Herbage utilisation method affects rumen fluid and milk fatty acid profile in Holstein and Montbéliarde cows.

Compared with maize silage- and concentrate-based diets, herbage-based diets were repeatedly shown to favourably influence the milk fatty acid (FA) profile. However, it is unclear how the herbage feeding mode (grazing vs indoor green-feeding) and conservation (fresh herbage vs hay vs silage) modify the milk FA profile. Therefore, the aim of the present experiment was to investigate the effect of different herbage utilisation methods (including herbage feeding mode and herbage conservation method) on the ruminal biohydrogenation of dietary FA and the consequences on the milk FA composition in cows of two breeds (Holstein and Montbéliarde). Concomitant effects of botanical composition and phenological stage of the herbage on milk FA profile were controlled for by harvesting barn-dried hay and silage simultaneously as first cut from the same ryegrass-dominated grassland in a semi-mountainous region. Seven weeks later, the first regrowth of the same plot was used as fresh herbage, either grazed or fed indoor (indoor green-feeding). Twenty-four Montbéliarde and 24 Holstein cows were randomly allocated to four groups of 12 cows balanced by breed, parity, and milk yield. In a free-stall barn, three groups were given ad libitum access to hay, silage, or fresh herbage, respectively. The fourth group was strip-grazing. All cows were supplemented with 3 kg DM/day of the same energy-rich concentrate. After 2 weeks of adaptation to the forage, samples of forage, concentrate, milk, blood, and rumen fluid were collected. Fatty acid composition of forages, rumen fluid, and milk was analysed by gas chromatography. Haymaking reduced total FA content of the herbage, in particular that of linoleic acid (LA) and α-linolenic acid (ALA). Still, rumen fluid lipids of hay-fed cows had the highest proportion of rumenic acid, LA, ALA, and total polyunsaturated fatty acids (PUFAs). Milk fat from hay-fed cows had the highest proportion of LA, and the apparent transfer rates from feed to milk of LA and ALA were higher in hay-fed cows than in silage-fed cows. The proportion of PUFAs was highest in milk fat from grazing and indoor green-fed Montbéliarde cows and lowest in silage-fed cows of both breeds. In conclusion, the herbage utilisation method affects the ruminal biohydrogenation of LA and ALA, whereby herbage drying particularly increases their transfer from herbage to milk.

Feeding heat-oxidized oil to dairy cows affects milk fat nutritional quality.

Heating oil and oilseeds results in oxidation products that affect ruminal biohydrogenation of polyunsaturated fatty acids, altering milk fatty acids profile, and could be transferred to milk. An experiment was conducted to investigate the effects of oil heating on rumen and milk fatty acids profile and the transfer of oxidation products to milk. Sunflower oil was heated at 150°C for 15 h and given to lactating dairy cows in a 2×2 arrangement: two groups of two cows, equipped with a ruminal cannula and receiving two diets (containing either heated or unheated oil) during two experimental periods. Oil heating generated hydroperoxides and/or hydroxyacids and aldehydes, in particular trans-2,trans-4-decadienal. In milk, heated oil only significantly decreased trans-11-C18:1 and cis-9,trans-11-CLA percentage compared to non-heated oil, and slightly increased cis-9,cis-12-C18:2 percentage, which was probably linked to an inhibition of the ruminal Δ12 isomerase by oxidative products in the rumen. However, feeding highly oxidized oil did not result in the appearance of hydroperoxides or hydroxyacids in milk and did not increase milk aldehydes content.

Rumen microbiota and dietary fat: a mutual shaping.

Although fat content in usual ruminant diets is very low, fat supplements can be given to farm ruminants to modulate rumen activity or the fatty acid (FA) profile of meat and milk. Unsaturated FAs, which are dominant in common fat sources for ruminants, have negative effects on microbial growth, especially protozoa and fibrolytic bacteria. In turn, the rumen microbiota detoxifies unsaturated FAs (UFAs) through a biohydrogenation (BH) process, transforming dietary UFAs with cis geometrical double-bonds into mainly trans UFAs and, finally, into saturated FAs. Culture studies have provided a large amount of data regarding bacterial species and strains that are affected by UFAs or involved in lipolysis or BH, with a major focus on the Butyrivibrio genus. More recent data using molecular approaches to rumen microbiota extend and challenge these data, but further research will be necessary to improve our understanding of fat and rumen microbiota interactions.

