Effects of Propionibacterium strains on ruminal fermentation,nutrient digestibility and methane emissions in beef cattle fed a corn grain finishing diet

Twenty ruminally cannulated beef heifers were fed a high corn grain diet in a randomized block design to determine the effect of three direct fed microbial (DFM) strains of Propionibacterium on ruminal fermentation, nutrient digestibility and methane (CH₄) emissions. The heifers were blocked in five groups on the basis of BW and used in five 28-day periods. Dietary treatments included (1) Control and three strains of Propionibacterium (2) P169, (3) P5, and (4) P54. Strains were administered directly into the rumen at 5×10⁹ CFU with 10 g of a maltodextrin carrier in a gel capsule; Control heifers received carrier only. All heifers were fed the basal diet (10 : 90 forage to concentrate, dry matter basis). Rumen contents were collected on days 15 and 18, ruminal pH was measured continuously between days 15 and 22, enteric CH₄ emissions were measured between days 19 and 22 and diet digestibility was measured from days 25 to 28. Mean ruminal pH was 5.91 and was not affected by treatments. Similarly, duration of time that pH<5.8 and 5.6 was not affected by treatment. Likewise, total and major volatile fatty acid profiles were similar among all treatments. No effects were observed on dry matter intake and total tract digestibility of nutrients. Total enteric CH₄ production (g/day) was not affected by Propionibacterium strains and averaged 139 g/day. Similarly, mean CH₄ yield (g CH₄/kg of dry matter intake) was similar for all the treatments. The relative abundance of total Propionibacteria in the rumen increased with administration of DFM and were greater 3 h post-dosing relative to Control, but returned to baseline levels before feeding. Populations of Propionibacterium P169 were higher at 3 and 9 h as compared with the levels at 0 h. In conclusion, moderate persistency of the inoculated strains within the ruminal microbiome and pre-existing high propionate production due to elevated levels of starch fermentation might have reduced the efficacy of Propionibacterium strains to increase molar proportion of propionate and subsequently reduce CH₄ emissions.     

Nitrous oxide emission factors for urine and dung from sheep fed either fresh forage rape (Brassica napus L.) or fresh perennial ryegrass (Lolium perenne L.)

In New Zealand, agriculture is predominantly based on pastoral grazing systems and animal excreta deposited on soil during grazing have been identified as a major source of nitrous oxide (N₂O) emissions. Forage brassicas (Brassica spp.) have been increasingly used to improve lamb performance. Compared with conventional forage perennial ryegrass (Lolium perenne L.), a common forage in New Zealand, forage brassicas have faster growth rates, higher dry matter production and higher nutritive value. The aim of this study was to determine the partitioning of dietary nitrogen (N) between urine and dung in the excreta from sheep fed forage brassica rape (B. napus subsp. oleifera L.) or ryegrass, and then to measure N₂O emissions when the excreta from the two different feed sources were applied to a pasture soil. A sheep metabolism study was conducted to determine urine and dung-N outputs from sheep fed forage rape or ryegrass, and N partitioning between urine and dung. Urine and dung were collected and then used in a field plot experiment for measuring N₂O emissions. The experimental site contained a perennial ryegrass/white clover pasture on a poorly drained silt-loam soil. The treatments included urine from sheep fed forage rape or ryegrass, dung from sheep fed forage rape or ryegrass, and a control without dung or urine applied. N₂O emission measurements were carried out using a static chamber technique. For each excreta type, the total N₂O emissions and emission factor (EF3; N₂O–N emitted during the 3- or 8-month measurement period as a per cent of animal urine or dung-N applied, respectively) were calculated. Our results indicate that, in terms of per unit of N intake, a similar amount of N was excreted in urine from sheep fed either forage rape or ryegrass, but less dung N was excreted from sheep fed forage rape than ryegrass. The EF3 for urine from sheep fed forage rape was lower compared with urine from sheep fed ryegrass. This may have been because of plant secondary metabolites, such as glucosinolates in forage rape and their degradation products, are transferred to urine and affect soil N transformation processes. However, the difference in the EF3 for dung from sheep fed ryegrass and forage rape was not significant.     

Does the diurnal pattern of enteric methane emissions from dairy cows change over time?

