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To meet the 1.5 °C target, methane (CH4) from ruminants must be reduced by 11 to 30% by 2030 and 24 to 47% by 2050 compared to 2010 levels. A meta-analysis identified strategies to decrease product-based (PB; CH4 per unit meat or milk) and absolute (ABS) enteric CH4 emissions while maintaining or increasing animal productivity (AP; weight gain or milk yield). Next, the potential of different adoption rates of one PB or one ABS strategy to contribute to the 1.5 °C target was estimated. The database included findings from 430 peer-reviewed studies, which reported 98 mitigation strategies that can be classified into three categories: animal and feed management, diet formulation, and rumen manipulation. A random-effects meta-analysis weighted by inverse variance was carried out. Three PB strategies—namely, increasing feeding level, decreasing grass maturity, and decreasing dietary forage-to-concentrate ratio—decreased CH4 per unit meat or milk by on average 12% and increased AP by a median of 17%. Five ABS strategies—namely CH4 inhibitors, tanniferous forages, electron sinks, oils and fats, and oilseeds—decreased daily methane by on average 21%. Globally, only 100% adoption of the most effective PB and ABS strategies can meet the 1.5 °C target by 2030 but not 2050, because mitigation effects are offset by projected increases in CH4 due to increasing milk and meat demand. Notably, by 2030 and 2050, low- and middle-income countries may not meet their contribution to the 1.5 °C target for this same reason, whereas high-income countries could meet their contributions due to only a minor projected increase in enteric CH4 emissions., Proceedings of the National Academy of Sciences of the United States of America, 119 (20), ISSN:0027-8424, ISSN:1091-6490

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

We examined the effects of 2 grass silage-based diets differing in forage:concentrate (FC) ratio and those of a red clover silage-based diet on intake, milk production, ruminal fatty acid (FA) biohydrogenation, milk FA composition, and milk fat globule (MFG) size distribution. Ten multiparous Nordic Red cows received the following treatments: grass silage-based diets containing high (70:30, HG) or low (30:70, LG) FC ratio or a red clover silage-based diet with an FC ratio of 50:50 (RC) on a dry matter basis. Determinations of MFG were performed from fresh milk samples without addition of EDTA so the results of fat globules >1 µm in diameter are emphasized instead of the entire globule population. Lower FC ratio in grass silage-based diets increased milk production with no effect on daily fat yield, leading to 13% lower milk fat concentration. The effect of FC ratio on MFG size was moderate. It did not affect the volume-weighted diameter in grass silage-based diets, although LG lowered the volume-surface diameter of MFG in the size class >1 µm compared with HG. Compared with HG, feeding LG moderately decreased the biohydrogenation of 18:2n-6, leading to a higher level of polyunsaturated fatty acids in milk fat. Feeding RC lowered milk fat concentration and daily milk fat yield compared with grass silage-based diets. The volume-weighted diameter of MFG in the size class >1 µm was smaller in RC milk compared with grass silage-based diets. Feeding RC increased the flow of 18:3n-3 at the omasum by 2.4-fold and decreased the apparent ruminal 18:3n-3 biohydrogenation compared with grass silage-based diets despite similar intake of 18:3n-3. It also resulted in the lowest amount of saturated FA and the highest amounts of cis-9 18:1, 18:3n-3, and polyunsaturated FA in milk. In conclusion, LG decreased milk fat content and induced minor changes in MFG size distribution compared with HG, whereas RC lowered milk fat production, altered milk FA composition to nutritionally more beneficial direction, and led to smaller MFG compared with grass silage-based diets.

It has been well established that milk yield is affected both by milking frequency and due to the removal of residual milk, but the influence of a combination of these factors is unclear. In this study, four mid-lactation cows were used in a 4 × 4 Latin square design to test the hypothesis that the effects of more frequent milking and residual milk removal on milk yield and composition are additive and alter milk fatty acid composition. Treatments comprised two or four times daily milking in combination with (or without) residual milk removal over a 96 h interval preceded by a 2 d pretreatment period and followed by a 8 d washout in each 14 d experimental period. Milk was sampled at each milking for the analysis of gross composition and SCC. Samples of available and residual milk collected on the last milking during each treatment period were collected and submitted for fatty acid composition analysis. Increases in milking frequency and residual milk removal alone or in combination had no effect on milk yield or on the secretion of lactose and protein in milk. However, residual milk removal during more frequent milking increased milk fat yield. Milking treatments had no major influence on the fatty acid composition of available milk, but resulted in rather small changes in the relative abundance of specific fatty acids, with no evidence that the additive effects of treatments were due to higher utilisation of preformed fatty acids relative to fatty acid synthesis de novo. For all treatments, fat composition of available and residual milk was rather similar indicating a highly uniform fatty acid composition of milk fat within the mammary gland.

