The effect of linseed oil (LO) supplementation on nutrient digestibility, forage (i.e., timothy hay) in sacco ruminal degradation, ruminal fermentation characteristics, protozoal populations, milk production, and milk fatty acid (FA) profile in dairy cows was investigated. Four ruminally cannulated, primiparous lactating cows were used in a 4 × 4 Latin square design (28-d periods). They were fed a total mixed ration (50:50 forage:concentrate (F:C) ratio [dry matter (DM) basis] without supplementation (control, CTL), or supplemented (wt/wt; DM basis) with LO at 2, 3, or 4%. Supplementation with LO had no effect on DM intake (19 kg/d) and apparent total-tract digestibility of nutrients (organic matter, neutral detergent fiber, acid detergent fiber, starch, and gross energy). Ruminal pH, ammonia, and total volatile FA concentrations were not changed by LO supplementation to diets. Extent of changes in volatile FA pattern and effective ruminal degradability of DM of timothy hay were minor. Neither the total numbers nor the genera distribution of protozoa was changed by the addition of increasing amounts of LO to the diet. Milk yield increased linearly (26.1, 27.3, 27.4, and 28.4 kg/d for CTL to LO4, respectively) as the amount of LO added to the diet increased. Milk fat content was not affected by LO supplementation, whereas milk protein content decreased linearly with increasing amounts of LO in the diet. Milk fat proportions of several intermediates of ruminal biohydrogenation of polyunsaturated FA (i.e., trans-10 18:1, trans-11 18:1, cis-9,trans-11 18:2, trans-11,cis-15 18:2, and cis-9,trans-11,cis-15 18:3) increased linearly with LO addition to the diet. The proportion of cis-9,cis-12 18:2 decreased linearly (2.06, 1.99, 1.91, and 1.83% for CTL to LO4, respectively) as the amount of LO in the diet increased. Milk fat content of cis-9,cis-12,cis-15 18:3 increased as the level of LO in the diet increased up to 3% but no further increase was observed when 4% of LO was fed (0.33, 0.79, 0.86, and 0.86% for CTL to LO4, respectively). A similar quadratic response to LO supplementation was also observed for cis-5,cis-8,cis-11,cis-14,cis-17 20:5 and cis-5,cis-7,cis-10,cis-13,cis-16 22:5. The results of the present study show that LO can be safely supplemented up to 4% in forage-based diets of dairy cows to enrich milk with potential health beneficial FA (i.e., n-3 FA) without causing any detrimental effects on rumen function, digestion, and milk production.

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 £ 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.

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.Key words: Lignans: Antioxidants: Flaxseed: mRNA abundance Dietary PUFA are perceived to be healthier than SFA. Research has shown several health benefits of n-3 fatty acids (FA) to humans including a decrease in the incidence of cancer, CVD, hypertension and arthritis and an improvement in visual acuity (1) . As a result, there has been a great deal of interest in manipulating the FA profile of ruminant milk fat to respond to consumer interest. However, feeding PUFA may contribute to increasing plasma lipoperoxidation as reported in steers (2) and rats (3) , which may increase oxidative stress in animals. It is known that in dairy cows the peripartum period, particularly postpartum, is characterised by a depleted antioxidant status (4) . Nevertheless, supplying n-3 PUFA in association with antioxidants such as vitamin E and plant polyphenols reduced lipoperoxidation in lactating cows, thereby contributing to protecting them against the deleterious consequences of lipoperoxidation (5) .Recent research on new antioxidants has focused on natural molecules to satisfy consumer concerns about safety and toxicity (3) . Plants lignans are naturally strong antioxidants and flax (Linum usitatissimum) is known as the richest dietary source of lignans, with glycosides of secoisolariciresinol and matairesinol as the major compounds (6) . Flax lignans are concentrated in the outer fibre-containing layers (7) , thus resulting in higher concentration of lignans in hulls than in seeds (8) . In nonruminant animals, the colonic microbiota converts plant into mammalian lignans, enterodiol and enterolactone (EL) (9) . In dairy co...

Four ruminally lactating Holstein cows averaging 602+/-25 kg of body weight and 64+/-6 d in milk at the beginning of the experiment were randomly assigned to a 4 x 4 Latin square design to determine the effects of feeding whole flaxseed and calcium salts of flaxseed oil on dry matter intake, digestibility, ruminal fermentation, milk production and composition, and milk fatty acid profile. The treatments were a control with no flaxseed products (CON) or a diet (on a dry matter basis) of 4.2% whole flaxseed (FLA), 1.9% calcium salts of flaxseed oil (SAL), or 2.3% whole flaxseed and 0.8% calcium salts of flaxseed oil (MIX). The 4 isonitrogenous and isoenergetic diets were fed for ad libitum intake. Experimental periods consisted of 21 d of diet adaptation and 7 d of data collection and sampling. Dry matter intake, digestibility, milk production, and milk concentrations of protein, lactose, urea N, and total solids did not differ among treatments. Ruminal pH was reduced for cows fed the CON diet compared with those fed the SAL diet. Propionate proportion was higher in ruminal fluid of cows fed CON than in that of those fed SAL, and cows fed the SAL and CON diets had ruminal propionate concentrations similar to those of cows fed the FLA and MIX diets. Butyrate concentration was numerically higher for cows fed the SAL diet compared with those fed the FLA diet. Milk fat concentration was lower for cows fed SAL than for those fed CON, and there was no difference between cows fed CON and those fed FLA and MIX. Milk yields of protein, fat, lactose, and total solids were similar among treatments. Concentrations of cis-9 18:1 and of intermediates of ruminal biohydrogenation of fatty acids such as trans-9 18:1 were higher in milk fat of cows fed SAL and MIX than for those fed the CON diet. Concentration of rumenic acid (cis-9, trans-11 18:2) in milk fat was increased by 63% when feeding SAL compared with FLA. Concentration of alpha-linolenic acid was higher in milk fat of cows fed SAL and MIX than in milk of cows fed CON (75 and 61%, respectively), whereas there was no difference between FLA and CON. Flaxseed products (FLA, SAL, and MIX diets) decreased the n-6 to n-3 fatty acid ratio in milk fat. Results confirm that flax products supplying 0.7 to 1.4% supplemental fat in the diet can slightly improve the nutritive value of milk fat for better human health.