Decreased CH4 Production Research Articles

Effects of phytonutrient-based encapsulation of Wolffia globosa on gas production, in vitro fermentation characteristics, and methane mitigation using in vitro study techniques

Microencapsulation, the advanced and newest coating technology, enhances the efficacy and stability of phytonutrients, particularly sensitive plant extracts, in animal feed. Phytonutrients have potential to mitigate methane emissions (CH4) and ammonia nitrogen (NH3-N) concentrations during rumen fermentation but require protection to maintain effectiveness. Therefore, this study evaluated the potential of duckweed powder (DWP) and microencapsulated duckweed extract (mDWE) at different levels of supplementation in an in vitro study using rumen fluid from Thai-crossbred dairy cows. Employing a 2 × 3 + 1 factorial design, the experiment tested two forms of duckweed (DWP and mDWE) at supplementation levels of 2%, 4%, and 6% + control (0%) of total dry matter substrate. Significant interactions were observed between the different forms and supplementation levels 4% mDWE (p < 0.05) notably enhanced NH3-N, total VFA (volatile fatty acid), and B. proteoclasticus, improved butyric acid (C4), Mathanobacteriales, while decreased microbial population including R. albus, R. flavefaciens and M. elsdenii and B. fibrisolvens at. mDWE increased fraction gas b, |a|+b, and cumulative gas production, dry matter (12 hours; h) and organic matter (24 h) degradability, and propionic acid (C3), while decreased CH4 production (12 and 24 h), acetate (C2) at 24 h, R. albus, R. flavefaciens, and Methanobacteriales (p < 0.05). Additionally, supplementation level significantly increased gas production kinetics and C3 (24 h), while decreased C2, C4, Methanobacteriales and CH4 production (p < 0.05). This study underscores the potential of microencapsulated duckweed extract 4% mDWE as an optimal ruminant feed additive for decreasing methane production and enhancing rumen fermentation. Further in vivo studies are recommended to validate these findings.

Potential use of Wolffia globosa powder supplementation on in vitro rumen fermentation characteristics, nutrient degradability, microbial population, and methane mitigation

This study examined the potential of duckweed powder (DWP) on in vitro fermentation characteristics, nutrient degradability, microbial change, and methane (CH4) production using in vitrostudy technique. This investigation used a 2 × 6 factorial arrangement in a completely randomized design (CRD) by different roughage-to-concentrate (R: C) ratios of 60:40 and 40:60 combined with DWP supplementation levels at 0, 2, 4, 6, 8, and 10% of the total dry matter (DM) substrate, respectively. There was an interaction effect by R: C ratios combined with DWP supplementations that changed gas production rate, pH value (4 h; h), volatile fatty acid (VFA) (8 h), in vitro dry matter degradability (IVDMD) at 12 h, and concentration of ammonium nitrogen (NH3-N) (p < 0.05). Furthermore, the R: C ratio (40:60) significantly decreased CH4 production (4 and 8 h), pH (8 h), and Ruminococcus albus (8 h) (p < 0.05), while it significantly increased total VFA (8 h), and nutrient degradability (p < 0.05). DWP 4% significantly increased to the highest of gas production, improved nutrient degradability (IVDMD at 24 h and in vitro organic matter degradability; IVOMD at 12 h), whereas significantly decreased Methanobacteriales (8 h) and CH4 production. DWP 4% has potential as a ruminant feed additive for reducing Methanobacteriales and CH4 emission and enhancing rumen fermentation.

Hydroponic fodders as alternative feeds for ruminants to reduce ruminal methane emissions: An in vitro study

Malate, a precursor in the ruminal propionate production pathway, competes with methanogenesis for metabolic hydrogen, offering a way to reduce ruminal methane (CH4) production in ruminants. However, cost considerations hinder widespread use of malate in ruminant diets. An alternative approach involves utilizing transient malate levels generated during seed germination via the glyoxylate cycle. This study investigated the methane-mitigating potential of malate-containing hydroponic fodder. Fodder samples with peak malate concentrations from alfalfa, forage pea, Italian ryegrass, rye, soybean, triticale, and wheat during germination were subjected to in vitro rumen fermentation using the Hohenheim gas test. The basal diet of in vitro fermentation comprised 40% grass silage, 40% maize silage, 15% hay, and 5% concentrate on a dry matter basis, with nutritional characteristics including 42.1% neutral detergent fiber (NDF), 25.0% acid detergent fiber, 14.0% starch, 12.7% crude protein, and 3.5% ether extract (EE), on a dry matter basis. Experimental treatments were fodder inclusion involved replacing 20% of the basal diet (20R), and additionally, 100% replacement of the silages with alfalfa d 10 and rye d 9 (SR), the 2 high-malate fodders. Reductions in CH4 production were observed with soybean (20R, 6.7% reduction), alfalfa (20R, 6.6% reduction), and increased with rye (20R, 6.3% increase). In the setup replacing silages with high-malate fodders (SR), alfalfa decreased CH4 production (17.7%) but increased ammonia (174%), while rye increased CH4 production (35.8%). Organic matter digestibility increased with SR rye (12.6%). Marginal effects of dietary variables were analyzed in a Generalized Additive Model. A negative relationship between dietary malate content and CH4 production was observed, whereas dietary NDF and starch content were positive correlated with CH4 production. In conclusion, malate within the hydroponic fodder could potentially reduce CH4 emissions in ruminants. However, achieving sufficient efficacy requires high malate content. Additionally, use of hydroponic fodder may increase the risk of nitrogen emissions. Animal studies are required for further investigation.

