Spot urine collection: A valid alternative to total urine collection for metabolomic studies in dairy cattle.

Urine is a highly suitable biological matrix for metabolomics studies. Total collection for 24-h periods is the gold standard as it ensures the presence of all metabolites excreted throughout the day. However, in animal studies, it presents limitations related to animal welfare and also due to alterations of the metabolome originating from the use of acid for preventing microbial growth or microbial contamination. In this study, we investigated whether spot urine collection is a practical alternative to total collection for metabolomic studies in lactating cows. For this purpose, we collected urine samples from 4 lactating Holstein cows fed 4 diets in a 4 × 4 Latin square design. Urine was collected for 24 h using a collecting device (i.e., total collection) or collected once per day 4 h after the morning feeding (i.e., spot urine collection). Dietary treatments differed by the amount of nitrogen content (high vs. low) and by the nature of the energy (starch vs. fiber). Urine metabolome was analyzed by 2 untargeted complementary methods, nuclear magnetic resonance and hydrophilic-interaction liquid chromatography (HILIC) coupled to a time-of-flight mass spectrometer, and by 1 targeted method, HILIC-tandem mass spectrometry. Although sampling technique had an effect on the abundance of metabolites detected, spot urine samples were equally capable of showing differences in urine metabolome than samples from total collection. When considering nitrogen levels in the diet, the robustness and precision for discriminating high- and low-nitrogen diets was equally achieved with both sampling techniques. A total of 22 discriminant metabolites associated with the N level of diets were identified from untargeted HILIC coupled to a time-of-flight mass spectrometer (n = 9) and nuclear magnetic resonance (n = 11), and 2 from targeted HILIC-tandem mass spectrometry. Alternatively, starch or fiber in the diet induced less changes in the metabolome that were not clearly discriminated independently of the sampling technique. We concluded that spot urine collection can successfully reveal differences in the urine metabolome elicited by dietary N levels and be used as a substitute of total urinary 24-h collection for metabolomic studies.

A new Tier 3 method to calculate methane emission inventory for ruminants.

Livestock is the main source of methane (CH4) emissions. It is important to accurately determine emissions from ruminants that meet standardized international guidelines for national greenhouse gas inventories. A new method to improve the accuracy of CH4 emissions that complies with IPCC rules for a Tier 3 method is described and evaluated. This method, developed by INRA (French Institute for Agricultural Research), was applied to the French inventory of CH4 emissions by ruminants and compared with the IPCC Tier 2 method. For enteric CH4, depending on the animal category, the INRA CH4 emission estimates lay between 88% and 114% of IPCC's. The INRA/IPCC ratio for enteric emission was close to unity and did not differ between methods (P = 0.43) for adult cows (i.e. most cattle). In France, feedlot manure is stored in aerobic conditions, and so the INRA/IPCC fit for manure emission was poorer (P < 0.05). The INRA/IPCC fit for enteric CH4 was very close between methods to that for total CH4 (P = 0.39), enteric CH4 representing 93% of total emissions. The main improvement is the use of a robust equation (from numerous data and diets), based on digestible organic matter intake (DOMI) corrected for the digestive interactions, to predict CH4 consistently from enteric and manure sources. It was developed for the French livestock inventory but is customizable for other countries. This new improved CH4 estimation method, based on equations from a large literature database, complies with IPCC rules for a Tier 3 method.

Effects of carbohydrate type or bicarbonate addition to grass silage-based diets on enteric methane emissions and milk fatty acid composition in dairy cows.