Diet manipulation and genetic selection are two important mitigation strategies for reducing enteric methane (CH₄) emissions from ruminant livestock. The aim of this study was to assess whether the diurnal pattern of CH₄ emissions from individual dairy cows changes over time when cows are fed on diets varying in forage composition. Emissions of CH₄ from 36 cows were measured during milking in an automatic (robotic) milking station in three consecutive feeding periods, for a total of 84 days. In Periods 1 and 2, the 36 cows were fed a high-forage partial mixed ration (PMR) containing 75% forage, with either a high grass silage or high maize silage content. In Period 3, cows were fed a commercial PMR containing 69% forage. Cows were offered PMR ad libitum plus concentrates during milking and CH₄ emitted by individual cows was sampled during 8662 milkings. A linear mixed model was used to assess differences among cows, feeding periods and time of day. Considerable variation was observed among cows in daily mean and diurnal patterns of CH₄ emissions. On average, cows produced less CH₄ when fed on the commercial PMR in feeding Period 3 than when the same cows were fed on high-forage diets in feeding Periods 1 and 2. The average diurnal pattern for CH₄ emissions did not significantly change between feeding periods and as lactation progressed. Emissions of CH₄ were positively associated with dry matter (DM) intake and forage DM intake. It is concluded that if the management of feed allocation remains constant then the diurnal pattern of CH₄ emissions from dairy cows will not necessarily alter over time. A change in diet composition may bring about an increase or decrease in absolute emissions over a 24-h period without significantly changing the diurnal pattern unless management of feed allocation changes. These findings are important for CH₄ monitoring techniques that involve taking measurements over short periods within a day rather than complete 24-h observations.     

Diet supplementation with cinnamon oil,cinnamaldehyde, or monensin does not reduce enteric methane production of dairy cows

This study was conducted to evaluate the effect of dietary addition of cinnamon oil (CIN), cinnamaldehyde (CDH), or monensin (MON) on enteric methane (CH₄) emission in dairy cows. Eight multiparous lactating Holstein cows fitted with ruminal cannulas were used in a replicated 4×4 Latin square design (28-day periods). Cows were fed (ad libitum) a total mixed ration ((TMR); 60 : 40 forage : concentrate ratio, on a dry matter (DM) basis) not supplemented (CTL), or supplemented with CIN (50 mg/kg DM intake), CDH (50 mg/kg DM intake), or monensin (24 mg/kg of DM intake). Dry matter intake (DMI), nutrient digestibility, N retention, and milk performance were measured over 6 consecutive days. Ruminal degradability of the basal diet (with no additive) was assessed using in sacco incubations (0, 2, 4, 8, 16, 24, 48, 72 and 96 h). Ruminal fermentation characteristics (pH, volatile fatty acids (VFA), and ammonia (NH₃)) and protozoa were determined over 2 days. Enteric CH₄ emissions were measured over 6 consecutive days using the sulfur hexafluoride (SF₆) tracer gas technique. Adding CIN, CDH or MON to the diet had no effects on DMI, N retention, in sacco ruminal degradation and nutrient digestibility of the diet. Ruminal fermentation characteristics and protozoa numbers were not modified by including the feed additives in the diet. Enteric CH₄ emission and CH₄ energy losses averaged 491 g/day and 6.59% of gross energy intake, respectively, and were not affected by adding CIN, CDH or MON to the diet. Results of this study indicate that CIN, CDH and MON are not viable CH₄ mitigation strategies in dairy cows.     

Behavioural adaptations of sheep to repeated acidosis challenges and effect of yeast supplementation

This study aims to determine whether sheep modify their feeding and general behaviour when they undergo acidosis challenge, whether these modifications are maintained when acidosis challenges are repeated and whether yeast supplementation affects these modifications. Twelve rumen-cannulated wethers fed concentrate (wheat) and forage (hay) were exposed to three 28-day periods consisting of a 23-day recovery phase (20% of wheat) followed by a 5-day acidosis challenge (60% of wheat). Both diets limited food intake to 90% of ad libitum intake. Six sheep received a daily supplementation of a live yeast product, six received a placebo. Ruminal pH was recorded continuously. Daily consumption of wheat, hay, water and weekly consumption of salt were monitored. Behavioural observations were performed twice in each period: once under the recovery phase and once under acidosis challenge. These observations included video recordings over 24 h (time budget), social tests (mixing with another sheep for 5 min) and nociception tests (CO₂ hot laser). As expected, sheep spent more time with a ruminal pH below 5.6 during challenges than during recovery phases (12.5 v. 4.7 h/day). Sheep drank more water (3.87 v. 3.27 l/day) and ingested more salt (16 v. 11 g/day) during challenges. They also spent more time standing than during recovery phases, adopting more frequent alarm postures and reacting more slowly to the hot stimulus. More severe behavioural modifications were observed during the first challenge than the two other challenges. Significant concentrate refusals were observed during challenge 1: from days 3 to 5 of this challenge, sheep ate only half of the distributed concentrate. Sheep were also more active and more aggressive towards each other in challenge 1. These behavioural modifications disappeared as the challenges were repeated: no behavioural modifications were observed between challenges and recovery phases during periods 2 and 3, and furthermore, sheep rapidly ate all the concentrate distributed during the third challenge. Focusing on the effects of yeast, the only differences registered between the two groups concerned ruminal pH, that is, mean ruminal pH values in the supplemented group were lower during the first challenge (5.11 v. 5.60) but higher during the third challenge (5.84 v. 5.28). In conclusion, our experiment suggests sheep can adapt to acidosis challenges, especially with yeast supplementation. Otherwise, ruminal pH values remained low during challenges, indicating that the modifications of general and feeding behaviour in subacute ruminal acidosis situations are not due exclusively to low ruminal pH values.     

