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A meta-analysis based on an individual-cow data set was conducted to investigate the effects of between-cow variation and related animal variables on predicted CH4 emissions from dairy cows. Data were taken from 40 change-over studies consisting of a total of 637 cow/period observations. Animal production and rumen fermentation characteristics were measured for 154 diets in 40 studies; diet digestibility was measured for 135 diets in 34 studies, and ruminal digestion kinetics was measured for 56 diets in 15 studies. The experimental diets were based on grass silage, with cereal grains or by-products as energy supplements, and soybean or canola meal as protein supplements. Average forage:concentrate ratio across all diets on a dry matter basis was 59:41. Methane production was predicted from apparently fermented substrate using stoichiometric principles. Data were analyzed by mixed-model regression using diet and period within experiment as random effects, thereby allowing the effect of experiment, diet, and period to be excluded. Dry matter intake and milk yield were more repeatable experimental measures than rumen fermentation, nutrient outflow, diet digestibility, or estimated CH4 yield. Between-cow coefficient of variation (CV) was 0.010 for stoichiometric CH4 per mol of volatile fatty acids and 0.067 for predicted CH4 yield (CH4/dry matter intake). Organic matter digestibility (OMD) also displayed little between-cow variation (CV = 0.013), indicating that between-cow variation in diet digestibility and rumen fermentation pattern do not markedly contribute to between cow-variation in CH4 yield. Digesta passage rate was much more variable (CV = 0.08) between cows than OMD or rumen fermentation pattern. Increased digesta passage rate is associated with improved energetic efficiency of microbial N synthesis, which partitions fermented substrate from volatile fatty acids and gases to microbial cells that are more reduced than fermented carbohydrates. Positive relationships were observed between CH4 per mol of volatile fatty acids versus OMD and rumen ammonia N concentration versus OMD; and negative relationships between the efficiency of microbial N synthesis versus OMD and digesta passage rate versus OMD, suggesting that the effects of these variables on CH4 yield were additive. It can be concluded that variations in OMD and efficiency in microbial N synthesis resulting from variations in digesta passage contribute more to between-animal variation in CH4 emissions than rumen fermentation pattern.

Simple Summary Methane produced by ruminants contributes to increased greenhouse gas effect. There are various nutritional strategies to reduce methane emission, such as supplementing fat or changing starch levels in the diet. Understanding the interactions of these strategies on methane emission, as well as performance, digestibility, and rumen fermentation is important. The present study aimed to assess the effects of starch level with or without a mixture of sunflower and fish oils on nutrient intake and digestibility, milk yield and composition, rumen fermentation, ruminal CH4 emissions and microbial ecology in dairy cows. Oil mixture rich in polyunsaturated fatty acids supplemented to low- or high-starch diets reduced dry matter intake and increased energy digestibility of lactating cows. High starch level improved nutrient digestibility and tended to reduce ruminal acetate:propionate ratio but did not affect rumen pH, molar propionate ratio, or ruminal CH4 emissions. Oil decreased absolute ruminal CH4 emission or tended to decrease CH4 per energy corrected milk. Abstract Four multiparous dairy cows were used in a 4 × 4 Latin square to examine how starch level and oil mixture impact dry matter (DM) intake and digestibility, milk yield and composition, rumen fermentation, ruminal methane (CH4) emissions, and microbial diversity. Experimental treatments comprised high (HS) or low (LS) levels of starch containing 0 or 30 g of a mixture of sunflower and fish oils (2:1 w/w) per kg diet DM (LSO and HSO, respectively). Intake of DM did not differ between cows fed LS and HS diets while oil supplementation reduced DM intake. Dietary treatments did not affect milk and energy corrected milk yields. There was a tendency to have a lower milk fat concentration due to HSO compared with other treatments. Both high starch level and oil supplementation increased digestibility of gross energy. Cows receiving HS diets had higher levels of total rumen VFA while acetate was lower than LS without any differences in rumen pH, or ruminal CH4 emissions. Although dietary oil supplementation had no impact on rumen fermentation, decreased CH4 emissions (g/day and g/kg milk) were observed with a concomitant increase in Anoplodinium-Diplodinium sp. and Epidinium sp. but a decrease in Christensenellaceae, Ruminococcus sp., Methanobrevibacter ruminantium and Mbb. gottschalkii clades.

Cow genes dictate environmentally friendly microbiomes: a step towards reducing greenhouse gas emissions., A 1000-cow study across four European countries was undertaken to understand to what extent ruminant microbiomes can be controlled by the host animal and to identify characteristics of the host rumen microbiome axis that determine productivity and methane emissions. A core rumen microbiome, phylogenetically linked and with a preserved hierarchical structure, was identified. A 39-member subset of the core formed hubs in co-occurrence networks linking microbiome structure to host genetics and phenotype (methane emissions, rumen and blood metabolites, and milk production efficiency). These phenotypes can be predicted from the core microbiome using machine learning algorithms. The heritable core microbes, therefore, present primary targets for rumen manipulation toward sustainable and environmentally friendly agriculture.

Public health policies recommend a population wide decrease in the consumption of saturated fatty acids (SFA) to lower the incidence of cardiovascular and metabolic diseases. In most developed countries, milk and dairy products are the major source of SFA in the human diet. Altering milk fat composition offers the opportunity to lower the consumption of SFA without requiring a change in eating habits. Supplementing the diet of lactating cows with oilseeds, plant oils and marine lipids can be used to replace the SFA in milk fat with monounsaturated fatty acids (MUFA), and to a lesser extent, polyunsaturated fatty acids (PUFA). Due to ruminal metabolism, the decreases in milk SFA are also accompanied by increases in trans fatty acids (TFA), including conjugated isomers. The potential to lower SFA, enrich cis MUFA and PUFA, and alter the abundance and distribution of individual TFA in milk differs according to oil source, form of lipid supplement and degree of oilseed processing, and the influence of other components in the diet. The present review summarises recent evidence on changes in milk fat composition that can be achieved using dietary lipid supplements and highlights the challenges to commercial production of modified milk and dairy products. A meta-analysis on the effects of oilseeds on milk fatty acid composition is also presented.

Studies on microbial biohydrogenation of fatty acids in the rumen are of importance as this process lowers the availability of nutritionally beneficial unsaturated fatty acids for incorporation into meat and milk but also might result in the accumulation of biologically active intermediates. The impact was studied of adsorption of 22:6n-3 (DHA) to particulate material on its disappearance during 24 h in vitro batch incubations with rumen inoculum. Four adsorbants were used in two doses (1 and 5 mg/ml of mucin, gum arabic, bentonite or silicic acid). In addition, the distribution of 22:6n-3 in the pellet and supernatant of diluted rumen fluid was measured. Bentonite and silicic acid did not alter the distribution of 22:6n-3 between pellet and supernatant nor increased the disappearance of 22:6n-3 during the incubation. Both mucin and gum arabic increased the recovery of 22:6n-3 in the supernatant, indicating that these compounds lowered the adsorption of the fatty acid to ruminal particles. This was associated with an increased disappearance of 22:6n-3, when initial 22:6n-3 was 0.06 or 0.10 mg/ml, and an increased formation of 22:0, when initial 22:6n-3 was 0.02 mg/ml, during the 24 h batch culture experiment. Addition of gum arabic to pure cultures of Butyrivibrio fibrisolvens or Butyrivibrio proteoclasticus did not negate the inhibitory effect of 22:6n-3 on growth. As both mucin and gum arabic provide fermentable substrate for ruminal bacteria, an additional experiment was performed in which mucin and gum arabic were replaced by equal amounts of starch, cellulose or xylan. No differences in disappearance of 22:6n-3 were observed, suggesting that the stimulatory effect of mucin and gum arabic on disappearance of 22:6n-3 most probably is not due to provision of an alternative site of adsorption but related to stimulation of bacterial growth. A relatively high proportion of 22:6n-3 can be reduced to 22:0 provided the initial concentration is low.

Forage type and management influences the nutritional quality and fatty acid composition of ruminant milk. Replacing grass silage with red clover (RC; L.) silage increases milk fat 18:3-3 concentration. Red clover has a higher polyphenol oxidase (PPO) activity compared with grasses, which has been suggested to decrease lipolysis and . The present study characterized the abundance and fatty acid composition of esterified lipid and NEFA before and after ensiling of grass and RC to investigate the influence of forage species, growth stage, and extent of fermentation on lipolysis. A randomized block design with a 2 × 3 × 4 factorial arrangement of treatments was used. Treatments comprised RC or a mixture of timothy ( L.) and meadow fescue ( Huds.) harvested at 3 growth stages and treated with 4 levels of formic acid (0, 2, 4, and 6 L/t). Lipid in silages treated with 0 or 6 L/t formic acid were extracted and separated into 4 fractions by TLC. Total PPO activity in fresh herbage and the content of soluble bound phenols in all silages were determined. Concentrations of 18:3-3 and total fatty acids (TFA) were higher ( < 0.001) for RC than for grass. For both forage species, 18:3-3 and TFA content decreased linearly ( < 0.001) with advancing growth stage, with the highest abundance at the vegetative stage. Most of lipid in fresh RC and grass herbage (97%) was esterified, whereas NEFA accounted for 71% of TFA in both silages. Ensiling resulted in marginal increases in TFA content and the amounts of individual fatty acids compared with fresh herbages. Herbage total PPO activity was higher ( < 0.001) for RC than grass (11 vs. 0.11 μkatal/g leaf fresh weight). Net lipolysis during ensiling was extensive for both forage species (660 to 759 g/kg fatty acid for grass and 563 to 737 g/kg fatty acid for RC). Formic acid application (0 vs. 6 L/t) resulted in a marked decrease ( = 0.026) in net lipolysis during the ensiling of RC, whereas the opposite was true ( = 0.026) for grass. In conclusion, results suggest that formic acid addition during the ensilage of RC decreases lipolysis . For both plant species, total PPO activity was not associated with the extent of lipolysis . However, bound phenols formed via PPO activity appear to have a role in protecting lipid and protein against degradation in grass and lowering proteolysis of RC during ensiling.

International audience; Based on the potential benefits for long-term human health, there is interest in developing sustainable nutritional strategies for lowering medium-chain saturated fatty acids (FA) and increasing specific unsaturated FA in ruminant milk. Dietary supplements of extruded linseeds (EL), fish oil (FO) or a mixture of EL and FO increase cis-9,trans-11 CLA and long-chain n-3 polyunsaturated FA in bovine milk. Supplements of FO cause milk fat depression in lactating cows, but information for dairy goats is limited. A total of 14 Alpine goats were used in a replicated 3×3 Latin square with 28-days experimental periods to examine the effects of EL alone or in combination with FO on animal performance, milk fat synthesis and milk FA composition. Treatments comprised diets based on natural grassland hay supplemented with no additional oil (control), 530 of EL or 340 g/day of EL and 39 g/day of FO (ELFO). Compared with the control, ELFO tended (P=0.08) to lower milk fat yield, whereas EL increased (P

The nutritional value of meat is an increasingly important factor influencing consumer preferences for poultry, red meat and processed meat products. Intramuscular fat content and composition, in addition to high quality protein, trace minerals and vitamins are important determinants of nutritional value. Fat content of meat at retail has decreased substantially over the past 40 years through advances in animal genetics, nutrition and management and changes in processing techniques. Evidence of the association between diet and the incidence of human non-communicable diseases has driven an interest in developing production systems for lowering total SFA andtransfatty acid (TFA) content and enrichment ofn-3 PUFA concentrations in meat and meat products. Typically, poultry and pork has a lower fat content, containing higher PUFA and lower TFA concentrations than lamb or beef. Animal genetics, nutrition and maturity, coupled with their rumen microbiome, are the main factors influencing tissue lipid content and relative proportions of SFA, MUFA and PUFA. Altering the fatty acid (FA) profile of lamb and beef is determined to a large extent by extensive plant and microbial lipolysis and subsequent microbial biohydrogenation of dietary lipid in the rumen, and one of the major reasons explaining the differences in lipid composition of meat from monogastrics and ruminants. Nutritional strategies can be used to align the fat content and FA composition of poultry, pork, lamb and beef with Public Health Guidelines for lowering the social and economic burden of chronic disease.

