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Ruminant livestock enteric fermentation contributes approximately one-third of the global anthropogenic methane (CH 4 ) emissions and is projected to increase significantly to meet the increasing demand for animal-sourced protein. Methane, a short-lived greenhouse gas, needs to be reduced -24 to -47% by 2050 relative to 2010 to meet the 2.0°C target. This study describes the results of a comprehensive meta-analysis to determine effective mitigation strategies. The database included findings from 425 peer-reviewed studies (1963 to 2018). Mitigation strategies were classified into three main categories [animal and feed management, diet formulation, and rumen manipulation (additives and methods used to modify the rumen)] and up to five subcategories (98 total mitigation strategy combinations). A random-effects meta-analysis weighted by inverse variance was carried out (Comprehensive Meta-Analysis, V3.3.070). Five feeding strategies, namely CH 4 inhibitors, oils and fats, oilseeds, electron sinks, and tanniferous forages, decreased absolute CH 4 emissions by on average -21% (range -12 to -35%) and CH 4 emissions per unit of product (CH 4 I; meat or milk) by on average -17% (range -12 to -32%) without negatively affecting animal production (weight gain or milk yield). Furthermore, three strategies, namely decreasing dietary forage-to-concentrate ratio, increasing feeding level, and decreasing grass maturity, decreased CH 4 I by on average -12% (range -9 to -17%) and increased animal production by on average 45% (range 9 to 162%). The latter strategies are central to meeting the increasing demand for animal-sourced food. All strategies, but CH 4 inhibitors, can be implemented now and offer immediate approaches for combating global warming.

Direct-fed microbials (DFM) are considered as a promising technique to improve animal productivity without affecting animal health or harming the environment. The potential of three bacterial DFM to reduce methane (CH4) emissions, modulate ruminal fermentation, milk production and composition of primiparous dairy cows was examined in this study. As previous reports have shown that DFM respond differently to different diets, two contrasting diets were used in this study. Eight lactating primiparous cows were randomly divided into two groups that were fed a corn silage-based, high-starch diet (HSD) or a grass silage-based, high-fiber diet (HFD). Cows in each dietary group were randomly assigned to four treatments in a 4 × 4 Latin square design. The bacterial DFM used were selected for their proven CH4-reducing effect in vitro. Treatments included control (without DFM) and 3 DFM treatments: Propionibacterium freudenreichii 53-W (2.9 × 1010 colony forming units (CFU)/cow per day), Lactobacillus pentosus D31 (3.6 × 1011 CFU/cow per day) and Lactobacillus bulgaricus D1 (4.6 × 1010 CFU/cow per day). Each experimental period included 4 weeks of treatment and 1 week of wash-out, with measures performed in the fourth week of the treatment period. Enteric CH4 emissions were measured during 3 consecutive days using respiration chambers. Rumen samples were collected for ruminal fermentation parameters and quantitative microbial analyses. Milk samples were collected for composition analysis. Body weight of cows were recorded at the end of each treatment period. Irrespective of diet, no mitigating effect of DFM was observed on CH4 emissions in dairy cows. In contrast, Propionibacterium increased CH4 intensity by 27% (g CH4/kg milk) in cows fed HSD. There was no effect of DFM on other fermentation parameters and on bacterial, archaeal and protozoal numbers. Similarly, the effect of DFM on milk fatty acid composition was negligible. Propionibacterium and L. pentosus DFM tended to increase body weight gain with HSD. We conclude that, contrary to the effect previously observed in vitro, bacterial DFM Propionibacterium freudenreichii 53-W, Lactobacillus pentosus D31 and Lactobacillus bulgaricus D1 did not alter ruminal fermentation and failed to reduce CH4 emissions in lactating primiparous cows fed high-starch or high-fiber diets. Electronic supplementary material The online version of this article (10.1186/s40104-019-0342-9) contains supplementary material, which is available to authorized users.

This study investigated the effects of bacterial direct fed microbials (DFM) on ruminal fermentation and microbial characteristics, methane (CH4) emission, diet digestibility, and milk fatty acid (FA) composition in dairy cows fed diets formulated to induce different ruminal volatile fatty acid (VFA) profiles. Eight ruminally cannulated dairy cows were divided into 2 groups based on parity, days in milk, milk production, and body weight. Cows in each group were fed either a high-starch (38%, HS) or a low-starch (2%, LS) diet in a 55:45 forage-to-concentrate ratio on a dry matter (DM) basis. For each diet, cows were randomly assigned to 1 of 4 treatments in a Latin square design of (1) control (CON); (2) Propionibacterium P63 (P63); (3) P63 plus Lactobacillus plantarum 115 (P63+Lp); (4) P63 plus Lactobacillus rhamnosus 32(P63+Lr). Strains of DFM were administered at 10(10) cfu/d. Methane emission (using the sulfur hexafluoride tracer technique), total-tract digestibility, dry matter intake, and milk production and composition were quantified in wk 3. Ruminal fermentation and microbial characteristics were measured in wk 4. Data were analyzed using the mixed procedure of SAS (SAS Institute Inc., Cary, NC). The 2 diets induced different ruminal VFA profiles, with a greater proportion of propionate at the expense of acetate and butyrate for the HS diet. Greater concentrations of total bacteria and selected bacterial species of methanogenic Archaea were reported for the HS diet, whereas the protozoa concentration in HS decreased. For both diets, bacterial DFM supplementation raised ruminal pH (+0.18 pH units, on average) compared with CON. Irrespective of diet, P63+Lp and P63+Lr increased ruminal cellulase activity (3.8-fold, on average) compared with CON, but this effect was not associated with variations in ruminal microbial numbers. Irrespective of diet, no effect of bacterial DFM on ruminal VFA was observed. For the LS diet, supplementing cows with P63+Lr tended to decrease CH4 emission (26.5%, on average, when expressed per kilogram of milk or 4% fat-corrected milk). Only P63 supplementation to cows fed the HS diet affected the concentration of some milk FA, such as cis isomers of 18:1 and intermediates of ruminal biohydrogenation of polyunsaturated FA. Overall, bacterial DFM could be useful to stabilize ruminal pH. Their effects on CH4 production mitigation and milk FA profile depended on DFM strain and diet and should be confirmed under a greater variation of dietary conditions.

Knowledge on the flux of nitrogen (N) excreted through urines is essential in animal nutrition since it may provide information on animal N metabolism and the extent of environmental N pollution. Both the 15N natural enrichment of animal proteins over the diet (Δ15Nanimal-diet) and the milk urea N (MUN) concentration have been previously linked to N partitioning. The objective of this meta-analysis was to test the effect of combination of Δ15Nanimal-diet in plasma and MUN for predicting the urinary N excretion (UNE) in dairy cows. Individual data on Δ15Nanimal-diet in plasma, MUN, and UNE of 177 cow/period observations were evaluated by meta-analysis. Data were analyzed by mixed-model regression analysis. When model was fitted with either Δ15Nanimal-diet or MUN, both biomarkers were capable to independently predict UNE. However, when Δ15Nanimal-diet and MUN were combined in one model, both biomarkers remained significant and the error of the model decreased. In conclusion, both Δ15Nanimal-diet and MUN can predict UNE and their combination can strengthen the prediction capacity of the model.