Lambs Fed Fresh Winter Forage Rape (Brassica napus L.) Emit Less Methane than Those Fed Perennial Ryegrass (Lolium perenne L.), and Possible Mechanisms behind the Difference

The objectives of this study were to examine long-term effects of feeding forage rape (Brassica napus L.) on methane yields (g methane per kg of feed dry matter intake), and to propose mechanisms that may be responsible for lower emissions from lambs fed forage rape compared to perennial ryegrass (Lolium perenne L.). The lambs were fed fresh winter forage rape or ryegrass as their sole diet for 15 weeks. Methane yields were measured using open circuit respiration chambers, and were 22-30% smaller from forage rape than from ryegrass (averages of 13.6 g versus 19.5 g after 7 weeks, and 17.8 g versus 22.9 g after 15 weeks). The difference therefore persisted consistently for at least 3 months. The smaller methane yields from forage rape were not related to nitrate or sulfate in the feed, which might act as alternative electron acceptors, or to the levels of the potential inhibitors glucosinolates and S-methyl L-cysteine sulfoxide. Ruminal microbial communities in forage rape-fed lambs were different from those in ryegrass-fed lambs, with greater proportions of potentially propionate-forming bacteria, and were consistent with less hydrogen and hence less methane being produced during fermentation. The molar proportions of ruminal acetate were smaller and those of propionate were greater in forage rape-fed lambs, consistent with the larger propionate-forming populations and less hydrogen production. Forage rape contained more readily fermentable carbohydrates and less structural carbohydrates than ryegrass, and was more rapidly degraded in the rumen, which might favour this fermentation profile. The ruminal pH was lower in forage rape-fed lambs, which might inhibit methanogenic activity, shifting the rumen fermentation to more propionate and less hydrogen and methane. The significance of these two mechanisms remains to be investigated. The results suggest that forage rape is a potential methane mitigation tool in pastoral-based sheep production systems.     

Effects of feed intake on enteric methane emissions from sheep fed fresh white clover (Trifolium repens) and perennial ryegrass (Lolium perenne) forages

Published analyses of enteric methane (CH₄) emissions from sheep and cattle show an inverse relationship between feed intake and CH₄ yield (g CH₄/kg dry matter (DM) intake), which suggests opportunities for reducing CH₄ emissions from feed eaten and per unit of animal production. Most relationships between feed intake and CH₄ yield have been based on animals fed conserved feeds, especially silages and grains. Our research is a series of experiments with fresh white clover (Trifolium repens) and perennial ryegrass (Lolium perenne; ryegrass) forages fed to sheep at a range of feed intake levels. This study was comprised of four experiments where good quality freshly harvested white clover or ryegrass were fed to sheep over a three-fold range in DM intake, and CH₄ emissions were measured in respiration chambers for two consecutive days in each experiment. Measurements were made from 16 sheep in Experiment 1 (fed at 1.6 × metabolizable energy requirements for maintenance; ME_m), 28 sheep in Experiment 2 (at 0.8 and 2.0 × ME_m), eight sheep and two measurement periods in Experiment 3 (at 1.6 × ME_m), and 30 sheep in Experiment 4 (fed at 0.8, 1.2, 1.6, 2.0 and 2.5 × ME_m). Prior to each experiment, sheep had a 10 d acclimatization period to diets. Apparent digestibility was measured over 7 d from sheep in Experiments 1, 3 and 4, along with collection of rumen digesta for volatile fatty acid (VFA) determination. Although CH₄ yields differed when sheep were fed white clover or ryegrass at similar intakes, the differences were inconsistent and mean values similar across all experiments. This, and a similar structure of all experiments, enabled combined analysis of data from all four experiments using the restricted maximum likelihood (REML) procedure to estimate effects of feed intake level on digestibility, digestible nutrient intake, gas emissions, and VFA concentrations in the rumen. The REML analysis showed that when DM intake increased from 0.40 to 1.60 kg/d, the predicted responses were an increase in CH₄ production (g/d) of 187% (12.4–35.6 g/d; P<0.001), and a decline in CH₄ yield of 21% (25.6–20.2 g/kg DM intake; P<0.001). High feed intake levels were associated with increased molar proportions (mM of total VFA) of propionate from 0.17 to 0.21 (P=0.038). Single and multiple regressions were completed on the data from all experiments, with organic matter (OM) intake predicting 0.87 of the variation in CH₄ production, and molar proportion of propionate predicting 0.60 of the variation in CH₄ yield. Increasing feed intakes by 1 kg/d of DM reduced CH₄ yield by 4.5 g/kg DM intake. Plant chemical composition was weakly related to CH₄ yield. High intakes of fresh forages will lower CH₄ yield from fermentation, but effects of feed composition on CH₄ emissions were minor. The interaction between effects of feed intake and rumen function requires further investigation to understand relationships with CH₄ emissions.     