Fish oil (FO) alters ruminal biohydrogenation causing trans fatty acid (FA) intermediates to accumulate, but the effects of 18-carbon polyunsaturated FA supply on ruminal long-chain FA metabolism and microbial communities in cattle fed FO are not well established. Four cows fitted with rumen cannula were used in a 4 × 4 Latin square with 21-d experimental periods to evaluate the effects of FO alone or in combination with plant oils high in 18:2n-6 or 18:3n-3 on rumen microbial ecology and flow of FA at the omasum. Treatments comprised a basal grass silage-based diet containing no additional oil (control) or supplements of FO (200 g/d) or FO (200 g/d) plus 500 g/d of sunflower oil (SFO) or linseed oil (LFO). Flow of FA was determined using the omasal sampling technique. The relative abundance of key biohydrogenating bacteria was assessed by quantitative PCR on 16S rRNA genes in omasal digesta. Fish oil-supplemented treatments increased the amounts of trans-18:1, trans-18:2, and 20- to 22-carbon polyunsaturated FA escaping the rumen. Relative to the control, oil supplements had no effect on the amount of 18:0 leaving the rumen, but LFO decreased the flow of 18:0 at the omasum compared with SFO. Both SFO and LFO increased trans-18:1 relative to FO, whereas LFO resulted in the highest trans-18:2 and 20- to 22-carbon FA flow. Supplements of FO plus plant oils shifted biohydrogenation toward trans-10 18:1 formation. Compared with FO alone, the ruminal metabolism of 22:6n-3 in the rumen of lactating cows is more extensive on diets containing higher amounts of 18-carbon polyunsaturated FA. However, the biohydrogenation of 22:5n-3 was less extensive in LFO than SFO, but showed no difference between FO and diets containing plant oils. Ruminal outflow of 20:5n-3 was not altered when plant oils were added to FO. Alterations in the amount of intermediates at the omasum or ruminal biohydrogenation pathways were not accompanied by major changes in analyzed bacterial populations. In conclusion, dietary supplements of FO alone or in combination with plant oils increase the amount of biohydrogenation intermediates containing 1 or more trans double bonds escaping the rumen, which may have implications for host metabolism and the nutritional quality of ruminant foods.

The objective of this experiment was to quantify the effects of graded replacement of late-harvested grass silage and barley with early-harvested silage on nutrient digestion and rumen fermentation. Four experimental diets were fed to 4 multiparous rumen-cannulated Nordic Red cows in 4 × 4 Latin square design with 21-d periods. Dietary treatments consisted of late-cut grass silage (LS) and rolled barley, which was gradually replaced with early-cut grass silage [ES; 0, 33, 67, and 100% of the forage component (ES + LS) of the diet]. With increased proportion of ES in the diet, the proportion of barley decreased from 47.2 to 26.6% on a dry matter basis. Early- and late-cut silages were harvested at 2-wk intervals (predicted concentrations of metabolizable energy 11.0 and 9.7 MJ/kg of dry matter). The 4 diets were formulated to support the same milk production. Nutrient flows were quantified using omasal sampling technique applying the triple-marker method (Cr, Yb, and indigestible neutral detergent fiber) and 15N as a microbial marker. Feed intake decreased with graded replacement of LS and barley with ES, but milk production was not influenced by diet. Digestibility of nutrients improved with graded addition of ES in the diet with the greatest difference observed in digestibility of neutral detergent fiber (NDF) and potentially digestible NDF (pdNDF). The results suggested that improved cell wall digestibility with graded level of ES in the diet was partly related to higher intrinsic digestibility of ES than LS, and partly due to negative associative effects with an increased proportion of LS and barley in the diet. Efficiency of microbial N synthesis was not influenced by the diet, but ruminal protein degradability increased with ES in the diet. Rumen fermentation pattern was not affected by the diet despite large difference in the profile of dietary carbohydrates. Rumen pool size of NDF and pdNDF, and ruminal turnover time of NDF decreased with graded addition of ES in the diet, whereas digestion rate of pdNDF improved. The results of this study indicate that increased CH4 yield in a parallel production study with graded addition of ES in the diet were more related to greater ruminal and total digestibility of organic matter than to the changes in rumen fermentation pattern.

12 páginas, 3 tablas, 1 figura., The modulation of milk fat nutritional quality through fish oil supplementation seems to be largely explained by the action of n-3 very long chain polyunsaturated fatty acids (PUFA) on ruminal biohydrogenation (BH) of C18 fatty acids (FA). However, relationships among this action, disappearance of those PUFA in the rumen, and potential detrimental consequences on ruminal fermentation remain uncertain. This study compared the effect of 20:5n-3 (eicosapentaenoic acid; EPA), 22:5n-3 (docosapentaenoic acid; DPA), and 22:6n-3 (docosahexaenoic acid; DHA) on rumen fermentation and BH of C18 FA and was conducted simultaneously in cows and sheep to provide novel insights into interspecies differences. The trial was performed in vitro using batch cultures of rumen microorganisms with inocula collected from cannulated cows and ewes. The PUFA were added at a dose of 2% incubated dry matter, and treatment effects on ruminal C18 FA concentrations, PUFA disappearances, and fermentation parameters (gas production, ammonia and volatile FA concentrations, and dry matter and neutral detergent fiber disappearances) were examined after 24 h of incubation. A principal component analysis suggested that responses to PUFA treatments explained most of the variability; those of ruminant species were of lower relevance. Overall, EPA and DHA were equally effective for inhibiting the saturation of trans-11 18:1 to 18:0 and had a similar influence on ruminal fermentation in cows and sheep (e.g., reductions in gas production and acetate:propionate ratio). Nevertheless, DHA further promoted alternative BH pathways that lead to trans-10 18:1 accumulation, and EPA seemed to have specific effects on 18:3n-3 metabolism. Only minor variations attributable to DPA were observed in the studied parameters, suggesting a low contribution of this FA to the action of marine lipids. Although most changes due to the added PUFA were comparable in bovine and ovine, there were also relevant specificities, such as a stronger inhibition of 18:0 formation in cows and a greater increase in 18:3n-3 metabolites in sheep. No direct relationship between in vitro disappearance of the incubated PUFA and effect on BH (in particular, inhibition of the last step) was found in either cows or ewes, calling into question a putative link between extent of disappearance and toxicity for microbiota. Conversely, an association between the influence of these PUFA on ruminal lipid metabolism and fermentation may exist in both species. In vivo verification of these findings would be advisable. Copyright © 2017 American Dairy Science Association. Published by Elsevier Inc. All rights reserved., We dedicate this article to the memory of Kevin J. Shingfield, a renowned expert in ruminant lipid metabolism and, above all, a good friend of ours. This work was supported by the Spanish Ministry of Economy and Competitiveness (MINECO; AGL2014-54587-R) and the Spanish National Research Council (CSIC; PIE201540E104). P. G. Toral benefits from a Ramón y Cajal research contract and D. Carreño benefits from an FPI predoctoral grant, both from MINECO. Cofunding by the European Regional Development Fund is also acknowledged.

The comparison of the bacterial profile of intracellular (iDNA) and extracellular DNA (eDNA) isolated from cow rumen content stored under different conditions was conducted. The influence of rumen fluid treatment (cheesecloth squeezed, centrifuged, filtered), storage temperature (RT, -80 °C) and cryoprotectants (PBS-glycerol, ethanol) on quality and quantity parameters of extracted DNA was evaluated by bacterial DGGE analysis, real-time PCR quantification and metabarcoding approach using high-throughput sequencing. Samples clustered according to the type of extracted DNA due to considerable differences between iDNA and eDNA bacterial profiles, while storage temperature and cryoprotectants additives had little effect on sample clustering. The numbers of Firmicutes and Bacteroidetes were lower (P 0.01) in eDNA samples. The qPCR indicated significantly higher amount of Firmicutes in iDNA sample frozen with glycerol (P 0.01). Deep sequencing analysis of iDNA samples revealed the prevalence of Bacteroidetes and similarity of samples frozen with and without cryoprotectants, which differed from sample stored with ethanol at room temperature. Centrifugation and consequent filtration of rumen fluid subjected to the eDNA isolation procedure considerably changed the ratio of molecular operational taxonomic units (MOTUs) of Bacteroidetes and Firmicutes. Intracellular DNA extraction using bead-beating method from cheesecloth sieved rumen content mixed with PBS-glycerol and stored at -80 °C was found as the optimal method to study ruminal bacterial profile.

Diets based on red clover silage (RCS) typically increase the concentration of polyunsaturated fatty acids (PUFA) in ruminant meat and milk and lower the efficiency of N utilization compared with grass silages (GS). Four multiparous Finnish Ayrshire cows (108 d postpartum) fitted with rumen cannulas were used in a 4 × 4 Latin square design with 21-d periods to evaluate the effect of incremental replacement of GS with RCS on milk production, nutrient digestion, whole-body N metabolism, and milk fatty acid composition. Treatments comprised total mixed rations offered ad libitum, containing 600 g of forage/kg of diet dry matter (DM), with RCS replacing GS in ratios of 0:100, 33:67, 67:33, and 100:0 on a DM basis. Intake of DM and milk yield tended to be higher when RCS and GS were offered as a mixture than when fed alone. Forage species had no influence on the concentration or secretion of total milk fat, whereas replacing GS with RCS tended to decrease milk protein concentration and yield. Substitution of GS with RCS decreased linearly whole-tract apparent organic matter, fiber, and N digestion. Forage species had no effect on total nonammonia N at the omasum, whereas the flow of most AA at the omasum was higher for diets based on a mixture of forages. Replacing GS with RCS progressively lowered protein degradation in the rumen, increased linearly ruminal escape of dietary protein, and decreased linearly microbial protein synthesis. Incremental inclusion of RCS in the diet tended to lower whole-body N balance, increased linearly the proportion of dietary N excreted in feces and urine, and decreased linearly the utilization of dietary N for milk protein synthesis. Furthermore, replacing GS with RCS decreased linearly milk fat 4:0 to 8:0, 14:0, and 16:0 concentrations and increased linearly 18:2n-6 and 18:3n-3 concentrations, in the absence of changes in cis-9 18:1, cis-9, trans-11 18:2, or total trans fatty acid concentration. Inclusion of RCS in the diet progressively increased the apparent transfer of 18-carbon PUFA from the diet into milk, but had no effect on the amount of 18:2n-6 or 18:3n-3 at the omasum recovered in milk. In conclusion, forage species modified ruminal N metabolism, the flow of AA at the omasum, and whole-body N partitioning. A lower efficiency of N utilization for milk protein synthesis with RCS relative to GS was associated with decreased availability of AA for absorption, with some evidence of an imbalance in the supply of AA relative to requirements. Higher enrichment of PUFA in milk for diets based on RCS was related to an increased supply for absorption, with no indication that forage species substantially altered PUFA bioavailability.