The objective of this study was to evaluate the performance of existing models predicting enteric methane (CH) emissions, using a large database (3183 individual data from 103 in vivo studies on dairy and beef cattle, sheep and goats fed diets from different countries). The impacts of dietary strategies to reduce CH emissions, and of diet quality (described by organic matter digestibility (dOM) and neutral-detergent fiber digestibility (dNDF)) on model performance were assessed by animal category. The models were first assessed based on the root mean square prediction error (RMSPE) to standard deviation of observed values ratio (RSR) to account for differences in data between models and then on the RMSPE. For dairy cattle, the CH (g/d) predicting model based on feeding level (dry matter intake (DMI)/body weight (BW)), energy digestibility (dGE) and ether extract (EE) had the smallest RSR (0.66) for all diets, as well as for the high-EE diets (RSR = 0.73). For mitigation strategies based on lowering NDF or improving dOM, the same model (RSR = 0.48 to 0.60) and the model using DMI and neutral- and acid-detergent fiber intakes (RSR = 0.53) had the smallest RSR, respectively. For diets with high starch (STA), the model based on nitrogen, ADF and STA intake presented the smallest RSR (0.84). For beef cattle, all evaluated models performed moderately compared with the models of dairy cattle. The smallest RSR (0.83) was obtained using variables of energy intake, BW, forage content and dietary fat, and also for the high-EE and the low-NDF diets (RSR = 0.84 to 0.86). The IPCC Tier 2 models performed better when dietary STA, dOM or dNDF were high. For sheep and goats, the smallest RSR was observed from a model for sheep based on dGE intake (RSR = 0.61). Both IPCC models had low predictive ability when dietary EE, NDF, dOM and dNDF varied (RSR = 0.57 to 1.31 in dairy, and 0.65 to 1.24 in beef cattle). The performance of models depends mostly on explanatory variables and not on the type of data (individual vs. treatment means) used in their development or evaluation. Some empirical models give satisfactory prediction error compared with the error associated with measurement methods. For better prediction, models should include feed intake, digestibility and additional information on dietary concentrations of EE and structural and nonstructural carbohydrates to account for different dietary mitigating strategies., This study is part of the Joint Programming Initiative on Agriculture, Food Security and Climate Change (FACCE-JPI)’s “GLOBAL NETWORK” project and the “Feeding and Nutrition Network” (http://animalscience.psu.edu/fnn) of the Livestock Research Group within the Global Research Alliance for Agricultural Greenhouse Gases (www.globalresearchalliance.org). Authors gratefully acknowledge funding for this project from: USDA National Institute of Food and Agriculture (Grant no. 2014-67003-21979) University of California, Davis Sesnon Endowed Chair Program, USDA, and Austin Eugene Lyons Fellowship (University of California, Davis); Funding from USDA National Institute of Food and Agriculture Federal Appropriations under Project PEN 04539 and Accession number 1000803, DSM Nutritional Products (Basel, Switzerland), Pennsylvania Soybean Board (Harrisburg, PA, USA), Northeast Sustainable Agriculture Research and Education (Burlington, VT, USA), and PMI Nutritional Additives (Shoreview, MN, USA); the Ministry of Economic Affairs (the Netherlands; project BO-20-007-006; Global Research Alliance on Agricultural Greenhouse Gases), the Product Board Animal Feed (Zoetermeer, the Netherlands) and the Dutch Dairy Board (Zoetermeer, the Netherlands); The Cofund for Monitoring & Mitigation of Greenhouse Gases from Agri- and Silvi-culture (FACCE ERA-GAS)’s project Capturing Effects of Diet on Emissions from Ruminant Systems and the Dutch Ministry of Agriculture, Nature and Food Quality (AF-EU-18010 & BO-4400159-01); USDA National Institute of Food and Agriculture (Hatch Multistate NC-1042 Project Number NH00616-R; Project Accession Number 1001855) and the New Hampshire Agricultural Experiment Station (Durham, NH); French National Research Agency through the FACCE-JPI program (ANR-13-JFAC-0003-01); the Department of Agriculture, Food and the Marine, Ireland Agricultural GHG Research Initiative for Ireland (AGRI-I) project; Academy of Finland (No. 281337), Helsinki, Finland; Swiss Federal Office of Agriculture, Berne, Switzerland; the Department for Environment, Food and Rural Affairs (Defra; UK); Defra, the Scottish Government, DARD, and the Welsh Government as part of the UK’s Agricultural GHG Research Platform projects (www.ghgplatform.org.uk); INIA (Spain, project MIT01-GLOBALNET-EEZ); German Federal Ministry of Food and Agriculture (BMBL) through the Federal Office for Agriculture and Food (BLE); Swedish Infrastructure for Ecosystem Science (SITES) at Röbäcksdalen Research Station; Comisión Nacional de Investigación Científica y Tecnológica, Fondo Nacional de Desarrollo Científico y Tecnológico (Grant Nos. 11110410 and 1151355) and Fondo Regional de Tecnología Agropecuaria (FTG/RF-1028-RG); European Commission through SMEthane (FP7-SME-262270).

International audience; The aim of this work was to investigate the effects of feeding sheep with silage mixtures containing bioactive legumes on intake and digestive parameters. The bioactive legumes used were sainfoin (SF, Onobrychis viciifolia) and red clover (RC, Trifolium pratense), which contain condensed tannins (CT) and polyphenol oxidase respectively. Five treatments were assigned to two groups of sheep according to a replicated 5 × 5 Latin square design. The five types of silages tested were, on a dry matter (DM) basis: pure timothy grass silage (Phleum pratense, control, T), three binary mixtures of T‐SF, T‐RC and RC‐SF (500 g/kg each) and a ternary mixture of T‐RC‐SF (500, 250 and 250 g/kg respectively). The daily voluntary DM intake of silage mixtures containing both SF and RC was greater than for pure T silage, while the presence of SF resulted in lower organic matter digestibility compared to pure T. The rumen disappearance rate measured in situ increased linearly with the presence of SF and RC in silage. The nitrogen (N) digestibility was greater for pure T and T‐RC than for T‐SF, and the amount of N retained daily by the animals was greater for RC‐containing silages than for T and T‐SF. The methane (CH4) yield was greater for pure T than for the silage mixtures containing SF. We conclude that the presence of RC in silage could boost performances through intake and N retention, while SF‐based mixtures appear to have reduced negative environmental impacts through the reduction of CH4 emissions.

