocities (mmol/l.hour) of total and individual short chain fatty acids from glucose, ispagula, wheat bran, and albumin in six and 24 hour faecal incubations showed no differences. The ratio of butyrate production to total short chain fatty acid production from fibre, however, was reduced in patients with colonic cancer and adenomas compared with healthy controls (ispagula, six hours: 6.4, 7.6, and 11.5% respectively, p = 0.005 and 24 hour: 9.1, 9.9, and 15.4%, p = 0.002; wheat bran, six hours: 9.9, 10.2, and 14.7% respectively, p = 0.06 and 24 hours: 15.1, 16.8, and 21.0%, p = 0.01). It may be that the low ratios of colonic butyrate formation combined with low fibre diets increase the risk of colonic neoplasia.

Dependence of forage quality and methanogenic potential of tropical plants on their phenolic fractions as determined by principal component analysis

The influence of addition of gallic acid, tannic acid, or quebracho tannins to alfalfa hay on in vitro rumen fermentation and microbial protein synthesis

This study evaluates the effect of supplementing Acanthopora muscoides (L.) Bory on Pennisetum purpureum cv. Gama Umami as a basal diet on methane gas production and protozoa population. In June 2022, natural tropical seaweed A. muscoides (L.) Bory was harvested from Drini and Sepanjang Beach, Gunungkidul District, Yogyakarta Province, Indonesia. The setup of this research design used a randomized complete block with four treatments and replications. The methane production was measured with Gas Chromatography and the protozoa population was counted using a microscope with 10 or 40x magnification. The outcome demonstrated that taking supplements of A. muscoides (L.) Bory at 2.5-7.5% organic matter (OM) on P. purpureum cv. Gama Umami reduced (P<0.01) methane production on 24 and 48 h of incubation. Higher supplementation levels of A. muscoides (L.) Bory significantly reduced (P<0.01) methane production. Reducing (P<0.01) methane up to 31.40% and 34.72% on 24 and 48 h of incubation with A. muscoides (L.) Bory supplementation at 7.5% OM. This level also declined (P<0.01) in 28.14% of protozoa populations by 5.01 to 3.60 cells x 104/mL. It is concluded that taking A. muscoides (L.) Bory supplements at 7.5% OM on P. purpureum cv. Gama Umami resulted in the lowest methane production and protozoa population. A. muscoides (L.) Bory, one of a tropical red seaweeds, potential to integrate as an feed additive into sustainable livestock feeding practices

In vitro screening of plant extracts to enhance the efficiency of utilization of energy and nitrogen in ruminant diets

Background and Aim:Herbal plants have the potential to reduce the population of metagonic bacteria and protozoa due to the bioactive compound contained in herbal plants. This study aimed to evaluate the effect of herbal plant supplementation on rumen fermentation characteristics, methane (CH4) gas emissions, in vitro nutrient digestibility, and protozoan populations.Materials and Methods:This study consisted of two stages. Stage I involved determining the potential of herbal plants to increase total gas production (Orskov and McDonald methods) and reduce the protozoan population (Hristov method). Three potential herbs were selected at this stage and used in Stage II as supplements in the palm kernel cake (PKC)-based diet (30% herbal plants + 70% PKC). Proximate and Van Soest analyses were used to determine the chemical composition. In vitro dry matter digestibility (IVDMD), organic matter (IVOMD), and rumen fermentation characteristics were determined using Theodorous method. Conway microdiffusion was used to determine ammonia concentration (NH3). Gas chromatography was used to determine the total and partial volatile fatty acid production.Results:The results of the first stage showed that seven herbal plants (Moringa oleifera, Rhodomyrtus tomentosa, Clerodendron serratum, Curcuma longa Linn., Urena lobata, Uncaria, and Parkia timoriana) significantly differed in terms of total gas production (p < 0.05). Herbal plants can increase gas production and reduce protozoan populations. The highest total gas production was observed using P. timoriana, M. oleifera, and C. longa Linn. Moringa oleifera plants were the most effective in lowering protozoa population. In Stage 2, the supplementation of herbal plants in PKC-based-diet significantly increased IVDMD, that was ranged from 56.72% to 65.77%, IVOMD that was ranged from 52.10% to 59.54%, and NH3, that was ranged from 13.20 mM to 17.91 mM. Volatile fatty acid partial and total gas production potential and CH4 gas emissions were also significantly different from those of the control (p < 0.05).Conclusion:Supplementation of M. oleifera, C. longa Linn., and P. timoriana in ruminant diet effectively increased total gas production, IVDMD percentage, and IVOMD, and reduced CH4 gas emissions and protozoa populations during rumen fermentation.

