Aila Vanhatalo

Determination of reticulo-rumen and whole-stomach digestion in lactating cows by omasal canal or duodenal sampling

Four ruminally and duodenally cannulated multiparous Finnish Ayrshire cows were fed on diets consisting of grass silage (0.6 kg/kg DM) and one of four concentrates: barley, barley + urea, barley + rapeseed meal and barley + rapeseed cake. The objective of the present study was to compare omasal canal and duodenal digesta flows. Values for digesta flow into the omasal canal and duodenum were determined using a triple-marker method based on Co-EDTA, Yb-acetate and indigestible neutral-detergent fibre (NDF) markers. Microbial non-NH3 N (NAN) flow was assessed by purine flow. Microbial samples to determine the bacterial purine:N ratio were harvested from the rumen, omasum and duodenum. Organic matter flow was significantly lower into the omasum than the duodenum, indicating an endogenous organic matter secretion into the abomasum. In contrast, NDF and acid-detergent fibre flows were significantly higher into the omasum indicating digestion of fibre in the omasum. Microbial NAN flows were significantly different (P < 0.001) when estimates were based on bacterial samples harvested from different sites. Differences in total NAN, microbial NAN and dietary NAN flows entering the omasal canal and duodenum were non-significant. The results indicated that the omasal sampling technique provides a promising alternative to the duodenal sampling technique to investigate forestomach digestion in dairy cows and offers an alternative means to study rumen N metabolism.

Effect of incremental levels of sunflower-seed oil in the diet on ruminal lipid metabolism in lactating cows

Based on the potential benefits of cis-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 of cis-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 x 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 < 0.01) milk yield and milk lactose output and shifted (P < 0.001) rumen fermentation towards propionate at the expense of acetate. SFO supplements increased linearly (P < 0.05) the flow of 18 : 0, 18 : 1, 18 : 2n-6 and total CLA at the omasum and enhanced ruminal cis-9-18 : 1, 18 : 2n-6 and 18 : 3n-3 metabolism. Flows of all-trans- (Delta4-16) and cis- (Delta9-16) 18 : 1 isomers were elevated, while increases in ruminal CLA outflow were confined to trans-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 of trans-11-18 : 1 and cis-9, trans-11-CLA in lactating cows.

Effects of feeding grass or red clover silage cut at two maturity stages in dairy cows. 1. Nitrogen metabolism and supply of amino acids.

This study investigated the effects of plant species (red clover vs. timothy-meadow fescue) and forage maturity at primary harvest (early vs. late cut silage) on rumen fermentation, nutrient digestion, and nitrogen metabolism including omasal canal AA flow and plasma AA concentration in lactating cows. Five dairy cows equipped with rumen cannulas were used in a study designed as a 5 x 5 Latin square with 21-d periods. The diets consisted of early-cut and late-cut grass and red clover silage, respectively, and a mixture of late-cut grass and early-cut red clover silages given ad libitum with 9 kg/d of a standard concentrate. Grass silage dry matter intake tended to decrease but that of red clover silages tended to increase with advancing maturity. Milk yields were unchanged among treatments, milk protein and fat concentrations being lower for red clover than for grass silage diets. Rumen fluid pH was unchanged but volatile fatty acid and ammonia concentrations were higher for red clover than for grass silage diets. Intake of N, and omasal canal flows of total nonammonia N (NAN), microbial NAN, and dietary NAN were higher for red clover than for grass silage diets but were not affected by forage maturity. However, microbial NAN flow and amount of N excreted in the feces decreased with advancing maturity for grass diets but increased for red clover diets. Apparent ruminal N degradability of the diets was unchanged, but true ruminal N degradability decreased and efficiency of microbial synthesis increased with red clover diets compared with grass silage diets. Omasal canal flows of AA, except those for Met and Cys, were on average 20% higher for red clover than grass silage diets. Omasal canal digesta concentrations of Leu, Phe, branched-chain, and essential AA were higher but those of Met lower for red clover than for grass silage diets. Plasma AA concentrations, except for His (unchanged) and Met (lower), were higher for red clover than for grass diets. However, none of these AA-related variables were affected by forage maturity. Total digestibility of N and excretion of N in the urine were higher for red clover than for grass diets and decreased with advancing maturity. It was concluded that despite the higher total AA supply of cows fed red clover versus grass silage diets, further milk production responses on red clover diets were compromised by an inadequate supply of Met as evidenced by lower Met concentration in the AA profile of omasal digesta and plasma.