Effects of supplementation with corn distillers’ dried grains on animal performance,nitrogen balance,and enteric CH4 emissions of young Nellore bulls fed a high-tropical forage diet

The inclusion of corn-dried distillers’ grains (DDG) could be an alternative supplement to increase animal performance, nitrogen efficiency usage (NEU), and decrease enteric methane (CH₄) emissions. Our goal was to determine whether DDG could replace a traditional supplement (cottonseed meal) without affecting animal performance, N balance, and CH₄ emissions. The experiment was conducted during the forage growing season (December to April), with 15 d adaptation, and a 112 d experimental period. The experimental design was completely randomized with four treatments: a mineral supplement (MS), cottonseed meal supplement (CS), 50% replacement of CS by DDG (50DDG), and 100% replacement of CS by DDG (100DDG). Cottonseed meal and DDG were used as protein supplement. A total of 12 paddocks, 3 per treatment, were used to measure forage mass: morphological and chemical composition of forage, forage allowance, and animal performance. Six animals per treatment were used to evaluate DM intake, digestibility, CH₄ emissions, microbial protein production (MCP), and NEU of each treatment. Eighty-one Young Nellore bulls (48 testers, 12 per treatments and 33 adjusters) with initial BW of 255 ± 5 kg (10–12 months old) were supplemented with each supplement type at a level of 0.3% of BW. Pasture management was continuous stocking with a variable stocking rate (put-and-take). Enteric CH₄ was measured using the gas tracer technique. The MCP was quantified using purine derivatives and the NEU mass balance. No differences were found in nutrient intake (P > 0.228). Individual animal performance and gain per area were higher in the treatments with concentrates compared with that of MS; however, there was no difference among treatments CS, 50DDG, and 100DDG. The ADG was 0.83 for MS and 1.08 kg/animal/d when supplemented (P < 0.05). Gain per hectare was 709 kg/ha for MS and 915 kg/ha when supplemented with concentrates (P < 0.05). There was no difference in CH₄ production among treatments that average 180 g/animal/d; however, CH₄ per kg of gain was reduced with CS. The CH₄ conversion factor averaged 5.91%. There was no difference in the synthesis of MCP and NEU. Corn DDG can replace 100% of cottonseed meal as a protein source for supplementation of young Nellore bulls grazing in tropical pastures without affecting animal performance, NEU, MCP, and CH₄ emissions.     

Effects of forage composition and growing season on methane emission from sheep in the Inner Mongolia steppe of China

Understanding the effects of dietary composition on methane (CH₄) production of sheep can help us to understand grassland degradation resulting in an increase of CH₄ emission from ruminant livestock and its resulting significance affecting CH₄ source/sink in the grazing ecosystem. The objective of this study was to investigate the effect of forage composition in the diet of sheep in July and August on CH₄ production by sheep in the Inner Mongolia steppe. The four diet treatments were: (1) Leymus chinensis and Cleistogenes squarrosa (LC), (2) Leymus chinensis, Cleistogenes squarrosa and concentrate supplementation (LCC), (3) Artemisia frigida and Cleistogenes squarrosa (AC), and (4) Artemisia frigida, Cleistogenes squarrosa and concentrate supplementation (ACC). CH₄ production was significantly lower in July than in August (31.4 and 36.2 g per sheep-unit per day, respectively). The daily average CH₄ production per unit of digestive dry matter (DM), organic matter (OM) and neutral detergent fiber (NDF) increased by 10.9, 11.2 and 42.1% for the AC diet compared with the LC diet, respectively. Although concentrate supplementation in both the AC and LC diets increased total CH₄ production per sheep per day, it improved sheep productivity and decreased CH₄ production by 14.8, 12.5 and 14.8% per unit of DM, OM and NDF digested by the sheep, respectively. Our results suggested that in degraded grassland CH₄ emission from sheep was increased and concentrate supplementation increased diet use efficiency. Sheep-grazing ecosystem seems to be a source of CH₄ when the stocking rate is over 0.5 sheep-units ha⁻¹ during the growing season in the Inner Mongolia steppe. Shiping Wang and He Zhou contributed equally to this work.     

Supplementation with non-fibrous carbohydrates reduced fiber digestibility and did not improve microbial protein synthesis in sheep fed fresh forage of two nutritive values

