Dietary effects of linseed on fatty acid composition of milk and on liver, adipose and mammary gland metabolism of periparturient dairy cows.

During the transition period in dairy cows, drastic adaptations within and between key tissues and cell types occur in a coordinated manner to support late gestation, the synthesis of large quantities of milk and metabolic homoeostasis. The start of lactation coincides with an increase of triacylglycerols in the liver, which has been associated with several economically important diseases in dairy cows (i.e. hepatic lipidiosis, mastitis). The polyunsaturated fatty acids have been used to improve liver metabolism and immune function in the mammary gland. Therefore, the effects of dietary linseed supplementation on milk quality and liver, adipose and mammary gland metabolism of periparturient dairy cows were studied in 14 cows that were randomly assigned to control or linseed supplementation. Animals were treated from 3 weeks antepartum until 6 weeks post-partum. Linseed did not modify dry matter intake, but increased milk yield and lactose yield, and decreased milk fat concentration, which coincided with lower proportion of C16 and higher proportions of stearic acid, conjugated linoleic acid and α-linolenic acid in milk fat. Linseed supplementation did not significantly change the expression of key lipid metabolism genes in liver and adipose tissues, except of glucose transporter 2 (GLUT2) in liver, which was increased in cows supplemented with linseed, suggesting that more glucose was secreted and probably available for lactose synthesis compared with cows fed control diet. Large adaptations of transcription occurred in the mammary gland when dairy cows were supplemented with linseed. The main affected functional modules were related to energy metabolism, cell proliferation and remodelling, as well as the immune system response.

Effect of rumen-protected choline supplementation on liver and adipose gene expression during the transition period in dairy cattle.

We previously reported that supplementation of rumen-protected choline (RPC) reduces the hepatic triacylglycerol concentration in periparturient dairy cows during early lactation. Here, we investigated the effect of RPC on the transcript levels of lipid metabolism-related genes in liver and adipose tissue biopsies, taken at wk -3, 1, 3, and 6 after calving, to elucidate the mechanisms underlying this RPC-induced reduction of hepatic lipidosis. Sixteen multiparous cows were blocked into 8 pairs and randomly allocated to either 1 of 2 treatments, with or without RPC. Treatments were applied from 3 wk before to 6 wk after calving. Both groups received a basal diet and concentrate mixture. One group received RPC supplementation, resulting in an intake of 14.4 g of choline per day, whereas controls received an isoenergetic mixture of palm oil and additional soybean meal. Liver and adipose tissue biopsies were taken at wk -3, 1, 3, and 6 to determine the mRNA abundance of 18 key genes involved in liver and adipose tissue lipid and energy metabolism. Milk samples were collected in wk 1, 2, 3, and 6 postpartum for analysis of milk fatty acid (FA) composition. The RPC-induced reduction in hepatic lipidosis could not be attributed to altered lipolysis in adipose tissue, as no treatment effect was observed on the expression of peroxisome proliferator-activated receptor γ, lipoprotein lipase, or FA synthase in adipose tissue, or on the milk FA composition. Rumen-protected choline supplementation increased the expression of FA transport protein 5 and carnitine transporter SLC22A5 in the liver, suggesting an increase in the capacity of FA uptake and intracellular transport, but no treatment effect was observed on carnitine palmitoyl transferase 1A, transporting long-chain FA into mitochondria. In the same organ, RPC appeared to promote apolipoprotein B-containing lipoprotein assembly, as shown by elevated microsomal triglyceride transfer protein expression and apolipoprotein B100 expression. Cows supplemented with RPC displayed elevated levels of glucose transporter 2 mRNA and a reduced peak in pyruvate carboxylase mRNA immediately after calving, showing that supplementation also resulted in improved carbohydrate metabolism. The results from this study suggest that RPC supplementation reduces liver triacylglycerol by improved FA processing and very-low-density lipoprotein synthesis, and RPC also benefits hepatic carbohydrate metabolism.

