Linoleic acid, α-linolenic acid and enterolactone affect lipid oxidation and expression of lipid metabolism and antioxidant-related genes in hepatic tissue of dairy cows.

Although beneficial effects have been attributed to PUFA supplementation in high-yielding dairy cows, diets rich in PUFA may also increase oxidative stress in tissues such as the liver. To fully exploit the health benefits of PUFA, we believe that the addition of natural antioxidants could help in preventing oxidative damage. Using an in vitro precision-cut liver slices (PCLS) tissue culture system, we investigated the effects of different linoleic acid (LA, n-6):α-linolenic acid (ALA, n-3) ratios (LA:ALA ratio of 4, LA:ALA ratio of 15 and LA:ALA ratio of 25) in the presence or absence of the antioxidant enterolactone (ENL) on (1) the mRNA abundance of genes with key roles in hepatic lipid metabolism, oxidative stress response and inflammatory processes, (2) oxidative damages to lipids and proteins and (3) superoxide dismutase activity in early-lactating dairy cows. The addition of LA and ALA to PCLS culture media increased oxidative damage to lipids as suggested by higher concentrations of thiobarbituric acid reactive substances and increased the expression of nuclear factor erythroid 2-related factor 2 target genes. The addition of ENL was effective in preventing lipid peroxidation caused by LA and ALA. Transcript abundance of sterol regulatory element-binding transcription factor 1 and its lipogenic target genes acetyl-CoA carboxylase α, fatty acid synthase (FASN) and stearoyl-CoA desaturase (SCD) was decreased with LA and ALA, whereas ENL decreased FASN and SCD gene expression. Our results show that addition of LA and ALA to PCLS culture media lowers hepatic lipogenic gene expression and increases oxidative damages to lipids. On the other hand, addition of ENL prevents oxidative damages provoked by these PUFA.

Effect of abomasal or ruminal administration of citrus pulp and soybean oil on milk fatty acid profile and antioxidant properties.

Soybean oil (SBO) is rich in polyunsaturated fatty acids (FA) and rumen bypass of SBO can contribute to increase the polyunsaturated FA proportion in milk fat. Citrus pulp (CPP) is a source of antioxidants but there is little information on the effects of CP administration on milk properties. This study was performed to determine the role of rumen microorganisms in the transfer of antioxidants from CPP into milk when cows receive SBO as a source of polyunsaturated FA. Four ruminally fistulated lactating Holstein cows were assigned to a 4 × 4 Latin square design with a 2 × 2 factorial arrangement of treatments: (1) SBO administered in the rumen; (2) SBO infused in the abomasum; (3) SBO + CPP administered in the rumen; and (4) SBO + CPP infused in the abomasum. Product and site of administration had no effect on yield of milk components. Concentrations of total polyphenols and flavonoids, reducing power and production of conjugated diene (CD) hydroperoxides in milk were not affected by products, but infusion in the abomasum compared with administration in the rumen increased production of CD. Milk fat FA profile was not affected by products. However, cows infused in the abomasum compared with those administered in the rumen showed lower proportions of short-chain and monounsaturated FA and higher proportions of polyunsaturated, omega 3 and omega 6 FA in milk fat, which resulted in enhanced health-promoting index of milk. Administration of SBO and CPP (0.2 + 1.0 kg/d) in the rumen or the abomasum resulted in similar milk antioxidant properties, thus suggesting that the rumen microbes have little involvement in the metabolism of antioxidants from CPP.

mRNA Expression of lipogenic enzymes in mammary tissue and fatty acid profile in milk of dairy cows fed flax hulls and infused with flax oil in the abomasum.

In the present study, the effect of flax hulls with or without flax oil bypassing the rumen on the expression of lipogenic genes in the mammary tissue of dairy cows was investigated. A total of eight dairy cows were used in a replicated 4 × 4 Latin square design. There were four periods of 21 d each and four treatments: control diet with no flax hulls (CONT); diet with 9·88 % flax hulls in the DM (HULL); control diet with 500 g flax oil/d infused in the abomasum (COFO); diet with 9·88 % flax hulls in the DM and 500 g flax oil/d infused in the abomasum (HUFO). A higher mRNA abundance of sterol regulatory element binding transcription factor, fatty acid (FA) synthase, lipoprotein lipase (LPL), PPARγ1, stearoyl-CoA desaturase (SCD) and acetyl-coenzyme A carboxylase-α was observed in cows fed HULL than in those fed CONT, and HUFO had the opposite effect. Compared with CONT, COFO and HUFO lowered the mRNA abundance of SCD, which may explain the lower proportions of MUFA in milk fat with flax oil infusion. The mRNA abundance of LPL in mammary tissue and proportions of long-chain FA in milk fat were higher in cows fed COFO than in those fed CONT. The highest proportions of trans FA were observed when cows were fed HULL. The present study demonstrates that flax hulls with or without flax oil infusion in the abomasum can affect the expression of lipogenic genes in the mammary tissue of dairy cows, which may contribute to the improvement of milk FA profile.