Milk and dairy products are major sources of fat in the human diet, but there are few detailed reports on the fatty acid composition of retail milk, trans fatty acids in particular, and how these change throughout the year. Semi-skimmed milk was collected monthly for one year from five supermarkets and analysed for fatty acid composition. Relative to winter, milk sold in the summer contained lower total saturated fatty acid (SFA; 67 vs 72 g/100g fatty acids) and higher cis-monounsaturated fatty acid (MUFA; 23 vs 21 g/100g fatty acids) and total trans fatty acid (6.5 vs 4.5 g/100g fatty acids) concentrations. Concentrations of most trans-18:1 and -18:2 isomers also exhibited seasonal variation. Results were applied to national dietary intakes, and indicated that monthly variation in the fatty acid composition of milk available at retail has limited influence on total dietary fatty acid consumption by UK adults.

Diets based on red clover silage (RCS) typically increase the concentration of polyunsaturated fatty acids (PUFA) in ruminant milk and meat compared with grass silages (GS), an effect that has been attributed to higher activity of polyphenol oxidase in red clover, promoting ruminal escape of dietary lipid. Four multiparous Finnish Ayrshire cows in mid lactation fitted with rumen cannulas were used in a 4×4 Latin Square design with 21-d experimental periods to evaluate the effects of incremental replacement of GS with RCS on ruminal lipid metabolism, using the omasal sampling technique in combination with Cr-EDTA, Yb acetate, and indigestible neutral detergent fiber as markers. Treatments comprised total mixed rations offered ad libitum containing 600 g of forage/kg of diet dry matter, with RCS replacing GS in a ratio of 0:100, 33:67, 67:33, and 100:0 on a dry matter basis. Silages contained a high proportion of lipid as nonesterified fatty acids (NEFA), with no difference between forage species (75 and 73% for GS and RCS, respectively). Substitution of GS with RCS had no influence on the intakes of NEFA, polar lipid, triacylglycerol, diacylglycerol, monoacylglycerol, or total fatty acids (FA), but altered the ingestion of specific FA. Replacing GS with RCS decreased linearly 18:3n-3 and increased linearly 18:2n-6 intakes. Changes in the proportion of RCS in the diet had no effect on the amounts or on the relative proportions of different lipid fractions at the omasum. On average, NEFA, polar lipid, triacylglycerol, diacylglycerol, and monoacylglycerol accounted for 80, 12, 4.4, 2.4, and 0.8% of total FA in omasal digesta, respectively. Replacement of GS with RCS increased linearly the amount of esterified and nonesterified 18:3n-3 at the omasum. Flows of cis -9 18:1 and 18:2n-6 were also increased linearly in response to RCS in the diet, whereas 3,7,11,15-tetramethyl-16:0 at the omasum was decreased. Replacing GS with RCS in the diet decreased linearly the lipolysis of dietary esterified lipids in the rumen from 85 to 70%. Effects on lipolysis due to forage species were also associated with linear decreases in apparent ruminal 18:3n-3 biohydrogenation from 93 to 85% and a trend toward lowered biohydrogenation of cis -9 18:1 and 18:2n-6 in the rumen. However, forage species had no effect on the flow of bound phenols formed as a consequence of polyphenol oxidase activity at the omasum. In conclusion, despite minimal differences in the extent of lipolysis in silo, lipid and constituent FA in RCS were less susceptible to ruminal lipolysis and biohydrogenation compared with GS.

Ruminants have a unique ability to derive energy from the degradation of plant polysaccharides through the activity of the rumen microbiota. Although this process is well studied in vitro, knowledge gaps remain regarding the relative contribution of the microbiota members and enzymes in vivo. The present study used RNA-sequencing to reveal both the expression of genes encoding carbohydrate-active enzymes (CAZymes) by the rumen microbiota of a lactating dairy cow and the microorganisms forming the fiber-degrading community. Functional analysis identified 12,237 CAZymes, accounting for 1% of the transcripts. The CAZyme profile was dominated by families GH94 (cellobiose-phosphorylase), GH13 (amylase), GH43 and GH10 (hemicellulases), GH9 and GH48 (cellulases), PL11 (pectinase) as well as GH2 and GH3 (oligosaccharidases). Our data support the pivotal role of the most characterized fibrolytic bacteria (Prevotella, Ruminocccus and Fibrobacter), and highlight a substantial, although most probably underestimated, contribution of fungi and ciliate protozoa to polysaccharide degradation. Particularly these results may motivate further exploration of the role and the functions of protozoa in the rumen. Moreover, an important part of the fibrolytic bacterial community remains to be characterized since one third of the CAZyme transcripts originated from distantly related strains. These findings are used to highlight limitations of current metatranscriptomics approaches to understand the functional rumen microbial community and opportunities to circumvent them.

The ruminal microbiome, comprising large numbers of bacteria, ciliate protozoa, archaea and fungi, responds to diet and dietary additives in a complex way. The aim of this study was to investigate the benefits of increasing the depth of the community analysis in describing and explaining responses to dietary changes. Quantitative PCR, ssu rRNA amplicon based taxa composition, diversity and co-occurrence network analyses were applied to ruminal digesta samples obtained from four multiparous Nordic Red dairy cows fitted with rumen cannulae. The cows received diets with forage:concentrate ratio either 35:65 (diet H) or 65:35 (L), supplemented or not with sunflower oil (SO) (0 or 50 g/kg diet dry matter), supplied in a 4 × 4 Latin square design with a 2 × 2 factorial arrangement of treatments and four 35-day periods. Digesta samples were collected on days 22 and 24 and combined. QPCR provided a broad picture in which a large fall in the abundance of fungi was seen with SO in the H but not the L diet. Amplicon sequencing showed higher community diversity indices in L as compared to H diets and revealed diet specific taxa abundance changes, highlighting large differences in protozoal and fungal composition. Methanobrevibacter ruminantium and Mbb. gottschalkii dominated archaeal communities, and their abundance correlated negatively with each other. Co-occurrence network analysis provided evidence that no microbial domain played a more central role in network formation, that some minor-abundance taxa were at nodes of highest centrality, and that microbial interactions were diet specific. Networks added new dimensions to our understanding of the diet effect on rumen microbial community interactions.

Camelina is an ancient oilseed crop that produces an oil rich in cis-9,cis-12 18:2 (linoleic acid, LA) and cis-9,cis-12,cis-15 18:3 (α-linolenic acid, ALA); however, reports on the use of camelina oil (CO) for ruminants are limited. The present study investigated the effects of incremental CO supplementation on animal performance, milk fatty acid (FA) composition, and milk sensory quality. Eight Finnish Ayrshire cows (91d in milk) were used in replicated 4×4 Latin squares with 21-d periods. Treatments comprised 4 concentrates (12kg/d on an air-dry basis) based on cereals and camelina expeller containing 0 (control), 2, 4, or 6% CO on an air-dry basis. Cows were offered a mixture of grass and red clover silage (RCS; 1:1 on a dry matter basis) ad libitum. Incremental CO supplementation linearly decreased silage and total dry matter intake, and linearly increased LA, ALA, and total FA intake. Treatments had no effect on whole-tract apparent organic matter or fiber digestibility and did not have a major influence on rumen fermentation. Supplements of CO quadratically decreased daily milk and lactose yields and linearly decreased milk protein yield and milk taste panel score from 4.2 to 3.6 [on a scale of 1 (poor) to 5 (excellent)], without altering milk fat yield. Inclusion of CO linearly decreased the proportions of saturated FA synthesized de novo (4:0 to 16:0), without altering milk fat 18:0, cis-9 18:1, LA, and ALA concentrations. Milk fat 18:0 was low (

Since ruminants are capable of utilizing fibrous feeds not digested by mono-gastrics, they represent a valuable natural resource for meeting future increases in global food supply. Ruminants have both local (nitrogen and phosphorus pollutions) and global (greenhouse gases, GHG) environmental footprints. It is estimated that the livestock sector is responsible for 18% of global anthropogenic GHG emissions. Losses of methane represent 30 to 50% of total GHG from livestock production, with the contribution from ruminants accounting for about 80%. Due to the concerns of increases in GHG emissions into the environment and potential effects on global warming, there is a need to develop strategies to lower methane emissions from ruminants as part of an overall requirement to improve the sustainability of ruminant food production systems. Methane is produced as a by-product of anaerobic fermentation in the reticulo-rumen, largely due to the activity of methanogenic archaea. Recent research has focused on the potential of novel feed ingredients (probiotics, ionophores, acetogen-based inoculants, bacteriocins, organic acids and plant extracts) or vaccines to lower hydrogen production and/or increase the transfer and utilization of metabolic hydrogen in the production of end-products other than methane in the rumen. Research to date has provided evidence that dietary supplements of plant or marine oils, oilseeds, specific fatty acids and condensed tannins, as well as defaunation, increases in production level or decreases in the proportion of forage in the diet may lower enteric methane production. Even though dietary lipid supplements can be used to lower methane output, in high amounts a decrease in intake and milk production can be expected. While further investigations have demonstrated the efficacy of specific agents on methanogenesis in vitro, the effects have not been substantiated in vivo. Altering the ratio of H2 /non-H2 producing fibrolytic bacteria to lower methanogenesis without altering fibre digestion has been demonstrated under experimental conditions. Furthermore, non-H2 producing communities have been characterized in the digesta of certain ruminant species. In contrast, stimulating acetogenesis by inoculation with rumen acetogens or non-rumen acetogens have met with limited success in vitro and in vivo. Research has also concentrated on stimulating the ultilisation of metabolic hydrogen by sulphate reducing bacteria, but there remains concern over the toxicity of H2S in the host ruminant. Investigations of nitrate reducing bacteria which produce more NH3 and less toxic nitrite, have indicated promising results. Increasing the number of capnophilic bacteria which use CO2 and H2 to produce organic acids, succinic acid in particular, may decrease methane production. In isolation, several approaches have been shown to decrease enteric methane emissions, but often part of the changes observed are related to lowered organic matter digestion in the rumen. However, lowering methane production per unit product over the lifetime of an animal should be regarded as the central goal to decrease GHG from ruminant livestock systems. This highlights the need for integrated solutions to improve digestive efficiency, as well as fertility and health. In conclusion, any prospective solution to lower on-farm GHG emissions must be practical, cost effective and have no adverse effect on the profitability of ruminant meat and milk production. Recent research has indicated significant potential, but none of the strategies tested thus far satisfy all of the necessary criteria for immediate implementation.

