Plasma transport of ergocalciferol and cholecalciferol and their 25-hydroxylated metabolites in dairy cows.

In cattle, there are 2 significant forms of vitamin D: ergocalciferol (ERG) from fungi on roughage and cholecalciferol (CHO) from vitamin supplements or endogenous synthesis in the skin. The hypothesis of the present study is that vitamin D from the 3 sources is transported in different plasma fractions in the body. This is hypothesized to explain the lower efficiency of ERG compared to CHO in securing a sufficient plasma status of 25-hydroxyvitamin D and explain the inefficient excretion of dietary CHO into milk compared to endogenous CHO. Twenty vitamin D-depleted cows were assigned to 5 treatments: D2, housed indoor and fed 625-µg/d (25.000 IU) ERG; D3, housed indoor and fed 625-µg/d CHO; D2+D3, housed indoor and fed 625-µg/d ERG and 625-µg/d CHO; SUN, let out for daily pasture to facilitate CHO synthesis from sunlight; and D2+SUN, fed 625-µg/d ERG and let out for daily pasture. Blood samples were taken twice weekly and plasma fractionated by ultracentrifugation into 3 fractions: light lipoprotein (LLP), heavy lipoprotein (HLP), and protein and analyzed for content of ERG and CHO and their liver derived metabolites 25-hydroxyergocalciferol (25ERG) and 25-hydroxycholecalciferol (25CHO), respectively. Liver biopsies were taken on the last day of the study to asses gene expression related to vitamin D metabolism. During 4 wk of study, the vitamin D status in plasma increased to 19.3 to 22.8 ng/mL 25ERG in ERG-treated cows with the highest concentration in D2 (P ≤ 0.05) and to 25.0 to 33.4 ng/mL 25CHO in pasture or CHO-treated cows with the highest concentration in SUN (P ≤ 0.01). In plasma fractions, CHO was mainly found in the HLP fraction, whereas 25CHO was almost exclusively found in the protein fraction, probably due to its reported high binding affinity to vitamin D-binding protein. About 70% to 90% of 25ERG was found in the protein fraction and the remaining 25ERG was found in HLP, whereas ERG was found in both HLP and LLP fractions. In liver tissue, the expression of vitamin D-25-hydroxylase was lower in D2+D3 (P ≤ 0.05) and SUN (P ≤ 0.05) than that in the remaining groups, and the vitamin D receptor was expressed in the liver to a larger extent in D2+SUN than that in D2+D3 (P ≤ 0.05) and SUN (P ≤ 0.05). In conclusion, different plasma transport mechanisms may explain the lower physiological efficiency of ERG compared to CHO in securing the vitamin D status in plasma but do not explain the lower efficiency of synthetic CHO compared to endogenous CHO from sunlight or UV light in securing a high CHO content in milk.

Physiological limit of the daily endogenous cholecalciferol synthesis from UV light in cattle.

The link between UV light (sunlight) and endogenous cholecalciferol (vitamin D3 ) synthesis in the skin of humans has been known for more than a 100 years, since doctors for the first time successfully used UV light to cure rickets in children. Years later, it was shown that UV light also had a significant effect on the cholecalciferol status in the body of cattle. The cholecalciferol status in the body is measured as the plasma concentration of 25-hydroxycholecalciferol, which in cattle and humans is the major circulating metabolite of cholecalciferol. Very little is, however, known about the quantitative efficiency of UV light as a source of cholecalciferol in cattle nutrition and physiology. Hence, the aim of this study was to determine the efficiency of using UV light for increasing the plasma 25-hydroxycholecalciferol concentration in cholecalciferol-deprived cattle. Twelve cows deprived of cholecalciferol for 6 months were divided into three treatment groups and exposed to UV light for 30, 90 or 120 min/day during 28 days. UV-light wavelengths ranged from 280 to 415 nm and 30-min exposure to the UV light was equivalent to 60-min average summer-sunlight exposure at 56 °N. Blood samples were collected every 3-4 days and analysed for 25-hydroxycholecalciferol and cholecalciferol. Results showed that increasing the exposure time from 90-120 min/day did not change the slope of the daily increase in plasma 25-hydroxycholecalciferol. Hence, it appears that cholecalciferol-deprived dairy cattle are able to increase their plasma 25-hydroxycholecalciferol concentration by a maximum of 1 ng/ml/day from UV-light exposure.