The effects of dietary omega fatty acids on pregnancy rate, plasma prostaglandin metabolite levels, serum progesterone levels, and milk fatty-acid profile in beef cows.

The objectives were to determine the effects of feeding supplements rich in omega-6 or omega-3 fatty acids (FA) during the late gestation to the early postpartum and breeding periods on reproduction and milk FA profile in beef cows. For each of two years, at the beginning of period 1 (mid-December), 72 beef cows, calving in January or February, were assigned to diets supplemented with roasted flaxseed (Flax) or roasted soybean (Soybean). For each of two years, after 11 wk (end of period 1), 18 cows of 36 in the Flax group were switched to the soybean supplement and 18 cows of 36 in the Soybean group were switched to the flax supplement (start of Period 2). Cows were bred by timed artificial insemination (TAI) in week 5 of period 2. The FA composition of the milk reflected the FA profile of the oilseed supplements. There were no differences in pregnancy rates among the 4 groups. The treatments had no effect on plasma prostaglandin metabolite levels or ratios at 4 to 11 d postpartum. At 5 to 6 d post- TAI, pregnant cows fed Flax in period 1 had lower (P < 0.05) plasma prostaglandin F metabolite (PGFM) levels and PGFM to prostaglandin E metabolite (PGEM) ratio than cows fed Soybean, but there were no significant differences at 19 to 20 d post-TAI. Cows pregnant from TAI and fed Flax in period 2 had higher (P < 0.05) serum progesterone levels at 5 to 6 d post-TAI than cows fed Soybean, but there was no difference at 19 to 20 d post-TAI. The dietary treatments had no effect on pregnancy rates, but there were some effects on plasma PGFM levels, PGFM to PGEM ratios, and serum progesterone levels. The FA supplements influenced the FA composition of milk.

Les objectifs de la présente étude étaient de déterminer les effets de suppléments alimentaires riches en acides gras (FA) omega-6 ou omega-3 lors de la période fin de gestation au début du postpartum et lors des périodes d'accouplement sur la reproduction et les profils de FA chez les vaches d'embouche. Pour chacune des deux années, au début de la période 1 (mi-décembre), 72 vaches d'embouche, devant vêler en janvier ou février, ont été assignées à des rations supplémentées avec de la graine de lin rôtie (Flax) ou des graines de soya rôties (Soya). Pour chacune des deux années, après 11 semaines (fin de la période 1), 18 des 36 vaches dans le groupe Flax ont été changées au supplément de soya et 18 des 36 vaches du groupe soya ont été changées pour le groupe Flax (Début de la période 2). Les vaches ont été saillies par insémination artificielle minutée (TAI) lors de la semaine 5 de la période 2. La composition en FA du lait représentait le profil de FA des suppléments alimentaires. Il n'y avait pas de différence dans les taux de gestation parmi les 4 groupes. Le traitement n'avait pas d'effet sur les niveaux ou ratios plasmatiques des métabolites des prostaglandines du jour 4 au jour 11 postpartum. Aux jours 5 à 6 post-TAI, les vaches gestantes nourries au Flax durant la période 1 avaient des niveaux significativement (P < 0,05) plus bas de métabolite de la prostaglandine F (PGFM) et des ratios de PGFM au métabolite de la prostaglandine E (PGEM) que les vaches nourries avec le Soya, mais il n'y avait pas de différence significative aux jours 19 à 20 post-TAI. Les vaches gestantes suite à la TAI et nourries avec Flax durant la période 2 avaient des niveaux sériques de progestérone plus élevés (P < 0,05) aux jours 5 à 6 post-TAI que les vaches nourries au Soya, mais il n'y avait plus de différence aux jours 19­20 post TAI. Les traitements alimentaires n'avaient aucun effet sur les taux de gestation, mais il y avait des différences sur les niveaux plasmatiques de PGFM, les ratios PGFM/PGEM, et les niveaux sériques de progestérone. Les FA des suppléments ont influencé la composition en FA du lait.(Traduit par Docteur Serge Messier).