In this study, a red macroalgae species, Acanthopora muscoides (L.) Bory was processed as a supplement Pennisetum purpureum cv. Gama Umami was used to evaluate their in vitro ruminant methane characteristics. This parameter will give us insight into the fermentation product from the feed degradation process condition in the rumen. A complete randomized block design with four treatments and four replications was used, with a time of rumen fluid collection as a block. The treatments consisted of control (only Pennisetum purpureum cv. Gama Umami as a basal diet) and supplementation Acanthopora muscoides (L.) Bory at 2.5, 5.0, and 7.5% organic matter (OM). The result showed that supplementing Acanthopora muscoides (L.) Bory up to 7.5.0% OM on Pennisetum purpureum cv. Gama Umami increased (P&lt;0.01) total volatile fatty acid (VFA) production, propionate proportion, and microbial protein synthesis. Those treatments decreased (P&lt;0.01) the proportion of acetate, butyrate, and acetate propionate ratio without disturbing pH value and NH3 concentration. The conclusion was Pennisetum purpureum cv. Gama Umami supplemented with Acanthopora muscoides (L.) Bory 7.5% OM is the best treatment in this study to increase rumen fermentation characteristics.

BackgroundThe awareness of environmental and socio-economic impacts caused by greenhouse gas emissions from the livestock sector leverages the adoption of strategies to counteract it. Feed supplements can play an important role in the reduction of the main greenhouse gas produced by ruminants—methane (CH4). In this context, this study aims to assess the effect of two biochar sources and inclusion levels on rumen fermentation parameters in vitro.MethodsTwo sources of biochar (agro-forestry residues, AFB, and potato peel, PPB) were added at two levels (5 and 10%, dry matter (DM) basis) to two basal substrates (haylage and corn silage) and incubated 24-h with rumen inocula to assess the effects on CH4 production and main rumen fermentation parameters in vitro.ResultsAFB and PPB were obtained at different carbonization conditions resulting in different apparent surface areas, ash content, pH at the point of zero charge (pHpzc), and elemental analysis. Relative to control (0% biochar), biochar supplementation kept unaffected total gas production and yield (mL and mL/g DM, p = 0.140 and p = 0.240, respectively) and fermentation pH (p = 0.666), increased CH4production and yield (mL and mL/g DM, respectively, p = 0.001) and ammonia-N (NH3-N, p = 0.040), and decreased total volatile fatty acids (VFA) production (p < 0.001) and H2 generated and consumed (p ≤ 0.001). Biochar sources and inclusion levels had no negative effect on most of the fermentation parameters and efficiency. Acetic:propionic acid ratio (p = 0.048) and H2 consumed (p = 0.019) were lower with AFB inclusion when compared to PPB. Biochar inclusion at 10% reduced H2 consumed (p < 0.001) and tended to reduce total gas production (p = 0.055). Total VFA production (p = 0.019), acetic acid proportion (p = 0.011) and H2 generated (p = 0.048) were the lowest with AFB supplemented at 10%, no differences being observed among the other treatments. The basal substrate affected most fermentation parameters independently of biochar source and level used.DiscussionBiochar supplementation increased NH3-N content, iso-butyric, iso-valeric and valeric acid proportions, and decreased VFA production suggesting a reduced energy supply for microbial growth, higher proteolysis and deamination of substrate N, and a decrease of NH3-N incorporation into microbial protein. No interaction was found between substrate and biochar source or level on any of the parameters measured. Although AFB and PPB had different textural and compositional characteristics, their effects on the rumen fermentation parameters were similar, the only observed effects being due to AFB included at 10%. Biochar supplementation promoted CH4 production regardless of the source and inclusion level, suggesting that there may be other effects beyond biomass and temperature of production of biochar, highlighting the need to consider other characteristics to better identify the mechanism by which biochar may influence CH4 production.