The aim of the study was to compare the effect of fiber- or starch-rich diets based on grass silage, supplemented or not with bicarbonate, on CH4 emissions and milk fatty acid (FA) profile in dairy cows. The experiment was conducted as a 4 × 4 Latin square design with a 2 × 2 factorial arrangement: carbohydrate type [starch- or fiber-rich diets with dietary starch level of 23.1 and 5.9% on a dry matter basis, respectively], without or with bicarbonate addition [0 and 1% of the dry matter intake, respectively]. Four multiparous lactating Holstein cows were fed 4 diets with 42% grass silage, 8% hay, and 50% concentrate in 4 consecutive 4-wk periods: (1) starch-rich diet, (2) starch-rich diet with bicarbonate, (3) fiber-rich diet, and (4) fiber-rich diet with bicarbonate. Intake and milk production were measured daily and milk composition was measured weekly; CH4 emission and total-tract digestibility were measured simultaneously (5 d, wk 4) when animals were in open-circuit respiration chambers. Sensors continuously monitored rumen pH (3 d, wk 4), and fermentation parameters were analyzed from rumen fluid samples taken before feeding (1 d, wk 3). Cows fed starch-rich diets had less CH4 emissions (on average, -18% in g/d; -15% in g/kg of dry matter intake; -19% in g/kg of milk) compared with fiber-rich diets. Carbohydrate type did not affect digestion of nutrients, except starch, which increased with starch-rich diets. The decrease in rumen protozoa number (-36%) and the shift in rumen fermentation toward propionate at the expense of butyrate for cows fed the starch-rich diets may be the main factor in reducing CH4 emissions. Milk of cows fed starch-rich diets had lower concentrations in trans-11 C18:1, sum of cis-C18, cis-9,trans-11 conjugated linoleic acid (CLA), and sum of CLA, along with greater concentration of some minor isomers of CLA and saturated FA in comparison to the fiber-rich diet. Bicarbonate addition did not influence CH4 emissions or nutrient digestibility regardless of the carbohydrate type in the diet. Rumen pH increased with bicarbonate addition, whereas other rumen parameters and milk FA composition were almost comparable between diets. Feeding dairy cows a starch-rich diet based on grass silage helps to limit the negative environmental effect of ruminants, but does not lead to greater milk nutritional value because milk saturated FA content is increased.

Tea saponin reduced methanogenesis in vitro but increased methane yield in lactating dairy cows.

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 (105/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.

Increasing linseed supply in dairy cow diets based on hay or corn silage: Effect on enteric methane emission, rumen microbial fermentation, and digestion.

We investigated the effects of increasing extruded linseed supply in diets based on hay (H; experiment 1) or corn silage (CS; experiment 2) on enteric methane (CH4) emission, rumen microbial and fermentation parameters, and rumen and total-tract digestibility. In each experiment, 4 lactating Holstein cows fitted with cannulas at the rumen and proximal duodenum were used in a 4×4 Latin square design (28-d periods). Cows were fed ad libitum a diet [50:50 and 60:40 forage:concentrate on a dry matter (DM) basis for experiments 1 and 2, respectively] without supplementation (H0, CS0) or supplemented with extruded linseed at 5% (H5, CS5), 10% (H10, CS10), and 15% (H15, CS15) of dietary DM (i.e., 1.8, 3.6 and 5.4% total fatty acids added, respectively). All measurements were carried out during the last 8 d of each period. Linseed supply linearly decreased daily CH4 emission in cows fed H diets (from 486 to 289g/d for H0 to H15, on average) and CS diets (from 354 to 207g/d for CS0 to CS15, on average). The average decrease in CH4 per kilogram of DM intake was, respectively, -7, -15, and -38% for H5, H10, H15 compared with the H0 diet, and -4, -8, and -34% for CS5, CS10, and CS15 compared with the CS0 diet. The same dose-response effect was observed on CH4 emission in percent of gross energy intake, per kilogram of nutrient digested, and per kilogram of 4% fat- and 3.3% protein-corrected milk (FPCM) in both experiments. Changes in the composition of rumen volatile fatty acids in response to increasing linseed supply resulted in a moderate or marked linear decrease in acetate:propionate ratio for H or CS diets, respectively. The depressive effect of linseed on total protozoa concentration was linear for H diets (-15 to -40%, on average, for H5 to H15 compared with H0) and quadratic for CS diets (-17 to -83%, on average, for CS5 to CS15 compared with CS0). Concentration of methanogens was similar among H or CS diets. The energetic benefits from the decreased CH4 emission with linseed supply in diets based on hay or corn silage did not improve digestibility or milk yield. Milk efficiency (kg of FPCM/kg of DM intake) was improved with linseed supply up to H10 in H diets and was unchanged in CS diets. Lower CH4 enteric emission from dairy cows fed linseed helps limit the environmental footprint of ruminant livestock.