Effects of feeding different linseed sources on omasal fatty acid flows and fatty acid profiles of plasma and milk fat in lactating dairy cows.

The aim of this experiment was to study the effects of feeding different linseed sources on omasal fatty acid (FA) flows, and plasma and milk FA profiles in dairy cows. Four ruminally cannulated lactating Holstein-Friesian cows were assigned to 4 dietary treatments in a 4×4 Latin square design. Dietary treatments consisted of supplementing crushed linseed (CL), extruded whole linseed (EL), formaldehyde-treated linseed oil (FL) and linseed oil in combination with marine algae rich in docosahexaenoic acid (DL). Each period in the Latin square design lasted 21 d, with the first 16 d for adaptation. Omasal flow was estimated by the omasal sampling technique using Cr-EDTA, Yb-acetate, and acid detergent lignin as digesta flow markers. The average DM intake was 20.6 ± 2.5 kg/d, C18:3n-3 intake was 341 ± 51 g/d, and milk yield was 32.0 ± 4.6 kg/d. Milk fat yield was lower for the DL treatment (0.96 kg/d) compared with the other linseed treatments (CL, 1.36 kg/d; EL, 1.49 kg/d; FL, 1.54 kg/d). Omasal flow of C18:3n-3 was higher and C18:3n-3 biohydrogenation was lower for the EL treatment (33.8 g/d; 90.9%) compared with the CL (21.8 g/d; 94.0%), FL (15.5 g/d; 95.4%), and DL (4.6 g/d; 98.5%) treatments, whereas whole-tract digestibility of crude fat was lower for the EL treatment (64.8%) compared with the CL (71.3%), FL (78.5%), and DL (80.4%) treatments. The proportion of C18:3n-3 (g/100 g of FA) was higher for the FL treatment compared with the other treatments in plasma triacylglycerols (FL, 3.60; CL, 1.22; EL, 1.35; DL, 1.12) and milk fat (FL, 3.19; CL, 0.87; EL, 0.83; DL, 0.46). Omasal flow and proportion of C18:0 in plasma and milk fat were lower, whereas omasal flow and proportions of biohydrogenation intermediates in plasma and milk fat were higher for the DL treatment compared with the other linseed treatments. The results demonstrate that feeding EL did not result in a higher C18:3n-3 proportion in plasma and milk fat despite the higher omasal C18:3n-3 flow. This was related to the decreased total-tract digestibility of crude fat. Feeding FL resulted in a higher C18:3n-3 proportion in plasma and milk fat, although the omasal C18:3n-3 flow was similar or lower than for the CL and EL treatment, respectively. Feeding DL inhibited biohydrogenation of trans-11,cis-15-C18:2 to C18:0, as indicated by the increased omasal flows and proportions of biohydrogenation intermediates in plasma and milk fat.

Effects of forage type, forage to concentrate ratio, and crushed linseed supplementation on milk fatty acid profile in lactating dairy cows.