The effects of supplementing high- or low-concentrate diets with sunflower oil (SO) on rumen fermentation, nutrient utilization, and ruminal methane (CH4) emissions in lactating cows were examined. Four multiparous Nordic Red dairy cows fitted with rumen cannulae were used in a 4 × 4 Latin square with a 2 × 2 factorial arrangement of treatments and 35-d periods. Experimental treatments comprised iso-nitrogenous total mixed rations based on grass silage with forage to concentrate ratio of 65:35 or 35:65 supplemented with 0 or 50 g/kg diet DM of SO. Apparent ruminal OM and starch digestibility was greater (P < 0.05) with high- than low-concentrate diets but was unaffected by SO. Inclusion of SO in high-concentrate diet decreased (P ≤ 0.05) apparent total tract OM, fiber, and GE, and apparent ruminal fiber digestibility. High-concentrate diets and SO shifted (P < 0.05) fiber digestion from rumen to the hindgut. High-concentrate diet resulted in a lower rumen pH and elevated total rumen VFA concentration compared with low-concentrate diet, whereas SO increased rumen pH and decreased rumen VFA concentration when included in high-, but not low-concentrate diet (P < 0.05 for interaction). High-concentrate diet reduced rumen ammonia-N (P < 0.01) and molar proportion of acetate to propionate (P < 0.01), and decreased (P < 0.05) ruminal CH4 emissions when expressed as g/d or g/kg OM digested in the rumen. With both low- and high-concentrate diets, SO reduced (P < 0.05) daily emissions of CH4 as g/d or g/kg OM digested in the rumen, but SO reduced CH4 emissions expressed as g/kg OM intake, OM digested in total digestive tract, energy-corrected milk or % of GE intake only with low-concentrate diet (P ≤ 0.05 for interaction). In conclusion, replacing grass silage with concentrates led to a reduction in daily ruminal CH4 emissions that were accompanied by a shift in rumen fermentation toward the synthesis of propionate, and decreases in rumen pH and fiber digestion. Sunflower oil was effective in reducing daily CH4 emissions in lactating cows which was accompanied by a noticeable lower feed intake with high- but not low-concentrate diet. Overall the effects of SO and greater proportion of concentrates in the diet on daily CH4 emissions were additive but the additivity declined or vanished when different indices of CH4 emission intensity were considered. Consequently, SO was more effective in reducing CH4 emissions when low-concentrate diet was fed.

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.

The biohydrogenation theory of milk fat depression (MFD) attributes decreases in milk fat in cows to the formation of specific fatty acids (FA) in the rumen.Trans-10,cis-12-CLA is the only biohydrogenation intermediate known to inhibit milk fat synthesis, but it is uncertain if increased ruminal synthesis is the sole explanation of MFD. Four lactating cows were used in a 4×4 Latin square with a 2×2 factorial arrangement of treatments and 35-d experimental periods to evaluate the effect of diets formulated to cause differences in ruminal lipid metabolism and milk fat synthesis on the flow of FA and dimethyl acetal at the omasum. Treatments comprised total mixed rations based on grass silage with a forage:concentrate ratio of 35:65 or 65:35 containing 0 or 50 g/kg sunflower oil (SO). Supplementing the high-concentrate diet with SO lowered milk fat synthesis from −20·2 to −31·9 % relative to other treatments. Decreases in milk fat were accompanied by alterations in ruminal biohydrogenation favouring thetrans-10 pathway and an increase in the formation of specific intermediates includingtrans-4 totrans-10-18 : 1,trans-8,trans-10-CLA,trans-9,cis-11-CLA andtrans-10,cis-15-18 : 2. Flow oftrans-10,cis-12-CLA at the omasum was greater on high- than low-concentrate diets but unaffected by SO. In conclusion, ruminaltrans-10,cis-12-CLA formation was not increased on a diet causing MFD suggesting that other biohydrogenation intermediates or additional mechanisms contribute to the regulation of fat synthesis in the bovine mammary gland.

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.