Assessing the effects of high-carvacrol oregano oil on rumen microbial fermentation, gas production, and methane production in vitro

The objective of this in vitro study was to assess the effects of high-carvacrol oregano oil (ORE; 50 g/kg oregano oil, 800–802 g/kg carvacrol), on microbial fermentation and CH4 production. In the experiment (a complete randomized block design), treatments included a negative control (CTL, no additive), a positive control (monensin, MON, 10 mg/L of culture fluid), and ORE (20, 40, 80, 120, 200, 160, and 1000 mg/L of culture fluid). Compared with CTL, MON shifted ( p &lt; 0.05) volatile fatty acid (VFA) profile from acetate and butyrate to propionate production, thereby reducing ( p &lt; 0.05) CH4 production (−26%). Monensin decreased ( p &lt; 0.05) NH3 and branched-chain VFA concentrations. Of the doses evaluated, only the highest dose (1000 mg/L) affected ruminal microbial fermentation and CH4 production. At this dose, ORE reduced ( p &lt; 0.05) gas production, total VFA, acetate, propionate and NH3 concentrations, and CH4 production (−22%). The reduced gas production and total VFA is an indication of feed digestion inhibition. These results suggest that ORE may decrease CH4 production and improve ruminal N utilization. However, these findings need to be validated in vivo to determine the optimal dose to benefit from the positive effects while avoiding the negative impact of ORE on feed digestion.

Oat Brewery Waste Decreased Methane Production and Alters Rumen Fermentation, Microbiota Composition, and CAZymes Profiles.

The transformation of oat brewery waste (OBW) into livestock feed could be a potential replacement for the expensive concentrate and one of the effective approaches for avoiding health hazards due to the accumulation of oat brewery waste in the environment. To explore the potential of OBW as a methane (CH4) mitigating agent, an in vitro study was undertaken to investigate the effect of graded replacement of concentrate with OBW on CH4 production, microbiota, feed fermentation, and CAZymes. A total of five treatments with variable proportions of OBW were formulated. The results indicated a linear decrease in the total gas production and a 38-52% decrease in CH4 production with a 60 and 100% replacement of concentrate with OBW. The inclusion of OBW also affected the abundance of microbes such as Firmicutes, Euryarchaeota, Methanobrevibacter, and protozoa numbers. This study demonstrated that OBW can partially replace the concentrate and effectively mitigate CH4 production; however, the concurrent decrease in fermentation cautioned for the partial replacement of concentrate with OBW at an appropriate level at which the fermentation remains unaffected while decreasing CH4 production. Therefore, waste from oat breweries can contribute to curtailing the accumulation of greenhouse gases (GHGs) in the atmosphere.

Exploring the combination of Asparagopsis taxiformis and phloroglucinol to decrease rumen methanogenesis and redirect hydrogen production in goats