Dietary supplementation with linseed, saponins and nitrate is a promising methane mitigation strategy in ruminant production. The main objective of this work was to assess the effects of these additives on the rumen microbiota in order to understand underlying microbial mechanisms of methane abatement. Two 2 × 2 factorial design studies were conducted simultaneously, which also allowed us to make a broad-based assessment of microbial responses. Eight non-lactating cows were fed diets supplemented with linseed or saponin in order to decrease hydrogen production and nitrate to deviate hydrogen consumption; also, combinations of linseed plus nitrate or saponin plus nitrate were used to explore the interaction between dietary treatments. Amplicon sequencing of 18S and 16S rRNA genes was employed to characterise rumen microbes. Nitrate fed alone or in combination in both studies dramatically affected the composition and structure of rumen microbiota, though impacts were more evident in one of the studies. Linseed moderately modified bacterial community structure with no effect on rumen methanogens and protozoa. Indicator OTU analysis revealed that both linseed and nitrate reduced the relative abundance of hydrogen-producingRuminococcaceae. Linseed increased the proportion of bacteria known to reduce succinate to propionate, whereas nitrate supplementation increased nitrate-reducing bacteria and decreased the metabolic activity of rumen methanogens. Saponins had no effect on the microbiota. Inconsistency found between the two studies, when nitrate was fed to the cows could be explained by changes in microbial ecosystem functioning rather than changes in microbial community structure.ImportanceThis study aimed at identifying the microbial mechanisms of enteric methane mitigation when linseed, nitrate and saponins were fed to non-lactating cows alone or in a combination. Hydrogen is a limiting factor in rumen methanogenesis. We hypothesised that linseed and saponins would affect hydrogen producers and nitrate would deviate hydrogen consumption thus leading to reduced methane production in the rumen. Contrary to what was foreseen, both linseed and nitrate had a deleterious effect on hydrogen producers; linseed also redirected hydrogen consumption towards propionate production, whereas nitrate stimulated the growth of nitrate reducing and hence hydrogen-consuming bacterial taxa. Fundamental knowledge of microbial mechanism involved in rumen methanogenesis, provides novel insights for the development of new or the optimisation of existing methane mitigation strategies.

Evaluation and mitigation of enteric methane (CH4) emissions from ruminant livestock, in particular from dairy cows, have acquired global importance for sustainable, climate-smart cattle production. Based on CH4 reference measurements obtained with the SF6 tracer technique to determine ruminal CH4 production, a current equation permits evaluation of individual daily CH4 emissions of dairy cows based on milk Fourier transform mid-infrared (FT-MIR) spectra. However, the respiration chamber (RC) technique is considered to be more accurate than SF6 to measure CH4 production from cattle. This study aimed to develop an equation that allows estimating CH4 emissions of lactating cows recorded in an RC from corresponding milk FT-MIR spectra and to challenge its robustness and relevance through validation processes and its application on a milk spectral database. This would permit confirming the conclusions drawn with the existing equation based on SF6 reference measurements regarding the potential to estimate daily CH4 emissions of dairy cows from milk FT-MIR spectra. A total of 584 RC reference CH4 measurements (mean ± standard deviation of 400 ± 72 g of CH4/d) and corresponding standardized milk mid-infrared spectra were obtained from 148 individual lactating cows between 7 and 321 d in milk in 5 European countries (Germany, Switzerland, Denmark, France, and Northern Ireland). The developed equation based on RC measurements showed calibration and cross-validation coefficients of determination of 0.65 and 0.57, respectively, which is lower than those obtained earlier by the equation based on 532 SF6 measurements (0.74 and 0.70, respectively). This means that the RC-based model is unable to explain the variability observed in the corresponding reference data as well as the SF6-based model. The standard errors of calibration and cross-validation were lower for the RC model (43 and 47 g/d vs. 66 and 70 g/d for the SF6 version, respectively), indicating that the model based on RC data was closer to actual values. The root mean squared error (RMSE) of calibration of 42 g/d represents only 10% of the overall daily CH4 production, which is 23 g/d lower than the RMSE for the SF6-based equation. During the external validation step an RMSE of 62 g/d was observed. When the RC equation was applied to a standardized spectral database of milk recordings collected in the Walloon region of Belgium between January 2012 and December 2017 (1,515,137 spectra from 132,658 lactating cows in 1,176 different herds), an average ± standard deviation of 446 ± 51 g of CH4/d was estimated, which is consistent with the range of the values measured using both RC and SF6 techniques. This study confirmed that milk FT-MIR spectra could be used as a potential proxy to estimate daily CH4 emissions from dairy cows provided that the variability to predict is covered by the model.

The effect of tea saponin supplementation in the ruminant diet on methane emissions, rumen fermentation, and digestive processes is still under debate. The objective of this study was to assess the effect of this plant extract on methanogenesis, total-tract digestibility, and lactating performances of dairy cows. The work included 2 independent and successive experiments. First, the effect of 7 tea saponin doses (from 0 to 0.50 g/L) on methane emissions and protozoa concentrations was tested in 2 repeated in vitro batch culture incubations using bovine rumen contents as inoculum and a cereal mixture as substrate. After 18 h of incubation, total gas production and composition as well as rumen fermentation parameters and protozoa concentration were analyzed. Increasing dosage of the plant extract reduced methane production and protozoa concentration, with a maximum reduction of 29% for CH4 (mL/g of substrate) and 51% for protozoa (10(5)/mL). Tea saponin did not affect volatile fatty acids concentration, but marginally decreased total gas production by 5% at the highest dose. Second, a 2-period crossover design experiment was carried out with 8 lactating dairy cows fed a basal diet (54% corn silage, 6% hay, and 40% pelleted concentrates on a dry matter basis) without (control) or with 0.52% tea saponin (TSP). Each experimental period lasted 5 wk. Animals were fed ad libitum during the first 3 wk of the period (wk 1, 2, and 3) and restricted (95% of ad libitum intake) during the last 2 wk (wk 4 and 5). Intake and milk production were recorded daily. Methane emissions were quantified using open chambers (2 d, wk 4). Total-tract digestibility and nitrogen balance were determined from total feces and urine collected separately (5 d, wk 5). Rumen fermentation parameters and protozoa concentration were analyzed from samples taken after morning feeding (1 d, wk 5). Milk production, dry matter intake, and feed-efficiency were reduced with TSP (-18, -12, and -8%, respectively). As daily methane production (g/d) was not affected, methane emissions (g/kg of dry matter intake) increased by 14% with TSP. Total-tract digestibility and nitrogen balance were similar between diets, except for acid detergent fiber digestibility, which tended to be improved with TSP (+4 percentage units). Rumen fermentation parameters and protozoa concentration were relatively unchanged by diets. Under the conditions of this experiment, tea saponin is not efficient to reduce methane emissions from dairy cows.