Diet-induced milk fat depression (MFD) in lactating cows has been attributed to alterations in ruminal lipid metabolism leading to the formation of specific fatty acid (FA) biohydrogenation intermediates that directly inhibit milk fat synthesis. However, the mechanisms responsible for decreased lipid synthesis in the mammary gland over time are not well defined. The aim of this study was to evaluate the effect of diet on milk FA composition and milk fat production over time, especially during MFD, and explore the associations between MFD and FA biohydrogenation intermediates in omasal digesta and milk. Four lactating Finnish Ayrshire cows used in a 4 × 4 Latin square with a 2 × 2 factorial arrangement of treatments and 35-d experimental periods were fed diets formulated to cause differences in ruminal and mammary lipid metabolism. Treatments consisted of an iso-nitrogenous total mixed ration based on grass silage with a forage to concentrate ratio of 65:35 or 35:65 without added oil, or with sunflower oil at 50 g/kg of diet dry matter. The high-concentrate diet with sunflower oil (HSO) induced a 2-stage drop in milk fat synthesis that was accompanied by specific temporal changes in the milk FA composition. The MFD on HSO was associated especially with trans-10 18:1 and also with trans-9,cis-11 conjugated linoleic acid (CLA) in milk and omasal digesta across all diets and was accompanied by the appearance of trans-10,cis-15 18:2. Trans-10,cis-12 CLA was increased in HSO, but milk fat secretion was not associated with omasal or milk trans-10,cis-12 CLA. The temporal changes in milk fat content and yield and milk FA composition reflect the shift from the predominant ruminal biohydrogenation pathway to an alternative pathway. The ambiguous role of trans-10,cis-12 CLA suggests that trans-10 18:1, trans-9,cis-11 CLA and trans-10,cis-15 18:2 or additional mechanisms contributed to the diet-induced MFD in lactating cows.

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

13 páginas, 4 figuras, 2 tablas., A great deal of uncertainty still exists about intermediate metabolites and pathways explaining the biohydrogenation (BH) of 20- and 22-carbon polyunsaturated fatty acids (PUFA). Therefore, this study was conducted to provide further insight into the ruminal metabolism of 20:5 n-3 (EPA), 22:5 n-3 (DPA), and 22:6 n-3 (DHA), the main n-3 PUFA present in the marine lipids used in dairy ruminant feeding, and to examine potential differences between bovine and ovine. To meet this aim, we investigated the 20- and 22-carbon metabolites accumulated during in vitro incubation of EPA, DPA, and DHA with rumen inocula from cows and ewes. The PUFA were added at a dose of 2% incubated dry matter and digesta samples were analyzed after 24 h of incubation using complementary gas-liquid chromatography of fatty acid methyl esters and gas chromatography-mass spectrometry of 4,4-dimethyloxazoline derivatives. Results suggested that the main BH pathway of EPA and DPA would proceed via the reduction of the double bond closest to the carboxyl group (cis-5 in EPA and cis-7 in DPA); curiously, this mechanism seemed of much lower importance for DHA. Thus, DPA would not be a major intermediate product of DHA and their BH might actually follow separate pathways, with the accumulation of numerous unique metabolites in each case. A principal component analysis supported this hypothesis, with a clear separation between PUFA treatments in the score and loading plots. Within EPA and DPA groups, cow and ewe samples loaded separately from each other but not distant. No conjugated 20:5, 22:5, or 22:6 isomer compatible with the initial product of EPA, DPA, or DHA metabolism, respectively, was identified in the ruminal digesta, although this would not unequivocally exclude their transient formation. In this regard, results from DPA incubations provided the first indication that the metabolism of this very long chain PUFA may involve the formation of conjugated double bond structures. The BH of EPA, DPA, and DHA resulted in the appearance of several tentative trans-10-containing metabolites, showing a general trend to be more abundant in the digesta of ewes than in that of cows. This finding was speculated to have some relationship with the susceptibility of dairy sheep to marine lipid-induced milk fat depression. Differences in the relative proportion of intermediate products would also suggest an influence of ruminant species on BH kinetics, with a process that would likely be slower and less complete in cows than in ewes. Copyright © 2018 American Dairy Science Association. Published by Elsevier Inc. All rights reserved., This work was supported by the Spanish Ministry of Economy and Competitiveness (MINECO; AGL2014- 54587-R). P. G. Toral benefited from a Ramón y Cajal research contract from the MINECO. Co-funding by the European Regional Development Fund is also acknowledged.

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.