Many strategies for mitigating enteric methane (CH4) emissions in ruminants have focused on suppressing the activity of rumen methanogens, but this often leads to excess dihydrogen (H2) accumulation in the rumen, which is subsequently expelled and represents a potential energy loss. We hypothesized that phloroglucinol could act as a H2 acceptor when rumen methanogenesis is inhibited and be potentially transformed into beneficial compounds for the animal. Eight adult goats were randomly assigned to a replicated 4 × 4 Latin square design with a 2 × 2 factorial arrangement of treatments: two levels of Asparagopsis taxiformis as CH4 inhibitor [0 vs. 5 g/kg on a dry matter (DM) basis; AT- and AT+, respectively] and two levels of phloroglucinol as alternative H2 acceptor (0 vs. 20 g/kg DM, PG- and PG+, respectively). Therefore, four dietary treatments were considered: i) basal diet (AT-PG-); ii) A. taxiformis alone (AT+PG-); iii) phloroglucinol alone (AT-PG+); and iv) the combination of A. taxiformis and phloroglucinol (AT+PG+). Animals were fed a maintenance diet with a 70:30 forage-to-concentrate ratio. After 10 d of adaptation to the diet, enteric gas emissions were measured in respiration chambers during 3 d prior to rumen content sampling on d 14. Dietary supplementation with A. taxiformis decreased CH4 production (-33.9 %) and increased H2 emissions (+3465 %), along with greater rumen propionate concentration. In contrast, phloroglucinol supplementation alone did not impact CH4 emissions or the rumen concentration of the main microbial groups but substantially increased acetate molar proportion (+10.2 %) which could act as an alternative H2 acceptor. Moreover, when A. taxiformis was combined with phloroglucinol, it resulted in a decrease in H2 emissions (-68.1 %). However, this decrease in H2 emissions was not fully explained by the increase in the acetate as phloroglucinol led to an increase in acetate both when methanogenesis was inhibited and when it was not. These findings suggest that the rumen fermentation of phloroglucinol may capture some of the additional H2 arising from the inhibition of methanogenesis by A. taxiformis through pathways other than acetate formation. Moreover, H2 emissions were not eliminated and most of the decrease occurred during the post-prandial stage, suggesting that the efficiency of H2 redirection could be further improved.

Turmeric rhizomes reduced in vitro methane production and improved gas production and nutrient degradability

The present study aimed to evaluate the effect of dry turmeric rhizomes on in vitro biogas production and diet fermentability. Turmeric rhizomes were included at gradually increased levels: 0, 0.5, 1, 1.5 and 2% of a diet containing per kg dr matter (DM): 500 g concentrate feed mixture, 400 g berseem hay and 100 g rice straw, and incubated for 48 h. Gas chromatography-mass spectrometry analysis showed that ar-turmerone, α-turmerone and β-turmerone were the major bioactive compounds in the rhizomes. Turmeric rhizomes increased (p < 0.01) asymptotic gas production (GP) and rate and lag of CH4 production and decreased (p < 0.01) rate of GP, lag of GP, asymptotic CH4 production and proportion of CH4 production. Turmeric rhizome administration linearly increased (p < 0.01) DM and fiber degradability and concentrations of total short-chain fatty acids, acetic and propionic acids and ammonia-N and quadratically (p < 0.05) decreased fermentation pH. It is concluded that including up to 2% turmeric rhizomes improved in vitro ruminal fermentation and decreased CH4 production.

Rumen metagenome reveals the mechanism of mitigation methane emissions by unsaturated fatty acid while maintaining the performance of dairy cows

Dietary fat content can reduce the methane production of dairy cows; however, the relevance fatty acid (FA) composition has towards this inhibitory effect is debatable. Furthermore, in-depth studies elucidating the effects of unsaturated fatty acids (UFA) on rumen function and the mechanism of reducing methane (CH4) production are lacking. This study exposed 10 Holstein cows with the same parity, similar milk yield to two total mixed rations: low unsaturated FA (LUFA) and high unsaturated FA (HUFA) with similar fat content. The LUFA group mainly added fat powder (C16:0 > 90%), and the HUFA group mainly replaced fat powder with extruded flaxseed. The experiment lasted 26 d, the last 5 d of which, gas exchange in respiratory chambers was conducted to measure gas emissions. We found that an increase in the UFA in diet did not affect milk production (P > 0.05) and could align the profile of milk FAs more closely with modern human nutritional requirements. Furthermore, we found that increasing the UFA content in the diet lead to a decrease in the abundance of Methanobrevibacter in the rumen (|linear discriminant analysis [LDA] score| > 2 and P < 0.05), which resulted in a decrease in the relative abundance of multiple enzymes (EC:1.2.7.12, EC:2.3.1.101, EC:3.5.4.27, EC:1.5.98.1, EC:1.5.98.2, EC:6.2.1.1, EC:2.1.1.86 and EC:2.8.4.1) during methanogenesis (P < 0.05). Compared with the LUFA group, the pathway of CH4 metabolism was inhibited in the HUFA group (|LDA| > 2 and P < 0.05), which ultimately decreased CH4 production (P < 0.05). Our results illustrated the mechanism involving decreased CH4 production when fed a UFA diet in dairy cows. We believe that our study provides new evidence to explore CH4 emission reduction measures for dairy cows.