Subacute ruminal acidosis (SARA) is usually characterized by abnormal and intermittent drops in rumen pH. Nevertheless, high individual animal variability in rumen pH and the difference in measurement methods for pH data acquisition decrease the sensitivity and accuracy of pH indicators for detecting SARA in ruminants. The aim of this study was to refine rumen pH indicators in long-term SARA based on individual dairy cow reticulo-rumen pH kinetics. Animal performances and rumen parameters were studied weekly in order to validate SARA syndrome and rumen pH was continuously measured using reticulo-rumen sensors. In total, 11 primiparous dairy cows were consecutively fed two different diets for 12 successive weeks: a control diet as low-starch diet (LSD; 13% starch for 4 weeks in period 1), an acidotic diet as high-starch diet (HSD; 32% starch for 4 weeks in period 2), and again the LSD diet (3 weeks in period 3). There was a 1-week dietary transition between LSD and HSD. Commonly used absolute SARA pH indicators such as daily average, area under the curve (AUC) and time spent below pH

International audience; The effect of supplementation of increasing amounts of extruded linseed in diets based on hay (H; experiment 1) or corn silage (CS; experiment 2) was investigated in regard to dairy performance and the milk fatty acid (FA) composition. In each experiment, 4 lactating multiparous Holstein cows were used in a 4 x 4 Latin square design (28-d periods). The cows were fed a diet (50:50 and 40:60 concentrate:forage ratio for experiments 1 and 2, respectively; dry matter basis) without supplementation (H0 or CS0) or supplemented with 5% (H5 or CS5), 10% (H10 or CS10), or 15% (H15 or CS15) of extruded linseed. Regardless of the forage type, diet supplementation with increasing amounts of extruded linseed had no effect on the dry matter intake, milk yield, or protein content or yield. In contrast, the milk fat content decreased progressively from H0 to H10 diets, and then decreased strongly with the H15 diet in response to increasing amounts of extruded linseed. For CS diets, the milk fat content initially decreased from CS0 to CS10, but then increased with the CS15 diet. For the H diets, the milk saturated FA decreased (-24.1 g/100 g of FA) linearly with increasing amounts of extruded linseed, whereas the milk monounsaturated FA (+19.0 g/100 g), polyunsaturated FA (+4.9 g/100 g), and total trans FA (+14.7 g/100 g) increased linearly. For the CS diets, the extent of the changes in the milk FA composition was generally lower than for the H diets. Milk 12:0 to 16:0 decreased in a similar manner in the 2 experiments with increasing amounts of extruded linseed intake, whereas 18:0 and cis-9 18:1 increased. The response of total trans 18:1 was slightly higher for the CS than H diets. The milk trans-10 18:1 content increased more with the CS than the H diets. The milk cis-9,trans-11 conjugated linoleic acid response to increasing amounts of extruded linseed intake was linear and curvilinear for the H diets, whereas it was only linear for the CS diets. The milk 18:3n-3 percentage increased in a similar logarithmic manner in the 2 experiments. It was concluded that the milk FA composition can be altered by extruded linseed supplementation with increasing concentrations of potentially health-beneficial FA (i.e., oleic acid, 18:3n-3, cis-9,trans-11 conjugated linoleic acid, and odd- and branched-chain FA) and decreasing concentrations of saturated FA. Extruded linseed supplementation increased the milk trans FA percentage.

The SF6 tracer technique (SF6) and GreenFeed system (GF) are two methods for measuring enteric methane (CH4) emissions from cattle. Both methods estimate individual daily CH4 emissions from expired gas samples collected either continuously over 24 h in a canister (SF6) or several times a day during short-term periods (3–8 min) when cattle visit an automated head chamber (GF). The objective of this work was to study repeatability (R) of each method according to duration of measurement period as an indicator of their precision. The R of CH4 measurements was evaluated in two different trials using cows. For Experiment 1, the SF6 technique was used for 20 days in six non-lactating dairy cows fed a hay-based diet; for Experiment 2, the GF system was used for 91 days in seven lactating dairy cows fed a maize silage-based diet. The CH4 data were grouped by periods of 1–10 days (SF6) and 1–45 days (GF). The CH4 emissions averaged 23.6 ± 3.9 g/kg dry matter intake (DMI) for the SF6 and 17.4 ± 3.3 g/kg DMI for the GF on the measurement period. To achieve an R value of 0.70 for CH4 emissions (g/kg DMI), 3-day periods were necessary for SF6 and 17-day periods for GF. The R did not increase after 4-day periods for SF6 (R = 0.73), but increased for GF until 45-day periods (R = 0.90). In our experimental conditions and R = 0.70, the total number of cows necessary to detect a significant difference in CH4 emissions (g/kg DMI) between two treatments (e.g. diet) was similar for SF6 and GF.

This study investigates the feasibility to predict individual methane (CH(4)) emissions from dairy cows using milk mid-infrared (MIR) spectra. To have a large variability of milk composition, two experiments were conducted on 11 lactating Holstein cows (two primiparous and nine multiparous). The first experiment aimed to induce a large variation in CH(4) emission by feeding two different diets: the first one was mainly composed of fresh grass and sugar beet pulp and the second one of maize silage and hay. The second experiment consisted of grass and corn silage with cracked corn, soybean meal and dried pulp. For each milking period, the milk yields were recorded twice daily and a milk sample of 50 ml was collected from each cow and analyzed by MIR spectrometry. Individual CH(4) emissions were measured daily using the sulfur hexafluoride method during a 7-day period. CH(4) daily emissions ranged from 10.2 to 47.1 g CH(4)/kg of milk. The spectral data were transformed to represent an average daily milk spectrum (AMS), which was related to the recorded daily CH(4) data. By assuming a delay before the production of fermentation products in the rumen and their use to produce milk components, five different calculations were used: AMS at days 0, 0.5, 1, 1.5 and 2 compared with the CH(4) measurement. The equations were built using Partial Least Squares regression. From the calculated R(2)(cv), it appears that the accuracy of CH(4) prediction by MIR changed in function of the milking days. In our experimental conditions, the AMS at day 1.5 compared with the measure of CH(4) emissions gave the best results. The R(2) and s.e. of the cross-validation were equal to 0.79 and 5.14 g of CH(4)/kg of milk. The multiple correlation analysis performed in this study showed the existence of a close relationship between milk fatty acid (FA) profile and CH(4) emission at day 1.5. The lower R(2) (R(2) = 0.76) obtained between FA profile and CH(4) emission compared with the one corresponding to the obtained calibration (R(2)(c) = 0.87) shows the interest to apply directly the developed CH(4) equation instead of the use of correlations between FA and CH(4). In conclusion, our preliminary results suggest the feasibility of direct CH(4) prediction from milk MIR spectra. Additional research has the potential to improve the calibrations even further. This alternative method could be useful to predict the individual CH(4) emissions at farm level or at the regional scale and it also could be used to identify low-CH(4)-emitting cows.

International audience; This study aimed to investigate whether sheep offered free-choice intake of forage and concentrate develop subacute ruminal acidosis (SARA) and to identify SARA-associated feeding behavior components. In a crossover design over two 28-d periods, 11 rumen-cannulated wethers received wheat and alfalfa hay in 2 separate compartments. Concentrate and forage were provided for ad libitum access or in a fixed amount corresponding to 80% of ad libitum hay intake with a concentrate: forage ratio of 60: 40 on a DM basis. In both diets, sheep were fed 2 equal portions at 0800 and 1600 h. Ruminal pH, voluntary intake, and feeding behavior were recorded continuously from d 1 to 9 and d 15 to 23 in each period. When no measurements were performed, the animals were housed in larger pens with straw bedding. When fed for ad libitum intake, the sheep ingested 1,340 g of DM/d consisting of 49.1% wheat, whereas with the fixed diet they ate 872 g of DM/d consisting of 58.4% wheat. Sheep fed for ad libitum intake spent more time with ruminal pH < 5.6 than when fed in fixed amounts (7.77 vs. 3.05 h/d, P < 0.001). The time spent with ruminal pH < 5.6 was mainly linked to the amount of feed ingested and especially the amount of wheat (P < 0.001). Our results suggest that when fed for ad libitum intake with free choice wheat, the achieved concentrate: forage ratio of near 50: 50 and a larger hay intake enable sheep to consume more wheat. When sheep were fed for ad libitum intake, feeding bouts were spread evenly throughout the day. Although ruminal pH reached the same minimum level in both diets after main meals, time to reach pH nadir was longer with ad libitum diet (P < 0.001). In addition, after reaching this minimum value, ruminal pH increased more slowly in this diet, inducing a decreased preprandial ruminal pH (P < 0.001). Consequently, the ad libitum diet led to a longer time below pH 5.6. A slow decrease in ruminal pH may enable sheep to consume larger quantities of food. However, free access to concentrate maintains continuously elevated content of ruminal fermentation end products and so requires more time for pH to return to neutral values. Thus, interval between feed distributions should be as large as possible to help resume the preprandial ruminal pH and to limit time spent with pH < 5.6.