Effects of the heating process of soybean oil and seeds on fatty acid biohydrogenation in vitro.

Heating fat is an efficient way to alter ruminal biohydrogenation (BH) and milk fat quality. Nevertheless, results are variable among studies and this could be due to various heating conditions differently affecting BH. The objectives of this study were to determine the effect of type and duration of heating of soybean oil or seeds on BH in vitro. Ruminal content cultures were incubated to first investigate the effects of roasting duration (no heating, and 0.5- and 6-h roasting) at 125°C and its interaction with fat source (soybean seeds vs. soybean oil), focusing on linoleic acid BH and its intermediates: conjugated linoleic acid (CLA) and trans-C18:1. Additionally, we compared the effects of seed extrusion with the 6 combinations of unheated and roasted oils and seeds. None of the treatments was efficient to protect linoleic acid from BH. Soybean oil resulted in higher trans-11 isomer production than seeds: 5.7 and 1.2 times higher for cis-9,trans-11 CLA and trans-11 C18:1, respectively. A 125°C, 0.5-h roasting increased trans-11 isomer production by 11% compared with no heating and 6-h roasted fat. Extrusion of seeds was more efficient to increase trans-11 C18:1 production than seed roasting, leading to values similar to oils. For other fatty acids, including cis-9,trans-11 CLA, extrusion resulted in similar balances to seeds (mainly 0.5-h-roasted seeds). Extruded oilseeds would be more efficient than roasted seeds to produce trans-11 C18:1; nevertheless, effects of conditions of extrusion need to be explored.

Establishment of ruminal bacterial community in dairy calves from birth to weaning is sequential.

AIM: Establishment of ruminal bacterial community in dairy calves. METHODS AND RESULTS: Rumen bacterial community was analysed on 6 calves bred according to commercial practices from day one to weaning at day 83 of age, using 454 16S rRNA-based pyrosequencing. Samples taken at day 1 did not produce amplicons. Analysis of data revealed a three-stage implantation process with a progressive but important shift of composition. At day 2, the bacterial community was mainly composed of Proteobacteria (70%) and Bacteroidetes (14%), and Pasteurellaceae was the dominant family (58%). The bacterial community abruptly changed between days 2 and 3, and until day 12, dominant genera were Bacteroides (21%), Prevotella (11%), Fusobacterium (5%) and Streptococcus (4%). From 15 to 83 days, when solid food intake rapidly increased, Prevotella became dominant (42%) and many genera strongly decreased or were no longer detected. A limited number of bacteria genera correlated with feed intake, rumen volatile fatty acids and enzymatic activities. CONCLUSION: The ruminal bacterial community is established before intake of solid food, but solid food arrival in turn shapes this community. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provides insight into the establishment of calves' rumen bacterial community and suggests a strong effect of diet.

Starch plus sunflower oil addition to the diet of dry dairy cows results in a trans-11 to trans-10 shift of biohydrogenation.