Milk production response to varying protein supply is independent of forage digestibility in dairy cows.

The aim of this experiment was to examine whether the positive response in milk production to increased crude protein (CP) supply in dairy cows was dependent on the digestibility of the forage. Forty-eight lactating Danish Holstein cows were used in a 4 × 4 Latin square design experiment with 4 rations: (1) high digestibility and high CP concentration (HdHp), (2) high digestibility and low CP concentration (HdLp), (3) low digestibility and high CP concentration (LdHp), and (4) low digestibility and low CP concentration (LdLp). All rations contained 30% corn silage, 25% grass-clover silage, and 45% concentrate on a dry matter (DM) basis. Different digestibilities were obtained by replacing a high-digestible grass-clover silage combined with a high-digestible corn silage with a low-digestible grass-clover silage combined with a low-digestible corn silage. Organic matter digestibilities were 79.8 and 74.7% in the high- and low-digestibility rations, respectively. Dietary CP concentration in the ration was increased by substituting barley and sugar beet pulp with rapeseed meal and soybean meal, whereby CP increased from 13.9 to 14.0% (Lp) to 15.7 to 16.0% (Hp). All cows were offered 3 kg of the same concentrate per day in the automatic milking system in addition to the mixed ration. Every feeding period lasted 3 wk, and DM intake and milk yield were measured in the last week in each period, and milk samples for determining milk composition, including fatty acid content, and blood samples were taken during the last 3d of each period. Dry matter intake increased by 2.2 kg/d on Hd compared with Ld and by 0.7 kg/d on Hp compared with Lp. The positive effect on DM intake was reflected in the energy-corrected milk (ECM) yield, as a higher ration digestibility increased the ECM yield by 1.7 kg/d and a higher CP concentration increased it by 1.2 kg/d. We detected no interaction between forage digestibility and CP concentration on milk production. Reduced digestibility was accompanied by an increase in the plasma level of glucose, suggesting that other nutrients were limiting to milk production. In conclusion, milk production responses to dietary CP supply appeared independent of forage digestibility.

High-quality forage can replace concentrate when cows enter the deposition phase without negative consequences for milk production.

Mobilization and deposition in cows are different strategies of metabolism; hence, the aim was to study the possibility of reducing the crude protein (CP) supply during deposition to limit the use of protein supplements and minimize the environmental impact. A total of 61 Jersey and 107 Holstein cows were assigned to 4 mixed rations in a 2 × 2 factorial design with 2 concentrate to forage ratios (CFR) and 2 CP levels: high CFR (40:60) and recommended CP [16% of dry matter (DM); HCFR-RP], high CFR (40:60) and low CP (14% of DM; HCFR-LP), low CFR (30:70) and recommended CP (16% of DM; LCFR-RP), and low CFR (30:70) and low CP (14% of DM; LCFR-LP), where RP met the Danish recommendations. Cows were fed concentrate in an automatic milking unit. After calving, cows were fed HCFR-RP until entering deposition, defined as 11 kg (Jersey) or 15 kg (Holstein) of weight gain from the lowest weight after calving. Subsequently, cows either remained on HCFR-RP or changed to one of the other mixed rations. Comparing strategies during wk 9 to 30 of lactation showed higher dry matter intake (DMI) of mixed ration on HCFR compared with LCFR and on RP compared with LP. The DMI of the concentrate was higher on LCFR than on HCFR and higher on LP than on RP, resulting in overall higher DMI on HCFR and RP than on LCFR and LP. Crude protein intakes were higher on RP than on LP and starch intakes were higher on HCFR than on LCFR. Intakes of neutral detergent fiber tended to be higher on LCFR than on HCFR. Intakes of net energy for lactation were affected by CFR and CP level, with a higher intake on HCFR and RP than on LCFR and LP. No interactions were found between CFR and CP level for any feed intake variables. Yields of milk and energy-corrected milk were higher on RP than on LP, with no difference in yield persistency after the ration change. Milk composition did not differ among strategies but the protein to fat ratio was higher on HCFR than on LCFR and tended to be lower on RP than on LP. Differences in fatty acid composition were small, and de novo synthesis was high (>60%). Energy efficiency was higher on LCFR than on HCFR and no interaction with breed or parity was found. The N efficiency was higher on LP than RP, but with an interaction with breed due to lower N efficiency in Jersey than Holstein cows on HCFR-RP but higher N efficiency in Jersey than Holstein on LCFR-LP. In dairy production, concentrate in the mixed ration can be substituted with high-quality forage during deposition without negative effects on milk yield and composition when a sufficient CP level is ensured.