International audience; The activities of bulls, their feeding behaviour and their ruminal pH were examined at several stages during the finishing period, according to the forage-to-concentrate ratio of their diet. Twenty-four bulls of the Blond d’Aquitaine breed (initial body weight = 326 ± 21 kg) were assigned to six balanced pens with a space allowance of 9.4 m2 per bull during the finishing period. They were fed three different diets with achieved forage-to-concentrate ratios of (i) 8% straw and 92% concentrate, (ii) 44% hay and 56% concentrate and (iii) 57% maize silage and 43% concentrate. Bulls had ad libitum access to feed dispensed once daily. Offered and refusals were weighed on 5 consecutive days per week. The bulls were slaughtered at the common final live weight of 650 kg and the finishing period lasted 138, 181 and 155 days for straw–concentrate, hay–concentrate and maize silage–concentrate diets, respectively. The time budget was estimated four times by scan sampling with a 10-min interval. Feeding behaviour was appraised using data from electronic feeding gates. Ruminal pH was measured from a ruminal fluid sample collected by rumenocentesis. On average, the bulls spent 78% of the time lying or standing still, and 11% of the time eating. The forage-to-concentrate ratio of the diet influenced only those activities directly linked to feeding, i.e. eating and drinking. Bulls fed a high-concentrate diet spent less time eating than the other bulls (47 min v. >2 h) and took shorter meals (7 min v. 17 min). The bulls fed the straw–concentrate diet spread their meals over the entire day, whereas the others maintained two major peaks of eating activity, the main one in the morning after feed dispensing, the other one at the end of the diurnal period. Intake rate ranged widely between diets, from 58 g/min on average for the diets based on hay or maize silage up to 173 g/min for the high-concentrate diet. The concentrate-diet bulls also had a lower ruminal pH during the first 2 months of the finishing period. The dispersion of meals based on a high-acidosis-risk diet may be a way to limit the decrease in ruminal pH.

We used six ruminally cannulated Texel wethers to study the relative role of protozoa and lactate-metabolizing bacteria in ruminal fermentative patterns during an induced latent acidosis. The sheep were fed an alfalfa hay diet (H) and latent acidosis was induced, following a short transition period of one week, with a grain-rich acidotic diet (W, 60% wheat + 40% alfalfa hay). Ruminal pH, ruminal volatile fatty acids (VFA), lactate and NH3 concentrations, protozoa and lactate-utilizing bacterial counts, the relative proportions of three main bacteria implicated in lactate metabolism (a lactate-producing species, Streptococcus bovis, and two lactate-utilizing species, Selenomonas ruminantium, and Megasphaera elsdenii) using specific 16S-rRNA-targeting oligonucleotide probes, and lactate dehydrogenase (LDH) activity were determined for both diets. The pH parameters (mean, minimum, maximum, time and area under pH 6.0 and 5.5) measured with the W diet were indicative of a latent (i.e., subacute and maintained) acidosis. However, a butyric rather than lactic latent acidosis was observed in this study. Total ruminal lactate concentration remained at low levels with the acidotic diet (< 4 mmol·L -1 ), but changes were observed in VFA composition, which was oriented towards butyrate at the expense of acetate (P < 0.05), while propionate remained constant. In agreement with the low ruminal lactate concentration, no changes in the proportion of S. bovis 16S-rRNA were observed. The lactate-metabolizing bacterial population also remained fairly constant in number, proportion and activity. The increase in butyrate concentration was accompanied by a proliferation of entodiniomorphs (P < 0.01). These results suggest that the protozoa limited lactate accumulation and possibly also the decrease in pH during latent acidosis. Experiments with defaunated and faunated sheep could provide further evidence of the role of protozoa in the development of rumen latent acidosis. acidosis / rumen fermentation / protozoa / bacteria / lactate

In order to examine the relationship between herbage maturity and methane (CH4) emission under conditions which minimized opportunities for selective grazing, six 8-yr-old, dry and early-pregnant Charolais cows [712 ± 22.7 kg liveweight (LW)] were grazed on a mono-specific pasture of timothy at four stages of maturity: early vegetative, heading, flowering, and senescence. Daily CH4 production was measured during 7 d using the sulfur hexafluoride (SF6) tracer-gas technique. Organic matter intake (OMI) was determined from fecal output estimated by Yb2O3 dosing and feed digestibility (OMD) estimated using a fecal N index. The OMD (%) decreased (P < 0.05) from 77.6 at early vegetative to 56.2 at senescence, but no cow effect upon OMD was observed. The OMI (14.8 g kg-1 LW) and CH4 production (273 g d-1) were higher (P < 0.05) at heading than at the other stages of maturity. The proportion of gross energy intake (GEI) lost in CH4 did not differ with maturity (mean 6.4% of GEI). Cow effects upon OMI and CH4 emission were significant (P < 0.05). CH4 production (g d-1) was not related to diet chemical composition or OMD, but was correlated with digestible neutral detergent fiber (NDF) intake (kg d-1) (r = 0.61, P < 0.01). Across all stages of maturity between-cow variation explained 54 to 70% of the total variation in CH4 production. Key words: Methane, cattle, animal variation, timothy, phenology, selective grazing

© 2012 Chaucheyras-Durand et al., licensee InTech. This is an open access chapter distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Use of Yeast Probiotics in Ruminants: Effects and Mechanisms of Action on Rumen pH, Fibre Degradation, and Microbiota According to the Diet

10 páginas, 4 tablas, 1 figura., Understanding the interactions between hydrogen producers and consumers in the rumen ecosystem is important for ruminant production and methane mitigation. The present study explored the relationships between rumen protozoa, methanogens and fermentation characteristics. A total of six donor sheep harbouring (F, faunated) or not (D, defaunated) protozoa in their rumens (D animals were kept without protozoa for a period of a few months (D2) or for more than 2 years (Dþ)) were used in in vitro and in vivo experiments. In vitro the absence of protozoa decreased NH3 and butyrate production and had no effect on methane. In contrast, the liquid-associated bacterial and methanogens fraction of Dþ inocula produced more methane than D2 and F inoculum (P,0·05). In vivo fermentation parameters of donor animals showed the same trend on NH3 and butyrate and showed that Dþ animals were high methane emitters, while D2 were the lowest (235 %). The concentration of dissolved dihydrogen measured after feeding followed the opposite trend. Methane emissions did not correlate with the relative abundance of methanogens in the rumen measured by quantitative PCR, but there was a trend for higher methanogens concentration in the solid-associated population of Dþ animals compared with D2 animals. In contrast, PCR-denaturing gradient gel electrophoresis profiles of methanogens’ methyl coenzyme-M reductase A gene showed a clear clustering in liquid-associated fractions for all three groups of donors but fewer differences in solid-associated fractions. These results show that the absence of protozoa may affect differently the methanogen community and methane emissions in wethers., The present study was supported by the French National Institute of Agricultural Research (INRA). D. P. M. and M. J. R. conceived and designed the study and carried out the primary data analysis and experimental work. D. P. M. wrote the manuscript. C. M. and J.-P. J. contributed to the statistical analysis and discussion of the results. All authors revised the manuscript. There are no conflicts of interest