Microwave pyrolysis-prepared engineering carbons from corn cobs and red mombin seeds towards xylene adsorption

High-quality biochars/activated carbons were prepared, optimizing individual parameters of energetically-save microwave pyrolysis (raw material loading - 20 vs. 60 g, nitrogen atmosphere - flow vs. batch, ZnCl2 activation) from two agricultural wastes - corn cobs, red mombin seeds. Most promising carbons were examined for gaseous xylene adsorption and showed higher sorption capacity (∼250–475 mgxylene· g−1) than commercial carbon (∼214 mgxylene· g−1). ZnCl2 activation of both raw materials reduces the fixed carbon content and increases volatiles in activated carbon, suggesting microwave pyrolysis of activated feedstock should take 25 min. While biochars are microporous materials with inhomogeneous low-surface mesopore/macropore network, activated carbons are highly microporous-mesoporous. ZnCl2 activation of both raw materials contributes to formation of extensive high-surface mesopore network (with pore-size < 20 nm) and enlargement of micropore-size, but does not affect the micropore volume. ZnCl2 activation increases H2 and decreases CH4 production. Microwave pyrolysis of larger raw material loading with ZnCl2 leads to CO2 increase. Best xylene adsorption capacity (475 mgxylene· g−1) was determined for activated carbon produced from 60 g loading of corn cobs in batch nitrogen atmosphere, showing the largest micropore volume, lowest surface polarity and medium rate of graphitization. Large micropore volume, low surface polarity and high rate of graphitization of carbon are xylene sorption capacity-determining factors.

Effect of buffer pH on methane production and fermentation characteristics of three forages tested in vitro.

Low rumen pH is proposed to be a major mechanism for low methane (CH4) emissions from sheep fed forage rape. However, it is difficult to separate this from other in vivo factors, such as rumen passage rate. The objective of this study was to determine the effect of pH alone on CH4 production in vitro using different pH buffers. Ryegrass, white clover and forage rape were incubated in vitro using three different incubation buffers with starting pH values of 5.5, 6.2 and 6.8. Decreasing pH reduced overall in vitro CH4 emission relative to fermented hexoses (CH4/FHex) by up to 54% and overall fermentation by 40%. pH also changed fermentation profiles where the acetate + butyrate to propionate + valerate ratio decreased when pH decreased. Within the three forages, forage rape led to the lowest CH4/FHex, but only in pH 5.5 and 6.2 buffer, and this was enhanced when the pH fell below 6. Reducing pH in vitro decreased CH4 production and overall fermentation across all forages. The lower pH reached by forage rape compared to ryegrass and white clover appears to drive the lower CH4 production relative to the extent of fermentation from forage rape compared to the other forages. © 2024 The Author(s). Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Scaling-up fungal pretreatment of lignocellulose biomass: Impact on nutritional value, ruminal degradability, methane production, and performance of lactating dairy cows

Scaling-up fungal pretreatment of lignocellulose biomass (LCB) for ruminant nutrition has become a major research challenge in recent years. This study systematically investigated the effectiveness of fungal (Pleurotus ostreatus) pretreatment (for 30 days under solid state fermentation (SSF)) of large quantities (8400 kg) of lime-pasteurized wheat straw, in terms of improvement in nutritional value, in vitro ruminal fermentation characteristics and methane (CH4) production potential. We further investigated the effects of stepwise replacement of untreated wheat straw (UTWS) with the lime pasteurized P. ostreatus treated wheat straw (PTWS) on dry matter intake (DMI), apparent total tract digestibility, production performance, CH4 emission and feed efficiency of lactating dairy cows. The results revealed that PTWS had lower lignin (P< 0.001), hemicellulose (P< 0.001) and cellulose (P< 0.05) contents and higher crude protein (CP; P< 0.001) content and cellulose to lignin ratio (P< 0.01), as compared to UTWS. The PTWS had higher in vitro DM digestibility (IVDMD; P< 0.001), total gas production (IVGP; P< 0.01), total volatile fatty acids (VFAs, P< 0.01) and propionate (P< 0.05) concentration, and lower (P< 0.05) pH and CH4 gas production during 72 h in vitro fermentation. Notably, the fungus degraded 33.3 % lignin at the expense of 6.56 % cellulose, and markedly increased CP content (46.3 %), IVDMD (20.2 %), IVGP (16.8 %) and VFAs (10.0 %) production, and decreased CH4 production (10.3 %). The aflatoxin B1 concentration of PTWS was <5.0 µg/kg, and mycelium concentration was 181.9 mg/kg DM, reflecting safe and effective tretment of the straw. Replacement of 32 % UTWS in total mixed ration with the PTWS, increased DMI (0.84 kg/day; P = 0.01), apparent total tract DM digestibility (5.5 g/100 g; P< 0.05) and milk yield (1.17 liter/day; P= 0.032), and decreased CH4 emission (1.45 g/kg DMI; P< 0.05). In conclusion, hydrated lime can replace traditional high-tech pasteurization methods for the fungal pretreatment of LCB, and present prospects for scaling up the process for ruminant nutrition.