Effects of dietary nitrogen levels and carbohydrate sources on apparent ruminal synthesis of some B vitamins in dairy cows.

Effects of nitrogen level and carbohydrate source on apparent ruminal synthesis (ARS) of thiamin, riboflavin, niacin, vitamin B6, folates, and vitamin B12 were evaluated using 4 lactating Holstein cows distributed in a 4 × 4 Latin square design with treatments following a 2 × 2 factorial arrangement. Cows were fitted with cannulas in the rumen and proximal duodenum. The treatments were 2 N levels and 2 carbohydrate sources. The diet with the high N level provided 14% crude protein, calculated to meet 110% of the protein requirements and an adequate supply in rumen-degradable protein, whereas the diet with the low N level contained 11% crude protein, calculated to meet 80% of the protein requirements with a shortage in rumen-degradable protein. Carbohydrate source treatments differed by their nature (i.e., high in starch from barley, corn, and wheat, or high in fiber from soybean hulls and dehydrated beet pulp). All 4 diets were isoenergetic, based on corn silage, and had the same forage-to-concentrate ratio (60:40, dry matter basis). Duodenal flow was determined using YbCl3 as a marker. Each B-vitamin ARS was calculated as duodenal flow minus daily intake. The intake of several B vitamins varied among treatments, but because the animals consumed a similar amount of feed every day (average of 20 kg of dry matter/d) the difference was mostly due to vitamin content of each ingredient and their relative proportion in the diets. Decreasing N concentration in the diet reduced vitamin B6 duodenal flow and increased its apparent ruminal degradation. It also decreased duodenal flow and ARS of folates. The high-starch diets increased duodenal flow and ruminal balance of riboflavin, vitamin B6, and folates, whereas the high-fiber diets increased vitamin B12 ARS and duodenal flow. These effects on apparent synthesis are possibly due to changes in ruminal fermentation.

Best available technology for European livestock farms: Availability, effectiveness and uptake.

Concerns over the negative environmental impact from livestock farming across Europe continue to make their mark resulting in new legislation and large research programs. However, despite a huge amount of published material and many available techniques, doubts over the success of national and European initiatives remain. Uptake of the more cost-effective and environmentally-friendly farming methods (such as dietary control, building design and good manure management) is already widespread but unlikely to be enough in itself to ensure that current environmental targets are fully met. Some of the abatement options available for intensive pig and poultry farming are brought together under the European IPPC/IED directive where they are listed as Best Available Techniques (BAT). This list is far from complete and other methods including many treatment options are currently excluded. However, the efficacies of many of the current BAT-listed options are modest, difficult to regulate and in some cases they may even be counterproductive with respect to other objectives ie pollution swapping. Evaluation of the existing and new BAT technologies is a key to a successful abatement of pollution from the sector and this in turn relies heavily on good measurement strategies. Consideration of the global effect of proposed techniques in the context of the whole farm will be essential for the development of a valid strategy.

Potential of tannin-rich plants, Leucaena leucocephala, Glyricidia sepium and Manihot esculenta, to reduce enteric methane emissions in sheep.

An in vivo trial was conducted in sheep to investigate the effect of three tropical tannin-rich plants (TRP) on methane emission, intake and digestibility. The TRP used were leaves of Glyricidia sepium, Leucaena leucocephala and Manihot esculenta that contained, respectively, 39, 75 and 92 g condensed tannins/kg DM. Methane was determined with the sulphur hexafluoride tracer technique. Eight rumen-cannulated sheep of two breeds (four Texel, four Blackbelly) were used in two 4 × 4 Latin square designs. Four experimental diets were tested. They consisted in a tropical natural grassland hay based on Dichanthium spp. fed alone (C) or in association with G. sepium (G), L. leucocephala (L) or M. esculenta (M) given as pellets at 44% of the daily ration. Daily organic matter intake was higher in TRP diets (686, 984, 1054 and 1186 g/day for C, G, L and M respectively; p < 0.05) while apparent organic matter total tract digestibility was not affected (69.9%, 62.8%, 65.3% and 64.7% for C, G, L and M respectively; p > 0.05). Methane emission was 47.1, 44.9, 33.3 and 33.5 g/kg digestible organic matter intake for C, G, L and M, respectively, and was significantly lower (p < 0.05) for L and M than for G and C. Our results confirm the potential of some TRP to reduce methane production. The strong decrease in methane and the increase in intake with TRPs may be due to their presentation as pellets.