Effect of plant oils and camelina expeller on milk fatty acid composition in lactating cows fed diets based on red clover silage

Five multiparous Finnish Ayrshire cows fed red clover silage-based diets were used in a 5 × 5 Latin square with 21-d experimental periods to evaluate the effects of various plant oils or camelina expeller on animal performance and milk fatty acid composition. Treatments consisted of 5 concentrate supplements containing no additional lipid (control), or 29 g/kg of lipid from rapeseed oil (RO), sunflower-seed oil (SFO), camelina-seed oil (CO), or camelina expeller (CE). Cows were offered red clover silage ad libitum and 12kg/d of experimental concentrates. Treatments had no effect on silage or total dry matter intake, whole-tract digestibility coefficients, milk yield, or milk composition. Plant oils in the diet decreased short- and medium-chain saturated fatty acid (6:0-16:0) concentrations, including odd- and branched-chain fatty acids and enhanced milk fat 18:0 and 18-carbon unsaturated fatty acid content. Increases in the relative proportions of cis 18:1, trans 18:1, nonconjugated 18:2, conjugated linoleic acid (CLA), and polyunsaturated fatty acids in milk fat were dependent on the fatty acid composition of oils in the diet. Rapeseed oil in the diet was associated with the enrichment of trans 18:1 (Δ4, 6, 7, 8, and 9), cis-9 18:1, and trans-7,cis-9 CLA, SFO resulted in the highest concentrations of trans-5, trans-10, and trans-11 18:1, Δ9,11 CLA, Δ10,12 CLA, and 18:2n-6, whereas CO enhanced trans-13-16 18:1, Δ11,15 18:2, Δ12,15 18:2, cis-9,trans-13 18:2, Δ11,13 CLA, Δ12,14 CLA, Δ13,15 CLA, Δ9,11,15 18:3, and 18:3n-3. Relative to CO, CE resulted in lower 18:0 and cis-9 18:1 concentrations and higher proportions of trans-10 18:1, trans-11 18:1, cis-9,trans-11 CLA, cis-9,trans-13 18:2, and trans-11,cis-15 18:2. Comparison of milk fat composition responses to CO and CE suggest that the biohydrogenation of unsaturated 18-carbon fatty acids to 18:0 in the rumen was less complete for camelina lipid supplied as an expeller than as free oil. In conclusion, moderate amounts of plant oils in diets based on red clover silage had no adverse effects on silage dry matter intake, nutrient digestion, or milk production, but altered milk fat composition, with changes characterized as a decrease in saturated fatty acids, an increase in trans fatty acids, and enrichment of specific unsaturated fatty acids depending on the fatty acid composition of lipid supplements.

Dietary Fish Oil Supplements Modify Ruminal Biohydrogenation, Alter the Flow of Fatty Acids at the Omasum, and Induce Changes in the Ruminal Butyrivibrio Population in Lactating Cows

Four lactating cows fitted with ruminal cannulae and fed a grass silage-based diet were used in a 4 × 4 Latin square with 28-d periods to investigate the effects of incremental dietary fish oil (FO) supplementation (0, 75, 150, or 300 g/d) on the flow of fatty acids at the omasum and populations of rumen bacteria capable of biohydrogenation. FO decreased silage intake and ruminal volatile fatty acid concentrations and promoted an increase in molar butyrate and propionate proportions at the expense of acetate. Extensive ruminal biohydrogenation of 20:5(n-3) and 22:6(n-3) resulted in corresponding increases in numerous 20- and 22-carbon unsaturated fatty acids at the omasum. Omasal flow of several 20-, 21-, and 22-carbon all-cis (n-3) PUFA exceeded the intake from FO. Supplements of FO also induced a dose-dependent decrease in 18:0 and increased trans 18:1 and trans 18:2 flow at the omasum. Trans-11 was the major 18:1 intermediate in digesta, while FO induced quadratic increases in trans-10 18:1 flow, reaching a maximum of 300 g/d. FO had no substantial influence on omasal flow of CLA. Results suggest that one or more fatty acids in FO inhibit the reduction of trans-18:1 and trans-18:2 intermediates by ruminal microorganisms. qPCR based on 16S rRNA genes in omasal digesta indicated that key Butyrivibrio spp. declined linearly in response to FO. Dose-dependent increases in ruminal outflow of biohydrogenation intermediates containing one or more trans double bonds in response to FO has major implications for host metabolism and the nutritional quality of ruminant foods.