Effects of 3-nitrooxypropanol (Bovaer10) and whole cottonseed on milk production and enteric methane emissions from dairy cows under Swiss management conditions

The objective of this study was to determine the potential effect and interaction of 3-nitrooxypropanol (3-NOP; Bovaer, DSM-Firmenich Nutrition Products Ltd.) and whole cottonseed (WCS) on lactational performance and enteric methane (CH4) emission of dairy cows. A total of 16 multiparous cows, including 8 Holstein Friesian (HF) and 8 Brown Swiss (BS; 224 ± 36 DIM, 26 ± 3.7 kg milk yield, mean ± SD), were used in a split-plot design, where the main plot was the breed of cows. Within each subplot, cows were randomly assigned to a treatment sequence in a replicated 4 × 4 Latin square design with 2 × 2 factorial arrangements of treatments with four 24-d periods. The experimental treatments were as follows: (1) control (basal TMR), (2) 3-NOP (60 mg/kg TMR DM), (3) WCS (5% TMR DM), and (4) 3-NOP + WCS. The treatment diets were balanced for ether extract, crude protein, and NDF contents (4%, 16%, and 43% of TMR DM, respectively). The basal diets were fed twice daily at 0800 and 1800 h. Dry matter intake and milk yield were measured daily, and enteric gas emissions were measured (using the GreenFeed System, C-Lock Inc.) during the last 3 d of each 24-d experimental period when animals were housed in tiestalls. There was no difference in DMI on treatment level, whereas the WCS treatment increased ECM yield and milk fat yield. No interaction of 3-NOP and WCS occurred for any of the enteric gas emission parameters, but 3-NOP decreased CH4 production (g/d), CH4 yield (g/kg DMI), and CH4 intensity (g/kg ECM) by 13%, 14%, and 13%, respectively. Further, an unexpected interaction of breed by 3-NOP was observed for different enteric CH4 emission metrics: HF cows had a greater CH4 mitigation effect compared with BS cows for CH4 production (g/d; 18% vs. 8%), CH4 intensity (g/kg milk yield; 19% vs. 3%), and CH4 intensity (g/kg ECM; 19% vs. 4%). Hydrogen production was increased by 2.85-fold in HF and 1.53-fold in BS cows receiving 3-NOP. Further, a 3-NOP × time interaction occurred for both breeds. In BS cows, 3-NOP tended to reduce CH4 production by 18% at approximately 4 h after morning feeding, but no effect was observed at other time points. In HF cows, the greatest mitigation effect of 3-NOP (29.6%) was observed immediately after morning feeding, and it persisted at around 23% to 26% for 10 h until the second feed provision, and 3 h thereafter, in the evening. In conclusion, supplementing 3-NOP at 60 mg/kg DM to a high-fiber diet resulted in 18% to 19% reduction in enteric CH4 emission in Swiss HF cows. The lower response to 3-NOP by BS cows was unexpected and has not been observed in other studies. These results should be interpreted with caution due to the low number of cows per breed. Finally, supplementing WCS at 5% of DM improved ECM and milk fat yield but did not enhance the CH4 inhibition effect of 3-NOP of dairy cows.

Ruminal methane emission and lactational performance of cows fed rapeseed cake and oats on a grass silage–based diet