Relationships between patterns of rumen fermentation measured in sheep and in situ degradability and the in vitro gas production profile of the diet

BackgroundThe objectives of this study were to determine the effect of commercial slow-release urea (SRU) on in vitro fermentation characteristics, nutrient digestibility, gas production, microbial protein synthesis and bacterial community using a rumen simulation technique (RUSITEC). The experiment was a completely randomized design with four treatments and four replications of each treatment. Treatments were: control diet (no SRU addition), control diet plus 0.28% SRU (U28), or plus 0.56% SRU (U56), and control diet that was modified substituting a part of soybean meal equivalent to 0.35% SRU (MU35; dry matter [DM] basis). The experiment consisted of 8 d of adaptation and 7 d of data and sample collection. Rumen inoculum was obtained from three ruminally fistulated Angus cows fed the same diet to the substrate incubated.ResultsDigestibility of DM, organic matter (OM), crude protein (CP), fibre and starch was not affected, but daily production of gas (P < 0.07) and methane (P < 0.05) was quadratically increased with increasing SRU supplementation. The increase of SRU addition did not affect fermentation pH and total volatile fatty acid (VFA) production, whereas linearly (P < 0.01) decreased proportion of propionate, and linearly (P < 0.01) increased acetate to propionate ratio and ammonia nitrogen (N) concentration. The microbial N efficiency was also linearly (P < 0.03) improved with increasing supplementation of SRU. In comparison with control diet, the dietary substitution of SRU for part of soybean meal increased (P < 0.05) the digestibility of DM, OM and CP and decreased (P < 0.02) the total gas production. The total VFA production and acetate to propionate ratio did not differ between control and MU35, whereas the proportion of butyrate was lower (P < 0.05) and that of branched-chain VFA was greater (P < 0.05) with MU35 than control diet. Total and liquid-associated microbial N production as well as ammonia N concentration were greater (P < 0.03) with MU35 than control diet. Observed operational taxonomic units (OTUs), Shannon diversity index, and beta diversity of the microbial community did not differ among treatments. Taxonomic analysis revealed no effect of adding SRU on the relative abundance of bacteria at the phylum level, while at the genus level, the beneficial impact of SRU on relative abundance of Rikenellaceae and Prevotellaceae in feed particle-associated bacteria, and the abundance of Roseburia in liquid associate bacteria was greater (P < 0.05) with MU35.ConclusionsSupplementation of a dairy cow diet with SRU showed potential of increase in ammonia N concentration and microbial protein production, and change fermentation pattern to more acetate production. Adding SRU in dairy cow diet also showed beneficial effect on improving digestibility of OM and fibre. The results suggest that SRU can partially substitute soybean meal in dairy cow diet to increase microbial protein production without impairing rumen fermentation.

Four crossbred dairy cows (50% Holstein-Friesian × 50% Thai native), 404 ± 50.0kg of body weight (4years old) and 90 ± 5day in milk with daily milk production of 9 ± 2.0kg/day, were randomly assigned according to a 4 × 4 Latin square design to study the effect of mangosteen (Garcinia mangostana) peel powder (MSP) supplementation on rumen microorganisms, methane production, and microbial protein synthesis fed concentrate containing yeast fermented cassava chip protein (YEFECAP). The treatments were different levels of MSP supplementation at 0, 100, 200, and 300g/head/day. Rice straw was used as a roughage source fed ad libitum, and concentrate containing YEFECAP at 200g/kg concentrate was offered corresponding to concentrate-to-milk-yield ratio at 1:2. A quantitative real-time PCR approach was used to determine the population densities of ruminal microorganisms. The results revealed that supplementation of MSP did not affect on Fibrobactor succinogenes, Ruminococcus flavefaciens, and Ruminococcus albus (P > 0.05). However, total bacteria was linearly increased (P < 0.01) while methanogens and protozoal population were linearly decreased (P < 0.01) with increasing level of MSP supplementation. Increasing level of MSP supplement could decrease rumen methane production from 27.5 to 23.7mmol/100ml(3). Furthermore, cows that received MSP at 300g/head/day had the highest microbial crude protein and efficiency of rumen microbial N synthesis (416.8g/day and 16.2g/kg organic matter truly digested in the rumen (OMDR), respectively). In conclusion, supplementation of MSP at 300g/head/day with YEFECAP as a protein source in the concentrate mixture revealed an enhancement of rumen fermentation and methane reduction in lactating dairy cows.