To determine whether non-fibrous carbohydrate (NFC) supplementation improves fiber digestibility and microbial protein synthesis, 18 Corriedale ewes with a fixed intake level (40 g dry matter (DM)/kg BW0.75) were assigned to three (n = 6) diets: F = 100% fresh temperate forage, FG = 70% forage + 30% barley grain and FGM = 70% forage + 15% barley grain + 15% molasses-based product (MBP, Kalori 3000). Two experimental periods were carried out, with late (P1) and early (P2) vegetative stage forage. For P2, ewes were fitted with ruminal catheters. Forage was distributed at 0900 h, 1300 h, 1800 h and 2300 h, and supplement added at 0900 h and 1800 h meals. Digestibility of the different components of the diets, retained N and rumen microbial protein synthesis were determined. At the end of P2, ruminal pH and N-NH3 concentration were determined hourly for 24 h. Supplementation increased digestibility of DM (P < 0.001) and organic matter (OM; P < 0.001) and reduced NDF digestibility (P = 0.043) in both periods, with greater values in P2 (P = 0.008) for the three diets. Daily mean ruminal pH differed (P < 0.05) among treatments: 6.33 (F), 6.15 (FG) and 6.51 (FGM). The high pH in FGM was attributed to Ca(OH)2 in MBP. Therefore, the decreased fiber digestibility in supplemented diets could not be attributed to pH changes. The mean ruminal concentration of N-NH3 was 18.0 mg/dl, without differences among treatments or sampling hours. Microbial protein synthesis was greater in P2 (8.0 g/day) than in P1 (6.1 g/day; P = 0.006), but treatments did not enhance this parameter. The efficiency of protein synthesis tended to be lower in supplemented groups (16.4, 13.9 and 13.4 in P1, and 20.8, 16.7 and 16.2 g N/kg digestible OM ingested in P2, for F, FG and FGM, respectively; P = 0.07) without differences between supplements. The same tendency was observed for retained N: 2.55, 1.38 and 1.98 in P1, and 2.28, 1.23 and 1.10 g/day in P2, for F, FG and FGM, respectively; P = 0.05). The efficiency of microbial protein synthesis was greater in P2 (P = 0.007). In conclusion, addition of feeds containing NFCs to fresh temperate forage reduced the digestibility of cell walls and did not improve microbial protein synthesis or its efficiency. An increase in these parameters was associated to the early phenological stage of the forage.     

Soybean oil and linseed oil supplementation affect profiles of ruminal microorganisms in dairy cows

The objective of this study was to evaluate changes in ruminal microorganisms and fermentation parameters due to dietary supplementation of soybean and linseed oil alone or in combination. Four dietary treatments were tested in a Latin square designed experiment using four primiparous rumen-cannulated dairy cows. Treatments were control (C, 60 : 40 forage to concentrate) or C with 4% soybean oil (S), 4% linseed oil (L) or 2% soybean oil plus 2% linseed oil (SL) in a 4 × 4 Latin square with four periods of 21 days. Forage and concentrate mixtures were fed at 0800 and 2000 h daily. Ruminal fluid was collected every 2 h over a 12-h period on day 19 of each experimental period and pH was measured immediately. Samples were prepared for analyses of concentrations of volatile fatty acids (VFA) by GLC and ammonia. Counts of total and individual bacterial groups (cellulolytic, proteolytic, amylolytic bacteria and total viable bacteria) were performed using the roll-tube technique, and protozoa counts were measured via microscopy in ruminal fluid collected at 0, 4 and 8 h after the morning feeding. Content of ruminal digesta was obtained via the rumen cannula before the morning feeding and used immediately for DNA extraction and quantity of specific bacterial species was obtained using real- time PCR. Ruminal pH did not differ but total VFA (110 v. 105 mmol/l) were lower (P < 0.05) with oil supplementation compared with C. Concentration of ruminal NH3-N (4.4 v. 5.6 mmol/l) was greater (P < 0.05) due to oil compared with C. Compared with C, oil supplementation resulted in lower (P < 0.05) cellulolytic bacteria (3.25 × 108 v. 4.66 × 108 colony-forming units (CFU)/ml) and protozoa (9.04 × 104 v. 12.92 × 104 cell/ml) colony counts. Proteolytic bacteria (7.01 × 108 v. 6.08 × 108 CFU/ml) counts, however, were greater in response to oil compared with C (P < 0.05). Among oil treatments, the amount of Butyrivibrio fibrisolvens, Fibrobacter succinogenes and Ruminococcus flavefaciens in ruminal fluid was substantially lower (P < 0.05) when L was included. Compared to C, the amount of Ruminococcus albus decreased by an average of 40% regardless of oil level or type. Overall, the results indicate that some ruminal microorganisms, except proteolytic bacteria, are highly susceptible to dietary unsaturated fatty acids supplementation, particularly when linolenic acid rich oils were fed. Dietary oil effects on ruminal fermentation parameters seemed associated with the profile of ruminal microorganisms.     