The objective of this experiment was to investigate the effect of lipid from rapeseed cake and oats on ruminal CH4 emission and lactational performance of dairy cows. Twelve lactating Nordic Red cows, of which 4 were primiparous, and averaging (±SD) 48 ± 22.9 DIM, 37.8 ± 7.14 kg/d milk yield were enrolled in a switch-back design experiment with 3 periods of 4 wk each. The cows were assigned into 6 pairs based on parity, DIM, milk yield, and BW at the beginning of the experiment. The experimental treatments were (1) rapeseed cake and oats (RSC+O), and (2) rapeseed meal and barley (RSM+B) as the concentrate feeds. Cows in each pair were randomly assigned to 1 of the 2 groups, which received the treatments in 2 different sequences (i.e., group 1 received RSC+O in period 1 and 3, and RSM+B in period 2, whereas group 2 was fed RSM+B in period 1 and 3, and RSC+O in period 2). The diets consisted of a partially mixed ration with grass silage mixed with either oats or barley, according to the treatment sequence, and the rapeseed cake or meal being mixed into a pellet with either oats or barley according to the treatments, and a mineral mix. The pellet was delivered at a fixed amount (i.e., 6 kg/d for multiparous and 5 kg/d for the primiparous cows) from the milking robot. The actual forage to concentrate ratios for RSC+O and RSM+B were 51:49 and 52:48, respectively, with NDF concentrations of 41.5% and 36.0% and CP concentrations of 17.0% and 16.7% of diet DM. Dry matter intake, milk yield, and gas exchange (with a GreenFeed system attached to the milking robot) were recorded daily, and milk composition and spot fecal samples were collected during the last week of each period. Based on feed analysis, and DMI of the cows during the experiment, the total fat content of the experimental diets was 4.1% and 2.7% of DM for RSC+O and RSM+B diets, respectively. Dry matter intake was 1.6 kg/d lower, and milk yield tended to be 1.0 kg/d greater for RSC+O versus RSM+B. There were no differences in ECM yield and milk composition between the treatments, whereas milk ME efficiency was greater for cows fed RSC+O than RSM+B. Methane yield (g/kg DMI) did not differ between treatments, but CH4 production (g/d) was 9.4% and CH4 intensity as g/kg ECM was 11.7% lower for RSC+O versus RSM+B. The lower CH4 production was likely caused by the lower DMI and fiber digestibility, observed with the RSC+O diet. In addition, the greater lipid intake also contributed to lower rate of fermentation and subsequent decrease in CH4 production. Overall, feeding rapeseed cake with oats in a grass silage-based diet increased feed efficiency while decreasing CH4 emission intensity in lactating cows. This provides a practical way of mitigating ruminal CH4 emission from dairy operations while maintaining milk production with commonly used feedstuffs in Nordic conditions.

Dietary supplementation of phytoncide and soybean oil increases milk conjugated linoleic acid and depresses methane emissions in Holstein dairy cows

The objective of this study was to determine whether adding phytoncide oil (PO) and soybean oil (SBO) to the dairy cow diet could increase milk conjugated linoleic acid (CLA) and depress methane (CH4) emissions in Holstein dairy cows. Rumen fermentation was conducted at four levels of SBO (0, 1, 2, and 4%, on DM basis) and two levels of PO (0 and 0.1%, on DM basis) with in vitro experiment. To evaluate blood parameters, fecal microbe population, milk yield and fatty acid compositions, and CH4 production, in vivo experiment was conducted using 38 Holstein dairy cows divided into two groups of control (fed TMR) and treatment (fed TMR with 0.1% PO and 2% SBO as DM basis). In the in vitro study (Experiment 1), PO or SBO did not affect rumen pH. However, SBO tended to decrease ruminal ammonia-N (p = 0.099). Additionally, PO or SBO significantly decreased total gas production (p = 0.041 and p = 0.034, respectively). Both PO and SBO significantly decreased CH4 production (p < 0.05). In addition, PO significantly increased both CLA isomers (c9, t11 and t10, c12 CLA) (p < 0.001). Collectively, 0.1% PO and 2% SBO were selected resulting in most effectively improved CLA and decreased CH4 production. In the in vivo study (Experiment 2), 0.1% PO with 2% SBO (PSO) did not affect complete blood count. However, it decreased blood urea nitrogen and magnesium levels in blood (p = 0.021 and p = 0.01, respectively). PSO treatment decreased pathogenic microbes (p < 0.05). It increased milk yield (p = 0.017) but decreased percentage of milk fat (p = 0.013) and MUN level (p < 0.01). In addition, PSO treatment increased both the concentration of CLA and PUFA in milk fat (p < 0.01). Finally, it decreased CH4 emissions from dairy cows. These results provide compelling evidence that a diet supplemented with PSO can simultaneously increase CLA concentration and decrease CH4 production with no influence on the amount of milk fat (kg/day) in Holstein dairy cows.

The Effect of Bismuth and Tin on Methane and Acetate Production in a Microbial Electrosynthesis Cell Fed with Carbon Dioxide.

This study investigates the impacts of bismuth and tin on the production of CH4 and volatile fatty acids in a microbial electrosynthesis cell with a continuous CO2 supply. First, the impact of several transition metal ions (Ni2+, Fe2+, Cu2+, Sn2+, Mn2+, MoO42-, and Bi3+) on hydrogenotrophic and acetoclastic methanogenic microbial activity was evaluated in a series of batch bottle tests incubated with anaerobic sludge and a pre-defined concentration of dissolved transition metals. While Cu is considered a promising catalyst for the electrocatalytic conversion of CO2 to short chain fatty acids such as acetate, its presence as a Cu2+ ion was demonstrated to significantly inhibit the microbial production of CH4 and acetate. At the same time, CH4 production increased in the presence of Bi3+ (0.1 g L-1) and remained unchanged at the same concentration of Sn2+. Since Sn is of interest due to its catalytic properties in the electrochemical CO2 conversion, Bi and Sn were added to the cathode compartment of a laboratory-scale microbial electrosynthesis cell (MESC) to achieve an initial concentration of 0.1 g L-1. While an initial increase in CH4 (and acetate for Sn2+) production was observed after the first injection of the metal ions, after the second injection, CH4 production declined. Acetate accumulation was indicative of the reduced activity of acetoclastic methanogens, likely due to the high partial pressure of H2. The modification of a carbon-felt electrode by the electrodeposition of Sn metal on its surface prior to cathode inoculation with anaerobic sludge showed a doubling of CH4 production in the MESC and a lower concentration of acetate, while the electrodeposition of Bi resulted in a decreased CH4 production.

