A.J.M. Fonseca

Rumen biohydrogenation-derived fatty acids in milk fat from grazing dairy cows supplemented with rapeseed, sunflower, or linseed oils

The effects of supplementation with rapeseed, sunflower, and linseed oils (0.5 kg/d; good sources of oleic, linoleic, and linolenic acids, respectively) on milk responses and milk fat fatty acid (FA) profile, with special emphasis on rumen-derived biohydrogenation intermediates (BI), were evaluated in a replicated 4 x 4 Latin square study using 16 grazing dairy cows. The dietary treatments were 1) control diet: 20-h access to grazing pasture supplemented with 5 kg/d of corn-based concentrate mixture (96% corn; CC); 2) RO diet: 20-h access to grazing supplemented with 4.5 kg/d of CC and 0.5 kg of rapeseed oil; 3) SO diet: 20-h access to grazing supplemented with 4.5 kg/d of CC and 0.5 kg of sunflower oil; and 4) LO diet: 20-h access to grazing supplemented with 4.5 kg/d of CC and 0.5 kg of linseed oil. Milk fatty acids were converted to methyl esters and analyzed by gas-liquid chromatography and silver-ion HPLC. Dietary treatments had no effect on milk production or on milk protein content and milk protein production. Supplementation with rapeseed and sunflower oils lowered milk fat content and milk fat production, but linseed oil had no effect. Inclusion of dietary vegetable oils promoted lower concentrations of short-chain (including 4:0) and medium-chain FA (including odd- and branched-chain FA) and 18:3n-3, and higher concentrations of C(18) FA (including stearic and oleic acids). The BI concentration was higher with the dietary inclusion of vegetable oils, although the magnitude of the concentration and its pattern differed between oils. The RO treatment resulted in moderate increases in BI, including trans 18:1 isomers and 18:2 trans-7,cis-9, but failed to increase 18:1 trans-11 and 18:2 cis-9,trans-11. Sunflower oil supplementation resulted in the highest concentrations of the 18:1 trans-10, 18:1 cis-12, and 18:2 trans-10,trans-12 isomers. Concentrations of 18:1 trans-11 and 18:2 cis-9,trans-11 were higher than with the control and RO treatments but were similar to the LO treatment. Concentration of BI in milk fat was maximal with LO, having the highest concentrations of some 18:1 isomers (i.e., trans-13/14, trans-15, cis-15, cis-16), most of the nonconjugated 18:2 isomers (i.e., trans-11,trans-15, trans-11,cis-15, cis-9,cis-15, and cis-12,cis-15), and conjugated 18:2 isomers (i.e., trans-11,cis-13, cis-12,trans-14, trans-11,trans-13, trans-12,trans-14, and trans-9,trans-11), and all conjugated 18:3 isomers. The LO treatment induced the highest amount and diversity of BI without decreasing milk fat concentration, as the RO and SO treatments had, suggesting that the BI associated with 18:3n-3 intake may not be the major contributors to inhibition of mammary milk fat synthesis.

Detailed dimethylacetal and fatty acid composition of rumen content from lambs fed lucerne or concentrate supplemented with soybean oil.

Lipid metabolism in the rumen is responsible for the complex fatty acid profile of rumen outflow compared with the dietary fatty acid composition, contributing to the lipid profile of ruminant products. A method for the detailed dimethylacetal and fatty acid analysis of rumen contents was developed and applied to rumen content collected from lambs fed lucerne or concentrate based diets supplemented with soybean oil. The methodological approach developed consisted on a basic/acid direct transesterification followed by thin-layer chromatography to isolate fatty acid methyl esters from dimethylacetal, oxo- fatty acid and fatty acid dimethylesters. The dimethylacetal composition was quite similar to the fatty acid composition, presenting even-, odd- and branched-chain structures. Total and individual odd- and branched-chain dimethylacetals were mostly affected by basal diet. The presence of 18∶1 dimethylacetals indicates that biohydrogenation intermediates might be incorporated in structural microbial lipids. Moreover, medium-chain fatty acid dimethylesters were identified for the first time in the rumen content despite their concentration being relatively low. The fatty acids containing 18 carbon-chain lengths comprise the majority of the fatty acids present in the rumen content, most of them being biohydrogenation intermediates of 18∶2n−6 and 18∶3n−3. Additionally, three oxo- fatty acids were identified in rumen samples, and 16-O-18∶0 might be produced during biohydrogenation of the 18∶3n−3.

Improved method for fatty acid analysis in herbage based on direct transesterification followed by solid-phase extraction

Direct transesterification (DT) and solvent extraction with acid or basic derivatization procedure for fatty acid (FA) analysis in herbage were compared. The highest total FA, alpha-linolenic and linoleic acid contents were obtained with DT. However, DT also produced the highest amount of interfering compounds, identified as phytadienes and sugar derivative products, which may co-elute with FA. An additional step based on solid-phase extraction to produce clean samples was proposed. This procedure is simple and gives good recoveries for the FA fortified samples. Additionally, structural characterization of 16:1 trans-3 was conducted by covalent adduct chemical ionization tandem mass spectrometry.