Milk production is unaffected by replacing barley or sodium hydroxide wheat with maize cob silage in rations for dairy cows.

Starch is an important energy-providing nutrient for dairy cows that is most commonly provided from cereal grains. However, ruminal fermentation of large amounts of easily degradable starch leads to excessive production and accumulation of volatile fatty acids (VFA). VFA not only play a vital role in the energy metabolism of dairy cows but are also the main cause of ruminal acidosis and depressed feed intake. The aim of the present study was to compare maize cob silage (MCS) as an energy supplement in rations for dairy cows with highly rumen-digestible rolled barley and with sodium hydroxide wheat (SHW), which has a higher proportion of by-pass starch than barley. Two studies were carried out: (1) a production study on 45 Danish Holstein cows and (2) an intensive study to determine digestibilities, rumen fermentation patterns and methane emission using three rumen-cannulated Danish Holstein cows. Both studies were organised as a 3×3 Latin square with three experimental periods and three different mixed rations. The rations consisted of grass-clover silage and maize silage (~60% of dry matter (DM)), rapeseed cake, soybean meal, sugar beet pulp and one of three different cereals as a major energy supplement: MCS, SHW or rolled barley (~25% of DM). When MCS replaced barley or SHW as an energy supplement in the mixed rations, it resulted in a lower dry matter intake; however, the apparent total tract digestibilities of DM, organic matter, NDF, starch and protein were not different between treatments. The energy-corrected milk yield was unaffected by treatment. The fat content of the milk on the MCS ration was not different from the SHW ration, whereas it was higher on the barley ration. The protein content of the milk decreased when MCS was used in the ration compared with barley and SHW. From ruminal VFA patterns and pH measures, it appeared that MCS possessed roughage qualities with respect to rumen environment, while at the same time being sufficiently energy rich to replace barley and SHW as a major energy supplement for milk production. The environmental impact, expressed as methane emissions, was not different when comparing MCS, SHW and barley.

Milk fatty acid composition and production performance of Danish Holstein and Danish Jersey cows fed different amounts of linseed and rapeseed.

Fat supplements are used in diets for dairy cows to increase energy intake and milk production and the fatty acid composition of the feed affects milk fatty acid composition. A total of 74 Danish Holstein and 41 Danish Jersey cows were divided into 4 groups and the cows within each group were fed a mixed ration supplemented with 0, 3.5, 6.8, or 10.2% of dry matter of a linseed:rapeseed (1:3) mixture during lactation wk 6 to 30. Milk yield, fat, and lactose contents were not affected by treatments for Danish Holsteins, whereas these parameters increased when increased amounts of oilseeds were fed to Danish Jerseys. For both breeds, milk protein content decreased when increased amounts of oilseeds were fed. The milk fatty acid composition showed higher concentrations of saturated fatty acids and lower concentrations of unsaturated fatty acids in milk fat from Danish Jerseys compared with Danish Holsteins. Increased amounts of oilseeds in feed increased milk fat concentration of all C18 fatty acids except C18:2 n-6, whereas the content of C6 to C14, C11 to C17, and in particular, C16, decreased. This effect was more pronounced for Danish Holsteins than for Danish Jerseys. The apparent recovery of C18:2 n-6 and C18:3 n-3 decreased when increased amounts of oilseeds were fed; however, this was most likely due to increased amounts of fatty acid from feed used for other energy demands than milk production. It was concluded that up to 6.8% of oilseed supplementation can be fed without production problems and, in many cases, with positive production responses, including an improved milk fatty acid profile.

Vitamin D2 impairs utilization of vitamin D3 in high-yielding dairy cows in a cross-over supplementation regimen.