Relationships between Dietary Chemical Components and Enteric Methane Production and Application to Diet Formulation in Beef Cattle

We used published data consisting of 263 treatment mean observations from beef cattle and dairy steers and heifers, in which CH4 was measured via chambers or head boxes, to evaluate relationships between enteric CH4 production and dry matter intake (DMI) and dietary components. Daily DMI was positively related (slope = 15.371, p &lt; 0.001) to total daily production (g/d) of CH4 (r2 = 0.821). Among chemical components, dietary neutral detergent fiber (NDF) concentration was the most highly related (r2 = 0.696; slope = 0.2001; p &lt; 0.001) to CH4 yield (g/kg of DMI), with strong relationships also noted for dietary starch:NDF ratio (r2 = 0.662; slope = −2.4587; p &lt; 0.001), starch (r2 = 0.495; slope = −0.106; p &lt; 0.001), and the proportion of metabolizable energy relative to gross energy (r2 = 0.561; slope = −23.663; p &lt; 0.001). The slope (−0.5871) and intercept (22.2295) for the dietary ether extract vs. CH4 yield were significant (p &lt; 0.001), but the relationship was highly variable (r2 = 0.150). For dietary crude protein concentration, the slope for CH4 yield was not significant (−0.0344; p &lt; 0.381) with an r2 value near zero. Decreasing DMI by programming body weight gain or restricting feed intake could decrease CH4 production in confined cattle, but these approaches might negatively affect growth performance and product quality, potentially negating positive effects on CH4 production. Feeding higher-quality forages or using grazing management systems that decrease dietary NDF concentrations or substituting grain (starch) for forage should decrease both CH4 yield from enteric production and manure CH4 production via increased digestibility. Effects of feeding management and diet formulation strategies should be additive with other mitigation approaches such as feed additives, allowing the cattle industry to achieve maximal decreases in enteric CH4 production, while concurrently maintaining optimal beef production.

Inoculum microbial mass is negatively related to microbial yield and positively to methane yield in vitro.

Ruminal microbes catabolise feed carbohydrates mainly into SCFA, methane (CH4), and carbon dioxide (CO2), with predictable relationships between fermentation end products and net microbial increase. We used a closed in vitro batch culture system, incubating grass and maize silages, and measured total gas production at 8 and 24 h, as well as the truly degraded substrate, the net production of SCFA, CH4, and microbial biomass at 24 h, and investigated the impact of silage type and inoculum microbial mass on fermentation direction. Net microbial yield was negatively correlated with total gas at 8 h (P < 0•001), but not at 24 h (P = 0•052), and negatively correlated with CH4 production (P < 0•001). Higher initial inoculum microbial mass was related to a lower net microbial yield (P < 0•001) but a higher CH4 production (P < 0•001). A significant difference between grass silage and maize silage was detected within the context of these relationships (P < 0•050). The metabolic hydrogen (2H) recovery was 102.8 ± 12.3 % for grass silages and 118.8 ± 13.3% for maize silages. Overall, grass silages favoured more substrate conversion to microbial biomass and less to fermentation end products than maize silage. Lower inoculum microbial mass facilitated more microbial growth and, because of the 2H sink by microbial synthesis, decreased CH4 production.

Investigating the effect of supplementing different levels of Isochrysis galbana on in vitro rumen fermentation parameters.

This study aimed to investigate the effect of supplementing Isochrysis galbana (I. galbana) at levels of 0 (control), 1, 2, 3, 4, and 5 (g/100 g DM) of the diet on the gas production kinetics, methane production, rumen fermentation parameters, and relative microbial population in vitro. Supplementation of I. galbana at high level (5g/100 g DM) caused a significant decrease in total gas production (p < 0.05). High supplementation rates (4 and 5g/100 g DM) decreased CH4 production relative to the control by 18.4% and 23.2%, respectively. Although rumen ammonia nitrogen (N-NH3) and total volatile fatty acids (VFA) concentrations were affected by dietary treatments, but the VFA profile did not changed. The relative proportion of protozoa and methanogenic archaea as well as Anaerovibrio lipolytica, Prevotella spp., Ruminococcus flavefaciens, and Fibrobacter succinogenes were decreased significantly as a result of microalgae supplementation. However, the relative abundance of Ruminococcus albus, Butyrivibrio fibrisolvens and Selenomonas ruminantium were significantly increased (p < 0.05), related to the control group. As well, the pH was not affected by dietary treatments. It was concluded that I. galbana reduced in vitro CH4 production and methanogenic archaea that its worth to be investigated further in in vivo studies.