Mammary gene expression and activity of antioxidant enzymes and oxidative indicators in the blood, milk, mammary tissue and ruminal fluid of dairy cows fed flax meal.

The effects of flax meal (FM) on the activity of antioxidant enzymes (superoxide dismutase (SOD), glutathione peroxidase (GPx) and catalase (CAT)) in the blood, mammary tissue and ruminal fluid, and oxidative stress indicators (thiobarbituric acid-reactive substances(TBARS) and 1,1-diphenyl-2-picrylhydrazyl-scavenging activity) in the milk, plasma and ruminal fluid of dairy cows were determined.The mRNA abundance of the antioxidant enzymes and oxidative stress-related genes was assessed in mammary tissue. A total of eight Holstein cows were used in a double 4 x 4 Latin square design. There were four treatments in the diet: control with no FM(CON) or 5% FM (5FM), 10% FM (10FM) and 15% FM (15FM). There was an interaction between treatment and time for plasma GPx and CAT activities. Cows supplemented with FM had a linear reduction in TBARS at 2 h after feeding, and there was no treatment effect at 0, 4 and 6 h after feeding. TBARS production decreased in the milk of cows fed the 5FM and 10FM diets. There was a linear increase in nuclear factor (erythroid-derived 2)-like 2 (NFE2L2) mRNA abundance in mammary tissue with FM supplementation.A linear trend for increased mRNA abundance of the CAT gene was observed with higher concentrations of FM. The mRNA abundance of CAT, GPx1, GPx3, SOD1, SOD2, SOD3 and nuclear factor of k light polypeptide gene enhancer in B-cells (NFKB) genes was not affected by the treatment. These findings suggest that FM supplementation can improve the oxidative status of Holstein cows as suggested by decreased TBARS production in ruminal fluid 2 h post-feeding and increased NFE2L2/nuclear factor-E2-related factor 2 (Nrf2) mRNA abundance in mammary tissue.

Effects of abomasal infusion of flaxseed (Linum usitatissimum) oil on microbial ß-glucuronidase activity and concentration of the mammalian lignan enterolactone in ruminal fluid, plasma, urine and milk of dairy cows.

Ruminal microbiota plays an important role in the conversion of plant lignans into mammalian lignans. The main mammalian lignan present in the milk of dairy cows fed flax products is enterolactone (EL). The objectives of the present study were to investigate the effects of abomasal infusion of flax oil on the metabolism of flax lignans and concentrations of EL in biological fluids of dairy cows. A total of six rumen-cannulated dairy cows were assigned within a 2 × 3 factorial arrangement of six treatments utilising flax hulls (0 and 15·9 % of DM) and abomasal infusion of flax oil (0, 250 and 500 g/d). There were six periods of 21 d each. Samples were collected during the last 7 d of each period and subjected to chemical analysis. Flax hull supplementation increased concentrations of EL in ruminal fluid, plasma, urine and milk, while flax oil infusion had no effect. Post-feeding, ß-glucuronidase activity in the ruminal fluid of cows infused with 250 g flax oil was significantly lower for cows fed hulls than for those fed the control diet. The present study demonstrated that the presence of a rich source of n-3 fatty acids such as flax oil in the small intestine does not interfere with the absorption of the mammalian lignan EL and that lower ruminal ß-glucuronidase activity had no effect on the conversion of flax lignans into EL in the rumen of dairy cows.

Mammary gene expression and activity of antioxidant enzymes and concentration of the mammalian lignan enterolactone in milk and plasma of dairy cows fed flax lignans and infused with flax oil in the abomasum.

The objectives of the study were to investigate the effects of dietary supplementation of flax hulls and/or flax oil on the activity of antioxidant enzymes (superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPX)) in plasma and the mammary gland and the relative mRNA abundance of antioxidant genes in the mammary gland of dairy cows. A total of eight dairy cows were used in a replicated 4 × 4 Latin square design. There were four treatments: control with no flax hulls (CONT), 9·88% flax hulls in the DM (HULL), control with 500 g flax oil/d infused in the abomasum (COFO), 9·88% flax hulls in the DM and 500 g flax oil/d infused in the abomasum (HUFO). Plasma GPX activity tended to decrease with flax oil supplementation. Cows fed HULL had higher levels of CAT, GPX1 and SOD1 mRNA in the mammary gland and lower mRNA abundance of GPX3, SOD2 and SOD3 compared with those fed CONT. Abundance of CAT, GPX1, GPX3, SOD2 and SOD3 mRNA was down-regulated in the mammary gland of cows fed HUFO compared to those fed CONT. The mRNA abundance of CAT, GPX1, GPX3 and SOD3 was lower in the mammary gland of cows fed COFO than in the mammary gland of cows fed CONT. The present study demonstrates that flax hulls contribute to increasing the abundance of some antioxidant genes, which can contribute to protecting against oxidative stress damage occurring in the mammary gland and other tissues of dairy cows.