The effects of an increasing proportion of crushed linseed (CL) in combination with varying forage type (grass or corn silage) and forage to concentrate ratio (F:C), and their interactions on milk fatty acid (FA) profile of high-producing dairy cows was studied using a 3-factor Box-Behnken design. Sixteen Holstein and 20 Swedish Red cows were blocked according to breed, parity, and milk yield, and randomly assigned to 4 groups. Groups were fed different treatment diets formulated from combinations of the 3 main factors each containing 3 levels. Forage type (fraction of total forage dry matter, DM) included 20, 50, and 80% grass silage, with the remainder being corn silage. The F:C (DM basis) were 35:65, 50:50, and 65:35, and CL was supplied at 1, 3, and 5% of diet DM. Starch and neutral detergent fiber content (DM basis) of the treatment diets ranged from 117 to 209 g/kg and 311 to 388 g/kg, respectively. Thirteen treatment diets were formulated according to the Box-Behnken design. During 4 experimental periods of 21 d each, all treatment diets were fed, including a repetition of the center point treatment (50% grass silage, 50:50F:C, 3% CL) during every period. Intake, production performance, and milk FA profile were measured, and response surface equations were derived for these variables. Shifting from 80% grass silage to 80% corn silage in the diet linearly increased dry matter intake (DMI), net energy for lactation (NE(L)) intake, cis-9,cis-12-C18:2 (C18:2n-6) intake, and milk yield, and linearly decreased cis-9,cis-12,cis-15-C18:3 (C18:3n-3) intake and milk fat content. Shifting from a high forage to a high concentrate diet linearly increased DMI, NE(L) intake, C18:2n-6 intake, and milk yield, and decreased milk fat content. Supplementation of CL linearly increased C18:3n-3 intake, but had no effect on DMI, NE(L) intake, milk yield, or milk fat content. Shifting from 80% grass silage to 80% corn silage linearly increased proportions of trans-10-C18:1 and C18:2n-6 in milk fat, whereas the proportions of trans-11,cis-15-C18:2 and C18:3n-3 linearly decreased. Significant interactions between CL supplementation and F:C were found for proportions of trans-10-C18:1, trans-15-C18:1, cis-15-C18:1, trans-11,cis-15-C18:2, and C18:3n-3 in milk fat, with the highest levels achieved when the diet contained 5% CL and a 35:65F:C ratio. The effect of supplementing CL on several milk FA proportions, including C18:2n-6 and C18:3n-3, depends significantly on the F:C ratio and forage type in the basal diet.

Effect of rumen-protected choline on performance, blood metabolites, and hepatic triacylglycerols of periparturient dairy cattle.

The effects of a dietary supplement of rumen-protected choline on feed intake, milk yield, milk composition, blood metabolites, and hepatic triacylglycerol were evaluated in periparturient dairy cows. Thirty-eight multiparous cows were blocked into 19 pairs and then randomly allocated to either one of 2 treatments. The treatments were supplementation either with or without (control) rumen-protected choline. Treatments were applied from 3 wk before until 6 wk after calving. Both groups received the same basal diet, being a mixed feed of grass silage, corn silage, straw, and soybean meal, and a concentrate mixture delivered through transponder-controlled feed dispensers. For all cows, the concentrate mixture was gradually increased from 0 kg/day (wk -3) to 0.9 kg of dry matter (DM)/d (day of calving) and up to 8.1 kg of DM/d on d 17 postcalving until the end of the experiment. Additionally, a mixture of 60 g of a rumen-protected choline supplement (providing 14.4 g of choline) and of 540 g of soybean meal or a (isoenergetic) mixture of 18 g of palm oil and 582 g of soybean meal (control) was offered individually in feed dispensers. Individual feed intake, milk yield, and body weight were recorded daily. Milk samples were analyzed weekly for fat, protein, and lactose content. Blood was sampled at wk -3, d 1, d 4, d 7, d 10, wk 2, wk 3, and wk 6 and analyzed for glucose, nonesterified fatty acids, and ß-hydroxybutyric acid. Liver biopsies were taken from 8 randomly selected pairs of cows at wk -3, wk 1, wk 4, and wk 6 and analyzed for triacylglycerol concentration. We found that choline supplementation increased DM intake from 14.4 to 16.0 kg/d and, hence, net energy intake from 98.2 to 109.1 MJ/d at the intercept of the lactation curve at 1 day in milk (DIM), but the effect of choline on milk protein yield gradually decreased during the course of the study. Choline supplementation had no effect on milk yield, milk fat yield, or lactose yield. Milk protein yield was increased from 1.13 to 1.26 kg/d at the intercept of the lactation curve at 1 DIM, but the effect of choline on milk protein yield gradually decreased during the course of the study. Choline supplementation was associated with decreased milk fat concentration at the intercept of the lactation curve at 1 DIM, but the effect of choline on milk fat concentration gradually decreased as lactation progressed. Choline supplementation had no effect on energy-corrected milk yield, energy balance, body weight, body condition score, and measured blood parameters. Choline supplementation decreased the concentration of liver triacylglycerol during the first 4 wk after parturition. Results from this study suggest that hepatic fat export in periparturient dairy cows is improved by choline supplementation during the transition period and this may potentially decrease the risk for metabolic disorders in the periparturient dairy cow.