The objective was to determine CH(4) production from bulls fed a feedlot diet rich in either fibre (F)or starch and lipid (SL) over the fattening period. Fifty six Charolais bulls (259 +/- 9.4 d of age and 339 +/- 8.2 kg live weight (LW)) were allocated randomly to one of two diets and blocked with 4 replicate pens/diet based on LW and age, and fattened for up to 18 months. Both treatments included barley straw with the appropriate concentrate mixture rich in fibre or starch and fat. The concentrate mixture and barley straw were available ad libitum, and the intake ratio (870:130: dry matter (DM)basis) for the concentrate mixture and barley straw was similar for both diets. Methane production was determined for each bull for 5 d using the sulfur hexafluoride tracer gas method at the beginning (24 d on diet +/- 3.4), middle (120 d +/- 8.2), and end (228 d +/- 11.1) of the fattening period. Feed intake was measured daily and bulls were weighed every 15 d. Ruminal fluid samples were collected on the last day of each CH(4) measurement period by rumenocentesis and measured for pH and concentrations of volatile fatty acids (VFA). Bulls fed SL had lower DM, organic matter and gross energy intake (P

This review gives an update of available data on the effect of nutrition on milk fatty acid (FA) composition in dairy cow and goat. It starts by an overview of the main digestive and metabolic pathways involved in the milk fat secretion processes. Used data are originated from literature and from two databases developed by INRA. Altogether, analyses confirm the wide plasticity of milk FA profiles in dairy ruminant, peculiarly for the saturated FA (10–18 carbon), oleic (9c-18:1) and vaccenic (11t-18:1) acids, and other trans isomers of 18:1 and 18:2. Comparing effects of diets with large amount of concentrate and/or maize silage with grass-based diets (grazed or correctly preserved) shows a decrease in saturated FA, at the expense of 9c- and 11t-18:1, and to a lesser extent, 18:3 n-3 and 9c11t-CLA. Dietary supplements of plant oil or oilseeds have similar effects, sometimes more marked, but they increase simultaneously other trans isomers of 18:1 and 18:2, especially when added to maize silage and/or high-concentrate diets. Oils or seeds rich in 18:2 n-6 (sunflower, soybean…) increase particularly 10t-18:1, and 10t12c-, 8t10c-, 7t9c- and 9t11c-CLA, whereas those rich in 18:3 n-3 (linseed) enhance notably 13t/14t-18:1, and 9ct12-, 9ct13- and 11tc15-18:2. When comparing goats’ with cows’ results, it seems that they are less sensitive to the 11t- to 10t- ruminal shift, thus explaining the good stability and the large magnitude of the responses of their milk 11t-18:1 and 9c11t-CLA content to lipid supplementation of high-concentrate diets. Furthermore, goats seem to respond better than cows to 18:3 n-3 rich supplements. Ongoing studies are evaluating the use of milk FA profile as biomarkers for the authentication of milk according to production zone and type of feeding, or to predict methane enteric emissions by dairy cows receiving oilseeds. Future studies should also integrate the expected progress of knowledge on the potential effects on consumers of the different major and minor FA found in standard or modified dairy products, as well as putative secondary effects of feeding conditions on sensory quality of dairy products and ruminant health.; La composition en acides gras (AG) du lait est une composante importante de sa qualité nutritionnelle pour l’homme, qualité qui est fortement modulable à court terme par l’alimentation des ruminants. Après un rappel des principaux phénomènes digestifs et métaboliques qui concourent à la synthèse des matières grasses du lait, cette revue présente les données récentes sur les effets de l’alimentation sur la composition en AG du lait de vache, ainsi que sur quelques particularités de la chèvre. Ces données confirment et précisent la forte plasticité du profil en AG du lait de ruminant, notamment pour les AG saturés (C10–C18), l’acide oléique (9c-18:1), l’acide vaccénique (11t-18:1) et d’autres isomères trans du 18:1 et du 18:2. Par rapport à des rations riches en concentrés et/ou en ensilage de maïs, les rations à base d’herbe (pâturée ou correctement conservée) diminuent les AG saturés au profit des 9c- et 11t-18:1, et dans une moindre mesure du 18:3 n-3 et du 9c11t-CLA. Les suppléments d’huiles ou graines oléagineuses ont des effets similaires, parfois plus marqués, mais ils augmentent aussi d’autres isomères trans du 18:1 et du 18:2, notamment lorsqu’ils sont ajoutés à des rations riches en concentrés et/ou en ensilage de maïs. Les huiles et graines riches en 18:2 n-6 (tournesol, soja…) accroissent en particulier le 10t-18:1 et les 10t12c-, 8t10c-, 7t9c- et 9t11c-CLA, alors que celles qui sont riches en 18:3 n-3 (lin) accroissent, notamment, le 13t/14t-18:1 et les 9c12t-, 9c13t- et 11t15c-18:2. Des différences marquées existent entre les réponses de la lipogenèse mammaire et du profil des AG du lait de la vache et de la chèvre laitière. La chèvre est moins sujette que la vache à la déviation du métabolisme ruminal du 11t- vers le 10t-18:1, ce qui explique probablement la stabilité et l’ampleur des réponses à la supplémentation lipidique des 11t-18:1 et 9c11t-CLA de son lait, y compris avec des rations riches en concentrés. En outre, l’effet inhibiteur du 10t12c-CLA sur la lipogenèse mammaire est beaucoup moins marqué chez cette espèce que chez la vache. Enfin, la chèvre semble mieux répondre que la vache à la supplémentation du régime en 18:3 n-3. Des études en cours évaluent une possible utilisation du profil en AG du lait pour tracer l’origine des produits, ou pour prédire la réduction de la production de méthane d’origine digestive chez la vache laitière recevant des graines oléagineuses. Les études futures devront aussi intégrer l’avancée attendue des connaissances en nutrition humaine sur les effets potentiels des différents AG majeurs et mineurs des produits laitiers standards ou modifiés, ainsi que les éventuels effets secondaires des pratiques alimentaires sur la qualité sensorielle des produits et la santé des animaux.