Postruminal synthesis modifies the odd- and branched-chain fatty acid profile from the duodenum to milk.

Milk odd- and branched-chain fatty acids (OBCFA) have been suggested as potential biomarkers for rumen function. The potential of milk OBCFA as a biomarker depends on whether their profile reflects the profile observed in the duodenum. The objective of this study was to evaluate whether the OBCFA profile in duodenum samples is reflected in plasma and milk. For this, 2 dairy cattle experiments were used. In experiment 1, 4 Holstein cows fitted with rumen and proximal duodenum cannulas were used in a 4×4 Latin square design. The treatments consisted of 2 nitrogen levels (143 vs. 110g of crude protein/kg of dry matter for high and low N, respectively) combined with either 1 of the 2 energy sources (i.e., starch from barley, corn, and wheat or fiber from soybean hulls and dehydrated beet pulp). In experiment 2, 4 Holstein cows fitted with rumen and proximal duodenum cannulas were used in a 3×3 Latin square design, with the treatments consisting of 3 diets: (1) RNB-, a diet with a crude protein content of 122g/kg of dry matter, predicted to provide protein digested in the small intestine according to the requirement of the animals, but with a shortage of rumen degradable protein; (2) RNB- to which 6g/d of niacin was added through inclusion in the mineral and vitamin premix, and (3) RNB- to which urea was added to balance rumen degradable N supply resulting in a CP content of 156g/kg of dry matter. In both experiments, samples of duodenal digesta, plasma, and milk were collected and analyzed for fatty acids. Additionally, lipids in plasma samples were separated in lipid classes and analyzed for fatty acids. The OBCFA profile in milk was enriched in 15:0, iso-17:0, anteiso-17:0, and cis-9-17:1 as compared with duodenal samples, and milk secretions even exceeded duodenal flows, which suggests occurrence of postruminal synthesis, such as de novo synthesis, desaturation, and elongation. The postruminal modification of the OBCFA profile might hamper the application of OBCFA as diagnostic tools of rumen function.

Effects of incremental amounts of extruded linseed on the milk fatty acid composition of dairy cows receiving hay or corn silage.

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 × 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 (-24.1g/100g 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.

Effect of dairy production system, breed and co-product handling methods on environmental impacts at farm level.

Six dairy farms with the same on-farm area and milk production were compared. One farm (G-No) used grass as the sole forage for a herd of Normande cows, a dual-purpose breed. Three farms, with Holstein cows, varied forage for the herd from grass only (G-Ho) to low (G/LM-Ho) or high (G/HM-Ho) proportion of maize silage in the total forage area. Finally, two farms based on G/LM-Ho and G/HM-Ho systems aimed to increase omega-3 fatty acids in the winter diets of cows (G/LM/O3-Ho, G/HM/O3-Ho). Allocation methods (biophysical, protein, economic allocation) and system expansion applied for co-product (milk and meat) handling were examined. The impact categories considered were climate change, climate change including the effects of land use and land use change (CC/LULUC), cumulative energy demand, eutrophication, acidification and land occupation. The impacts per kg of fat-and-protein-corrected milk (FPCM) of G-No were highest, followed by those of G-Ho, G/LM-Ho and G/HM-Ho, regardless co-product handling methods and impact categories (except for eutrophication). CC/LULUC per kg FPCM of G/LM/O3-Ho and G/HM/O3-Ho were both 1% and 3% lower than those of G/LM-Ho and G/HM-Ho, respectively, but other impacts were higher. With system expansion, impacts per kg FPCM were lower than when allocation methods were used. Enteric fermentation was the greatest contributor (45-50%) to CC/LULUC, while grass production was the most important contributor to other impacts. The highest CC/LULUC (for G-No) can be explained by (1) G-No having the lowest milk yield/cow (though it produced the most meat) and (2) the fact that grass required more N fertiliser, but had lower yields than silage maize, even though grassland sequestered C. Among Holstein systems, increasing cow productivity by increasing feed intake (including maize silage and supplementing with concentrate) decreased impacts of milk. Reducing replacement rate and age of first calving also decreased impacts of milk. Increasing cow productivity reduced the amount of on-farm area required to produce a given amount of milk. Thus, the "liberated" on-farm area of Holstein systems was used to produce cash crops, and total impacts of these systems were lower than those of G-No (except for eutrophication and land occupation).