Effects of feeding rapeseed oil, soybean oil, or linseed oil on stearoyl-CoA desaturase expression in the mammary gland of dairy cows.

Stearoyl-CoA desaturase (SCD) is an important enzyme in the bovine mammary gland, and it introduces a double bond at the Δ(9) location of primarily myristoyl-, palmitoyl-, and stearoyl-CoA. The main objective of this study was to compare the effects of various fatty acids (FA) typically present in dairy cow rations on the expression of SCD1 and SCD5 in the mammary gland of dairy cows. Twenty-eight Holstein-Friesian cows were randomly assigned to 1 of 4 dietary treatments. The dietary treatments were a basal diet supplemented (dry matter basis) with 2.7% rapeseed oil as a source of C18:1 cis-9; 2.7% soybean oil as a source of C18:2 cis-9,12; 2.7% linseed oil as a source of C18:3 cis-9,12,15; or 2.7% of a 1:1:1 mixture of the 3 oils. The oil supplements were included in the concentrate, which was fed together with corn silage and grass silage. In addition, cows were grazing on pasture, consisting mainly of perennial ryegrass, during the day. Biopsies from the mammary gland were taken and analyzed for mRNA expression of SCD1 and SCD5 by using quantitative real-time PCR. Milk yield as well as milk protein and fat contents did not differ among the 4 dietary treatments. Dietary supplementation with rapeseed oil and linseed oil increased proportions of C18:1 cis-9 and C18:3 cis-9,12,15 in blood plasma, respectively, compared with the other treatments. Supplementation with soybean oil and linseed oil increased milk FA proportions of C18:2 cis-9,12 and C18:3 cis-9,12,15, respectively, but supplementation with rapeseed oil did not increase C18:1 cis-9 in milk. Mammary SCD1 expression was reduced by supplementation of soybean oil compared with rapeseed oil and linseed oil. In contrast, SCD5 expression did not differ among the 4 treatments. The C16 and C18 desaturation indices, representing proxies for SCD activity, were lower for the soybean oil diet compared with the diet supplemented with a mixture of the 3 oils. In conclusion, our study shows that mammary SCD1 expression is significantly downregulated in dairy cows by feeding unprotected soybean oil compared with rapeseed oil or linseed oil, and this is partially reflected by the lower desaturase indices in the milk. Furthermore, mammary SCD5 expression appears to be differently regulated than expression of SCD1.

Effect of feeding rumen protected rice bran on calcium homeostasis of non-lactating multiparous cows.

Milk fever in dairy cows can be prevented by activating Ca homeostasis before calving. Homeostatic adaptation can be achieved by reducing dietary Ca availability. Formaldehyde-treated rice bran was studied to supply rumen protected phytic acid to reduce Ca availability. Twelve multiparous dry cows were used in a 3×3 Latin square change-over design with 5-day periods to test three dietary treatments. Diets consisted of a forage mix (maize silage, grass silage and hay), being 77% of ration dry matter, supplemented with three concentrates: Control (no formaldehyde-treated rice bran), T1 (100% formaldehyde-treated rice bran) and T2 (99.5% formaldehyde-treated rice bran with 0.6% Ca carbonate, to equal Ca content of Control). Dietary treatments did not affect urine pH (8.14, 8.13 and 8.11 for Control, T1 and T2 respectively) or dry matter intake (13.9, 13.7 and 13.8 kg for Control, T1 and T2 respectively). Including formaldehyde-treated rice bran in the diet resulted in lower urinary Ca/creatinine ratio (0.970, 0.457 and 0.618 for Control, T1 and T2 respectively). A sudden increase of urinary Ca excretion took place after withdrawal of T1 and T2 at introduction of Control, peaking on the first day and coming back down progressively in the second and third days. Peak was greatest after T1 and was not observed in transitions between rice bran treatments. This is understood as indirect evidence of activation of intestinal Ca absorption during formaldehyde-treated rice bran feeding, because renal adaptations to changes in blood Ca clearance are immediate and intestinal adaptations delay 2 days. It was concluded that including formaldehyde-treated rice bran in rations before calving may represent a dietary strategy to prevent milk fever without reducing dry matter intake.