Elimination of protozoa or defaunation is known to reduce methane emission by ruminants. However, the long-term efficacy of this practice is not well documented. In this work methane and other end products of fermentation were monitored in five adult, rumen-cannulated wethers that had been kept defaunated for two years (Def1) and successively refaunated (Fau, 12 weeks) and defaunated (Def2) during a 6-month-long experiment. Methane was measured for periods of 4 days at about 4-weekly intervals using the SF6 tracer technique, and rumen content samples were taken regularly throughout the experiment. Refaunation was performed by single administration of a mixed protozoal inoculum (103 cells). A noticeable protozoal population was observed at 12 days after inoculation, which was followed by an exponential increase that peaked at 12 ± 3 × 105 cells/mL rumen fluid at 4 weeks and decreased thereafter to 8 ± 4 × 105 cells/mL at the end of the 12-week Fau period. Production of methane, which was 33 ± 8 L/animal.day in Def1, increased (P 0.05). Even though protozoal numbers were comparable to conventional animals 20 days after inoculation, the increase in methane production was observed later, at 8 weeks, suggesting that protozoa are not the only microbial factor favouring methanogenesis and that the ecosystem needs a relatively long adaptation time following protozoal introduction to optimise this function. The presence of protozoa did not affect total volatile fatty acids and acetate concentration in the rumen but increased butyrate, while decreasing propionate concentration (Fau v. Def1, Def2, P

International audience; Four ruminal cannulated Texel wethers were used to study the long-term effects of an acidotic diet on ruminal parameters and blood acid-base status. The short-term events around feeding and the recovery of the animals were followed after this nutritional disturbance. Sheep were limit-fed consecutively a control diet H (100% hay) (one week), an acidotic diet W (60% wheat + 40% hay) (2 weeks), and again the control diet (2 weeks). Mean, minimum and maximum ruminal pH were lower, and the time and area under pH 6.0 were higher ($P < 0.001$) with the W diet than with the H diet. These pH parameters indicate a latent acidosis defined here as a subacute and maintained acidosis. Before feeding, the drop in ruminal pH with the W diet was correlated with an increase in the VFA buffering capacity (BC) (R$^2 = 0.70$) and with a decrease in the BC of both carbonic acid functions (R$^2 = 0.52$ for H$_2$CO$_3$ and 0.55 for HCO$_3^-$). After feeding, the acidotic diet effect on ruminal pH was not explained by variations in the BC of either of these chemical species. Ruminal lactate concentration was higher with the W diet compared to the H diet ($P < 0.001$) but remained low ($ 0.05$). All ruminal parameters, except for ammonia, recovered to initial levels during the H diet redistribution ($P < 0.05$) while blood parameters decreased (pH, $P < 0.05$; bicarbonates (HCO$_3^-$), total CO$_2$ content (TCO$_2$), base excess in whole blood (Beb) and in extra cellular fluid (Beecf), $P < 0.01$). This decrease, initiated during the distribution of the acidotic diet, suggests a mobilization of body alkaline reserves and a longer recovery time in blood than in the rumen. We observed a non-lactic but butyric latent acidosis, linked to Entodiniomorph proliferation, suggesting an intermediate stage before the onset of acute lactic acidosis.; Modifications des paramètres fermentaires ruminaux et de l'équilibre acido-basique sanguin durant le développement d'une acidose latente provoquée sur moutons et lors de leur rétablissement. Quatre moutons Texels castrés, équipés d'une canule ruminale, ont été utilisés pour étudier les effets à long terme d'un régime acidogène sur les paramètres ruminaux et l'équilibre acido-basique sanguin. Les effets à court terme autour du repas ainsi que le rétablissement des animaux après cette perturbation nutritionnelle ont aussi été examinés. Les moutons ont reçu successivement en quantité limitée un régime témoin H (100 % foin) (une semaine), un régime acidogène W (60 % de blé + 40 % de foin) (2 semaines), et à nouveau le régime témoin (2 semaines). Les pH moyen, minimum et maximum étaient plus faibles et le temps et l'aire sous pH 6,0 plus élevés ($P < 0,001$) avec le régime W qu'avec le régime H. Les valeurs observées pour ces paramètres de pH sont caractéristiques d'une acidose latente définie ici comme une acidose subaiguë et prolongée. Avant repas, la chute du pH ruminal avec le régime W est corrélée avec une augmentation du pouvoir tampon (BC) des AGV (R$^2 = 0,70$) et avec une diminution du BC des deux fonctions de l'acide carbonique (R$^2 = 0,52$ pour H$_2$CO$_3$ et 0,55 pour HCO$_3^-$). Après repas, l'effet du régime acidogène sur le pH ruminal n'est pas expliqué par les variations du BC de l'une ou l'autre de ces espèces chimiques. La concentration du lactate ruminal est plus élevée avec le régime W qu'avec le régime H ($P < 0,001$) mais est restée à des valeurs faibles (< 2 mmol$\cdot$L$^{-1}$). La concentration en AGV totaux ($P < 0,001$), les proportions en acétate ($P < 0,001$) et en propionate ($P < 0,01$) dans le rumen ont diminué avec le régime W, alors que la proportion de butyrate a augmenté ($P < 0,001$). Parallèlement, le nombre d'Entodiniomorphes a augmenté avec le régime W ($P < 0,001$). La plupart des paramètres n'ont pas montré de variations significatives entre les 2 semaines de régime W ($P > 0,05$). Tous les paramètres ruminaux, à l'exception de l'ammoniaque, sont revenus à leur niveau initial dès la redistribution du régime H ($P < 0,05$), alors que les paramètres sanguins ont diminué (pH, $P < 0,05$ ; taux de bicarbonates (HCO$_3^-$) et de CO$_2$ total (TCO$_2$), excès de base dans le sang (Beb) et dans le fluide extracellulaire (Beecf), $P < 0,01$). Cette baisse, initiée durant la distribution du régime acidogène, suggère une mobilisation des réserves alcalines corporelles et un temps de rétablissement plus long dans le sang que dans le rumen. L'acidose latente butyrique et la prolifération des Entodiniomorphes observés dans cet essai pourraient être une étape intermédiaire avant le développement d'une acidose lactique aiguë.

ruminally cannulated cows were fed a forage diet (93% hay + 7% straw) and a mixed diet (33 % hay + 7% straw + 40% barley) in a 2 ¥ 2 crossover experimental design. In sacco degra- dation of forage, fibrolytic activities (polysaccharidases and glycosidases) of the solid-associated bacteria (SAB), and distribution of the 3 main cellulolytic bacterial species (Fibrobacter succinogenes, Ruminococcus albus, Ruminococcus flavefaciens) were determined for both diets. Barley supple- mentation decreased the hay degradation rate and mainly the polysaccharidase activities of the SAB (30% on average). The sum of rRNA of the 3 cellulolytic bacterial species represented on average 17% of the total bacterial signal and R. albus was the dominant cellulolytic bacterial species of the 3 stud- ied. Barley supplementation did not modify the proportion of the 3 cellulolytic bacteria attached to plant particles. The negative effect of barley on the ruminal hay degradation rate is due to a decrease in fibrolytic activity of the SAB, and not to a modification of the balance of the three cellulolytic bac- terial species examined.