Microbial colonisation of tannin-rich tropical plants: Interplay between degradability, methane production and tannin disappearance in the rumen.

Condensed tannins in plants are found free and attached to protein and fibre but it is not known whether these fractions influence rumen degradation and microbial colonisation. This study explored the rumen degradation of tropical tannin-rich plants and the relationship between the disappearance of free and bound condensed tannin fractions and microbial communities colonising plant particles using in situ and in vitro experiments. Leaves from Calliandra calothyrsus, Gliricidia sepium, and Leucaena leucocephala, pods from Acacia nilotica and the leaves of two agricultural by-products: Manihot esculenta and Musa spp. were incubated in situ in the rumen of three dairy cows to determine their degradability for up to 96 h. Tannin disappearance was determined at 24 h of incubation, and adherent microbial communities were examined at 3 and 12 h of incubation using a metataxonomic approach. An in vitro approach was also used to assess the effects of these plants on rumen fermentation parameters. All plants contained more than 100 g/kg of condensed tannins with a large proportion (32-61%) bound to proteins. Calliandra calothyrsus had the highest concentration of condensed tannins at 361 g/kg, whereas Acacia nilotica was particularly rich in hydrolysable tannins (350 g/kg). Free condensed tannins from all plants completely disappeared after 24-h incubation in the rumen. Disappearance of protein-bound condensed tannins was variable with values ranging from 93% for Gliricidia sepium to 21% for Acacia nilotica. In contrast, fibre-bound condensed tannin disappearance averaged âˆ¼ 82% and did not vary between plants. Disappearance of bound fractions of condensed tannins was not associated with the degradability of plant fractions. The presence of tannins interfered with the microbial colonisation of plants. Each plant had distinct bacterial and archaeal communities after 3 and 12 h of incubation in the rumen and distinct protozoal communities at 3 h. Adherent communities in tannin-rich plants had a lower relative abundance of fibrolytic microbes, notably Fibrobacter spp. whereas, archaea diversity was reduced in high-tannin-containing Calliandra calothyrsus and Acacia nilotica at 12 h of incubation. Concurrently, in vitro methane production was lower for Calliandra calothyrsus, Acacia nilotica and Leucaena leucocephala although for the latter total volatile fatty acids production was not affected and was similar to control. Here, we show that the total amount of hydrolysable and condensed tannins contained in a plant govern the interaction with rumen microbes affecting degradability and fermentation. The effect of protein- and fibre-bound condensed tannins on degradability is less important.

Simultaneous quantification of carotenoids, retinol, and tocopherols in forages, bovine plasma, and milk: validation of a novel UPLC method.

Simultaneous quantification of various liposoluble micronutrients is not a new area of interest since these compounds participate in the nutritional quality of feeds that is largely explored in human, and also in animal diet. However, the development of related methods is still under concern, especially when the carotenoid composition is complex such as in forages given to ruminants or in lipid-rich matrices like milk. In this paper, an original method for simultaneous extraction and quantification of all carotenoids, vitamins E, and A in milk was proposed. Moreover, a new UPLC method allowing simultaneous determination of carotenoids and vitamins A and E in forage, plasma and milk, and separation of 23 peaks of carotenoids in forages was described. This UPLC method using a HSS T3 column and a gradient solvent system was compared to a previously published reverse-phase HPLC using two C18 columns in series and an isocratic solvent system. The UPLC method gave similar concentrations of carotenoids and vitamins A and E than the HPLC method. Moreover, UPLC allowed a better resolution for xanthophylls, especially lutein and zeaxanthin, for the three isomers of beta-carotene (all-E-, 9Z- and 13Z-) and for vitamins A, an equal or better sensitivity according to gradient, and a better reproducibility of peak areas and retention times, but did not reduce the time required for analysis.

Effects of increasing amounts of extruded linseed in the diet on apparent ruminal synthesis of some B vitamins in dairy cows.