Trans fatty acids (FA), exhibit different biological properties. Among them, cis-9,trans-11 conjugated linoleic acid has some interesting putative health properties, whereas trans-10,cis-12 conjugated linoleic acid has negative effects on cow milk fat production and would negatively affect human health. In high-yielding dairy cows, a shift from trans-11 to trans-10 pathway of biohydrogenation (BH) can occur in the rumen of cows receiving high-concentrate diets, especially when the diet is supplemented with unsaturated fat sources. To study this shift, 4 rumen-fistulated nonlactating Holstein cows were assigned to a 4×4 Latin square design with 4 different diets during 4 periods. Cows received 12 kg of dry matter per day of 4 diets based on corn silage during 4 successive periods: a control diet (22% starch, <3% crude fat on DM basis), a high-starch diet supplemented with wheat plus barley (35% starch, <3% crude fat), a sunflower oil diet supplemented with 5% of sunflower oil (20% starch, 7.6% crude fat), and a high-starch plus sunflower oil diet (33% starch, 7.3% crude fat). Five hours after feeding, proportions of trans-11 BH isomers greatly increased in the rumen content with the addition of sunflower oil, without change in ruminal pH compared with the control diet. Addition of starch to the control diet had no effect on BH pathways but decreased ruminal pH. The addition of a large amount of starch in association with sunflower oil increased trans-10 FA at the expense of trans-11 FA in the rumen content, revealing a trans-11 to trans-10 shift. Interestingly, with this latter diet, ruminal pH did not change compared with a single addition of starch. This trans-11 to trans-10 shift occurred progressively, after a decrease in the proportion of trans-11 FA in the rumen, suggesting that this shift could result from a dysbiosis in the rumen in favor of trans-10-producing bacteria at the expense of those producing trans-11 or a modification of bacterial activities.

Effects of oil and natural or synthetic vitamin E on ruminal and milk fatty acid profiles in cows receiving a high-starch diet.

Among trans fatty acids, trans-10,cis-12 CLA has negative effects on cow milk fat production and can affect human health. In high-yielding dairy cows, a shift from the trans-11 to the trans-10 pathway of biohydrogenation (BH) can occur in the rumen of cows receiving high-concentrate diets, especially when the diet is supplemented with unsaturated fat sources. In some but not all experiments, vitamin E has been shown to control this shift. To ascertain the effects of vitamin E on this shift of BH pathway, 2 studies were conducted. The first study explored in vitro the effects of addition of natural (RRR-α-tocopherol acetate) and synthetic (dl-α-tocopherol acetate) vitamin E. Compared with control and synthetic vitamin E, the natural form resulted in a greater trans-10/trans-11 ratio; however, the effect was very low, suggesting that vitamin E was neither a limiting factor for rumen BH nor a modulator of the BH pathway. An in vivo study investigated the effect of natural vitamin E (RRR-α-tocopherol) on this shift and subsequent milk fat depression. Six rumen-fistulated lactating Holstein cows were assigned to a 2×2 crossover design. Cows received 20-kg DM of a control diet based on corn silage with 22% of wheat, and after 2 wk of adaptation, the diet was supplemented with 600 g of sunflower oil for 2 more weeks. During the last week of this 4-wk experimental period, cows were divided into 2 groups: an unsupplemented control group and a group receiving 11 g of RRR-α-tocopherol acetate per day. A trans-10 shift of ruminal BH associated with milk fat depression due to oil supplementation of a high-wheat diet was observed, but vitamin E supplementation of dairy cows did not result in a reversal toward a trans-11 BH pathway, and did not restore milk fat content.

In vitro study of dietary factors affecting the biohydrogenation shift from trans-11 to trans-10 fatty acids in the rumen of dairy cows.

On the basis of the isomer-specific effects of trans fatty acids (FA) on human health, and the detrimental effect of t10,c12-conjugated linoleic acid (CLA) on cows' milk fat production, there is a need to identify factors that affect the shift from trans-11 to trans-10 pathway during ruminal biohydrogenation of FA. This experiment was conducted in vitro and aimed at separating the effects of the diet of the donor cows from those of the fermentative substrate, which is necessary to prevent this shift. A total of four dry Holstein dairy cows were used in a 4 × 4 Latin square design. They received 12 kg of dry matter per day of four diets based on maize silage during four successive periods: the control diet (22% starch, <3% fat); the high-starch diet, supplemented with wheat plus barley (35% starch, <3% crude fat); the sunflower oil diet, supplemented with 5% of sunflower oil (20% starch, 7.6% crude fat); and the high-starch plus oil diet (33% starch, 7.3% crude fat). Ruminal fluid of each donor cow was incubated for 5 h with four substrates having similar chemical composition to the diets, replacing sunflower oil by pure linoleic acid (LA). The efficiency of isomerisation of LA to CLA was the highest when rumen fluids from cows receiving dietary oil were incubated with added LA. The shift from trans-11 to trans-10 isomers was induced in vitro by high-starch diets and the addition of LA. Oil supplementation to the diet of the donor cows increased this shift. Conversely, the trans-10 isomer balance was always low when no LA was added to incubation cultures. These results showed that a large accumulation of trans-10 FA was only observed with an adapted microflora, as well as an addition of non-esterified LA to the incubation substrate.