Intake and digestibility of fatty acids in late-lactating dairy cows fed flaxseed hulls supplemented with monensin.

Flaxseed hull, a co-product obtained from flax processing, is a rich source of n-3 fatty acids but there is little information on digestibility of its nutrients by dairy cows. Four rumen-cannulated multiparous Holstein cows averaging 665 ± 21 kg of body weight and 190 ± 5 d in milk at the beginning of the experiment were assigned to a 4 × 4 Latin square design with four 28-d experimental periods to determine the effects of feeding monensin and flaxseed hulls on total tract apparent digestibility of nutrients and fatty acids. The four treatments were: (1) diet CO: control with neither flaxseed hulls nor monensin added; (2) diet FH containing 19·8 g flaxseed hulls/100 g dry matter (DM); (3) diet MO with 16 mg monensin/kg DM; (4) diet HM containing 19·8 g flaxseed hulls/100 g DM and 16 mg monensin/kg DM. Diets provided similar amounts of protein and net energy of lactation. Digestibility of crude protein was higher for diets containing flaxseed hulls and for diets supplemented with monensin. Flaxseed hulls supplementation decreased digestibility of acid and neutral detergent fibre. Significantly higher digestibility of ether extract and individual fatty acids was observed for treatments with flaxseed hulls compared with treatments without flaxseed hulls. A combination of flaxseed hulls and monensin did not result in better fatty acid digestibility than when feeding only flaxseed hulls.

Digestion, milk production and milk fatty acid profile of dairy cows fed flax hulls and infused with flax oil in the abomasum.

Flax hull, a co-product obtained from flax processing, is a rich source of n-3 fatty acids (FA) but there is little information on digestion of flax hull based diets and nutritive value of flax hull for dairy production. Flax oil is rich in α-linolenic acid (LNA) and rumen bypass of flax oil contributes to increase n-3 FA proportions in milk. Therefore, the main objective of the experiment was to determine the effects of abomasal infusion of increasing amounts of flax oil on apparent digestibility, dry matter (DM) intake, milk production, milk composition, and milk FA profile with emphasis on the proportion of LNA when cows were supplemented or not with another source of LNA such as flax hull. Six multiparous Holstein cows averaging 650±36 kg body weight and 95±20 d in milk were assigned to a 6×6 Latin square design (21-d experimental periods) with a 2×3 factorial arrangement of treatments. Treatments were: 1) control, neither flax hull nor flax oil (CON), 2) diet containing (DM basis) 15·9% flaxseed hull (FHU); 3) CON with abomasal infusion of 250 g/d flax oil; 4) CON with abomasal infusion of 500 g/d flax oil; 5) FHU with abomasal infusion of 250 g/d flax oil; 6) FHU with abomasal infusion of 500 g/d flax oil. Infusion of flax oil in the abomasum resulted in a more pronounce decrease in DM intake for cows fed the CON diets than for those fed the FHU diets. Abomasal infusion of flax oil had little effect on digestibility and FHU supplementation increased digestibility of DM and crude protein. Milk yield was not changed by abomasal infusion of flax oil where it was decreased with FHU supplementation. Cows fed FHU had higher proportions of 18:0, cis9-18:1, trans dienes, trans monoenes and total trans in milk fat than those fed CON. Proportion of LNA was similar in milk fat of cows infused with 250 and 500 g/d flax oil in the abomasum. Independently of the basal diet, abomasal infusion of flax oil resulted in the lowest n-6:n-3 FA ratio in milk fat, suggesting that the most important factor for modification of milk FA profile was the amount of n-3 FA bypassing the rumen and not the amount of flax hull fed to dairy cows. Moreover, these data suggest that there is no advantage to supply more than 250 g/d of flax oil in the abomasum to increase the proportion of LNA in milk fat.

Effect of flaxseed lignans added to milk or fed to cows on oxidative degradation of dairy beverages enriched with polyunsaturated fatty acids.