Many studies have shown that metabolic efficiency of ruminants can be significantly decreased when B-vitamin supply is insufficient. Under the present state of knowledge, the amounts of B vitamins available for intestinal absorption cannot be predicted based on diet composition. Therefore, in an attempt to increase our understanding of the effects of dietary factors, on B-vitamin supply for dairy cows, the effects of increasing amounts of extruded linseed in diets based on hay (permanent grassland hay, H; Experiment 1) or corn silage (CS; Experiment 2) on apparent ruminal synthesis (ARS) of thiamin, riboflavin, niacin, vitamin B6, folates and vitamin B12 were evaluated. In each experiment, four lactating Holstein cows fitted with cannulas in the rumen and the proximal duodenum were used in a 4 × 4 Latin square design. In both experiments, the dietary treatments consisted of an increasing supply of extruded linseed representing 0%, 5%, 10% or 15% of diet DM. The forage : concentrate ratios were 50 : 50 and 60 : 40 for Experiments 1 and 2, respectively. Duodenal flow was determined using YbCl3 as a marker. The ARS of each B vitamin was calculated as duodenal flow - daily intake. In both experiments, treatments did not affect thiamin, riboflavin, niacin and vitamin B12 duodenal flow or ARS. Increasing the dietary concentration of extruded linseed decreased folate intake in Experiment 1 and vitamin B6 intake in Experiment 2 but resulted in a greater duodenal flow of vitamin B6 and folates regardless of the forage used in basal diet. Greater dietary linseed concentrations decreased vitamin B6 apparent degradation in the rumen in CS-based diet only and increased folate ARS in both H- and CS-based diets. Increasing linseed concentration of isonitrogenous and isoenergetic diets increased vitamin B6 and folate supply to dairy cows, both with H- and CS-based diets.

Milk fatty acids in dairy cows fed whole crude linseed, extruded linseed, or linseed oil, and their relationship with methane output.

This experiment studied the effect of 3 different physical forms of linseed fatty acids (FA) on cow dairy performance, milk FA secretion and composition, and their relationship with methane output. Eight multiparous, lactating Holstein cows were assigned to 1 of 4 dietary treatments in a replicated 4 x 4 Latin square design: a control diet (C) based on corn silage (59%) and concentrate (35%), and the same diet supplemented with whole crude linseed (CLS), extruded linseed (ELS), or linseed oil (LSO) at the same FA level (5% of dietary dry matter). Each experimental period lasted 4 wk. Dry matter intake was not modified with CLS but was lowered with both ELS and LSO (-3.1 and -5.1 kg/d, respectively) compared with C. Milk yield and milk fat content were similar for LSO and ELS but lower than for C and CLS (19.9 vs. 22.3 kg/d and 33.8 vs. 43.2 g/kg, on average, respectively). Compared with diet C, CLS changed the concentrations of a small number of FA; the main effects were decreases in 8:0 to 16:0 and increases in 18:0 and cis-9 18:1. Compared with diet C (and CLS in most cases), LSO appreciably changed the concentrations of almost all the FA measured; the main effects were decreases in FA from 4:0 to 16:0 and increases in 18:0, trans-11 16:1, all cis and trans 18:1 (except trans-11 18:1), and nonconjugated trans 18:2 isomers. The effect of ELS was either intermediate between those of CLS and LSO or similar to LSO with a few significant exceptions: increases in 17:0 iso; 18:3n-3; trans-11 18:1; cis-9, trans-11 conjugated linoleic acid; and trans-11, trans-13 conjugated linoleic acid and a smaller increase in cis-9 18:1. The most positive correlations (r = 0.87 to 0.91) between milk FA concentrations and methane output were observed for saturated FA from 6:0 to 16:0 and for 10:1, and the most negative correlations (r = -0.86 to -0.90) were observed for trans-16+cis-14 18:1; cis-9, trans-13 18:2; trans-11 16:1; and trans-12 18:1. Thus, milk FA profile can be considered a potential indicator of in vivo methane output in ruminants.

Effects of starch-rich or lipid-supplemented diets that induce milk fat depression on rumen biohydrogenation of fatty acids and methanogenesis in lactating dairy cows.