Because of the potential benefits to hu- man health, there is interest in increasing 18:3n-3, 20:5n- 3, 22:6n-6, and cis-9,trans-11 CLA in ruminant foods. Four Aberdeen Angus steers (406 ± 8.2 kg of BW) fit- ted with ruminal and duodenal cannulas were used in a 4 × 4 Latin square experiment with 21-d periods to ex- amine the potential of fish oil (FO) and linseed oil (LO) in the diet to increase ruminal outflow of trans-11 18:1 and total n-3 PUFA in growing cattle. Treatments con- sisted of a control diet (60:40; forage:concentrate ratio, on a DM basis, respectively) based on maize silage, or the same basal ration containing 30 g/kg of DM of FO, LO, or a mixture (1:1, wt/wt) of FO and LO (LFO). Diets were offered as total mixed rations and fed at a rate of 85 g of DM/(kg of BW 0.75 /d). Oils had no ef- fect (P = 0.52) on DMI. Linseed oil had no effect (P > 0.05) on ruminal pH or VFA concentrations, whereas FO shifted rumen fermentation toward propionate at the expense of acetate. Compared with the control, LO increased (P < 0.05) 18:0, cis 18:1 (Δ9, 12-15), trans 18:1 (Δ4-9, 11-16), trans 18:2, geometric isomers of Δ9,11, Δ11,13, and Δ13,15 CLA, trans-8,cis-10 CLA, trans-10,trans-12 CLA, trans-12,trans-14 CLA, and 18:3n-3 flow at the duodenum. Inclusion of FO in the diet resulted in greater (P < 0.05) flows of cis-9 16:1, trans 16:1 (Δ6-13), cis 18:1 (Δ9, 11, and 13), trans 18:1 (Δ6-15), trans 18:2, 20:5n-3, 22:5n-3, and 22:6n-3, and decreased (P < 0.001) 18:0 at the duodenum relative to the control. For most fatty acids at the duodenum, re- sponses to LFO were intermediate of FO and LO. How- ever, LFO resulted in greater (P = 0.04) flows of total trans 18:1 than LO and increased (P < 0.01) trans-6 16:1 and trans-12 18:1 at the duodenum compared with FO or LO. Biohydrogenation of cis-9 18:1 and 18:2n-6 in the rumen was independent of treatment, but both FO and LO increased (P < 0.001) the extent of 18:3n-3 biohydrogenation compared with the control. Ruminal 18:3n-3 biohydrogenation was greater (P < 0.001) for LO and LFO than FO, whereas biohydrogenation of 20:5n-3 and 22:6n-3 in the rumen was marginally less (P = 0.05) for LFO than FO. In conclusion, LO and FO at 30 g/kg of DM altered the biohydrogenation of un- saturated fatty acids in the rumen, causing an increase in the flow of specific intermediates at the duodenum, but the potential of these oils fed alone or as a mixture to increase n-3 PUFA at the duodenum in cattle ap- pears limited.

Based on the potential benefits to human health, there is interest in developing sustainable nutritional strategies to enhance the concentration of long-chainn-3 fatty acids in ruminant-derived foods. Four Aberdeen Angus steers fitted with rumen and duodenal cannulae were used in a 4 × 4 Latin square experiment with 21 d experimental periods to examine the potential of fish oil (FO) in the diet to enhance the supply of 20 : 5n-3 and 22 : 6n-3 available for absorption in growing cattle. Treatments consisted of total mixed rations based on maize silage fed at a rate of 85 g DM/kg live weight0·75/d containing 0, 8, 16 and 24 g FO/kg diet DM. Supplements of FO reduced linearly (P P P trans-16 : 1,trans-18 : 1,trans-18 : 2, 20 : 5n-3 and 22 : 6n-3, and decreased linearly (P n-3 at the duodenum. Increases in the flow oftrans-18 : 1 were isomer dependent and were determined primarily by higher amounts oftrans-11 reaching the duodenum. In conclusion, FO alters ruminal lipid metabolism of growing cattle in a dose-dependent manner consistent with an inhibition of ruminal biohydrogenation, and enhances the amount of long-chainn-3 fatty acids at the duodenum, but the increases are marginal due to extensive biohydrogenation in the rumen.

Maitorasvan koostumusta voidaan muuttaa ihmisterveydelle suotuisammaksi lehmien ruokintaa muuttamalla. Puna-apilasäilörehun tiedetään lisäävän monityydyttymättömien rasvahappojen, erityisesti α-linoleenihapon (18:3n-3), osuutta maitorasvassa. Kasviöljyt ovat heinäkasvi- ja maissisäilörehuun perustuvassa ruokinnassa tyypillisesti vähentäneet maitorasvan tyydyttyneiden ja lisänneet tyydyttymättömien rasvahappojen pitoisuuksia. Rypsiöljy sisältää runsaasti öljyhappoa (18:1n-9), auringonkukkaöljy linolihappoa (18:2n-6) ja camelinaöljy α-linoleenihappoa. Camelinapuriste vastaa raakarasva- ja valkuaispitoisuudeltaan rypsipuris-tetta, mutta sen käytöstä märehtijöiden rehuna on olemassa niukasti tutkimustietoa. Tämän tutkimuksen tarkoituksena oli selvittää maltillisen kasviöljylisäyksen (rypsiöljy, auringonkukkaöljy, camelinaöljy ja camelinapuriste) vaikutus maidon rasvahappokoostumukseen puna-apilaan pohjautuvassa ruokinnassa. Koe tehtiin Helsingin yliopiston Viikin opetus- ja tutkimustilan navetassa keväällä 2008. Kokeessa oli viisi kaksi kertaa poikinutta ay-lehmää ja se toteutettiin 5 x 5 latinalaisena neliönä. Koejakson pituus oli 21 päivää. Kokeen alkaessa lehmien poikimisesta oli kulunut keskimäärin 115 ± 5,0 päivää ja ne lypsivät 33,5 ± 1,62 kg maitoa/pv. Koekäsittelyinä olivat väkirehut, jotka kontrollia lukuun ottamatta sisälsivät lipidilisän (29 g/kg väkirehua) rypsiöljystä, auringonkukkaöljystä, camelinaöljystä tai camelinapuristeesta. Lehmät saivat väkirehua 12 kg/pv ja apilasäilörehua vapaasti. Lehmät söivät koko väkirehuannoksensa. Lipidilisä ei vaikuttanut säilörehun syöntiin, maitotuotokseen eikä maidon rasva- tai valkuaispitoisuuteen. Lipidilisä vähensi maitorauhasen de novo –synteesistä peräisin olevien lyhytketjuisten, tyydyttyneiden rasvahappojen pitoisuutta ja lisäsi tyydyttymättömien pitoisuutta maitorasvassa. Rypsiöljyruokinta lisäsi maitorasvan öljyhappopitoisuutta, kun taas linolihappopitoisuus oli suurempi ja α-linoleenihappopitoisuus pienempi auringonkukkaruokinnassa camelinaruokintoihin nähden, muutosten heijastaessa lipidilisien rasvahappokoostumusta. Lipidilisä, erityisesti camelinapuriste, lisäsi trans-rasvahappojen osuutta maitorasvassa. Camelinaöljyn antotavalla oli suuri vaikutus maitorasvan koostumukseen. Alfalinoleenihapon pitoisuus oli suurempi annettaessa camelinan lipidit öljynä kuin puristeena. Camelinaöljyn antaminen puristeena lisäsi kaksin-kolminkertaisesti C18-rasvahappojen biohydrogenaation välituotteiden pitoisuuksia maitorasvassa öljyruokintaan verrattuna (trans-10 ja trans-11 18:1, cis-9,trans-11 CLA ja trans-11,cis-15 18:2) ja vähensi steariinihapon (18:0) ja öljyhapon pitoisuuksia. Tämä viittaa rasvahappojen epätäydellisempään tyydyttymiseen pötsissä, kun camelinan lipidit annetaan puristeena eikä öljynä. On mahdollista, että murskattu camelinankuori suojaa siemenen sisällä olevia rasvahappoja pötsin biohydrogenaatiolta ja/tai camelinan siemenessä oleva jokin muu ainesosa inhiboi biohydrogenaatiota.

National audience; Fat is an important constituent contributing to the organoleptic, processing and physical properties of ruminant milk. Understanding the regulation of milk fat synthesis is central to the development of nutritional strategies to enhance the nutritional value of milk, decrease milk energy secretion and improve the energy balance of lactating ruminants. Nutrition is the major environmental factor regulating the concentration and composition of fat in ruminant milk. Feeding low-fibre/high-starch diets and/or lipid supplements rich in polyunsaturated fatty acids induce milk fat depression (MFD) in the bovine, typically increase milk fat secretion in the caprine, whereas limited data in sheep suggest that the responses are more similar to the goat than the cow. Following the observation that reductions in milk fat synthesis during diet-induced MFD are associated with increases in the concentration of specific trans fatty acids in milk, the biohydrogenation theory of MFD was proposed, which attributes the causal mechanism to altered ruminal lipid metabolism leading to increased formation of specific biohydrogenation intermediates that exert anti-lipogenic effects. Trans-10, cis-12 conjugated linoleic acid (CLA) is the only biohydrogenation intermediate to have been infused at the abomasum over a range of experimental doses (1.25 to 14.0 g/day) and shown unequivocally to inhibit milk fat synthesis in ruminants. However, increases in ruminal trans-10, cis-12 CLA formation do not explain entirely diet-induced MFD, suggesting that other biohydrogenation intermediates and/or other mechanisms may also be involved. Experiments involving abomasal infusions (g/day) in lactating cows have provided evidence that cis-10, trans-12 CLA (1.2), trans-9, cis-11 CLA (5.0) and trans-10 18:1 (92.1) may also exert anti-lipogenic effects. Use of molecular-based approaches have demonstrated that mammary abundance of transcripts encoding for key lipogenic genes are reduced during MFD in the bovine, changes that are accompanied by decrease in sterol response element binding protein 1 (SREBP1) and alterations in the expression of genes related to the SREBP1 pathway. Recent studies indicate that transcription of one or more adipogenic genes is increased in subcutaneous adipose tissue in cows during acute or chronic MFD. Feeding diets of similar composition do not induce MFD or substantially alter mammary lipogenic gene expression in the goat. The available data suggests that variation in mammary fatty acid secretion and lipogenic responses to changes in diet composition between ruminants reflect inherent interspecies differences in ruminal lipid metabolism and mammary specific regulation of cellular processes and key lipogenic enzymes involved in the synthesis of milk fat triacylglycerides.

This experiment (replicated 3 x 3 Latin square design) was conducted to investigate the effects of lauric acid (LA) or coconut oil (CO) on ruminal fermentation, nutrient digestibility, ammonia losses from manure, and milk fatty acid (FA) composition in lactating cows. Treatments consisted of intraruminal doses of 240 g of stearic acid/d (SA; control), 240 g of LA/d, or 530 g of CO/d administered once daily, before feeding. Between periods, cows were inoculated with ruminal contents from donor cows and allowed a 7-d recovery period. Treatment did not affect dry matter intake, milk yield, or milk composition. Ruminal pH was slightly increased by CO compared with the other treatments, whereas LA and CO decreased ruminal ammonia concentration compared with SA. Both LA and CO decreased protozoal counts by 80% or more compared with SA. Methane production rate in the rumen was reduced by CO compared with LA and SA, with no differences between LA and SA. Treatments had no effect on total tract apparent dry matter, organic matter, N, and neutral detergent fiber digestibility coefficients or on cumulative (15 d) in vitro ammonia losses from manure. Compared with SA, LA and CO increased milk fat 12:0, cis-9 12:1, and trans-9 12:1 content and decreased 6:0, 8:0, 10:0, cis-9 10:1, 16:0, 18:0, cis 18:1, total 18:2, 18:3 n-3 and total polyunsaturated FA concentrations. Administration of LA and 14:0 (as CO) in the rumen were apparently transferred into milk fat with a mean efficiency of 18 and 15%, respectively. In conclusion, current data confirmed that LA and CO exhibit strong antiprotozoal activity when dosed intraruminally, an effect that is accompanied by decreases in ammonia concentration and, for CO, lowered methane production. Administration of LA and CO in the rumen also altered milk FA composition.