Effects of chemically or technologically treated linseed products and docosahexaenoic acid addition to linseed oil on biohydrogenation of C18:3n-3 in vitro.

Rumen biohydrogenation kinetics of C18:3n-3 from several chemically or technologically treated linseed products and docosahexaenoic acid (DHA; C22:6n-3) addition to linseed oil were evaluated in vitro. Linseed products evaluated were linseed oil, crushed linseed, formaldehyde treated crushed linseed, sodium hydroxide/formaldehyde treated crushed linseed, extruded whole linseed (2 processing variants), extruded crushed linseed (2 processing variants), micronized crushed linseed, commercially available extruded linseed, lipid encapsulated linseed oil, and DHA addition to linseed oil. Each product was incubated with rumen liquid using equal amounts of supplemented C18:3n-3 and fermentable substrate (freeze-dried total mixed ration) for 0, 0.5, 1, 2, 4, 6, 12, and 24h using a batch culture technique. Disappearance of C18:3n-3 was measured to estimate the fractional biohydrogenation rate and lag time according to an exponential model and to calculate effective biohydrogenation of C18:3n-3, assuming a fractional passage rate of 0.060/h. Treatments showed no differences in rumen fermentation parameters, including gas production rate and volatile fatty acid concentration. Technological pretreatment (crushing) followed by chemical treatment applied as formaldehyde of linseed resulted in effective protection of C18:3n-3 against biohydrogenation. Additional chemical pretreatment (sodium hydroxide) before applying formaldehyde treatment did not further improve the effectiveness of protection. Extrusion of whole linseed compared with extrusion of crushed linseed was effective in reducing C18:3n-3 biohydrogenation, whereas the processing variants were not different in C18:3n-3 biohydrogenation. Crushed linseed, micronized crushed linseed, lipid encapsulated linseed oil, and DHA addition to linseed oil did not reduce C18:3n-3 biohydrogenation. Compared with the other treatments, docosahexaenoic acid addition to linseed oil resulted in a comparable trans11,cis15-C18:2 biohydrogenation but a lesser trans10+11-C18:1 biohydrogenation. This suggests that addition of DHA in combination with linseed oil was effective only in inhibiting the last step of biohydrogenation from trans10+11-C18:1 to C18:0.

Effects of supplementing concentrates differing in carbohydrate composition in veal calf diets: II. Rumen development.

The objective of this experiment was to examine the effects of concentrates in feed, differing in carbohydrate source, on the rumen development of veal calves. For this purpose, 160 male Holstein Friesian x Dutch Friesian crossbred calves were used in a complete randomized block design with a 5 x 2 factorial arrangement. Dietary treatments consisted of 1) a milk replacer control, 2) a pectin-based concentrate, 3) a neutral detergent fiber-based concentrate, 4) a starch-based concentrate, and 5) a mixed concentrate (equal amounts of the concentrates in treatments 2, 3, and 4). Concentrate diets were provided as pellets in addition to a commercial milk replacer. Calves were euthanized at either 8 or 12 wk of age. Plasma acetate and beta-hydroxybutyrate (BHBA) were measured as indicators of rumen development. Empty rumen weight was determined, and wall samples were taken at slaughter. In most calves, a poorly developed rumen mucosa was observed. Coalescing rumen papillae with embedded hair, feed particles, and cell debris were found in all calves fed the concentrate diets. Calves fed concentrates had significantly heavier rumens than calves fed the control diet. In the dorsal location of the rumen, calves fed concentrate diets showed an increased ratio of mucosa to serosa length compared with calves fed the control diet, whereas in the ventral location only, calves fed the pectin and mixed diets showed larger ratios of mucosa to serosa length. Mucosa thickness and muscle thickness were greater in the ventral and dorsal locations of the rumen, respectively. In both locations, the NDF diet resulted numerically in the lowest mucosa thickness and highest muscle thickness among the concentrate treatments. At 8 wk, calves fed the concentrate diets had higher plasma acetate concentrations than calves on the control treatment. However, at 12 wk, only NDF-fed calves showed significantly higher plasma acetate concentrations. The plasma BHBA concentrations of calves at 8 wk of age fed the pectin and mixed diets were higher than those of the control diet-fed calves. At 12 wk, no differences in BHBA concentrations were observed among treatments. Results of a principal component analysis indicated that, in addition to rumen volatile fatty acid concentrations, other factors were likely to affect rumen development, and that the relationships between rumen development and individual types of volatile fatty acids present in the rumen liquor were similar. Also, variations in rumen development coincided with variations in plasma acetate and BHBA concentrations.