Optimizing milk production efficiency implies diets allowing low methane (CH4) emissions and high dairy performance. We hypothesize that nature of energy (starch v. lipids) and lipid supplement types (monounsaturated fatty acid (MUFA) v. polyunsaturated fatty acid (PUFA) mitigate CH4 emissions and can induce low milk fat content via different pathways. The main objective of this experiment was to study the effects of starch-rich or lipid-supplemented diets that induce milk fat depression (MFD) on rumen biohydrogenation (RBH) of unsaturated fatty acids (FA) and enteric CH4 emissions in dairy cows. Four multiparous lactating Holstein cows (days in milk=61±11 days) were used in a 4×4 Latin square design with four periods of 28 days. Four dietary treatments, three of which are likely to induce MFD, were based (dry matter basis) on 56% maize silage, 4% hay and 40% concentrates rich in: (1) saturated fatty acid (SFA) from Ca salts of palm oil (PALM); (2) starch from maize grain and wheat (MFD-Starch); (3) MUFA (cis-9 C18:1) from extruded rapeseeds (MFD-RS); and (4) PUFA (C18:2n-6) from extruded sunflower seeds (MFD-SF). Intake and milk production were measured daily. Milk composition and FA profile, CH4 emissions and total-tract digestibility were measured simultaneously when animals were in open-circuit respiration chambers. Fermentation parameters were analysed from rumen fluid samples taken before feeding. Dry matter intake, milk production, fat and protein contents, and CH4 emissions were similar among the four diets. We observed a higher milk SFA concentration with PALM and MFD-Starch, and lower milk MUFA and trans-10 C18:1 concentrations in comparison to MFD-RS and MFD-SF diets, while trans-11 C18:1 remained unchanged among diets. Milk total trans FA concentration was greater for MFD-SF than for PALM and MFD-Starch, with the value for MFD-RS being intermediate. Milk C18:3n-3 content was higher for MFD-RS than MFD-SF. The MFD seems more severe with MFD-SF and MFD-RS than PALM and MFD-Starch diets, because of a decrease in milk SFA concentration and a stronger shift from trans-11 C18:1 to trans-10 C18:1 in milk. The MFD-SF diet increased milk trans FA (+60%), trans-10 C18:1 (+31%), trans-10,cis-12 CLA (+27%) and PUFA (+36%) concentrations more than MFD-RS, which explains the numerically lowest milk fat yield and indicates that RBH pathways of PUFA differ between these two diets. Maize silage-based diets rich in starch or different unsaturated FA induced MFD with changes in milk FA profiles, but did not modify CH4 emissions.

Long-chain fatty acid metabolism in dairy cows: a meta-analysis of milk fatty acid yield in relation to duodenal flows and de novo synthesis.

This study is a meta-analysis of the response of milk long-chain fatty acid (FA) yield and composition to lipid supply, based on published experiments reporting duodenal FA flows or duodenal lipid infusions and milk FA composition (i.e., 39 experiments reporting 139 experimental treatments). Analysis of these data underlined the interdependence between milk yields of C18 and short- and medium-chain (C4 to C16) FA. Lipid supplementation (producing an increase in duodenal C18 flow) decreased linearly milk C4 to C16 yield (-0.26 g of C4 to C16 produced per gram of duodenal C18 flow increase) and increased quadratically milk C18 yield. When these 2 effects increased the percentage of C18 in milk FA up to a threshold value (around 52% of total FA), then milk C18 yield was limited by C4 to C16 yield, decreasing the C18 transfer efficiency from duodenum to milk with high-lipid diets. Moreover, for a given duodenal C18 flow, a decrease in milk C4 to C16 yield induced a decrease in milk C18 yield. Despite high variations in C18 transfer efficiency between duodenum and milk, for a given experimental condition, the percentages of C18 FA in milk total C18 could be predicted from their percentages in duodenal C18, and the percentages at the duodenum and in milk were very similar when mammary desaturation was taken into account (i.e., considering the sums of substrates and products of mammary desaturase). The estimated amounts of 18:0, trans-11-, and trans-13-18:1 desaturated by the mammary gland were a linear function of their mammary uptake, and mammary desaturation was responsible for 80, 95, and 81%, respectively, of the yield of their products (i.e., cis-9-18:1; cis-9, trans-11-, and cis-9, trans-13-18:2). Thus, mammary FA desaturation capacity did not seem to be a limiting factor in the experimental conditions published so far.