Effect of ensiling and silage additives on fatty acid composition of ryegrass and corn experimental silages

Two experiments were conducted using laboratory mini-silos to study the effect of ensiling and silage additives on fatty acid (FA) composition, including minor or unusual FA, of ryegrass and corn silages. Ryegrass was ensiled for 12 wk with no additives, with the addition of a bacterial inoculant or formic acid. Corn was ensiled for 9 wk without additives, with the addition of a bacterial inoculant or calcium formate. Ensiling affected both total FA content and FA composition of ryegrass silages. Total FA concentration increased (P < 0.001) during ryegrass ensiling. The proportions (g/100 g of total FA) of the major unsaturated FA, 18:3n-3 and 18:2n-6, were not affected (P > 0.05) by ensiling. However, their concentration (mg/g of DM) in silages was greater (P=0.017 and P=0.001, respectively) than in fresh ryegrass. Two 18:2 FA (trans-11,cis-15 and cis-9,cis-15) that were not originally present in the fresh ryegrass were detected in silages. Silage additives affected the FA composition of ryegrass silages, mostly by increasing the proportions of SFA, but not on total FA concentration. Ensiling did not affect (P=0.83) total FA content of corn silages; however, FA composition was affected, mostly by decreasing the proportions of 18:2n-6 and 18:3n-3. Silage additives had no effect on corn silage FA composition. Exposing corn silages to air resulted in no oxidation of FA or reduction in total FA content or composition.

Effects of dietary protein concentration and balance of absorbable amino acids on productive responses of dairy cows fed corn silage-based diets

A cyclical changeover design experiment (3-wk periods; 12-wk total) was conducted to evaluate whether improving the balance of absorbable AA would allow the feeding of less crude protein (CP) without compromising production, thereby reducing the potential environmental pollution from dairy farms. Sixteen multiparous Holstein cows were assigned to 1 of 8 dietary treatments as total mixed rations (TMR) containing [dry matter (DM) basis] 45% corn silage, 5% coarsely chopped wheat straw, and 50% concentrate mixture. The 8 treatments were formulated to differ in dietary CP (14 and 16%; DM basis) and in the balance of absorbable AA achieved by changing the main protein source (MPS) of the concentrate mixtures [replacing soybean meal (SBM) with corn byproducts (CBP), dried corn distillers grains (DDG), and some corn gluten meal], and by adding a mixture of rumen-protected Lys and Met (RPLM). Feeding lactating dairy cows corn silage-based diets with 16% CP promoted significantly higher DM intakes and milk yields, and lower feed N-use efficiency than feeding diets with 14% CP. Replacing SBM with CBP significantly increased milk yields and decreased milk fat and protein concentrations, but had no effect on the efficiency of conversion of feed N into milk N. With 16% CP diets, the addition of RPLM decreased feed N use efficiency. A significant effect was observed for the MPS × RPLM interaction on milk protein concentrations. Plasma Lys concentration was lower with diets based on CBP, and plasma Met increased with RPLM. We did not find clear benefits of RPLM in facilitating a reduction of dietary protein without loss of production.

Tracing seaweeds as mineral sources for farm-animals

This study characterized the mineral composition of 15 common Portuguese seaweed (green, brown, and red) species. Total measured mineral content ranged from 10.9xa0gxa0kg−1 dry matter (DM) in Gracilaria vermiculophylla to 71.0xa0gxa0kg−1 DM in Codium adhaerens, calcium being the mineral generally found in higher amounts. Overall, the results suggest that seaweeds have great potential as mineral sources for animal feeding, but a great variability between species was observed regarding their mineral profile. Compared to common animal feed ingredients, the studied seaweeds can be considered as good sources of calcium, magnesium, iron, iodine, copper, manganese, and selenium but are poor sources of phosphorous and zinc. The maximum level of dietary inclusion will be strongly dependent on the mineral profile of the seaweeds. Depending on the seaweed, the upper level of inclusion in poultry and swine diets may reach more than 40xa0%. The high iodine content of studied seaweeds limits their use in diets for horses, and, to a lesser extent, for ruminants. This work constitutes a paramount contribution regarding the use of seaweeds as mineral sources in animal diets, allowing a more precise choice of the algae species and level of inclusion to be used, thus assuring animal health and strengthening the seaweed industry through this underexploited application field.

European marketable grain legume seeds: Further insight into phenolic compounds profiles.

Twenty-nine mature raw varieties of grain legume seeds (chickpeas, field peas, faba beans, common vetch and lupins) produced in Europe were investigated for their phenolic profile by means of high performance liquid chromatography coupled to diode array detection (HPLC-DAD). To the best of our knowledge, this study reported for the first time the phenolic composition of mature raw seeds of chickpea type Desi, field pea and common vetch. Phenolic acids were predominant compounds in chickpeas, field peas and common vetch compared to flavonoids, whereas the opposite was observed for lupin seeds. Yellow lupins presented the highest levels of total phenolic compounds followed by narrow-leafed lupins (in average 960 and 679mg/kg, dry basis, respectively), whereas Kabuli chickpeas got the lowest ones (in average 47mg/kg, dry basis). Principal component analysis revealed that flavones and total levels of phenolic compounds were responsible for nearly 51% of total data variability.