This study evaluated the effects of gradual replacement of a mixture of late-cut grass silage (LS) and barley with early-cut grass silage (ES) on milk production, CH4 emissions, and N utilization in Swedish Red cows. Two grass silages were prepared from the same primary growth of timothy grass sward but harvested 2 wk apart [11.0 and 9.7 MJ of metabolizable energy/kg of dry matter (DM)]. Four diets, fed as a total mixed ration, were formulated to meet the metabolizable energy and protein requirements of 35 kg of energy-corrected milk (ECM) by gradually replacing a mixture of LS and barley with ES (0, 33, 67, and 100% of the forage component of the diet), whereas the proportion of barley decreased from 47.2 to 26.6% of diet DM. Expeller canola meal was used as a protein supplement. Sixteen Swedish Red cows were used in 4 replicated 4 × 4 Latin squares. Cows were offered diets ad libitum and milked twice daily. Each period of 28 d comprised 14 d of diet adaptation followed by 14 d of data collection. Intake and milk yield were recorded daily, and milk samples were collected on d 19 to 21 and d 26 to 28 of each period. Diet digestibility was determined by grab sampling using indigestible neutral detergent fiber as an internal marker. Gas emissions were measured using the GreenFeed system (C-Lock Inc., Rapid City, SD). Dry matter intake (DMI) linearly decreased from 22.6 to 19.3 kg/d as the proportion of ES increased in the diet. The ECM yield did not differ among treatments, but milk protein yield decreased with increasing proportion of ES in the diet. Because of reduced DMI with increasing ES, feed efficiency (ECM/DMI) improved with an increased proportion of ES in the diet. Nitrogen efficiency (milk N/N intake) did not change despite a linear increase in milk urea N concentration from 9.7 (LS alone) to 11.9 mg/dL (ES alone) with graded replacement of LS and barley by ES in the diet. Lower DMI responses in ES diets were partly compensated for by increased organic matter digestibility (656 g/kg of DM for LS alone; 715 g/kg of DM for ES alone) related to improved forage digestibility at early harvesting. Total CH4 emissions and CH4 intensity (CH4/ECM) were not influenced by diet, but CH4 yield (CH4/DMI) increased linearly from 19.5 to 23.0 g/kg of DMI with greater inclusion of ES in the diet. In conclusion, replacing LS and barley with ES improved the conversion of feed to milk without increasing CH4 emissions or compromising N efficiency.

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 (

Description ofα-linolenic acid (cis-9,cis-12,cis-15-18 : 3, ALA) metabolism in the rumen is incomplete. Ruminal digesta samples were incubated with ALA and buffer containing water or deuterium oxide to investigate the products and mechanisms of ALA biohydrogenation. Geometric Δ9,11,15-18 : 3 isomers were the main intermediates formed from ALA. An increase in then+1 isotopomers of Δ9,11,15-18 : 3 was due to2H labelling at C-13. Isomers of Δ9,11,13-18 : 3,cis-7,cis-12,cis-15-18 : 3 andcis-8,cis-12,cis-15-18 : 3 were also formed. No increase inn+1 isotopomers of Δ7,12,15-18 : 3 or Δ8,12,15-18 : 3 was detected. Enrichment inn+2 isotopomers of 18 : 2 products indicated that ALA metabolism continued via the reduction of 18 : 3 intermediates. Isomers of Δ9,11,15-18 : 3 were reduced to Δ11,15-18 : 2 labelled at C-9 and C-13. ALA resulted in the formation of Δ11,13-18 : 2 and Δ12,14-18 : 2 containing multiple2H labels. Enrichment of then+3 isotopomer of Δ12,15-18 : 2 was also detected. Metabolism of ALA during incubations with rumen contents occurs by one of three distinct pathways. Formation of Δ9,11,15-18 : 3 appears to be initiated by H abstraction on C-13. Octadecatrienoic intermediates containingcis-12 andcis-15 double bonds are formed without an apparent H exchange with water. Labelling of Δ9,11,13-18 : 3 was inconclusive, suggesting formation by an alternative mechanism. These findings explain the appearance of several bioactive fatty acids in muscle and milk that influence the nutritional value of ruminant-derived foods.