Starch and oil in the donor cow diet and starch in substrate differently affect the in vitro ruminal biohydrogenation of linoleic and linolenic acids.

Trans isomers of fatty acids exhibit different health properties. Among them, trans-10,cis-12 conjugated linoleic acid has negative effects on milk fat production and can affect human health. A shift from the trans-11 to the trans-10 pathway of biohydrogenation (BH) can occur in the rumen of dairy cows receiving high-concentrate diets, especially when the diet is supplemented with highly unsaturated fat sources. The differences of BH patterns between linoleic acid (LeA) and linolenic acid (LnA) in such ruminal conditions remain unknown; thus, the aim of this work was to investigate in vitro the effects of starch and sunflower oil in the diet of the donor cows and starch level in the incubates on the BH patterns and efficiencies of LeA and LnA. The design was a 4 × 4 Latin square design with 4 cows, 4 periods, and 4 diets with combinations of 21 or 34% starch and 0 or 5% sunflower oil. The rumen content of each cow during each period was incubated with 4 substrates, combining 2 starch levels and either LeA or LnA addition. Capillary electrophoresis single-strand conformation polymorphism of incubates showed that dietary starch decreased the diversity of the bacterial community and the high-starch plus oil diet modified its structure. High-starch diets poorly affected isomerization and first reduction of LeA and LnA, but decreased the efficiencies of trans-11,cis-15-C18:2 and trans C18:1 reduction. Dietary sunflower oil increased the efficiency of LeA isomerization but decreased the efficiency of trans C18:1 reduction. An interaction between dietary starch and dietary oil resulted in the highest trans-10 isomers production in incubates when the donor cow received the high-starch plus oil diet. The partition between trans-10 and trans-11 isomers was also affected by an interaction between starch level and the fatty acid added to the incubates, showing that the trans-10 shift only occurred with LeA, whereas LnA was mainly hydrogenated via the more usual trans-11 pathway, whatever the starch level in the substrate, although the bacterial communities were not different between LeA and LnA incubates. In LeA incubates, trans-10 isomer production was significantly related to the structure of the bacterial community.

Temperature and duration of heating of sunflower oil affect ruminal biohydrogenation of linoleic acid in vitro.

Sunflower oil heated at 110 or 150 degrees C for 1, 3, or 6h was incubated with ruminal content in order to investigate the effects of temperature and duration of heating of oil on the ruminal biohydrogenation of linoleic acid in vitro. When increased, these 2 parameters acted together to decrease the disappearance of linoleic acid in the media by inhibiting the isomerization of linoleic acid, which led to a decrease in conjugated linoleic acids and trans-C18:1 production. Nevertheless, trans-10 isomer production increased with heating temperature, suggesting an activation of Delta(9)-isomerization, whereas trans-11 isomer production decreased, traducing an inhibition of Delta(12)-isomerization. The amount of peroxides generated during heating was correlated with the proportions of biohydrogenation intermediates so that they might explain, at least in part, the observed effects. The effects of heating temperature and duration on ruminal bacteria community was assessed using capillary electrophoresis single-strand conformation polymorphism. Ruminal bacterial population significantly differed according to heating temperature, but was not affected by heating duration. Heating of fat affected ruminal biohydrogenation, at least in part because of oxidative products generated during heating, by altering enzymatic reactions and bacterial population.

Conjugated linolenic acid (CLnA), conjugated linoleic acid (CLA) and other biohydrogenation intermediates in plasma and milk fat of cows fed raw or extruded linseed.