The aim of the present work was to estimate volatile fatty acid (VFA) production rate in the rumen of sheep fed two levels of intake using both a tracer (TM; by isotope dilution) and a non-tracer method (NTM; by supplementary infusion) in steady-state conditions. Six wethers received a diet containing 700 g lucerne hay and 300 g ground maize/kg in eight equal meals at 3 h intervals per d. The diet (9·8 MJ metabolizable energy (ME)/kg DM) was offered at 90 %ad libitumconsumption (high intake, HI) or 45 %ad libitumconsumption (low intake, LI) in a 2×2 crossover design. Each sheep received five intrarumen VFA solutions infused continuously for 24 h at rates of 250 ml and 165 ml/h for the HI and LI respectively. The first infusion, considered as a control treatment (Con), consisted of a solution of [1-13C]propionate (7 mmol/d). The four other solutions were isoenergetic (1·9 MJ ME/kg DM intake) mixtures of unlabelled propionate (C3) and butyrate (C4) at different levels: 0·90 mol C4/kg DM intake; 0·60 mol C4+0·45 mol C3/kg DM intake; 0·30 mol C4+0·90 mol C3/kg DM intake; 1·35 mol C3/kg DM intake. The VFA infusions did not affect rumen fermentation of the basal diet (pH, osmotic pressure, protozoa numbers), and comparable DM digestibility of the diet among the different treatments was observed. Both estimation methods demonstrated a similar increase (1·7-fold) in the rumen VFA production rate of sheep fed at intakes varying between 0·9 to 1·7 times maintenance. Irrespective of the intake level, the rumen production rate of individual VFA was on average 1·5-fold higher when estimated by the TM compared with the NTM. Rumen VFA production rates estimated by the NTM and TM represented 80 % and 120 % ME intake respectively. The difference between NTM and TM estimates seems likely to be caused mainly by overestimation of the VFA production rates by the TM.

Four ewes, each fitted with a rumen cannula and with catheters in the mesenteric artery and portal and mesenteric veins, received continuous intrarumen infusions of water or of short-chain fatty acids (SCFA). SCFA infusions were isoenergetic (83 kJ/h) and provided rumen molar proportions (acetate : propionate : butyrate) of 70 : 20 : 10, 50 : 40 : 10 or 50 : 20 : 30. The rumen SCFA production rate with the basal diet was 90·0, 23·1 and 8·8 mmol/h for acetate, propionate and butyrate respectively. Portal net fluxes indicated that 74, 67 and 22–30 % of infused acetate, propionate and butyrate respectively, reached the portal vein. Portal net release of β-hydroxybutyrate increased with SCFA infusions, irrespective of the amount of butyrate infused. Portal net release of lactate decreased with high-butyrate infusion. Portal net uptake of glucose increased with the SCFA infusions. In ewes infused with water, a portal net uptake of total amino acids (AA) was observed. SCFA infusions decreased the uptake of nonessential AA (glutamate, glycine, but not glutamine) and increased the net release of tyrosine and essential AA (isoleucine, leucine). Portal net fluxes of AA were similar with both high-acetate and high-propionate infusions. Lower net uptake of glutamine and net release of most essential AA and some nonessential AA were observed with the high-butyrate infusion. Energetic summation of portal net release was not significantly different between the three SCFA infusions, although it tended to be lower with high-butyrate infusion. This may be related to the higher trophic effect of butyrate on the digestive mucosa.

Four ruminally cannulated cows were used to compare amino acid (AA) composition of protozoal and bacterial fractions as affected by sampling time and diet. Cows were given once a day restricted feed (80% of ad libitum intake) of 7 kg DM with two successive diets. Diet HB was 65% Cocksfoot hay and 35% pelleted ground barley, and Diet H was 100% Cocksfoot hay. Samples of whole ruminal contents were taken 2, 5, 8, 11, and 23 h after feeding for Diet HB and 2 h after feeding for Diet H to isolate the liquid-associated protozoa and bacteria (LAP, LAB) and particle-associated bacteria (PAB). At each sampling time, the AA compositions of the different microbial populations were determined. The AA profiles of the LAP were different from those of the bacteria for 13 AA out of 17 studied. Differences between AA compositions of LAB and PAB were also observed for 10 AA out of 17 studied. Irrespective of the microbial population, AA composition did not vary with sampling time after feeding diet HB (P > .05; except for arginine, glutamate, and glycine). The AA contents of none of the three microbial populations were affected (P > .05) by the diet except for leucine and glutamate (P < .01). The differences in AA profiles between LAP and bacteria and between LAB and PAB confirm the importance of the representativeness of the microbial reference sample for correctly estimating microbial AA flow into the small intestine.

Four ruminally cannulated cows were used to investigate effect of barley with or without buffer on modifications of mass and enzyme activity in rumen microorganisms. Animals were given restricted feed (80% of ad libitum intake) of 7 kg DM day−1 and with three successive diets. The diet H consisted of 100% Cocksfoot hay. The diet HB was 65% hay with 35% pelleted ground barley with infusion of bicarbonate salt for the diet HBB. The rumen was sampled before (-1 h) and after feeding (+ 5 h) to isolate liquid-and solid-associated microbial population. 15(NH4)2SO4 was continuously infused into the rumen as a microbial marker. For all diets microorganisms associated with particles constituted a mean of 74% of the total rumen microbial mass. The density of microorganisms adherent to particles was similar (P > 0.05) for the different diets and the two sampling times, but fibrolytic enzymes activities (xylanase, CMCase, glycosidases) in this microbial population were maximal (P < 0.001) 23 h after feed intake. Xylanase (P < 0.05) and CMCase (P < 0.01) activities just before feeding varied according to diet; they were highest with diet H and similar between barley supplemented diets. Higher glycosidase activities were measured with diet H and HBB than with diet HB 5 h after feeding.

Background Ruminal disbiosis induced by feeding is the cause of ruminal acidosis, a digestive disorder prevalent in high-producing ruminants. Because probiotic microorganisms can modulate the gastrointestinal microbiota, propionibacteria- and lactobacilli-based probiotics were tested for their effectiveness in preventing different forms of acidosis. Results Lactic acidosis, butyric and propionic subacute ruminal acidosis (SARA) were induced by feed chalenges in three groups of four wethers intraruminally dosed with wheat, corn or beet pulp. In each group, wethers were either not supplemented (C) or supplemented with Propionibacterium P63 alone (P) or combined with L. plantarum (Lp + P) or L. rhamnosus (Lr + P). Compared with C, all the probiotics stimulated lactobacilli proliferation, which reached up to 25% of total bacteria during wheat-induced lactic acidosis. This induced a large increase in lactate concentration, which decreased ruminal pH. During the corn-induced butyric SARA, Lp + P decreased Prevotella spp. proportion with a concomitant decrease in microbial amylase activity and total volatile fatty acids concentration, and an increase in xylanase activity and pH. Relative to the beet pulp-induced propionic SARA, P and Lr + P improved ruminal pH without affecting the microbial or fermentation characteristics. Regardless of acidosis type, denaturing gradient gel electrophoresis revealed that probiotic supplementations modified the bacterial community structure. Conclusion This work showed that the effectiveness of the bacterial probiotics tested depended on the acidosis type. Although these probiotics were ineffective in lactic acidosis because of a deeply disturbed rumen microbiota, some of the probiotics tested may be useful to minimize the occurrence of butyric and propionic SARA in sheep. However, their modes of action need to be further investigated.