Effects of grass silage and soybean meal supplementation on milk production and milk fatty acid profiles of grazing dairy cows.

The effects of supplementation with grass silage and replacement of some corn in the concentrate with soybean meal (SBM) on milk production, and milk fatty acid (FA) profiles were evaluated in a replicated 4 x 4 Latin square study using 16 dairy cows grazing pasture composed of ryegrass, Kentucky bluegrass, and white clover. Each experimental period lasted for 3 wk. The 4 dietary treatments were PC, 20 h of access to grazing pasture, supplemented with 6 kg/d of corn-based concentrate mixture (96% corn; C); PCSB, 20 h of access to grazing pasture, supplemented with 6 kg/d of corn- and SBM-based concentrate mixture (78% corn and 18% SBM; CSB); SC, 7 h of access to grazing pasture during the day and 13 h of ad libitum access to grass silage at night, supplemented with 6 kg/d of C concentrate; and SCSB, 7 h of access to grazing pasture during the day and 13 h of ad libitum access to grass silage at night, supplemented with 6 kg/d of CSB concentrate. The concentrate mixtures were offered twice each day in the milking parlor and were consumed completely. Grass silage supplementation reduced dietary crude protein and concentration of total sugars, and dietary SBM inclusion increased dietary crude protein concentration and decreased dietary starch concentration. Milk yield and energy-corrected milk were increased by SBM supplementation of cows with access to grass silage. Milk protein concentration was lower in cows offered grass silage, regardless of whether SBM was fed. Dietary SBM inclusion tended to increase milk fat concentration. Plasma urea N was reduced by silage feeding and increased by SBM supplementation. Supplementation with grass silage overnight could represent a useful strategy for periods of lower pasture availability. Dietary inclusion of SBM in solely grazing cows had no effects on milk production and composition, exacerbated the inefficient capture of dietary N, and increased diet cost. Grass silage supplementation affected milk FA profiles, increasing both the FA derived from de novo synthesis and those derived from rumen microbial biomass, and decreasing the sum of C18 FA (mostly derived from diet or from mobilization of adipose tissue reserves). Milk fat concentrations of conjugated linoleic acid cis-9, trans-11, vaccenic acid (18:1 trans-11), and linolenic acid (18:3n-3) were unaffected by grass silage supplementation, suggesting that partial replacement of pasture by unwilted grass silage does not compromise the dietary quality of milk fat for humans.

Evaluation of the chemical composition and the particle size of maize silages produced in north-west of Portugal

Abstract The chemical composition and the particle size distribution of 37 maize silages produced in north-west of Portugal were determined. Mean dry matter (DM), crude protein (CP), neutral detergent fibre (NDF) and starch were 288±33, 81±9, 502±45 and 229±54xa0gxa0kg−1 DM, respectively. Particle size was determined by manual separation of fresh silage into three fractions: material >30xa0mm in length; material between 30 and 10xa0mm in length; and material The results of this study suggested that most farmers chopped the maize plants very coarsely prior to storage. In fact, the type of chopping–harvesting machine had no effect on median particle size, but the percentage of blades used affected significantly (p

Technical note: Stearidonic acid metabolism by mixed ruminal microorganisms in vitro

Dietary supplementation of stearidonic acid (SDA; 18:4n-3) has been considered a possible strategy to increase n-3 unsaturated fatty acid content in ruminant products; however, little is known about its metabolism in the rumen. In vitro batch incubations were carried out with bovine ruminal digesta to investigate the metabolism of SDA and its biohydrogenation products. Incubation mixtures (4.5 mL) that contained 0 (control), 0.25, 0.50, 0.75, 1.00, 1.25, or 1.50 mg of SDA supplemented to 33 mg (DM basis) of commercial total mixed ration based on corn silage, for dairy cows, were incubated for 72 h at 39°C. The content of most fatty acids in whole freeze-dried cultures was affected by SDA supplementation. Branched-chain fatty acids decreased linearly (P < 0.01), and odd-chain fatty acids decreased quadratically (P < 0.01), particularly from 1.00 mg of SDA and above, whereas most C18 fatty acids increased linearly or quadratically (P ≤ 0.04). Stearidonic acid concentrations at 72 h of incubation were very small (<0.6% of total fatty acids and ≤0.9% of added SDA) in all treatments. The apparent biohydrogenation of SDA was extensive, but it was not affected by SDA concentration (P > 0.05). Biohydrogenation followed a pattern similar to that of other C18 unsaturated fatty acids up to 1.00 mg of SDA. Stearic acid (18:0) and vaccenic acid (18:1 trans-11) were the major fatty acids formed, with the latter increasing 9-fold in the 1.00 mg of SDA treatment. At greater inclusion rates, 18:0 and 18:1 trans isomers decreased (P ≤ 0.03), accompanied by increases in unidentified 18:3 and 18:4 isomers (P = 0.02), suggesting that the biohydrogenation pathway was inhibited. The present results clearly indicate that SDA was metabolized extensively, with numerous 18:4 and 18:3 products formed en route to further conversion to 18:2, 18:1 isomers, and 18:0.