Dietary unsaturated fatty acids are extensively hydrogenated in the rumen, resulting in the formation of numerous intermediates that may exert physiological effects and alter the fat composition of ruminant-derived foods. A batch culture method was used to characterize the hydrogenation of linoleic acid (LeA) by strained rumen fluid in vitro. Incubations (n=5) were performed in 100-mL flasks maintained at 39°C containing 400mg of grass hay, 50mL of buffered rumen fluid, and incremental amounts of LeA (0, 1.0, 2.5, 5.0, or 10.0mg) for 0, 1.5, 3.0, 4.5, 6.0, and 9.0h. The fatty acid composition of flask contents was determined using complimentary silver-ion thin-layer chromatography, gas chromatography mass-spectrometry, and silver-ion high-performance liquid chromatography. Linoleic acid was extensively (98.1, 97.6, 98.0, and 89.8% for additions of 1.0, 2.5, 5.0, and 10.0mg of LeA, respectively) hydrogenated over time. Complete reduction of LeA to 18:0 was inhibited in direct relation to the amount of added substrate, the extent of which was greatest for the highest amount of LeA addition. Recoveries of 1.0, 2.5, 5.0, and 10.0mg of added LeA as 18:0 averaged 73.6, 65.0, 57.3, and 10.7%, respectively. Incubation of incremental amounts of LeA resulted in a time-dependent accumulation of geometric isomers of 9,11 and 10,12 conjugated linoleic acid, several nonconjugated 18:2 isomers, and a wide range of cis 18:1 and trans 18:1 intermediates. Several unusual intermediates including cis -6, cis -12 18:2; cis -7, cis -12 18:2; and cis -8, cis -12 18:2, were found to accumulate in direct relation to the amount of added LeA, providing the first indications that hydrogenation of LeA by ruminal bacteria may also involve mechanisms other than hydrogen abstraction or isomerization of the cis -12 double bond. Fitting of single-pool, first-order kinetic models to experimental data indicated that the rate of LeA disappearance decreased with increases in substrate availability. Reduction of 18:1 and 18:2 intermediates occurred at much lower rates compared with conjugated linoleic acid and nonconjugated 18:2 isomer formation. In conclusion, the extent of LeA biohydrogenation in vitro was shown to be time- and dose-dependent with evidence that LeA is hydrogenated by ruminal bacteria via several distinct metabolic pathways. The accumulation of several unusual 18:2 isomers indicates that biohydrogenation of LeA also proceeds via mechanisms other than isomerization of the cis -12 double bond.

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.

The objective of this experiment was to investigate the effects of replacing conventional, solvent-extracted canola meal (control; CTRL) with high oil content; conventional, mechanically extracted canola meal (CMEC); high-oleic, low polyunsaturated fatty acid (FA) canola meal (HOLL); and high-erucic acid, low-glucosinolate rapeseed meal (RPS) on rumen function, digestibility, milk production, and milk FA composition in lactating dairy cows. The experimental design was a replicated 4×4 Latin square with 8 lactating dairy cows. Four of the cows were ruminally cannulated. All oilseed meals were included at approximately 12 to 13% of dietary dry matter (DM). Crude protein and fat concentrations (% of DM) of the meals were 43 and 3.1%, 32.8 and 16.1%, 45.2 and 13.7%, and 34.3 and 17.9% for CTRL, CMEC, HOLL, and RPS, respectively. All diets were formulated to supply net energy of lactation in excess of requirements. The CMEC and RPS diets were predicted to be about 1% deficient in metabolizable protein. Relative to the CTRL, inclusion of high-oil seed meals in the diet lowered ruminal acetate concentration and the molar acetate:propionate ratio and decreased DM intake. Milk yield generally followed DM intake and was lower for CMEC and RPS than the CTRL. Treatments had no effect on milk composition, other than an increase in milk urea nitrogen concentration for HOLL. Fat-corrected milk (3.5%) feed efficiency was increased by HOLL and RPS compared with CTRL. Urinary urea nitrogen losses were increased by HOLL, which, as a consequence, increased the ammonia-emitting potential of manure. The ratio of milk N-to-N intake was greater for CMEC and RPS. Replacing solvent-extracted canola meal with the high-oil meal decreased milk fat 12:0, 14:0, 16:0, and total saturated FA content and enhanced cis-9 18:1 and total monounsaturated FA concentrations. Relative to the CTRL, canola increased total trans FA in milk, whereas inclusion of HOLL in the diet increased trans-11 18:1 and cis-9, trans-11 CLA content. The RPS increased milk fat cis-13 22:1 content from 0.07 to 2.33 g/100g of FA. In conclusion, HOLL or RPS, which are likely to come from small-scale biodiesel plants where oil is cold pressed without hexane extraction, fed at levels at or above 12 to 13% of dietary DM may decrease feed intake and milk production, but can be used to alter milk FA composition in lactating dairy cows.