Determination of digesta flow entering the omasal canal of dairy cows using different marker systems

Four studies were conducted to compare the effect of four indigestible markers (LiCoEDTA, Yb-acetate, Cr-mordanted straw and indigestible neutral-detergent fibre (INDF)) and three marker systems on the flow of digesta entering the omasal canal of lactating dairy cows. Samples of digesta aspirated from the omasal canal were pooled and separated using filtration and high-speed centrifugation into three fractions defined as the liquid phase, small particulate and large particulate matter. Co was primarily associated with the liquid phase, Yb was concentrated in small particulate matter, whilst Cr and INDF were associated with large particles. Digesta flow was calculated based on single markers or using the reconstitution system based on combinations of two (Co + Yb, Co + Cr and Co + INDF) or three markers (Co + Yb + Cr and Co + Yb + INDF). Use of single markers resulted in large differences between estimates of organic matter (OM) flow entering the omasal canal suggesting that samples were not representative of true digesta. Digesta appeared to consist of at least three phases that tended to separate during sampling. OM was concentrated in particulate matter, whilst the liquid phase consisted mainly of volatile fatty acids and inorganic matter. Yb was intimately associated with nitrogenous compounds, whereas Cr and INDF were concentrated in fibrous material. Current data indicated that marker systems based on Yb in combination with Cr or INDF are required for the accurate determination of OM, N and neutral-detergent fibre flow. In cases where the flow of water-soluble nutrients entering the omasal canal is also required, the marker system should also include Co.

Effects of Supplementation of a Grass Silage and Barley Diet with Urea, Rapeseed Meal and Heat-moisture-treated Rapeseed Cake on Omasal Digesta Flow and Milk Production in Lactating Dairy Cows

Four multiparous dairy cows, fitted with rumen and duodenal cannulas, were fed grass silage and one of four concentrates of barley (B), barley+urea (BU), barley+solvent-extracted rapeseed meal (BRM) or barley+heat-moisture-treated rapeseed cake (BRC). Despite marginal N deficiencies for the control diet (B), BU did not affect milk yield, ruminal fibre digestion or omasal total non-ammonia-N (NAN) or microbial NAN flow. In contrast, BRM and BRC diets significantly increased milk and milk protein yields and omasal dietary NAN flows. Effects of rapeseed meal (RSM) and rapeseed cake (RSC) on milk yield and omasal NAN flow were similar. The diets had no effect on rumen microbial protein synthesis. In general, non-protein N and soluble true protein comprised 0.20 of omasal total NAN flow, for all diets. Supplementary AAT from rapeseed feeds appeared to escape from the rumen as insoluble feed particles.

Effects of feeding grass or red clover silage cut at two maturity stages in dairy cows. 2. Dry matter intake and cell wall digestion kinetics.

This study examined the effects of red clover or grass silages cut at 2 stages of growth on feed intake, cell wall digestion, and ruminal passage kinetics in lactating dairy cows. Five dairy cows equipped with rumen cannulas were used in a study designed as a 5 x 5 Latin square with 21-d periods. Diets consisted of early-cut and late-cut grass and red clover silages and a mixture of late-cut grass and early-cut red clover silages offered ad libitum. All diets were supplemented with 9 kg/d of concentrate. Ruminal digestion and passage kinetics were assessed by the rumen evacuation technique. Apparent total-tract digestibility was determined by total fecal collection. The silage dry matter intake was highest when the mixed forage diet was fed and lowest with the early-cut red clover diet. Delaying the harvest tended to decrease DMI of grass and increase that of red clover. The intake of neutral detergent fiber (NDF) and potentially digestible NDF (pdNDF) was lower but the intake of indigestible NDF (iNDF) was higher for red clover diets than for grass diets. The rumen pool size of iNDF and the ratio of iNDF to pdNDF in the rumen contents were larger, and pool sizes of NDF and pdNDF were smaller for red clover than for grass silage diets. Outflow of iNDF and the ratio of iNDF to pdNDF in digesta entering the omasal canal were larger, and the outflow of pdNDF was smaller for red clover than for grass silage diets. The digestion rate (k(d)) of pdNDF was faster for red clover diets than for grass silage diets. Delaying the harvest decreased k(d) for grass but increased it for red clover silage diets. Observed differences in fiber characteristics of red clover and grass silages were reflected in ruminal digestion and passage kinetics of these forages. The low intake of early-cut red clover silage could not be explained by silage digestibility, fermentation quality, or rumen fill, but was most likely related to nutritionally suboptimal composition because inclusion of moderate quality grass silage improved silage intake. Increasing the maturity of ensiled red clover does not seem to affect silage dry matter intake as consistently as that of grasses.