Measurement and prediction of enteric methane emission

The greenhouse gas (GHG) emissions from the agricultural sector account for about 25.5% of total global anthropogenic emission. While CO₂ receives the most attention as a factor relative to global warming, CH₄, N₂O and chlorofluorocarbons (CFCs) also cause significant radiative forcing. With the relative global warming potential of 25 compared with CO₂, CH₄ is one of the most important GHGs. This article reviews the prediction models, estimation methodology and strategies for reducing enteric CH₄ emissions. Emission of CH₄ in ruminants differs among developed and developing countries, depending on factors like animal species, breed, pH of rumen fluid, ratio of acetate:propionate, methanogen population, composition of diet and amount of concentrate fed. Among the ruminant animals, cattle contribute the most towards the greenhouse effect through methane emission followed by sheep, goats and buffalos, respectively. The estimated CH₄ emission rate per cattle, buffaloe, sheep and goat in developed countries are 150.7, 137, 21.9 and 13.7 (g/animal/day) respectively. However, the estimated rates in developing countries are significantly lower at 95.9 and 13.7 (g/animal/day) per cattle and sheep, respectively. There exists a strong interest in developing new and improving the existing CH₄ prediction models to identify mitigation strategies for reducing the overall CH₄ emissions. A synthesis of the available literature suggests that the mechanistic models are superior to empirical models in accurately predicting the CH₄ emission from dairy farms. The latest development in prediction model is the integrated farm system model which is a process-based whole-farm simulation technique. Several techniques are used to quantify enteric CH₄ emissions starting from whole animal chambers to sulfur hexafluoride (SF6) tracer techniques. The latest technology developed to estimate CH₄ more accurately is the micrometeorological mass difference technique. Because the conditions under which animals are managed vary greatly by country, CH₄ emissions reduction strategies must be tailored to country-specific circumstances. Strategies that are cost effective, improve productivity, and have limited potential negative effects on livestock production hold a greater chance of being adopted by producers. It is also important to evaluate CH₄ mitigation strategies in terms of the total GHG budget and to consider the economics of various strategies. Although reductions in GHG emissions from livestock industries are seen as high priorities, strategies for reducing emissions should not reduce the economic viability of enterprises.     

Effect of increasing the proportion of chicory in forage-based diets on intake and digestion by sheep

There is a lot of evidence that chicory could be a highly palatable and nutritious source of forage for ruminants, well adapted to climate change and dry conditions in summer, thanks to its resistance to drought and high water content. This study aimed to describe the effect of incorporating chicory to ryegrass or to a ryegrass–white clover mixture on feeding behaviour, digestive parameters, nitrogen (N) balance and methane (CH₄) emissions in sheep. In total, three swards of ryegrass, white clover and chicory were established and managed in a manner ensuring the forage use at a constant vegetative stage throughout the experiment. In all, four dietary treatments (pure ryegrass; binary mixture: 50% ryegrass–50% chicory; ternary mixture: 50% ryegrass–25% white clover–25% chicory; and pure chicory) were evaluated in a 4×4 replicated Latin square design with eight young castrated Texel sheep. Each experimental period consisted of an 8-day diet adaptation phase, followed by a 6-day measuring phase during which intake dynamics, chewing activity, digestibility, rumen liquid passage rate, fermentation end-products, N balance and CH₄ emissions were determined. Data were analysed using a mixed model and orthogonal contrasts were used to detect the potential associative effects between ryegrass and chicory. The daily voluntary dry matter intake was lower for pure ryegrass than for diets containing chicory (P<0.001) and increased quadratically from 1.39 to 1.74 kg/day with increasing proportion of chicory. Huge positive quadratic effects (P<0.001) between ryegrass and chicory were detected on eating time and eating rate just after feeding indicating an increase of the motivation to eat with mixtures, whereas rumination activity decreased linearly with the proportion of chicory (P<0.001). The organic matter digestibility was similar among treatments (around 80%), but a strong positive quadratic P<0.001) effect was observed on liquid passage rate suggesting that chicory allowed fast particle breakdown in the rumen. Animals fed with the ryegrass–white clover–chicory mixture had the higher urinary N losses (P<0.001), whereas retained N per day or per g N intake was greater when the proportion of chicory was at least 50% (P<0.001) being ~40% greater than for the other treatments. The CH₄ yield was lower with pure chicory than with the other treatments (P<0.001) for which emissions were similar. In conclusion, mixing ryegrass and chicory in equal proportions produces a synergy on voluntary intake and an improved N use efficiency likely due to complementarity in chemical composition, increased motivation to eat and faster ruminal particle breakdown.     

Effect of fumarate reducing bacteria on in vitro rumen fermentation,methane mitigation and microbial diversity

The metabolic pathways involved in hydrogen (H₂) production, utilization and the activity of methanogens are the important factors that should be considered in controlling methane (CH₄) emissions by ruminants. H₂ as one of the major substrate for CH₄ production is therefore should be controlled. One of the strategies on reducing CH₄ is through the use of hydrogenotrophic microorganisms such as fumarate reducing bacteria. This study determined the effect of fumarate reducing bacteria, Mitsuokella jalaludinii, supplementation on in vitro rumen fermentation, CH4 production, diversity and quantity. M. jalaludinii significantly reduced CH₄ at 48 and 72 h of incubation and significantly increased succinate at 24 h. Although not significantly different, propionate was found to be highest in treatment containing M. jalaludinii at 12 and 48 h of incubation. These results suggest that supplementation of fumarate reducing bacteria to ruminal fermentation reduces CH₄ production and quantity, increases succinate and changes the rumen microbial diversity.     