Based on potential benefits to human health, there is increasing interest in altering the composition of ruminant-derived foods. Including rapeseeds in the dairy cow diet is an effective strategy for replacing medium-chain saturated fatty acids (SFA) with cis-monounsaturated fatty acids (MUFA) in bovine milk, but there is limited information on the optimum level of supplementation. Decreases in SFA due to plant oils are also accompanied by increases in milk trans fatty acid (FA) content and it is possible that high oleic acid rapeseeds may result in a higher enrichment of cis-9 18:1 and lower increases in trans FAs in milk compared with conventional varieties. Seven multiparous lactating Holstein–Friesian cows were allocated to one of seven treatments in an incomplete Latin square design with five 28-day experimental periods, to evaluate the effect of replacing calcium salts of palm oil distillate (CPO; 41 g/kg diet dry matter, DM) with 128, 168 or 207 g/kg diet DM of conventional (COR) or a high oleic acid (HOR) rapeseed fed as a supplement milled with wheat. Rapeseed variety and inclusion level had no effect (P > 0.05) on DM intake, milk yield and composition. Both rapeseed varieties decreased linearly (P < 0.001) milk fat SFA content, which was partially compensated for by a linear increase (P < 0.001) in cis-9 18:1 concentration. Reductions in milk SFA were also associated with increases (P < 0.05) in trans 18:1 and total trans FA content, with no difference (P > 0.05) between rapeseed varieties. Replacing CPO in the diet with milled rapeseeds had no effect (P > 0.05) on total milk conjugated linoleic acid (CLA) concentration. Relative to a COR, inclusion of a high oleic acid variant in the diet increased (P = 0.01) the ratio of trans-MUFA : trans-polyunsaturated fatty acids in milk that may have implications with respect to cardiovascular disease risk in humans. In conclusion, data indicated that replacing CPO with milled rapeseeds at levels up to 1150 g oil/day could be used as a nutritional strategy to lower milk SFA content without inducing adverse effects on DM intake and milk production. HOR reduced milk fat SFA content to a greater extent than a conventional variety, but did not minimise associated increases in trans FA concentrations. However, the high oleic acid variant did alter the relative abundance of specific trans 18:1, CLA and trans 18:2 isomers compared with conventional rapeseeds.

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.

Inclusion of rapeseed feeds in dairy cow diets has the potential to reduce milk fat saturated fatty acid (SFA) and increase cis-monounsaturated fatty acid (cis-MUFA) content, but effectiveness may depend on the form in which the rapeseed is presented. Four mid-lactation Holstein dairy cows were allocated to four maize silage-based dietary treatments according to a 4 × 4 Latin Square design, with 28-day experimental periods. Treatments consisted of a control diet (C) containing 49 g/kg dry matter (DM) of calcium salts of palm oil distillate (CPO), or 49 g/kg DM of oil supplied as whole rapeseeds (WR), rapeseeds milled with wheat (MR) or rapeseed oil (RO). Replacing CPO with rapeseed feeds had no effect (P > 0.05) on milk fat and protein content, while milk yields were higher (P < 0.05) for RO and MR compared with WR (37.1, 38.1 and 34.3 kg/day, respectively). Substituting CPO with RO or MR reduced (P < 0.05) milk fat total SFA content (69.6, 55.6, 71.7 and 61.5 g/100 g fatty acids for C, RO, WR and MR, respectively) and enhanced (P < 0.05) milk cis-9 18:1 MUFA concentrations (corresponding values 18.6, 24.3, 17.0 and 23.0 g/100 g fatty acids) compared with C and WR. Treatments RO and MR also increased (P < 0.05) milk trans-MUFA content (4.4, 6.8, 10.5 g/100 g fatty acids, C, MR and RO, respectively). A lack of significant changes in milk fat composition when replacing CPO with WR suggests limited bioavailability of fatty acids in intact rapeseeds. In conclusion, replacing a commercial palm oil-based fat supplement in the diet with milled rapeseeds or rapeseed oil represented an effective strategy to alter milk fatty acid composition with the potential to improve human health. Inclusion of processed rapeseeds offered a good compromise for reducing milk SFA and increasing cis-MUFA, whilst minimising milk trans-MUFA and negative effects on animal performance.

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.