Rapeseed or linseed in grass-based diets: Effects on conjugated linoleic and conjugated linolenic acid isomers in milk fat from Holstein cows over 2 consecutive lactations

Changes in the distribution of conjugated linoleic (CLA) and conjugated linolenic (CLnA) acid isomers in milk from Holstein cows in response to 4 different oilseed supplements rich in either cis-9 18:1 or 18:3n-3 were determined over 2 consecutive lactations in 58 and 35 cows during the first and second years, respectively. For the first 5 wk of the first lactation, all cows were fed the same diet. Thereafter, cows received 1 of 5 treatments for 2 consecutive lactations, including the prepartum period. Treatments comprised the basal diet with no additional lipid, or supplements of extruded linseeds (EL), extruded rapeseeds (ER), cold-pressed fat-rich rapeseed meal, or whole unprocessed rapeseeds to provide 2.5 to 3.0% of additional oil in diet dry matter. During indoor periods, cows were housed and received a mixture (3:1, wt/wt) of grass silage and hay, whereas cows were at pasture during outdoor periods. Over the entire study, EL resulted in the enrichment of ∆11,13 CLA, ∆12,14 CLA, trans-9,trans-11 CLA, trans-13,trans-15 CLA, ∆9,11,15 CLnA, and cis-9,trans-11,trans-13 CLnA (identified for the first time in bovine milk fat) in milk fat, whereas ER and cold-pressed fat-rich rapeseed meal in particular, increased milk fat trans-7,cis-9 CLA concentration. With the exception of the first indoor period, whole unprocessed rapeseeds decreased cis-9,trans-11 CLA, trans-9,cis-11 CLA, and trans-10,trans-12 CLA abundance. During the second indoor period, EL increased milk trans-9,cis-11 CLA and trans-10,cis-12 CLA concentrations, but the increases in cis-9,trans-11 CLA, cis-12,trans-14 CLA, trans-11,cis-13 CLA, and cis-9,trans-11,cis-15 CLnA concentrations to EL and ER were lower for the second than first indoor period. In contrast to the indoor periods, EL and ER decreased milk cis-9,trans-11 CLA, trans-9,cis-11 CLA, and trans-10,cis-12 CLA concentrations at pasture. The extent of changes in the relative distribution and abundance of CLA and CLnA isomers in milk fat were related to the nature (rapeseed or linseed) and form of oilseed (extruded, cold-pressed fat-rich meal or whole unprocessed) supplement and their interactions with the composition of the basal diet (conserved grass or pasture and dietary starch content). Furthermore, milk fat CLA and CLnA responses to treatments were repeatable between both outdoor periods. Variations in milk fat content and yield measured during the entire study were significantly and inversely associated with milk trans-10 18:1, trans-10,cis-12 CLA, and in particular, trans-9,cis-11 CLA concentrations.

Equol in milk of dairy cows is derived from forage legumes such as red clover

The intake of isoflavones and the resulting equol contents of both plasma and milk of the same red clover-fed cows are reported for the first time in cyclic change-over design study. Cows were fed four different red clover silages and two timothy-meadow fescue silages as controls. The red clover silages contained daidzein, formononetin, biochanin A and genistein, whereas the timothy-meadow fescue silages contained no isoflavones. We found a strong association (y = 0.071x+2.75, R 2 0.71) between the formononetin intake (x) and equol concentration (y) in the plasma, while the formononetin intake and milk equol concentration were weakly associated (y = 0.0035x+0.358, R 2 0.20). This means that a small part of the total formononetin in the silage is secreted into milk as equol. The mean equol contents in plasma and milk of cows fed red clover silage diets were in the range of 4.6-8.4 mg/l and 458-643 microg/l, respectively, while the respective values for the control diets were in the range of 0.8-1.5 mg/l and 171-287 microg/l. We showed that shorter growing periods of red clover resulted in higher silage formononetin contents and plasma and milk equol contents, suggesting that the equol content of milk can be manipulated by varying the harvesting strategy of red clover. We conclude that milk equol is derived from the formononetin of red clover silage and that milk from red clover-fed cows can be considered as a source of equol in human nutrition.