Clinical and biomedical studies have provided evidence for the critical role of n-3 fatty acids on the reduction of chronic disease risk in humans, including cardiovascular disease. In the current experiment, the potential to enhance milk n-3 content in two breeds with inherent genetic differences in mammary lipogenesis and de novo fatty acid synthesis was examined using extruded linseeds. Six lactating cows (three Holstein and three Jersey) were used in a two-treatment switchback design with 3 × 21-day experimental periods to evaluate the effect of iso-energetic replacement of calcium salts of palm oil distillate (CPO) in the diet (34 g/kg dry matter (DM)) with 100 g/kg DM extruded linseeds (LIN). For both breeds, replacing CPO with LIN had no effect (P > 0.05) on DM intake or milk yield, but reduced (P < 0.05) milk fat and protein yield (on average, from 760 to 706 and 573 to 552 g/day, respectively). Relative to CPO, the LIN treatment reduced (P < 0.01) total saturated fatty acid content and enhanced (P < 0.001) 18:3n-3 in milk, whereas breed by diet interactions were significant for milk fat 16:0, total trans fatty acid and conjugated linoleic acid concentrations. Increases in 18:3n-3 intake derived from LIN in the diet were transferred into milk with a mean marginal transfer efficiency of 1.8%. Proportionate changes in milk fatty acid composition were greater in the Jersey, highlighting the importance of diet–genotype interactions on mammary lipogenesis. More extensive studies are required to determine the role of genotype on milk fat composition responses to oilseeds in the diet.

Faecal bacteria from four human donors and six species of human intestinal bacteria known to metabolize linoleic acid (LA) were incubated with LA in deuterium oxide-enriched medium to investigate the mechanisms of conjugated linoleic acid (CLA) and vaccenic acid (VA) formation. The main CLA products in faecal suspensions, rumenic acid (cis-9,trans-11-CLA; RA) andtrans-9,trans-11-CLA, were labelled at C-13, as were other 9,11 geometric isomers. Traces oftrans-10,cis-12-CLA formed were labelled to a much lower extent. In pure culture,Bifidobacterium breveNCFB 2258 formed labelled RA andtrans-9,trans-11-CLA, whileButyrivibrio fibrisolvens16.4,Roseburia hominisA2-183T,Roseburia inulinivoransA2-192TandRuminococcus obeum-like strain A2-162 converted LA to VA, labelled in a manner indicating that VA was formed via C-13-labelled RA.Propionibacterium freudenreichiisubsp.shermaniiDSM 4902T, a possible probiotic, formed mainly RA with smaller amounts oftrans-10,cis-12-CLA andtrans-9,trans-11-CLA, labelled the same as in the mixed microbiota. Ricinoleic acid (12-OH-cis-9-18 : 1) did not form CLA in the mixed microbiota, in contrast to CLA formation described forLactobacillus plantarum. These results were similar to those reported for the mixed microbiota of the rumen. Thus, although the bacterial genera and species responsible for biohydrogenation in the rumen and the human intestine differ, and a second route of RA formation via a 10-OH-18 : 1 is present in the intestine, the overall labelling patterns of different CLA isomers formation are common to both gut ecosystems. A hydrogen-abstraction enzymic mechanism is proposed that may explain the role of a 10-OH-18 : 1 intermediate in 9,11-CLA formation in pure and mixed cultures.

Based on the potential benefits ofcis-9,trans-11-conjugated linoleic acid (CLA) for human health there is interest in developing sustainable nutritional strategies for enhancing the concentration of this fatty acid in ruminant-derived foods. Most evidence to date suggests that endogenous synthesis is the major source ofcis-9,trans-11 in milk fat and ruminal outflow is limited and largely independent of dietary 18 : 2n-6 supply. Four lactating cows fitted with a rumen cannula were used in a 4 × 4 Latin square with 14 d experimental periods to examine the effects of sunflower-seed oil (SFO) as a source of 18 : 2n-6 on ruminal lipid metabolism. Cows were offered grass silage-based diets supplemented with 0, 250, 500 or 750 g SFO/d. Supplements of SFO had no effect on DM intake, milk fat or protein secretion, but increased linearly (P P P n-6 and total CLA at the omasum and enhanced ruminalcis-9-18 : 1, 18 : 2n-6 and 18 : 3n-3 metabolism. Flows of all-trans- (Δ4–16) andcis- (Δ9–16) 18 : 1 isomers were elevated, while increases in ruminal CLA outflow were confined totrans-8,trans-10 and geometric 9,11 and 10,12 isomers. It is concluded that supplementing grass silage-based diets with plant oils rich in 18 : 2n-6 enhances ruminal outflow oftrans-11-18 : 1 andcis-9,trans-11-CLA in lactating cows.

Ruminal administration of a triple indigestible marker system comprised of cobalt EDTA (CoEDTA), ytterbium acetate (YbAc), and chromium-mordanted straw (CrS) decreases product:substrate ratios for Delta9-desaturase in bovine milk fat. This experiment was designed to identify the marker(s) responsible and develop an alternative system for simultaneous determination of nutrient flow in the gastro-intestinal tract and milk fatty acid composition. Five lactating dairy cows were used in a 5 x 5 Latin square with 21-d periods to evaluate the effects of YbAc, CoEDTA, and CrS independently or as part of a triple marker system (TMS), and CrEDTA as an alternative to CoEDTA on milk fat composition. Markers were administered in the rumen over a 7-d interval and samples of milk were collected on d -1, 3, 7, and 11. Both TMS and CoEDTA alone reduced the concentrations of milk fatty acids containing a cis-9 double bond, whereas YbAc, CrS, and CrEDTA had no effect. Reductions in product:substrate ratios for Delta9-desaturase were time dependent and evident within 3 d of administration. Ruminal infusion of CoEDTA for 7 d induced mean decreases in milk cis-9 14:1/14:0, cis-9 16:1/16:0, cis-9 18:1/18:0, and cis-9, trans-11 conjugated linoleic acid/trans-11 18:1 concentration ratios of 47.7, 26.7, 40.3, and 42.6%, respectively. In conclusion, ruminal infusion of CoEDTA alters milk fatty acid composition and appears to inhibit Delta9-desaturase activity in the bovine mammary gland. Results indicate that a TMS based on CrEDTA, YbAc, and indigestible neutral detergent fiber can be used for estimating nutrient flow without altering milk fat composition in lactating cows.

Aims: All members of the ruminal Butyrivibrio group convert linoleic acid (cis-9,cis-12-18 : 2) via conjugated 18 : 2 metabolites (mainly cis-9,trans-11-18 : 2, conjugated linoleic acid) to vaccenic acid (trans-11-18 : 1), but only members of a small branch, which includes Clostridium proteoclasticum, of this heterogeneous group further reduce vaccenic acid to stearic acid (18 : 0, SA). The aims of this study were to develop a real-time polymerase chain reaction (PCR) assay that would detect and quantify these key SA producers and to use this method to detect diet-associated changes in their populations in ruminal digesta of lactating cows. Methods and Results: The use of primers targeting the 16S rRNA gene of Cl. proteoclasticum was not sufficiently specific when only binding dyes were used for detection in real-time PCR. Their sequences were too similar to some nonproducing strains. A molecular beacon probe was designed specifically to detect and quantify the 16S rRNA genes of the Cl. proteoclasticum subgroup. The probe was characterized by its melting curve and validated using five SA-producing and ten nonproducing Butyrivibrio-like strains and 13 other common ruminal bacteria. Analysis of ruminal digesta collected from dairy cows fed different proportions of starch and fibre indicated a Cl. proteoclasticum population of 2–9% of the eubacterial community. The influence of diet on numbers of these bacteria was less than variations between individual cows. Conclusions: A molecular beacon approach in qPCR enables the detection of Cl. proteoclasticum in ruminal digesta. Their numbers are highly variable between individual animals. Significance and Impact of the Study: SA producers are fundamental to the flow of polyunsaturated fatty acid and vaccenic acid from the rumen. The method described here enabled preliminary information to be obtained about the size of this population. Further application of the method to digesta samples from cows fed diets of more variable composition should enable us to understand how to control these bacteria in order to enhance the nutritional characteristics of ruminant-derived foods, including milk and beef.

Digesta samples from the ovine rumen and pure ruminal bacteria were incubated with linoleic acid (LA) in deuterium oxide-containing buffer to investigate the mechanisms of the formation of conjugated linoleic acids (CLAs). Rumenic acid (RA; cis-9,trans-11-18:2), trans-9,trans-11-18:2, and trans-10,cis-12-18:2 were the major CLA intermediates formed from LA in ruminal digesta, with traces of trans-9,cis-11-18:2, cis-9,cis-11-18:2, and cis-10,cis-12-18:2. Mass spectrometry indicated an increase in the n+1 isotopomers of RA and other 9,11-CLA isomers, as a result of labeling at C-13, whereas 10,12 isomers contained minimal enrichment. In pure culture, Butyrivibrio fibrisolvens and Clostridium proteoclasticum produced mostly RA with minor amounts of other 9,11 isomers, all labeled at C-13. Increasing the deuterium enrichment in water led to an isotope effect, whereby (1)H was incorporated in preference to (2)H. In contrast, the type strain and a ruminal isolate of Propionibacterium acnes produced trans-10,cis-12-18:2 and other 10,12 isomers that were minimally labeled. Incubations with ruminal digesta provided no support for ricinoleic acid (12-OH,cis-9-18:1) as an intermediate of RA synthesis. We conclude that geometric isomers of 10,12-CLA are synthesized by a mechanism that differs from the synthesis of 9,11 isomers, the latter possibly initiated by hydrogen abstraction on C-11 catalyzed by a radical intermediate enzyme.

Five cows were used in a 5 x 5 Latin square to evaluate the effects of forage type and stage of herbage maturity on milk fatty acid (FA) composition. Four silages were prepared from primary growths of timothy-meadow fescue and red clover swards at an early and late stage of growth. Treatments consisted of these silages and an additional treatment comprised of an equal mixture of late-cut grass silage and early-cut red clover silage. Forages were fed ad libitum and supplemented with 9 kg/day of a standard concentrate. Compared with grass silage, red clover silage diets enhanced milk fat monounsaturated FA (MUFA) and polyunsaturated FA (PUFA) content, 18:3 n-3 in particular, responses that were associated with concomitant reductions in 10:0 to 16:0 concentrations. Decreases in 16:0 and increases in PUFA concentrations were more pronounced for early than late growths of red clover. Even though red clover resulted in marginal increases in total milk fat trans 18:1 content, milk trans-11 18:1 concentrations were lower for red clover than for grass silage diets. It is concluded that replacing grass with red clover silage prepared at an early stage of maturity represents a sustainable nutritional strategy for enhancing the MUFA and PUFA content of milk.