Maitorasvan rasvahappokoostumusta voidaan muuttaa varsin tehokkaasti lehmien ruokintaa muuttamalla. Lisäämällä rehuun kasvirasvaa vähennetään lyhytketjuisten, tyydyttyneiden rasvahappojen (C12:0, C14:0 ja C16:0) pitoisuutta maidossa ja lisätään pitkäketjuisten, kerta (MUFA)- ja monityydyttymättömien (PUFA) rasvahappojen osuutta. Lisättäessä rehuun suuria annoksia rasvaa, esimerkiksi öljynä, rehun monityydyttymättömät rasvahapot pelkistyvät pötsissä epätäydellisesti ja pötsiin kertyy erilaisia reaktiovälituotteita, kuten trans-rasvahappoja. Öljykasvinsiemenien kuoret suojaavat monityydyttymättömiä rasvahappoja pötsimikrobien toiminnalta ja saattavat siten ehkäistä myös trans-rasvahappojen muodostusta pötsissä. Tämän tutkimuksen tavoitteena oli selvittää kuinka hyvin rypsinsiemenen kuori suojaa rypsin monityydyttymättömiä rasvahappoja pelkistykseltä pötsissä ja mikä on kokonaisten öljykasvinsiemenien vaikutus maidon rasvahappokoostumukseen. Koe suoritettiin 4 x 4 latinalaisen neliön koemallin mukaan. Koekäsittelyn mukaan (kontrolli, ei rasvalisää; C) lehmien rehuseokseen sekoitettiin rypsiöljyä (RO), kokonaisia rypsinsiemeniä (WR) tai murskattuja rypsinsiemeniä (MR) siten, että rehuseokset sisälsivät rasvaa noin 50 g/kg KA. Rypsirasvojen lisäys ei vaikuttanut merkitsevästi kokonaiskuiva-aineen syöntiin (19.3, 18.8, 21.8 ja 20.5 kg/pv koekäsittelyissä C, RO, WR ja MR vastaavasti), eikä lehmien maitotuotokseen (28.5, 28.0, 28.3 ja 26.9 kg/pv), valkuais- (31.3, 31.0, 31.1 ja 30.5 g/kg), rasva- (41.2, 42.2, 43.0 ja 43.0 g/kg) ja laktoosipitoisuuksiin (48.9, 49.0, 49.6 ja 48.6 g/kg). Rypsirasvojen lisäys muutti maitorasvan koostumusta merkitsevästi. Tyydyttyneiden rasvahappojen osuus maidon kokonaisrasvahapoista väheni (72.1, 58.1, 65.3 ja 61.1 g/100g rasvahappoja; RA) ja kertatyydyttymättömien lisääntyi (26.4, 39.9, 33.2 ja 37.4 g/100g RA) rypsirasvoja lisättäessä. Tyydyttyneiden rasvahappojen osuuden väheneminen johtui erityisesti maitorauhasen de novo-synteesissä muodostuvien lyhytketjuisten rasvahappojen määrän vähentymisestä. Öljyhappopitoisuuden lisääntyminen (18.4, 30.2, 25.5 ja 29.4 g/100g RA) selittää lähes yksinomaan kertatyydyttymättömien rasvahappojen kokonaismäärän lisääntymisen maidon kokonaisrasvapoista. Rypsiöljylisäys nosti merkitsevästi maidon trans-rasvahappojen pitoisuutta muihin rypsirasvoihin verrattuna (3.91, 7.12, 4.51 ja 5.08 g/100g RA). Linolihapon (C18:2n-6) pitoisuus pieneni merkitsevästi (1.11, 0.98, 0.98 ja 0.92 g/100g RA). Cis-9, trans-11 CLA (engl. conjugated linoleic acid) oli maidon pääasiallisin CLA-isomeeri ja sen pitoisuus oli suurimmillaan lisättäessä rypsiöljyä lehmien rehuun (351, 597, 285 ja 346 mg/100g RA). Vaikka kaikki rasvalisät vähensivät maitorasvan tyydyttyneiden rasvahappojen pitoisuutta ja lisäsivät MUFA:n ja PUFA:n osuutta kontrolliruokintaan verrattuna, ravitsemuksellisesti paras muutos maidon rasvahappokoostumukseen saatiin lisättäessä lehmien rehuun murskattuja rypsinsiemeniä.