Effects of In-Feed Inclusion of Clinoptilolite on Blood Serum Concentrations of Aluminium and Inorganic Phosphorus and on Ruminal Ph and Volatile Fatty Acid Concentrations in Dairy Cows

The experiment investigated the effects of the dietary inclusion of 200 g of the natural zeolite, clinoptilolite on the blood serum concentrations of aluminium (Al) and inorganic phosphorus (P) as well as on the ruminal pH and the ruminal concentrations of Al and P and of certain volatile fatty acids. Sixteen Holstein cows with a rumen fistula were randomly assigned to one of two groups. Group A cows (n = 8) were fed the basal ration supplemented with 200 g of clinoptilolite per day, and group B cows (n = 8) were fed the basal ration and served as controls. Blood and rumen fluid samples were collected at the same day of each week and at the same time (5 h after morning feeding) for 12 weeks. Clinoptilolite supplementation had no significant effect on ruminal and blood serum concentrations of Al and P. However, clinoptilolite significantly increased ruminal pH and acetate, and decreased ruminal propionate and valerate.     

Relationship between rumen methanogens and methane production in dairy cows fed diets supplemented with a feed enzyme additive

Aims: To investigate the relationship between ruminal methanogen community and host enteric methane (CH₄) production in lactating dairy cows fed diets supplemented with an exogenous fibrolytic enzyme additive. Methods and Results: Ecology of ruminal methanogens from dairy cows fed with or without exogenous fibrolytic enzymes was examined using PCR–denaturing gradient gel electrophoresis (PCR–DGGE) analyses and quantitative real‐time PCR (qRT‐PCR). The density of methanogens was not affected by the enzyme additive or sampling times, and no relationship was observed between the total methanogen population and CH₄ yield (as g per head per day or g kg^?1 DMI). The PCR–DGGE profiles consisted of 26 distinctive bands, with two bands similar to Methanogenic archaeon CH1270 negatively correlated, and one band similar to Methanobrevibacter gottschalkii strain HO positively correlated, with CH₄ yield. Three bands similar to Methanogenic archaeon CH1270 or Methanobrevibacter smithii ATCC 35061 appeared after enzyme was added. Conclusions: Supplementing a dairy cow diet with an exogenous fibrolytic enzyme additive increased CH₄ yield and altered the composition of the rumen methanogen community, but not the overall density of methanogens. Significance and Impact of the Study: This is the first study to identify the correlation between methanogen ecology and host CH₄ yield from lactating dairy cows.     

Nitrogen utilisation,energy utilisation and methane emissions of sheep grazing in two types of pasture

Livestock grazing plays a significant role in maintaining grasslands and promoting animal production globally. To understand the livestock performance in sown pasture (SP) vs native pasture (NP) is important to ensure more effective grassland-livestock interactions with minimal environmental impact. A 2 (treatment) * 2 (period) Latin Square design experiment was conducted with 10 growing Hu sheep ♂ × thin-tailed Han sheep ♀ rams grazed perennially SP vs NP in an inland arid region of China. The objectives were to evaluate the effects of grazing management on nutrient digestibility, nitrogen (N) and energy utilisation and methane (CH₄) emission. The N intake, N retained and energy intake (gross energy (GE), and digestible and metabolisable energy) of sheep grazing in SP were significantly increased compared with those grazing in NP. There were significant linear relationships between DM intake (DMI) (g/kg BW or g/kg BW^0.75) or CH₄ (g/kg BW or g/kg BW^0.75) emissions and forage nutrient and GE concentrations within each grassland type. The linear regression analysis indicated that forage CP or ether extract concentration was a good predictor for DMI (g/kg BW or g/kg BW^0.75) (R² = 0.756 or 0.752), and CH₄ emission could be predicted using forage nutrient and GE concentrations (R² = 0.381–0.503). These results suggest that DMI and CH₄ emissions per unit metabolic BW were accurately predicted by multiple-factor combinations of forage nutrients, including ether extract and CP paired with GE. The present output could provide useful information for the development of sustainable sheep grazing systems in the inland arid regions of the world.     

Prediction of enteric methane emissions from Holstein dairy cows fed various forage sources