A meta-analysis on the relationship between rumen fermentation parameters and protozoa counts in in vitro batch experiments

In vitro evaluation of fermentation characteristics of type 3 resistant starch

Simple SummaryMethane from ruminants is a major contributor to total greenhouse gases. Therefore, ruminant nutritionists have proposed strategies to mitigate methane emissions, such as chemical inhibitors and ionophores. However, dietary manipulation including natural feed additives is more practical, considering consumer preferences. Therefore, the current experiment screened 137 plant species, indigenous to East Asian countries, to select novel anti-methanogenic candidates as natural feed additives. Among these species, an extract from the seeds of Pharbitis nil exhibited a maximum 37% reduction of methane in a conformation assay. Identification of active compounds present in the seeds of Pharbitis nil revealed enrichment of polyunsaturated fatty acids, which were dominated by linoleic acid (18:2). The extract had negative effects on the populations of ciliated protozoa and H2-producing Ruminococcus flavefaciens, thereby increasing the proportion of propionate, similar to the effect of monensin. This is the first report to suggest that the seeds of P. nil could be a promising anti-methanogenic alternative to ionophores or oil seeds.Indiscriminate use of antibiotics can result in antibiotic residues in animal products; thus, plant compounds may be better alternative sources for mitigating methane (CH4) production. An in vitro screening experiment was conducted to evaluate the potential application of 152 dry methanolic or ethanolic extracts from 137 plant species distributed in East Asian countries as anti-methanogenic additives in ruminant feed. The experimental material consisted of 200 mg total mixed ration, 20 mg plant extract, and 30 mL diluted ruminal fluid-buffer mixture in 60 mL serum bottles that were sealed with rubber stoppers and incubated at 39 °C for 24 h. Among the tested extracts, eight extracts decreased CH4 production by >20%, compared to the corresponding controls: stems of Vitex negundo var. incisa, stems of Amelanchier asiatica, fruit of Reynoutria sachalinensis, seeds of Tribulus terrestris, seeds of Pharbitis nil, leaves of Alnus japonica, stem and bark of Carpinus tschonoskii, and stems of Acer truncatum. A confirmation assay of the eight plant extracts at a dosage of 10 mg with four replications repeated on 3 different days revealed that the extracts decreased CH4 concentration in the total gas (7–15%) and total CH4 production (17–37%), compared to the control. This is the first report to identify the anti-methanogenic activities of eight potential plant extracts. All extracts decreased ammonia (NH3-N) concentrations. Negative effects on total gas and volatile fatty acid (VFA) production were also noted for all extracts that were rich in hydrolysable tannins and total saponins or fatty acids. The underlying modes of action differed among plants: extracts from P. nil, V. negundo var. incisa, A. asiatica, and R. sachalinensis resulted in a decrease in total methanogen or the protozoan population (p < 0.05) but extracts from other plants did not. Furthermore, extracts from P. nil decreased the population of total protozoa and increased the proportion of propionate among VFAs (p < 0.05). Identifying bioactive compounds in seeds of P. nil by gas chromatography-mass spectrometry analysis revealed enrichment of linoleic acid (18:2). Overall, seeds of P. nil could be a possible alternative to ionophores or oil seeds to mitigate ruminal CH4 production.

The increase in global temperatures over the past few decades due to greenhouse gas emissions has raised concerns and necessitated further research in climate change mitigation and adaptation. Methane is a prominent greenhouse gas that significantly contributes to climate change, with a substantial amount generated through fermentation processes occurring in the rumen of ruminant animals. The potential of plant secondary metabolites, especially those derived from tannin-rich plants, warrants investigation to modify rumen fermentation and mitigate methane emissions in livestock diets. The objective of this study was to assess the impact of extracts obtained from green and black tea waste on rumen fermentation dynamics and gas (methane) production, utilizing in vitro methods. For this purpose, rumen fluid was collected from two fistulated sheep and subjected to three treatments: (1) a basal diet (control), (2) a basal diet + green tea waste extract (5% of dry matter), (3) a basal diet + black tea waste extract (5% of dry matter). The study assessed the effects of incorporating extracts from green and black tea waste on various parameters, including digestibility, protozoa population, ammonia nitrogen levels, volatile fatty acids, and methane gas production following a 24-h incubation period. Statistical analysis of the data was conducted using SAS software within a completely randomized design framework. The findings indicated that the addition of green and black tea waste extracts significantly decreased methane gas production (p &lt; 0.05), protozoa count (p &lt; 0.05), and ammonia nitrogen concentrations in rumen fluid (p &lt; 0.05) when compared to the control group. The addition of green and black tea waste extracts has significantly altered the concentration of VFAs in rumen fluid (p &lt; 0.05). Specifically, the addition of green tea waste extract has led to a highly significant reduction in acetic acid, (p &lt; 0.01) and the addition of both extracts has resulted in a significant increase in propionic acid (p &lt; 0.05). Consequently, the results suggest that the inclusion of green and black tea waste extracts in livestock diets may effectively mitigate methane emissions in the rumen, thereby reducing feed costs and reducing environmental pollution.