There is increasing evidence that nutrition plays an important role in the development of chronic diseases in the human population including cancer, cardiovascular disease, insulin resistance and obesity. Developing foods that enhance human health is central to dietary approaches for preventing and reducing the economic and social impact of chronic disease. Numerous studies in human subjects have implicated a high consumption of medium-chain (12:0-16:0) saturated fatty acids (SFA) and trans fatty acids (TFA) as risk factors for cardiovascular disease risk, with emerging evidence of a possible role in the development of insulin resistance and inflammation. Milk and dairy products are a major source of 12:0, 14:0, 16:0 and TFA in the human diet. However, developing public health policies promoting a decrease in milk, cheese and butter consumption ignores the value of these foods as a versatile source of nutrients and bioactive lipids, including 4:0, branch-chain fatty acids, cis-9, trans-11 conjugated linoleic acid (CLA), vitamins A and D, β-carotene and sphingomyelin. Therefore, altering milk fatty acid composition through sustainable, environmental and welfare acceptable means is an integral component of an overall strategy for preventing human chronic disease. Nutrition is the major environmental factor regulating milk fat composition. Strategies for enhancing the nutritional value of milk fat have been directed towards reducing the proportion of 12:0, 14:0 and 16:0, increasing cis monounsaturated fatty acids (MUFA) and polyunsaturated fatty acid (PUFA) content and/or enhancing the concentration of bioactive lipids. Formulation of diets to alter milk fat composition to meet these targets typically involves 1) inclusion of plant oils and oilseeds, 2) supplements of marine lipids, 3) increasing the proportion of dietary energy derived from fresh grass, 4) replacing ensiled grass, maize or whole-crop cereal with forage legumes or 5) inclusion of rumen-protected lipids in the diet. Nutritional strategies for reducing 12:0, 14:0 and 16:0 and enhancing cis-MUFA in milk fat are dependent on increasing the supply of C18 or longer-chain fatty acids to inhibit mammary de novo fatty acid synthesis. Due to incomplete metabolism of dietary unsaturated fatty acids in the rumen, inclusion of oils or oilseeds in the diet also increases milk fat TFA content. Both the concentration and distribution of TFA isomers in milk is dependent on the amount and type of lipid supplement fed, composition of the basal diet and interactions between these factors. Altering the diet of lactating cows can be used as a means to significantly enrich milk fat cis-9, trans-11 CLA content. Increases in the concentration of CLA in milk are reliant, in the most part, on enhancing ruminal trans-11 18:1 outflow for endogenous cis-9, trans-11 CLA synthesis in the mammary gland. The potential to increase n-3 fatty acids in milk is relatively limited. Inclusion of fresh or ensiled red clover in the ruminant diet significantly increases 18:3 n-3 content, while increases in milk 20:5 n-3 and 22:6 n-3 concentrations to marine lipid supplements is relatively limited due to extensive ruminal metabolism of n-3 PUFA. Greater enrichment of 20:5 n-3, 22:5 n-3 and 22:6 n-3 in milk can be achieved using rumen-protected fish oil supplements. Overall, recent research has highlighted the important role of nutrition in attempts to modify milk fat composition for improved long-term human health.

Even though extensive research has examined the role of nutrition on milk fat composition, there is less information on the impact of forages on milk fatty acid (FA) composition. In the current study, the effect of replacing grass silage (GS) with maize silage (MS) as part of a total mixed ration on animal performance and milk FA composition was examined using eight multiparous mid-lactation cows in a replicated 4 × 4 Latin square with 28-day experimental periods. Four treatments comprised the stepwise replacement of GS with MS (0, 160, 334 and 500 g/kg dry matter (DM)) in diets containing a 54 : 46 forage : concentrate ratio on a DM basis. Replacing GS with MS increased (P < 0.001) the DM intake, milk yield and milk protein content. Incremental replacement of GS with MS in the diet enhanced linearly (P < 0.001) the proportions of 6:0–14:0, decreased (P < 0.01) the 16:0 concentrations, but had no effect on the total milk fat saturated fatty acid content. Inclusion of MS altered the distribution of trans-18:1 isomers and enhanced (P < 0.05) total trans monounsaturated fatty acid and total conjugated linoleic acid content. Milk total n-3 polyunsaturated fatty acid (PUFA) content decreased with higher amounts of MS in the diet and n-6 PUFA concentration increased, leading to an elevated n-6 : n-3 PUFA ratio. Despite some beneficial changes associated with the replacement of GS with MS, the overall effects on milk FA composition would not be expected to substantially improve long-term human health. However, the role of forages on milk fat composition must also be balanced against the increases in total milk and protein yield on diets containing higher proportions of MS.