Milk fatty acid (FA) profile has been previously used as a predictor of enteric CH₄output in dairy cows fed diets supplemented with plant oils, which can potentially impact ruminal fermentation. The objective of this study was to investigate the relationships between milk FA and enteric CH₄ emissions in lactating dairy cows fed different types of forages in the context of commonly fed diets. A total of 81 observations from three separate 3×3 Latin square design (32-day periods) experiments including a total of 27 lactating cows (96±27 days in milk; mean±SD) were used. Dietary forages were included at 60% of ration dry matter and were as follows: (1) 100% corn silage, (2) 100% alfalfa silage, (3) 100% barley silage, (4) 100% timothy silage, (5) 50 : 50 mix of corn and alfalfa silages, (6) 50 : 50 mix of barley and corn silages and (7) 50 : 50 mix of timothy and alfalfa silages. Enteric CH₄output was measured using respiration chambers during 3 consecutive days. Milk was sampled during the last 7 days of each period and analyzed for components and FA profile. Test variables included dry matter intake (DMI; kg/day), NDF (%), ether extract (%), milk yield (kg/day), milk components (%) and individual milk FA (% of total FA). Candidate multivariate models were obtained using the Least Absolute Shrinkage and Selection Operator and Least-Angle Regression methods based on the Schwarz Bayesian Criterion. Data were then fitted into a random regression using the MIXED procedure including the random effects of cow, period and study. A positive correlation was observed between CH₄ and DMI (r=0.59,P<0.001), whereas negative associations were observed between CH₄ and cis9-17:1 (r=−0.58, P<0.001), and trans8, cis13-18:2 (r=−0.51,P<0.001). Three different candidate models were selected and the best fit candidate model predicted CH₄ with a coefficient of determination of 0.84 after correction for cow, period and study effects and was: CH₄ (g/day)=319.7−57.4×15:0−13.8×cis9-17:1−39.5×trans10-18:1−59.9×cis11-18:1−253.1×trans8, cis12-18:2−642.7×trans8, cis13-18:2−195.7×trans11, cis15-18:2+16.5×DMI. Overall and linear prediction biases of all models were not significant (P>0.19). Milk FA profile and DMI can be used to predict CH₄emissions in dairy cows across a wide range of dietary forage sources     

Effects of Dietary Linseed Oil and Propionate Precursors on Ruminal Microbial Community,Composition, and Diversity in Yanbian Yellow Cattle

The rumen microbial ecosystem is a complex system where rumen fermentation processes involve interactions among microorganisms. There are important relationships between diet and the ruminal bacterial composition. Thus, we investigated the ruminal fermentation characteristics and compared ruminal bacterial communities using tag amplicon pyrosequencing analysis in Yanbian yellow steers, which were fed linseed oil (LO) and propionate precursors. We used eight ruminally cannulated Yanbian yellow steers (510 ± 5.8 kg) in a replicated 4 × 4 Latin square design with four dietary treatments. Steers were fed a basal diet that comprised 80% concentrate and 20% rice straw (DM basis, CON). The CON diet was supplemented with LO at 4%. The LO diet was also supplemented with 2% dl-malate or 2% fumarate as ruminal precursors of propionate. Dietary supplementation with LO and propionate precursors increased ruminal pH, total volatile fatty acid concentrations, and the molar proportion of propionate. The most abundant bacterial operational taxonomic units in the rumen were related to dietary treatments. Bacteroidetes dominated the ruminal bacterial community and the genus Prevotella was highly represented when steers were fed LO plus propionate precursors. However, with the CON and LO diet plus malate or fumarate, Firmicutes was the most abundant phylum and the genus Ruminococcus was predominant. In summary, supplementing the diets of ruminants with a moderate level of LO plus propionate precursors modified the ruminal fermentation pattern. The most positive responses to LO and propionate precursors supplementation were in the phyla Bacteriodetes and Firmicutes, and in the genus Ruminococcus and Prevotella. Thus, diets containing LO plus malate or fumarate have significant effects on the composition of the rumen microbial community.     

Metabolisable energy of grass and red clover silages fed to sheep at maintenance level

This study investigated the effect of forage type (grass or red clover) and harvesting time (primary growth or regrowth) of silage on energy and N utilisation by sheep fed at maintenance level. Specifically, the assumption of constant loss of energy of digestible organic matter from energy losses in urine and CH₄ applied in evaluation of silage metabolisable energy (ME) was investigated. Urinary excretion of high-energy phenolic compounds related to solubilisation of lignin was assumed to affect urinary energy (UE) losses from sheep fed highly digestible grass silage (GS). A total of 25 primary growth and regrowth silages of timothy (Phleum pratense) and meadow fescue (Festuca pratensis) grass mixtures and red clover (Trifolium pratense) samples collected in digestibility trials with sheep, including faecal and urine samples, were used for energy and N determinations. Urinary concentration of monophenolic compounds and CH₄ emissions in vitro were also analysed. Daily faecal N output, CH₄ yield (MJ/kg DM intake), proportion of CH₄ energy in digestible energy (DE) and proportion of UE in DE were greater (P ≤ 0.03) in sheep fed red clover silage (RCS) than GS. Furthermore, less (P = 0.01) energy was lost as UE of DE in sheep fed primary growth GS compared with the other treatments. The relationship between UE and silage N intake or urinary N output for both silage types (i.e. grass v. red clover) was strong, but the fit of the regressions was better for GS than RCS. The CH₄/DE ratio decreased (P < 0.05) and the UE/DE ratio increased (P < 0.05) with increasing organic matter digestibility in RCS. These relationships were not significant (P < 0.05) for the GS diets. The regression coefficient was higher (P < 0.05) for GS than RCS when regressing ME concentration on digestible organic matter. The results of this study imply that ME/DE ratio is not constant across first-cut GS of different maturities. The ME production response may be smaller from highly digestible first-cut GS but could not be clearly related to urinary excretion of monophenols derived from solubilisation of lignin. Furthermore, energy lost in urine was not clearly defined for RCS and was much more predictable for GS from silage N concentration.