Fat is the most variable constituent in milk and is reduced on low-fiber/high-concentrate diets or on rations containing lipid supplements rich in polyunsaturated fatty acids. The biohydrogenation theory attributes the causal mechanism underlying diet-induced milk fat depression (MFD) to changes in ruminal lipid metabolism leading to increased formation of specific biohydrogenation intermediates that exert anti-lipogenic effects. Trans-10, cis-12 conjugated linoleic acid (CLA) is the only intermediate shown unequivocally to inhibit milk fat synthesis. However, increases in ruminal trans-10, cis-12 CLA formation do not provide a universal explanation for the reductions in milk fat during diet-induced MFD, suggesting that other biohydrogenation intermediates may also be involved. Post-ruminal infusion experiments have provided tentative evidence that cis-10, trans-12 CLA and trans-9, cis-11 CLA also exert anti-lipogenic effects. Diet-induced MFD is consistently associated with an increase in milk fat trans-10 18:1 concentrations, but recent direct evidence offers little support for a role of this intermediate in the regulation of milk fat synthesis. Further research is required to characterize the structure and function of other biohydrogenation intermediates, as well as considering the contribution of more global changes in ruminal lipid metabolism to provide a more universal explanation of diet-induced MFD.

Cis-9, trans-11, the major isomer of conjugated linoleic acid (CLA) in bovine milk fat, is derived from ruminal biohydrogenation of 18:2 (n-6) and endogenous conversion of trans-11 18:1 (vaccenic acid; VA) in the mammary gland. Most evidence to date suggests that endogenous synthesis is the major source of cis-9, trans-11 CLA, but the extent of VA desaturation is less well defined. Four lactating cows were used in consecutive 4 x 4 Latin squares to examine changes in milk fatty acid composition and secretion in response to abomasal infusions of lipid supplements enriched with cis-9, trans-11 CLA (88.8%) or VA (29.4%). Treatments were infused over 4-d, followed by a 3-d washout, during 7 d experimental periods and administered to deliver 0, 3, 6, and 12 g cis-9, trans-11 CLA/d (Expt. 1) or 0, 7.5, 15 and 30 g VA/d (Expt. 2). Infusions of cis-9, trans-11 CLA increased linearly milk cis-9, trans-11 CLA concentrations from 0.68 to 1.46 g/100 g fatty acids. Abomasal infusions of VA increased linearly milk VA and cis-9, trans-11 CLA content from 1.22 to 2.72 and 0.61 to 1.24 g/100 g fatty acids, respectively. Changes in milk fatty acid secretion indicated that 28.9% of VA was converted to cis-9, trans-11 CLA. Results provide evidence that conversion by Delta9-desaturase to cis-9, trans-11 CLA in the lactating cow is independent of postruminal VA supply. In conclusion, endogenous synthesis via VA was equivalent to approximately 21% of the response to increases in cis-9, trans-11 CLA available for absorption.

International audience; Background: Intravenous or ruminal infusion of lithium salt of cobalt EDTA (Co-EDTA) or Cobalt-acetate alters milk fat composition in cattle, but the mechanisms involved are not known. Objective: The present study evaluated the effect of ruminal Co-EDTA infusion on milk FA composition, mammary lipid metabolism, and mammary lipogenic gene expression. Methods: For the experiment, 4 cows in midlactation and fitted with rumen cannulae were used in a 4 x 4 Latin square with 28-d periods. Co-EDTA was administered in the rumen to supply 0, 1.5, 3.0, or 4.5 g Co/d over an 18-d interval with a 10-d washout between experimental periods. Milk production was recorded daily, and milk FA composition was determined on alternate days. Mammary tissue was biopsied on day 16, and arteriovenous differences of circulating lipid fractions and FA uptake across the mammary gland were measured on day 18. Results: Co-EDTA had no effect on intake, proportions of rumen volatile FA, or milk production but caused dose-dependent changes in milk FA composition. Alterations in milk fat composition were evident within 3 d of infusion and characterized by linear or quadratic decreases (P < 0.05) in FAs containing a cis-9 double bond, an increase in 4:0 and 16:0, and linear decreases in milk 8:0, 10:0, 12:0, and 14:0 concentrations. Co-EDTA progressively decreased (P < 0.05) the stearoyl-CoA desaturase (SCD)-catalyzed desaturation of FAs in the mammary gland by up to 72% but had no effect on mammary SCD1 mRNA or SCD protein abundance. Changes in milk FA composition were accompanied by altered expression of specific genes involved in de novo FA and triacylglycerol synthesis. Conclusion: Ruminal infusion of Co-EDTA alters milk FA composition in cattle via a mechanism that involves decreases in the desaturation of FAs synthesized de novo or extracted from blood and alterations in mammary lipogenic gene expression, without affecting milk fat yield.

21 páginas, 8 tablas, 2 figuras, A study with 2 ruminant species (goats and cows) with inherent differences in lipid metabolism was performed to test the hypothesis that milk fat depression (MFD) due to marine lipid supplements or diets containing high amounts of starch and plant oil is caused by different mechanisms and that each ruminant species responds differently. Cows and goats were allocated to 1 of 3 groups (4 cows and 5 goats per group) and fed diets containing no additional oil (control) or supplemented with fish oil (FO) or sunflower oil and wheat starch (SOS) according to a 3 x 3 Latin square design with 26-d experimental periods. In cows, milk fat content was lowered by FO and SOS (-31%), whereas only FO decreased milk fat content in goats (-21%) compared with the control. Furthermore, FO and SOS decreased milk fat yield in cows, but not in goats. In both species, FO and SOS decreased the secretion of C16 FA output. However, SOS increased milk secretion of >C16 FA in goats. Compared with the control, SOS resulted in similar increases in milk trans-10, cis-12 conjugated linoleic acid (CLA) in both species, but caused a 2-fold larger increase in trans-10 18:1 concentration in cows than for goats. Relative to the control, responses to FO in both species were characterized by a marked decrease in milk concentration of 18:0 (-74%) and cis-9 18:1 (-62%), together with a similar to 5-fold increase in total trans 18:1, but the proportionate changes in trans-10 18:1 were lower for goats. Direct comparison of animal performance and milk FA responses to FO and SOS treatments demonstrated interspecies differences in mammary lipogenesis, suggesting a lower sensitivity to the inhibitory effects of trans-10, cis-12 CLA in goats and that ruminal biohydrogenation pathways are more stable and less prone to diet-induced shifts toward the formation of trans-10-containing intermediates in goats compared with cows. Even though a direct cause and effect could not be established, results suggest that regulation of milk fat synthesis during FO-induced MFD may be related to a shortage of 18:0 for endogenous mammary cis-9 18:1 synthesis, increase in the incorporation of trans FA in milk triacylglycerols, and limitations in the synthesis of FA de novo to maintain milk fat melting point. However, the possible contribution of biohydrogenation intermediates with putative antilipogenic effects in the mammary gland, including trans-9,cis-11 CLA, trans-10 18:1, or cis-11 18:1 to FO-induced MFD cannot be excluded., P. G. Toral was granted a post-doctoral fellowship from Fundacion Alfonso Martin Escudero (Madrid, Spain). The authors gratefully acknowledge the staff of Unite Experimentale des Ruminants de Theix (INRA, Saint-Genes-Champanelle, France) for the diligent care of the experimental animals, and D. Bany, C. Delavaud, E. Tixier, and M. Tourret from UMR 1213 Herbivores (INRA) for assistance in sampling collection and laboratory analysis. The contribution of Laura Ventto and Minna Aalto (Natural Resources Institute Finland, Jokioinen, Finland, formerly MTT Agrifood Research Finland) to the analysis of milk fat composition is acknowledged and appreciated.

Accurate determination of the flow of nutrients at the omasum requires the use of a triple marker system. Typically, a system based on ruminal administration of the lithium salt of CoEDTA, ytterbium acetate (Yb-Ac), and chromium-mordanted straw (Cr-S) has been used. However, there is evidence to suggest that product:substrate ratios for stearoyl-coenzyme A desaturase (Δ 9 -desaturase) are lower in milk fat from cows administered a combination of CoEDTA, Yb-Ac, and Cr-S, indicating reduced Δ 9 -desaturase activity. To evaluate this hypothesis, samples of milk were collected 1 d before, and on d 2, 6, and 9 of administering the CoEDTA, Yb-Ac, and Cr-S triple marker system into the rumen of 4 cows. A 4 × 4 Latin square with 28-d experimental periods was used to assess the effects of 0, 75, 150, and 300 g/d of fish oil in the diet on ruminal and mammary lipid metabolism. Irrespective of the amount of fish oil in the diet, concentrations of all milk fatty acids containing a cis -9 double bond were reduced after markers were given. Milk fatty acid pairs dependent on Δ 9 -desaturase were decreased over time, with responses reaching a nadir within 6 d of marker administration. Overall, administering markers into the rumen was associated with a mean decrease in milk cis -9 10:1/ 10:0, cis -9 12:1/12:0, cis -9 14:1/14:0, cis -9 16:1/16:0, cis -9 17:1/17:0, cis -9 18:1/18:0, and cis -9, trans -11 conjugated linoleic acid/ trans -11 18:1 concentration ratios of 44.6, 52.7, 58.7, 36.8, 37.2, 44.3, and 43.0%, respectively. In conclusion, one or more of the markers administered altered milk fatty acid composition and may act as an inhibitor of Δ 9 -desaturase in the bovine mammary gland.

Dietarycis-9,trans-11-conjugated linoleic acid (CLA) is generally thought to be beneficial for human health. Fish oil added to ruminant diets increases the CLA concentration of milk and meat, an increase thought to arise from alterations in ruminal biohydrogenation of unsaturated fatty acids. To investigate the mechanism for this effect,in vitroincubations were carried out with ruminal digesta and the main biohydrogenating ruminal bacterium,Butyrivibrio fibrisolvens. Linoleic acid (LA) or α-linolenic acid (LNA) was incubated (1·67g/l) with strained ruminal digesta from sheep receiving a 50:50 grass hay–concentrate ration. Adding fish oil (up to 4·17g/l) tended to decrease the initial rate of LA (P=0·025) and LNA (P=0·137) disappearance, decreased (PPtrans-11-18:1. Concentrations of EPA (20:5n-3) or DHA (22:6n-3), the major fatty acids in fish oil, were low (100mg/l or less) after incubation of fish oil with ruminal digesta. Addition of EPA or DHA (50mg/l) to pure cultures inhibited the growth and isomerase activity ofB. fibrisolvens, while fish oil had no effect. In contrast, similar concentrations of EPA and DHA had no effect on biohydrogenation of LA by mixed digesta, while the addition of LA prevented metabolism of EPA and DHA. Neither EPA nor DHA was metabolised byB. fibrisolvensin pure culture. Thus, fish oil inhibits ruminal biohydrogenation by a mechanism which can be interpreted partly, but not entirely, in terms of its effects onB. fibrisolvens.