Thirty lactating dairy cows were used in a 3 × 3 Latin-square design to investigate the effects of a raw or extruded blend of linseed and wheat bran (70:30) on plasma and milk fatty-acids (FA). Linseed diets, containing 16.6% linseed blend on a dry-matter basis, decreased milk yield and protein percentage. They decreased the proportions of FA with less than 18 carbons in plasma and milk and resulted in cis-9, cis-12, cis-15 18:3 proportions that were more than three and four times higher in plasma and milk, respectively, whereas cis-9, cis-12 18:2 proportions were decreased by 10-15%. The cis-9, trans-11, cis-15 18:3 isomer of conjugated linolenic acid was not detected in the milk of control cows, but was over 0.15% of total FA in the milk fat of linseed-supplemented cows. Similarly, linseed increased plasma and milk proportions of all biohydrogenation (BH) intermediates in plasma and milk, including the main isomer of conjugated linoleic acid cis-9, trans-11 18:2, except trans-4 18:1 and cis-11, trans-15 18:2 in plasma lipids. In milk fat, compared with raw linseed, extruded linseed further reduced 6:0-16:0 even-chain FA, did not significantly affect the proportions of 18:0, cis-9 18:1 and cis-9, cis-12 18:2, tended to increase cis-9, cis-12, cis-15 18:3, and resulted in an additional increase in the proportions of most BH intermediates. It was concluded that linseed addition can improve the proportion of conjugated linoleic and linolenic acids, and that extrusion further increases the proportions of intermediates of ruminal BH in milk fat.

Effects of copper, zinc and selenium status on performance and health in commercial dairy and beef herds: Retrospective study.

A retrospective study using analysis of plasma copper and zinc, and erythrocyte glutathione peroxidase from 2 080 dairy and beef cow herds was conducted to evaluate the relationship between trace-element status and production, reproduction and health in cows and their calves. Classification of the herd status as deficient, marginal, low-adequate or high-adequate was based on the lower tercile of individual values. Odds ratios for each disorder in herds were calculated by multivariable stepwise logistic regression. Inadequate copper status was not associated with adult disorders, but was an important risk factor for poor calf performance or health. Selenium deficient status was associated with most studied disorders in cows, and both deficient and marginal herd status were strongly associated with poor health of calves, particularly with increased risks of myopathy and infectious diseases. Zinc insufficiency was strongly associated with low milk production and impaired locomotion in dairy herds, and was also associated with diarrhoea and poor growth in calves. Because a low-adequate status increased the risk of many disorders in adults and calves, we propose to classify herds as deficient and marginal when the lower terciles of plasma zinc concentration are below 12 and between 12 and 14 mumol/l respectively.

In vitro versus in situ ruminal biohydrogenation of unsaturated fatty acids from a raw or extruded mixture of ground canola seed/canola meal.

Raw or extruded blends of ground canola seeds and canola meal were used to compare in vitro and in situ lag times and rates of disappearance due to ruminal biohydrogenation of unsaturated fatty acids. The in situ study resulted in higher lag times for biohydrogenation for polyunsaturated fatty acids and lower rates of biohydrogenation of unsaturated fatty acids than the in vitro study, so the in situ biohydrogenation of polyunsaturated fatty acids was not complete at 24 h of incubation. With both methods, rates of biohydrogenation of polyunsaturated fatty acids were higher than for cis-delta9C18:1. Extrusion did not affect the rate of biohydrogenation of cis-delta9C18:1, but resulted in higher rates of biohydrogenation of polyunsaturated fatty acids with higher proportions of trans intermediates of biohydrogenation at 4 h of incubation in vitro and at 8 h of incubation in situ. These results suggest that extrusion affects the isomerization of polyunsaturated fatty acids, rather than the hydrogenation steps. In conclusion, in vitro and in situ methods can both show differences of ruminal metabolism of unsaturated fatty acids due to processing, but the methods provide very different estimates of the rates of disappearance due to biohydrogenation.

Effects of copper supplementation on the copper status of peripartum beef cows and their calves.