Effect of Padina gymnospora biowaste inclusion on in vitro methane production, feed fermentation, and microbial diversity.

In vitro studies were undertaken aiming to study the methane (CH4) mitigation potential of biowaste (BW) of Padina gymnospora at the graded inclusion of 0% (C), 2% (A2), 5% (A5), and 10% (A10) of the diet composed of straw and concentrate in 40:60 ratio. The chemical composition analysis revealed that the BW contained higher crude protein (CP), neutral detergent fiber (NDF), acid detergent fiber (ADF), and ether extract (EE) than the PF (fresh seaweed, P. gymnospora). The concentration of cinnamic acid, sinapic acid, kaempferol, fisetin p-coumaric acid, ellagic acid, and luteolin in BW was 1.5-6-folds less than the PF. Inclusion of BW decreased (P < 0.0001) CH4 production by 34%, 38%, and 45% in A2, A5, and A10 treatments, respectively. A decrease (P < 0.0001) of 7.5%-8% in dry matter (DM) and organic matter (OM) digestibility was also recorded with the BW supplementation. The BW inclusion also decreased the numbers of total (P = 0.007), Entodinomorphs (P = 0.011), and Holotrichs (P = 0.004) protozoa. Metagenome data revealed the dominance of Bacteroidetes, Proteobacteria, Firmicutes, Actinobacteria, and Fibrobacter microbial phyla. At the phylum level, Euryarchaeota dominated the archaeal community, whereas Methanobrevibacter was most abundant at the genus level. It can be concluded that the inclusion of BW in straw and concentrate based diet by affecting rumen fermentation, protozoal numbers, and compositional shift in the archaeal community significantly decreased CH4 production. Utilization of biowaste of P. gymnospora as a CH4 mitigating agent will ensure its efficient utilization rather than dumping, which shall cause environmental pollution and health hazards.

Rumen microbial degradation of bromoform from red seaweed (Asparagopsis taxiformis) and the impact on rumen fermentation and methanogenic archaea

BackgroundThe red macroalgae Asparagopsis is an effective methanogenesis inhibitor due to the presence of halogenated methane (CH4) analogues, primarily bromoform (CHBr3). This study aimed to investigate the degradation process of CHBr3 from A. taxiformis in the rumen and whether this process is diet-dependent. An in vitro batch culture system was used according to a 2 × 2 factorial design, assessing two A. taxiformis inclusion rates [0 (CTL) and 2% DM diet (AT)] and two diets [high-concentrate (HC) and high-forage diet (HF)]. Incubations lasted for 72 h and samples of headspace and fermentation liquid were taken at 0, 0.5, 1, 3, 6, 8, 12, 16, 24, 48 and 72 h to assess the pattern of degradation of CHBr3 into dibromomethane (CH2Br2) and fermentation parameters. Additionally, an in vitro experiment with pure cultures of seven methanogens strains (Methanobrevibacter smithii, Methanobrevibacter ruminantium, Methanosphaera stadtmanae, Methanosarcina barkeri, Methanobrevibacter millerae, Methanothermobacter wolfei and Methanobacterium mobile) was conducted to test the effects of increasing concentrations of CHBr3 (0.4, 2, 10 and 50 µmol/L).ResultsThe addition of AT significantly decreased CH4 production (P = 0.002) and the acetate:propionate ratio (P = 0.003) during a 72-h incubation. The concentrations of CHBr3 showed a rapid decrease with nearly 90% degraded within the first 3 h of incubation. On the contrary, CH2Br2 concentration quickly increased during the first 6 h and then gradually decreased towards the end of the incubation. Neither CHBr3 degradation nor CH2Br2 synthesis were affected by the type of diet used as substrate, suggesting that the fermentation rate is not a driving factor involved in CHBr3 degradation. The in vitro culture of methanogens showed a dose-response effect of CHBr3 by inhibiting the growth of M. smithii, M. ruminantium, M. stadtmanae, M. barkeri, M. millerae, M. wolfei, and M. mobile.ConclusionsThe present work demonstrated that CHBr3 from A. taxiformis is quickly degraded to CH2Br2 in the rumen and that the fermentation rate promoted by different diets is not a driving factor involved in CHBr3 degradation.