PMID: 16428640 [PubMed], Previously published as proceeding at the XXXVII Jornadas de Estudio AIDA (XI Jornadas sobre Producción Animal) (Zaragoza, Spain, 2005). https://digital.csic.es/handle/10261/5104, Based on the potential benefits of cis-9, trans-11 conjugated linoleic acid (CLA) for human health, there is a need to develop effective strategies for enhancing milk fat CLA concentrations. Levels of cis-9, trans-11 CLA in milk can be increased by supplements of fish oil (FO) and sunflower oil (SO), but there is considerable variation in the response. Part of this variance may reflect time-dependent ruminal adaptations to high levels of lipid in the diet, which lead to alterations in the formation of specific biohydrogenation intermediates. To test this hypothesis, 16 late lactation Holstein-British Friesian cows were used in a repeated measures randomized block design to examine milk fatty acid composition responses to FO and SO in the diet over a 28-d period. Cows were allocated at random to corn silage-based rations (8 per treatment) containing 0 (control) or 45 g of oil supplement/kg of dry matter consisting (1:2; wt/wt) of FO and SO (FSO), and milk composition was determined on alternate days from d 1. Compared with the control, the FSO diet decreased mean dry matter intake (21.1 vs. 17.9 kg/d), milk fat (47.7 vs. 32.6 g/kg), and protein content (36.1 vs. 33.3 g/kg), but had no effect on milk yield (27.1 vs. 26.4 kg/d). Reductions in milk fat content relative to the FSO diet were associated with increases in milk trans-10 18:1, trans-10, cis-12 CLA, and trans-9, cis-11 CLA concentrations (r2 = 0.74, 0.57, and 0.80, respectively). Compared with the control, the FSO diet reduced milk 4:0 to 18:0 and cis 18:1 content and increased trans 18:1, trans 18:2, cis-9, trans-11 CLA, 20:5 n-3, and 22:6 n-3 concentrations. The FSO diet caused a rapid elevation in milk cis-9, trans-11 CLA content, reaching a maximum of 5.37 g/100 g of fatty acids on d 5, but these increases were transient, declining to 2.35 g/100 g of fatty acids by d 15. They remained relatively constant thereafter. Even though concentrations of trans-11 18:1 followed the same pattern of temporal changes as cis-9, trans-11 CLA, the total trans 18:1 content of FSO milk was unchanged because of the concomitant increases in the concentration of other isomers (4–10 and 12–15), predominantely trans-10 18:1. In conclusion, supplementing diets with FSO enhances milk fat cis-9, trans-11 CLA content, but the high level of enrichment declines because of changes in ruminal biohydrogenation that result in trans-10 replacing trans-11 as the major 18:1 biohydrogenation intermediate formed in the rumen., Financial support from the Biotechnology and Biological Sciences Research Council, Department for Environment, Food, and Rural Affairs, The Scottish Executive Environment and Rural Affairs Department and the Milk Development Council under the Eating, Food and Health LINK scheme is gratefully acknowledged. The authors thank the staff of the Center for Dairy Research Animal Production Research Unit for diligent care of experimental animals and sample collection and Vesa Toivonen for assistance with the interpretation of mass-spectral data.

Opportunities for the production of milk and dairy products enriched with cis-9, trans-11 conjugated linoleic acid (CLA) were investigated. Eighteen mid-lactation cows were used in a continuous-design for 7 weeks. During the first week, cows received grass silage ad libitum supplemented with 10 kg per day of a cereal-based concentrate (control) that was replaced with a concentrate containing 50 g kg−1 of rapeseed oil (RO). Changes in milk fatty acid composition were monitored on a weekly basis and milk produced was used to manufacture Edam cheese and butter. Inclusion of RO in the concentrate supplement increased the mean levels of trans-octadecanoic, monounsaturated, CLA and polyunsaturated fatty acid in the milk fat from 1.6, 25.7, 0.46 and 2.8 to 4.3, 35.3, 1.02 and 3.9 g 100 g−1 total fatty acids, respectively. In contrast, the mean level of saturated fatty acids decreased from 71.4 to 60.7 g 100 g−1 total fatty acids. Changes in milk fatty acid composition due to RO occurred within 7 days, with responses reaching a plateau after 21 days. Furthermore, the CLA concentrations in the milk fat from individual cows ranged between 0.37 and 0.65 and 0.43 and 2.06 g 100 g−1 total fatty acids for the control and RO diet, respectively. CLA enriched milk was used successfully to manufacture of Edam cheese and butter with softer textures but with acceptable organoleptic and storage properties. Processing milk into butter or cheese had no effect on the CLA concentrations indicating that enrichment of dairy products is dependent on the content in raw milk fat.

Foods derived from domestic animals are a significant source of nutrients in the UK diet. However, certain aspects of some animal-derived foods, notably levels of saturated fatty acids, have given rise to concerns that these foods may contribute to the risk of cardiovascular disease, the metabolic syndrome and other conditions. However, the composition of the many animal-derived foods is not constant and can often be enhanced by manipulating the nutrition of the animal. This paper reviews these possibilities with particular attention to lipids, and draws attention to the fact that milk in particular, contains a number of compounds which may, for example, exert anti-carcinogenic effects. It is clear that the role of animal nutrition in creating foods closer to the optimum composition for long-term human health will not only be more relevant in the future, but will be vital in attempts to improve the health of the human population.

Mechanisms underlying milk fat conjugated linoleic acid (CLA) responses to supplements of fish oil were investigated using five lactating cows each fitted with a rumen cannula in a simple experiment consisting of two consecutive 14-day experimental periods. During the first period cows were offered 18 kg dry matter (DM) per day of a basal (B) diet formulated from grass silage and a cereal based-concentrate (0·6 : 0·4; forage : concentrate ratio, on a DM basis) followed by the same diet supplemented with 250 g fish oil per day (FO) in the second period. The flow of non-esterified fatty acids leaving the rumen was measured using the omasal sampling technique in combination with a triple indigestible marker method based on Li-Co-EDTA, Yb-acetate and Cr-mordanted straw. Fish oil decreased DM intake and milk yield, but had no effect on milk constituent content. Milk fat trans-11 C18:1, total trans-C18 : 1, cis-9 trans-11 CLA, total CLA, C18 : 2(n-6) and total C18 : 2content were increased in response to fish oil from 1·80, 4·51, 0·39, 0·56, 0·90 and 1·41 to 9·39, 14·39, 1·66, 1·85, 1·25 and 4·00 g/100 g total fatty acids, respectively. Increases in the cis-9, trans-11 isomer accounted for proportionately 0·89 of the CLA response to fish oil. Furthermore, fish oil decreased the flow of C18 : 0(283 and 47 g/day for B and FO, respectively) and increased that of trans-C18 : 1fatty acids entering the omasal canal (38 and 182 g/day). Omasal flows of trans-C18 : 1acids with double bonds in positions from delta-4 to -15 inclusive were enhanced, but the effects were isomer dependent and primarily associated with an increase in trans-11 C18 : 1 leaving the rumen (17·1 and 121·1 g/day for B and FO, respectively). Fish oil had no effect on total (4·36 and 3·50 g/day) or cis-9, trans-11 CLA (2·86 and 2·08 g/day) entering the omasal canal. Flows of cis-9, trans-11 CLA were lower than the secretion of this isomer in milk. Comparison with the transfer of the trans-9, trans-11 isomer synthesized in the rumen suggested that proportionately 0·66 and 0·97 of cis-9, trans-11 CLA was derived from endogenous conversion of trans-11 C18 : 1in the mammary gland for B and FO, respectively. It is concluded that fish oil enhances milk fat cis-9, trans-11 CLA content in response to increased supply of trans-11 C18:1that arises from an inhibition of trans-C18 : 1reduction in the rumen.

The current study was conducted to establish if variations in nutrient supply due to forage conservation method could be compensated for by increases in concentrate feeding or supplements of a glucogenic substrate. Thirty two Finnish Ayrshire dairy cows were used in a cyclic change-over experiment with four 21-day experimental periods and a 4×2×2 factorial arrangement of treatments to evaluate the effects of forage conservation method, concentrate level and propylene glycol (PG), and their interactions, on digestibility, rumen fermentation, blood metabolite concentrations and nutrient utilisation. Experimental treatments consisted of four conserved forages offered ad libitum, supplemented with two levels of a cereal-based concentrate (7 or 10 kg per day) and PG (0 and 210 g per day) fed as three equal meals. Forages were prepared from primary growths of timothy and meadow fescue swards and ensiled using no additive (NA), an inoculant enzyme (IE) preparation or a formic acid (FA)-based additive or conserved as hay 1 week later. Forage conservation method had only minor effects on digestibility. Increases in concentrate feeding reduced the digestibility of neutral detergent fibre (NDF) while PG supplements tended to decrease that of nitrogen. Rumen fermentation of grass silage-based diets was characterised by higher molar proportions of butyrate and branched chain volatile fatty acids (BCVFA) and lower proportions of acetate compared with hay-based diets. Use of an ensiling additive reduced molar proportions of butyrate, while relative to FA silage, rumen fermentation of IE silage was associated with lower proportions of acetate and higher proportions of propionate and BCVFA. Increases in concentrate feeding resulted in a shift towards butyrate at the expense of acetate, while PG supplementation resulted in a rumen fermentation with higher proportions of propionate and lower proportions of acetate. Relative to silage fed animals, cows receiving hay-based diets had higher plasma concentrations of glucose and lower concentrations of insulin and beta-hydroxybutyrate (β-OHB). Increases in concentrate feeding tended to increase glucose concentrations while PG supplements elevated glucose but lowered plasma β-OHB concentrations. Differences in rumen fermentation associated with the ingestion of forages conserved using different methods were sufficient to suggest an important role on subsequent nutrient utilisation. Increases in concentrate feeding or supplements of a glucogenic precursor were unable to compensate for variations in nutrient supply due to forage conservation method.

The current study was conducted to establish if effects on animal performance due to differences in forage composition resulting from conservation method could be compensated for by increases in concentrate feeding or supplements of a gluconeogenic substrate. Thirty-two Finnish Ayrshire dairy cows were used in a cyclic changeover experiment with four 21-day experimental periods and a 4 ✕ 2 ✕ 2 factorial arrangement of treatments to evaluate the effects of forage conservation method, concentrate level and propylene glycol (PG), and their interactions, on intake, feeding behaviour and milk production. Experimental treatments consisted of four conserved forages offered ad libitum, supplemented with two levels of a cereal-based concentrate (7 or 10 kg/day) and PG (0 and 210 g/day) offered as three meals of equal size. Forages were prepared from primary growths of timothy and meadow fescue swards and ensiled using no additive (NA), an inoculant enzyme preparation (IE) or a formic-acid based (FA) additive or conserved as hay 1 week later. Cows given silage-based diets had higher (P 0·001) forage dry-matter (DM) intakes (mean increase 0·76 kg/day), spent less (P 0·001) time eating and chewing (mean response -159 and -119 min/day, respectively) and produced more (P 0·05) energy-corrected milk (ECM), milk fat and milk lactose (respective mean responses 1·52, 0·098 and 0·033 kg/day) than animals given hay-based diets. Use of an additive during ensiling further improved (P 0·05) silage DM intake, ECM yield and milk protein secretion (mean 0.72, 0.70 and 0.038 kg/day, respectively). Dietary inclusion of PG decreased forage DM intake for hay, IE and FA silage-based diets (mean –0·14, –0·16 and –0·42 kg/day, respectively) but elicited positive responses (mean 0·57 kg/day) for cows given NA silage. Furthermore, PG supplementation had no (P > 0·05) effects on ECM yield or milk protein output but depressed (P 0·05) mean milk fat content from 46·6 to 45·6 g/kg. Increases in concentrate feeding were associated with a reduction in the total amount of time cows spent eating, chewing and ruminating and elicited (P 0·001) mean DM intake, ECM yield, milk fat and milk protein responses of 1·5, 1·62, 0·061 and 0·064 kg/day, respectively. Use of a gluconeogenic substrate or increases in concentrate feeding were unable to compensate for variations in animal performance due to forage conservation method.