The effects of copper supplementation on the copper status of 40 late-pregnant Aubrac beef cows grazing a copper-deficient pasture and later fed a marginally deficient diet were studied for five months. They were divided into four equal groups; the control group received no copper supplement, groups 1 and 2 received copper as copper sulphate at 10 and 30 mg/kg of diet dry matter (DM), respectively, for five months, and group 3 received 120 mg/kg of diet dry matter for 10 days. Plasma copper concentration and the activity of erythrocyte superoxide dismutase (eSOD) were measured at the beginning of the experiment, in the cows and calves during weeks 1 and 3 after calving, and in the calves before they were turned out to pasture at a mean (sd) age of 51 (26) days. In spite of the low dietary copper content (4.2 mg/kg of DM), the plasma copper concentration of the control cows increased during the winter. All the copper supplements resulted in normal and similar plasma copper concentrations in the cows after calving, but the concentration decreased slightly between weeks 1 and 3 after calving in the group supplemented for 10 days. The treatments did not affect the eSOD of the cows. The calves born to the four groups showed the same patterns of plasma copper and eSOD. Compared with the cows, the calves had low plasma copper concentrations at week 1 and values in the normal range at week 3; their eSOD was high at weeks 1 and 3 but decreased after week 3.

Effects of different forms of canola oil fatty acids plus canola meal on milk composition and physical properties of butter.

Twenty multiparous Holstein cows were used in a 16-wk trial. A block of 10 cows received a control diet, based on corn silage, and the other block of 10 cows successively received four diets with 1) an extruded blend of canola meal and canola seeds, 2) canola meal and whole canola seeds, 3) canola meal and ground canola seeds, or 4) canola meal and calcium salts of canola oil fatty acids. Canola fat represented about 2% of dietary dry matter. Compared to control cows, treated cows had similar dry matter intake, milk production, and daily milk output of true protein or fat. Protein contents of milk was decreased by all treatments, with a lower effect of extruded or whole canola seeds. Milk fat contents was lowered by all treatments, extruded seeds and calcium salts resulting in most important effects. All treatments lowered the percentage of fatty acids with 12 to 16 carbons in milk fat, increased C18:0 and cis-C18:1 percentages, and the proportion of liquid fat in butter between 0 and 12 degrees C. Calcium salts and, to a lesser extent extruded seeds, resulted in most important improvements of milk fatty acid profile and butter softness, whereas whole seeds had low effects.

Effects of pre- or postpartum selenium supplementation on selenium status in beef cows and their calves.

The effect of Se supplementation before or after calving on Se status in deficient cows and their calves was studied using 72 beef cows in two experiments. In Exp. 1, cows calving in February or March 1997 were supplemented orally for 15 d in late pregnancy with 13.0, 32.5, or 45.5 mg of Se/d as sodium selenite. Glutathione peroxidase (GSH-Px) activities were measured in red blood cells (RBC) or plasma of cows and calves at d 15 and between d 17 and 88 after calving. In Exp. 2, cows calving in January 1997 were supplemented orally with .0, 13.0, or 32.5 mg of Se/d for 15 d postpartum, and calves were injected with 1.38 mg of Se when 2 d old and at an average age of 49 d. The GSH-Px activities were measured in 30-d-old calves and in cows and calves between d 77 and 115 after calving. In both experiments, Se supplementation resulted in adequate Se status for the dams. The increase in RBC GSH-Px activity was faster with 45.5 mg of Se/d, and GSH-Px activities remained high for up to 98 d after the end of supplementation. The improvement in Se status in calves as a result of maternal supplementation was greater in Exp. 1 than in Exp. 2, suggesting that the placental transfer of Se is more efficient than milk transfer. Prepartum oral Se supplementation of deficient beef cows with 13.0 mg of Se/d for 15 d allowed adequate Se status of dams and calves, and 45.5 mg of Se/d resulted in a faster improvement of Se status. Parenteral administration of 1.38 mg of Se to newborn calves did not sustain normal Se status in calves issued from deficient cows.