Nutritional value is a priority in new product development. Using vegetable or marine oils, rich in polyunsaturated fatty acids, in dairy beverage formulations is an option to provide the consumers with healthier products. However, these formulations are prone to oxidation, which is responsible for rapid flavour degradation and the development of potentially toxic reaction products during storage. Flaxseed lignans, secoisolariciresinol diglucoside (SDG), and its mammalian metabolites have antioxidant activity and could be used in beverage formulations to prevent oxidation. Commercially available SDG extract was added to the formulation of dairy beverages enriched with flaxseed oil. As an alternative approach, dairy beverages were produced from milk naturally rich in SDG metabolites obtained through the alteration of cow diet. Resistance to oxidation was determined from the kinetics of hexanal and propanal production during heat and light exposure treatments. Increasing SDG concentration in dairy beverage slightly reduced redox potential but had no effect on oxygen consumption during oxidation treatments. The presence of SDG in dairy beverage significantly improved resistance to heat- and light-induced oxidation. However, purified enterolactone, a mammalian metabolite from SDG, prevented oxidation at much lower concentrations. The use of milk from dairy cow fed flaxseed meal did not improve resistance to oxidation in dairy beverage. Enterolactone concentration in milk was increased by the experimental diet but it remained too low to observe any significant effect on dairy beverage oxidation.

Ruminal fermentation characteristics and fatty acid profile of ruminal fluid and milk of dairy cows fed flaxseed hulls supplemented with monensin.

Flaxseed hull, a co-product obtained from flax processing, is a rich source of n-3 fatty acids (FA) but there is little information on its value for dairy production. Monensin supplementation is known to modify biohydrogenation of FA by rumen microbes. Therefore, the main objective of the experiment was to determine the effect of feeding a combination of monensin and flaxseed hulls on ruminal fermentation characteristics and FA profile of ruminal fluid and milk. Four ruminally fistulated multiparous Holstein cows averaging 665 ± 21 kg body weight and 190 ± 5 d in milk were assigned to a 4×4 Latin square design (28-d experimental periods) with a 2×2 factorial arrangement of treatments. Treatments were: 1) control, neither flaxseed hulls nor monensin; 2) diet containing (dry matter basis) 19·8% flaxseed hulls; 3) diet with monensin (16 mg/kg dry matter); 4) diet containing 19·8% (dry matter basis) flaxseed hulls and 16 mg monensin/kg. Flaxseed hull supplementation decreased the acetate to propionate ratio in ruminal fluid and monensin had no effect. Concentrations of trans-18:1 isomers (trans9,trans11,trans13/14+6/8) and cis9,12,15-18:3 in ruminal fluid and milk fat were higher and those of cis9,12-18:2 in milk fat tended (P=0·07) to be higher for cows supplemented with flaxseed hulls than for cows fed no flaxseed hulls. Monensin had little effect on milk fatty acid profile. A combination of flaxseed hulls and monensin did not result in better milk fatty acid profile than when feeding only flaxseed hulls.

Ruminal metabolism of flaxseed ( Linum usitatissimum) lignans to the mammalian lignan enterolactone and its concentration in ruminal fluid, plasma, urine and milk of dairy cows.

Secoisolariciresinol diglucoside is the main flax (Linum usitatissimum) lignan that is converted to the mammalian lignans enterodiol (ED) and enterolactone (EL) by gastrointestinal microbiota. The objectives of the present study were to investigate the role of ruminal microbiota and the effects of flax oil on in vivo metabolism of flax lignans and concentration of EL in biological fluids. Four rumen-cannulated dairy cows were used in a 4 x 4 Latin square design. There were four periods of 21 d each and four treatments utilising flax hulls (1800 g/d) and oil (400 g/d) supplements. The treatments were: (1) oil and hulls administered in the rumen and abomasal infusion of water; (2) oil and hulls administered in the abomasum; (3) oil infused in the abomasum and hulls placed in the rumen; (4) oil placed in the rumen and hulls administered in the abomasum. Samples were collected during the last week of each period and subjected to chemical analysis. The site of supplementation of oil and hulls had no effect on ruminal EL concentration. Supplementing flax oil in the rumen and the abomasum led to similar EL concentrations in urine, plasma and milk. Concentrations of EL were higher in the urine, plasma and milk of cows supplemented with hulls in the rumen than in those placed with hulls in the abomasum. The present study demonstrated that ruminal microbiota play an important role in the metabolism of flax lignans.