Concentrations of milk urea N (MUN) are influenced by dietary crude protein concentration and intake and could therefore be used as a biomarker of the efficiency of N utilization for milk production (milk N/N intake; MNE) in lactating cows. In the present investigation, data from milk-production trials (production data set; n=1,804 cow/period observations from 21 change-over studies) and metabolic studies involving measurements of nutrient flow at the omasum in lactating cows (flow data set; n=450 cow/period observations from 29 studies) were used to evaluate the influence of between-cow variation on the relationship of MUN with MNE, urinary N (UN) output, and diet digestibility. All measurements were made on cows fed diets based on grass silage supplemented with a range of protein supplements. Data were analyzed by mixed-model regression analysis with diet within experiment and period within experiment as random effects, allowing the effect of diet and period to be excluded. Between-cow coefficient of variation in MUN concentration and MNE was 0.13 and 0.07 in the production data set and 0.11 and 0.08 in the flow data set, respectively. Based on residual variance, the best model for predicting MNE developed from the production data set was MNE (g/kg)=238 + 7.0 × milk yield (MY; kg/d) - 0.064 × MY(2) - 2.7 × MUN (mg/dL) - 0.10 body weight (kg). For the flow data set, including both MUN and rumen ammonia N concentration with MY in the model accounted for more variation in MNE than when either term was used with MY alone. The best model for predicting UN excretion developed from the production data set (n=443) was UN (g/d)=-29 + 4.3 × dry matter intake (kg/d) + 4.3 × MUN + 0.14 × body weight. Between-cow variation had a smaller influence on the association of MUN with MNE and UN output than published estimates of these relationships based on treatment means, in which differences in MUN generally arise from variation in dietary crude protein concentration. For the flow data set, between-cow variation in MUN and rumen ammonia N concentrations was positively associated with total-tract organic matter digestibility. In conclusion, evaluation of phenotypic variation in MUN indicated that between-cow variation in MUN had a smaller effect on MNE compared with published responses of MUN to dietary crude protein concentration, suggesting that a closer control over diet composition relative to requirements has greater potential to improve MNE and lower UN on farm than genetic selection.

7 páginas, 1 tabla, 5 figuras., The current short review summarizes recent data on the specificities of goats compared with cows, of milk fatty acid (FA) secretion and milk fat lipolysis responses to physiological and nutritional factors. The influence of lactation stage on milk fat yield and FA composition is similar between goats and cows. In contrast, changes in milk fat yield and composition to diet, lipid supplements in particular, differs between the two ruminant species. In almost all cases, dietary lipid supplements increase milk fat content in goats, but not in cows. The goat is much less sensitive to diet-induced alterations in ruminal biohydrogenation pathways causing trans-10 18:1 to replace trans-11 18:1 as the major intermediate relative to the cow. Mammary lipid secretion in the goat is also less sensitive to the anti-lipogenic effect of trans-10,. cis-12 conjugated linoleic acid (CLA) compared with the cow. Consistent with these observations, mammary lipogenic gene expression is less affected by diets rich in starch and polyunsaturated FA (PUFA) in goats than cows. However, diets containing PUFA induce much greater changes in delta-9 desaturase gene expression in goats compared with cows, that may be related to differences in the availability of biohydrogenation intermediates at the mammary glands (e.g. trans-9,. trans-11-CLA). The development of either goat flavour or rancidity is related to the inherent peculiarities of milk FA composition and lipolytic system in this species. In contrast with cows, milk LPL activity and lipolysis are low during early and late lactation in goats, and are decreased when animals are underfed or receive a diet supplemented with plant oils. In goats the alpha-s1-casein (CSN1S1) gene polymorphism is associated with a decrease in milk fat content and 8:0-12:0 concentrations in the low CSN1S1 genotype. Conversely, milk fat product/substrate concentration ratios for delta-9 desaturation and spontaneous lipolysis are increased in the low genotype. © 2014 Elsevier B.V., P.G. Toral was granted a fellowship from the Fundación Alfonso Martín Escudero (Madrid, Spain).

11 páginas, 3 tablas, 2 figuras., Previous investigations have shown that cobalt (Co) modifies milk fat composition in cattle, consistent with an inhibition of stearoyl-coenzyme A desaturase (SCD) activity, but it remains unclear whether other ruminant species are also affected. The present study examined the effects of oral administration of Co acetate on intake, rumen function, and milk production and fatty acid (FA) composition in sheep. Twenty lactating Assaf ewes were allocated into 1 of 4 groups and used in a continuous randomized block design that involved a 15-d adaptation, a 6-d treatment, and a 10-d posttreatment period. During the treatment period, animals received an oral drench supplying 0 (control), 3 (Co3), 6 (Co6), and 9 (Co9) mg of Co/kg of BW per day, administered in 3 equal doses at 8-h intervals. Cobalt acetate had no influence on intake or milk fat and protein concentrations, whereas treatments Co6 and Co9 tended to lower milk yield. Results on rumen parameters showed no effects on rumen fermentation, FA composition, or bacterial community structure. Administration of Co acetate decreased milk concentrations of FA containing a cis -9 double bond and SCD product:substrate ratios, consistent with an inhibition of SCD activity in the ovine mammary gland. Temporal changes in milk fat composition indicated that the effects of treatments were evident within 3d of dosing, with further changes being apparent after 6d and reverting to pretreatment values by d 6 after administration. Effect on milk FA composition did not differ substantially in response to incremental doses of Co acetate. On average, Co decreased milk cis-9 10:1/10:0, cis-9 12:1/12:0, cis- 9 14:1/14:0, cis-9 16:1/16:0, cis-9 17:1/17:0, cis-9 18:1/18:0, and cis-9,trans-11 18:2/trans-11 18:1 concentration ratios by 30, 32, 38, 33, 21, 24, and 25%, respectively. Changes in milk fat cis-9 10:1, cis-9 12:1, and cis-9 14:1 concentrations to Co treatment indicated that 51% of cis-9 18:1 and cis-9,trans-11 18:2 secreted in milk originated from δ9-desaturation. In conclusion, results demonstrated the potential of oral Co administration for the estimation of endogenous synthesis of FA containing a cis-9 double bond in the mammary gland of lactating ruminants. Indirect comparisons suggest that the effects of Co differ between sheep and cattle., This work was supported by the Spanish Ministry of Economy and Competitiveness (MINECO, Madrid, Spain; AGL2011-23700).

The effects of forage conservation method on plasma lipids, mammary lipogenesis, and milk fat were examined in 2 complementary experiments. Treatments comprised fresh grass, hay, or untreated (UTS) or formic acid treated silage (FAS) prepared from the same grass sward. Preparation of conserved forages coincided with the collection of samples from cows fed fresh grass. In the first experiment, 5 multiparous Finnish Ayrshire cows (229 d in milk) were used to compare a diet based on fresh grass followed by hay during 2 consecutive 14-d periods, separated by a 5-d transition during which extensively wilted grass was fed. In the second experiment, 5 multiparous Finnish Ayrshire cows (53 d in milk) were assigned to 1 of 2 blocks and allocated treatments according to a replicated 3 × 3 Latin square design, with 14-d periods to compare hay, UTS, and FAS. Cows received 7 or 9 kg/d of the same concentrate in experiments 1 and 2, respectively. Arterial concentrations of triacylglycerol (TAG) and phospholipid were higher in cows fed fresh grass, UTS, and FAS compared with hay. Nonesterified fatty acid (NEFA) concentrations and the relative abundance of 18:2n-6 and 18:3n-3 in TAG of arterial blood were also higher in cows fed fresh grass than conserved forages. On all diets, TAG was the principle source of fatty acids (FA) for milk fat synthesis, whereas mammary extraction of NEFA was negligible, except during zero-grazing, which was associated with a lower, albeit positive calculated energy balance. Mammary FA uptake was higher and the synthesis of 16:0 lower in cows fed fresh grass than hay. Conservation of grass by drying or ensiling had no influence on mammary extraction of TAG and NEFA, despite an increase in milk fat secretion for silages compared with hay and for FAS than UTS. Relative to hay, milk fat from fresh grass contained lower 12:0, 14:0, and 16:0 and higher S3,R7,R11,15-tetramethyl-16:0, cis-9 18:1, trans-11 18:1, cis-9,trans-11 18:2, 18:2n-6, and 18:3n-3 concentrations. Even though conserved forages altered mammary lipogenesis, differences in milk FA composition were relatively minor, other than a higher enrichment of S3,R7,R11,15-tetramethyl-16:0 in milk from silages compared with hay. In conclusion, differences in milk fat composition on fresh grass relative to conserved forages were associated with a lower energy balance, increased uptake of preformed FA, and decreased synthesis of 16:0 de novo in the mammary glands, in the absence of alterations in stearoyl-coenzyme A desaturase activity.

The effect of forage conservation method on ruminal lipid metabolism and microbial ecology was examined in 2 complementary experiments in cows. Treatments comprised fresh chopped grass, barn-dried hay, or untreated (UTS) or formic acid-treated silage (FAS) prepared from the same grass sward. Preparation of conserved forages coincided with the collection of samples from cows offered fresh grass. In the first experiment, 5 multiparous Finnish Ayrshire cows (229d in milk) were used to compare the effects of feeding diets based on grass followed by hay during 2 consecutive 14-d periods separated by a 5-d transition during which extensively wilted grass was fed. In the second experiment, 5 multiparous Finnish Ayrshire cows (53d in milk) were assigned to 1 of 2 blocks and allocated treatments according to a replicated 3×3 Latin square design with 14-d periods to compare the effects of hay, UTS, and FAS. Cows received 7 or 9kg/d of the same concentrate in experiments 1 and 2, respectively. Conservation of grass by drying, but not ensiling, decreased forage fatty acid content primarily due to losses of 18:2n-6 and 18:3n-3. Compared with grass, feeding hay had no effect on dry matter intake (DMI), rumen pH, or fermentation characteristics, other than increasing ammonia content, but lowered whole-tract organic matter and fiber digestibility (experiment 1). Relative to hay, silage increased DMI, rumen volatile fatty acid (VFA) concentrations, and molar proportions of butyrate, and decreased molar acetate proportions (experiment 2). Compared with UTS, FAS increased DMI, had no effect on rumen ammonia or VFA concentrations, but tended to lower rumen pH and the molar ratio of lipogenic to glucogenic VFA. Conservation method had no substantial effect on ruminal or whole-tract digestibility coefficients. Compared with fresh grass and silages, hay decreased lipolysis and biohydrogenation (BH) of dietary unsaturates in the rumen, resulting in similar flows of 18:2n-6 and 18:3n-3, but lower amounts of trans -11 18:1 and Δ11,13 18:2 at the omasum. The extent of silage fermentation had minimal influence on ruminal lipid metabolism. Treatments were not associated with changes in the relative abundance of specific bacteria known to be capable of BH or rumen protozoal numbers. In conclusion, conservation method altered forage lipids, the extent of lipolysis and BH in the rumen, and the flow of fatty acids at the omasum, in the absence of substantial changes in ruminal Butyrivibrio populations.