Starch supplementation of grass harvested at two stages of maturity prior to ensiling: intake, digestion, and degradability by dairy cows.

The effects of the maturity of grass prior to ensiling and the supplementation of starch to grass silage on apparent digestibility, degradability, rumen content, and feed intake by dairy cows were investigated using a Latin square design. Treatments were silages from early or late cut grass with or without 4 kg of supplemental flaked corn starch. The silage from early cut grass contained more N and sugars than did the silage from late cut grass but was lower in neutral detergent fiber (NDF). Degradation characteristics were not different between the two silages. Apparent digestibilities of dry matter (DM), organic matter (OM), and N in the silage from late cut grass were lower than those in the silage from early cut grass, but NDF digestibility was not affected. Starch supplementation increased the lag phase for DM and OM in both silages, but the rate of degradation was decreased for NDF. Starch supplementation did not influence digestibilities of DM and OM in silage from early cut grass but decreased the digestibilities of DM and OM in silage from late cut grass. Crude protein and NDF digestibilities were decreased for silages from early and late cut grass. Starch supplementation increased NDF in the rumen of cows fed the silage from early cut grass, but NDF was not affected by starch supplementation for cows fed the silage from late cut grass. Rumen-degradable starch negatively influences degradability and apparent OM digestibility; the extent of the decrease is related to the maturity of the NDF.

Influence of maturity of grass silage and flaked corn starch on the production and metabolism of volatile fatty acids in dairy cows.

An experiment employing a Latin square design was used to quantify the effects of two stages of maturity of grass silage (early cut and late cut) and three concentrations of flaked corn starch (0, 2, and 4 kg) on the molar proportion of rumen volatile fatty acids (VFA), the production of rumen VFA, and the net fluxes of VFA in the splanchnic tissue of cows. The molar proportions of VFA in rumen fluid were similar for cows fed both silages. When the silage diets were supplemented with starch, the proportion of propionic acid increased for cows fed diets containing early cut grass silage, but no effects were found for cows fed diets containing late cut grass silage. Estimated gastrointestinal production of acetate, propionate, butyrate, and branched-chain fatty acids plus valerate was related to intake of metabolizable energy and organic matter fermented into VFA. The portal release of acetate was approximately 14% lower than the estimated production of acetate by cows fed diets containing early cut grass silage, but cows fed diets containing late cut grass silage showed a variable difference between estimated production and portal release (31, 24, and 15%, respectively) as starch supplementation increased. The portal release of butyrate plus beta-hydroxybutyrate and the release of branched-chain fatty acids plus valerate were approximately 70 and 25%, respectively, of the estimated gastrointestinal production. Propionate production was similar to the portal release of propionate. Net flux measurements in splanchnic tissue in combination with gastrointestinal digestion and kinetics provide information that increases the knowledge of pathways and metabolism and quantifies the availability of individual nutrients for milk production in dairy cows.