Vitamin D exists in 2 forms that are important regarding vitamin D status and supply in cattle: vitamin D(2) (D(2)) and vitamin D(3) (D(3)). To become physiologically active, both D(2) and D(3) must undergo 25-hydroxylation in the liver. The resulting 25-hydroxyvitamin D(2) [25(OH)D(2)] and 25-hydroxyvitamin D(3) [25(OH)D(3)] are measured as indicators of the physiological vitamin D status of cattle. The study used 14 Danish Holstein cows housed without access to sunlight. The cows were orally administered 250 mg (1.0 × 10(7)IU) of D(2) and D(3) in a cross-over design with 2 treatment groups and 2 study periods, rendering 4 treatments when carryover effects were taken into account: D(2) given first, D(2) given last after D(3), D(3) given first, and D(3) given last after D(2). Two weeks elapsed between the treatment in the first study period and the treatment in the second study period. Blood samples were collected 0, 3, 6, 14, 17, 20, 23, 26, 40, 48, 70, 94, 166, and 214 h after providing the oral bolus of vitamin to the cows. Comparisons between plasma levels of the metabolites D(2), D(3), 25(OH)D(2), and 25(OH)D(3) over time were made by comparing areas under the plasma concentration curves. Oral administration of D(3) increased plasma D(3) (182.6±17.1 ng/mL; mean ± SEM) and 25(OH)D(3) (103.5±10.0 ng/mL) more efficiently than oral administration of D(2) increased plasma D(2) (49.1±32.6 ng/mL) and 25(OH)D(2) (27.9±2.1 ng/mL). The D(3) given after an oral dose of D(2) was less efficient for increasing plasma concentrations of 25(OH)D(3) (61.2±12.0 ng/mL) compared with D(3) given without previous D(2) administration (103.5±10.0 ng/mL), whereas the plasma concentrations of D(3) itself were the same when given first (182.6±17.1 ng/mL) as when given after D(2) (200.0±123.9 ng/mL). The same occurred for plasma concentrations of D(2) metabolites both if D(2) was given first (49.1±32.6 ng/mL) and after D(3) (54.7±7.7 ng/mL). In conclusion, D(3) given after D(2) is less efficient at increasing the plasma status of 25(OH)D(3) than D(3) given without previous D(2) administration.

Influence of vitamins A, D3 and E status on post-mortem meat quality in steers under winter housing or pasture finishing systems.

We investigated the influence of Swedish recommended vitamins A, D3 and E supplementation levels on muscle tenderness and fatty acid (FA) composition under indoor or outdoor finishing programmes. Swedish Red breed steer calves were divided into vitamin supplemented (n = 12) and non-supplemented (n = 15) groups while on pasture prior to the finishing period. This trial began at the beginning of the winter housing period during which the steers were fed a 55 : 45 dry matter barley : grass silage diet indoors. The indoor finished group was comprised of vitamin supplemented (n = 6) and non-supplemented (n = 8) steers slaughtered after about 155 days on feed. Vitamin supplemented steers were provided with 100 g mineral supplement providing 400 000 IU vitamin A, 100 000 IU D3 and 3000 IU E daily as recommended for Swedish production practices. In spring, outdoor finished vitamin supplemented (n = 6) and non-supplemented (n = 7) steers grazed semi-natural grassland for an additional 120 days before slaughter. During pasture, vitamin supplemented steers had free-choice access to a mineral supplement containing vitamins A, D3 and E. The mineral supplement for the non-supplemented steers did not contain vitamins A, D3 and E and was provided at the same amount as the vitamin supplemented steers. Shear force values were similar between vitamin supplemented and non-supplemented steers after ageing 2, 7 and 14 days within indoor and outdoor finishing programmes. The shear force values had decreased by 14 days of ageing within all programmes. The µ- and m-calpain activity did not differ between vitamin supplemented and non-supplemented steers for either the indoor or outdoor finishing programmes. The calpastatin activity was higher for the indoor, vitamin supplemented steers. Indoor finished vitamin supplemented steers had a greater proportion of C18:1c-9 and total monounsaturated fatty acids, whereas the non-supplemented steers had a greater proportion of total saturated fatty acids. We concluded that the meat quality from steers not receiving vitamin supplementation was similar to that of steers receiving vitamins A, D3 and E supplementation at Swedish recommended levels under indoor and outdoor finishing programmes.

Stability in the rumen and effect on plasma status of single oral doses of vitamin D and vitamin E in high-yielding dairy cows.