Linseed plus nitrate in the diet for fattening bulls: effects on methane emission, animal health and residues in offal.

The combination of linseed and nitrate is known to decrease enteric methane emission in dairy cows but few studies have been carried out in fattening cattle for animal liveweight gain, enteric methane emission, animal health and presence of residues in beef products. To address this gap, 16 young bulls received a control (C) diet between weaning at 9 months and 14 months, then were split into two groups of eight balanced on feed intake, BW gain and methane emission to receive either the C diet or a diet moderately supplemented with extruded linseed and calcium nitrate (LN) for 2 months before being slaughtered. On a dry matter (DM) basis, the C diet contained 70% baled grass silage and 30% concentrate mainly made of maize, wheat and rapeseed meal. In the LN diet, rapeseed meal and a fraction of cereals were replaced by 35% extruded linseed and 6% calcium nitrate; linseed fatty acids and nitrate supply in the LN diet were 1.9% and 1.0%, respectively. Methane emission was measured continuously using the GreenFeed system. Methaemoglobin was determined every week in peripheral blood from bulls receiving the LN diet. Nitrate and nitrite concentrations were determined in rumen, liver and tongue sampled at slaughter. Dry matter intake tended to be lower for LN diet (P=0.10). Body weight gain was lower for LN diet (P=0.01; 1.60 and 1.26 kg/day for C and LN diet, respectively). Daily methane emission was 9% lower (P<0.001) for LN than C diet (249 and 271 g/day, respectively) but methane yield did not differ between diets (24.1 and 23.2 g/kg DM intake for C and LN diet, respectively, P=0.34). Methaemoglobin was under the limit of detection (<2% of total haemoglobin) for most animals and was always lower than 5.6%, suggesting an absence of risk to animal health. Nitrite and nitrate concentrations in offal did not differ between C and LN diets. In conclusion, a moderate supply of linseed and nitrate in bull feed failed to decrease enteric methane yield and impaired bull liveweight gain but without adverse effects for animal health and food safety.

Comparison of 3 methods for estimating enteric methane and carbon dioxide emission in nonlactating cows.

Among techniques for estimating enteric methane (CH4) emission by ruminants, open-circuit respiration chambers (OC), the use of a gas tracer (SF6), and the GreenFeed (GF) device are the most commonly used. In this study, we compared these techniques in 8 dry cows receiving a diet made of 70% hay and 30% concentrates given in limited and constant amounts, in a 15-wk experiment. Two periods in free stalls for SF6 and GF and in chambers for OC were used; in addition, SF6 was determined in chambers for 1 period. Methane emission (g/d) and CH4 yield (g/kg DMI) were higher (P < 0.0001) for OC than for SF6 and GF (367, 310, and 319 g/d for OC, SF6, and GF, respectively). The difference between OC and GF was related to a difference in post-prandial rate of gas emission. The between-animal coefficient of variation of CH4 emission was higher for SF6 than for OC and GF (20.8, 13.5, and 12.0% on average, respectively). Correlation coefficients between OC and SF6 were high and significant for CH4 emission and CH4 yield (r = 0.782 and r = 0.717, respectively; P < 0.05), but not significant between OC and GF, or between SF6 and GF. Correlation coefficients were highly significant for SF6 determined either in free stalls or in chambers (r = 0.908 and 0.903 for CH4 in g/d and g/kg DMI, respectively; P < 0.01). Carbon dioxide (CO2) emission and CO2 yield were similar for GF and OC (10,003 and 9,887 g/d, 752 and 746 g/kg DMI, respectively); CO2 data obtained with SF6 were lower (7,718 g/d and 606 g/kg DMI; P < 0.0001), but this technique is not relevant for CO2 emission determination. Correlation coefficients between OC and GF were not significant for CO2 emission and CO2 yield. This set of results shows that differences between methods are minor for average values, but that individual correlations may limit their interchangeability for determining gas emissions of individual animals. This study also shows the reliability of GF on-farm determination of CH4 and CO2 emissions for groups of animals.