The ruminal fate of the fat-soluble vitamins D and E was studied in dairy cows. Ten to 15 kg of ruminal contents was taken from each cow through a ruminal fistula. A sample was taken out (0-h sample) and the rest of the contents were mixed with 4,360 mg of all-rac-α-tocopheryl acetate (vitamin E; study 1) or 4,360 mg of all-rac-α-tocopheryl acetate, 250 mg of ergocalciferol (vitamin D(2)), and 250 mg of cholecalciferol (vitamin D(3); study 2). After mixing, the ruminal contents were returned to the respective cows. Blood was collected 0, 6, 24, and 30 h after introducing the vitamins into the rumen. Samples of ruminal contents were collected at 0, 1, 2, 4, 6, 24, and 30 h (in vivo). From the 1-h sample, 6 subsamples from each cow were incubated at 37 °C and taken out at 2, 4, 6, 12, 24, and 30 h (in vitro). In vivo concentrations of added α-tocopherol, ergocalciferol, and cholecalciferol in the rumen first increased and subsequently declined due to dilution effects of eating and passage out of the rumen. The level of the free-alcohol form of α-tocopherol from the natural content in feed was constant throughout the in vivo study, in contrast to the content of total α-tocopherol, which indicated that no hydrolysis of the acetate form into alcohol form happened in the rumen. In vitro, all added vitamins were found at constant levels; hence, none of the added vitamins were degraded in ruminal contents. The concentration of α-tocopherol in plasma increased at a rate per milligram of ruminally introduced α-tocopherol below the rate of the increase in plasma ergocalciferol or cholecalciferol metabolites per milligram of introduced ergocalciferol or cholecalciferol, respectively, over 24h. In conclusion, ergocalciferol, cholecalciferol, and α-tocopheryl acetate proved to be stable in the rumen and in ruminal contents from high-yielding dairy cows. Changes in plasma concentrations of the vitamins relative to the amount of vitamin introduced to the rumen indicated a lower effect on plasma status of ergocalciferol than of cholecalciferol, and an even lower effect of α-tocopherol. The limited plasma response after a single dose of α-tocopheryl acetate led to the conclusion that oral single dose therapy with all-rac-α-tocopheryl acetate is of limited physiological value.

Vitamin D(3) synthesis in the entire skin surface of dairy cows despite hair coverage.

How hair-coated animals such as dairy cows synthesize endogenous vitamin D(3) during exposure to summer sunlight has been unclear since vitamin D(3) and its relation to sunlight was discovered. The fur of fur-bearing animals is thought to be comparable to clothing in humans, which prevents vitamin D(3) synthesis in the skin during exposure to sunlight. Different scenarios have been suggested but never tested in cows; for example, that vitamin D(3) is synthesized from sebum on the hair and ingested by cows during grooming or that body areas such as the udder and muzzle that have scant hair exclusively produce the vitamin. To test different scenarios, 16 Danish Holstein dairy cows were subjected to 4 degrees of coverage of their bodies with fabric that prevented vitamin D(3) synthesis in the covered skin areas. The treatments were horse blanket (cows fitted with horse blankets), udder cover (cows fitted with udder covers, horse blanket+udder cover (cows fitted with both horse blankets and udder covers), and natural (cows without any coverage fitted). The cows were let out to pasture daily between 1000 and 1500h for 4 wk in July and August 2009. Blood samples were collected 15 times during the study and analyzed for content of 25-hydroxyvitamin D(3) [25(OH)D(3)] indicative of the animals' vitamin D(3) status. Results showed that uncovered cows had a higher 25(OH)D(3) concentration in plasma after 28 d of access to sunlight compared with covered cows and that the plasma concentration of 25(OH)D(3) was strongly inversely correlated to the body surface area covered. These results are consistent with findings in humans, wherein the vitamin D(3) status of different individuals was inversely proportional to the amount of clothing worn during exposure to artificial sunlight. Hence, it appears that human clothing and cow hair are not comparable with respect to prevention of vitamin D(3) synthesis and that cows, like humans, synthesize vitamin D(3) evenly over their body surface. That vitamin D(3) should be synthesized from sebum on the hair and obtained by cows as a result of grooming is not supported by the findings in the present study either, because large differences were found between the treatment groups. If grooming were the source of vitamin D(3), then a relatively even 25(OH)D(3) concentration between treatments would be expected, because covered cows would obtain vitamin D(3) by grooming uncovered herdmates.