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Vitamin D exists in 2 forms that are important regarding vitamin D status and supply in cattle: vitamin D2 (D2) and vitamin D3 (D3). To become physiologically active, both D2 and D3 must undergo 25-hydroxylation in the liver. The resulting 25-hydroxyvitamin D2 [25(OH)D2] and 25-hydroxyvitamin D2 [25(OH)D2] 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 x 107 IU) of D2 and D3 in a cross-over design with 2 treatment groups and 2 study periods, rendering 4 treatments when carryover effects were taken into account: D2 given first, D2 given last after D3, D3 given first, and D3 given last after D3. 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 D2, D3, 25(OH) D2, and 25(OH)D3 over time were made by comparing areas under the plasma concentration curves. Oral administration of D3 increased plasma D3 (182.6 ± 17.1 ng/mL; mean ± SEM) and 25(OH)D3 (103.5 ± 10.0 ng/mL) more efficiently than oral administration of D2 increased plasma D3 (49.1 ± 32.6 ng/mL) and 25(OH) D3 (27.9 ± 2.1 ng/mL). The D3 given after an oral dose of D2 was less efficient for increasing plasma concentrations of 25(OH)D3 (61.2 ± 12.0 ng/mL) compared with D3 given without previous D2 administration (103.5 ± 10.0 ng/mL), whereas the plasma concentrations of D3 itself were the same when given first (182.6 ± 17.1 ng/ mL) as when given after D2 (200.0 ± 123.9 ng/mL). The same occurred for plasma concentrations of D3 metabolites both if D2 was given first (49.1 ± 32.6 ng/ mL) and after D3 (54.7 ± 7.7 ng/mL). In conclusion, D3 given after D2 is less efficient at increasing the plasma status of 25(OH)D3 than D3 given without previous D2 administration. [ABSTRACT FROM AUTHOR]

How hair-coated animals such as dairy cows synthesize endogenous vitamin D3 during exposure to summer sunlight has been unclear since vitamin D3 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 D3 synthesis in the skin during exposure to sunlight. Different scenarios have been suggested but never tested in cows; for example, that vitamin D3 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 D3 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 1500 h for 4 wk in July and August 2009. Blood samples were collected 15 times during the study and analyzed for content of 25-hydroxyvitamin D3 [25(OH)D3] indicative of the animals' vitamin D3 status. Results showed that uncovered cows had a higher 25(OH)D3 concentration in plasma after 28 d of access to sunlight compared with covered cows and that the plasma concentration of 25(OH)D3 was strongly inversely correlated to the body surface area covered. These results are consistent with findings in humans, wherein the vitamin D3 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 D3 synthesis and that cows, like humans, synthesize vitamin D3 evenly over their body surface. That vitamin D3 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 D3, then a relatively even 25(OH)D3 concentration between treatments would be expected, because covered cows would obtain vitamin D3 by grooming uncovered herdmates. [ABSTRACT FROM AUTHOR]

Synthetic α-tocopherol has eight isomeric configurations including four 2R (RSS, RRS, RSR, RRR) and four 2S (SRR, SSR, SRS, SSS). Only the RRR stereoisomer is naturally synthesised by plants. A ratio of 1·36:1 in biopotency of RRR-α-tocopheryl acetate to all-rac-α-tocopheryl acetate is generally accepted; however, studies indicate that neither biopotency of α-tocopherol stereoisomers nor bioavailability between them is constant, but depend on dose, time, animal species and organs. A total of forty growing young male mink were, after weaning, assigned one of the following treatments for 90 d: no α-tocopherol in diet (ALFA_0), 40 mg/kg RRR-α-tocopheryl acetate (NAT_40), 40 mg/kg all-rac-α-tocopheryl acetate (SYN_40) and 80 mg/kg feed all-rac-α-tocopheryl acetate (SYN_80). Mink were euthanised in CO2 and blood was collected by heart puncture. Mink were pelted and liver, heart, lungs, brain and abdominal fat were collected for α-tocopherol stereoisomer analysis. The proportion of RRR-α-tocopherol decreased in all organs and plasma with increasing amount of synthetic α-tocopherol stereoisomers in the diet (P≤0·05), whereas the proportion of all synthetic α-tocopherol stereoisomers increased with increasing amount of synthetic α-tocopherol stereoisomers in the diet (P≤0·05). The proportion of α-tocopherol stereoisomers in plasma, brain, heart, lungs and abdominal fat showed the following order: RRR>RRS, RSR, RSS>Σ2S, regardless of α-tocopherol supplement. The liver had the highest proportion of Σ2S stereoisomers, and lowest proportion of RRR-α-tocopherol. In conclusion, distribution of α-tocopherol stereoisomers differs with dose and form of α-tocopherol supplementation. The results did also reveal the liver's role as the major organ for accumulation of Σ2S α-tocopherol stereoisomers.

The objective of this study was to investigate the possible beneficial effect of vitamin D repletion on certain immune parameters of vitamin D insufficient dairy cows. Twenty dairy cows in late lactation were treated daily with vitamin D in five different ways: sunlight exposure (SUN), D 2 supplementation combined with sunlight exposure (D2SUN), D 2 supplementation (D2), D 3 supplementation (D3), and D 2 and D 3 supplementation combined (D2D3). The cows had very low vitamin D levels at d 0 because of the vitamin D deprivation before the study. After 1 month of vitamin D repletion, all cows had plasma 25(OH)D levels within the normal range. Total 25(OH)D concentration was significantly higher in SUN, D2SUN and D2D3 than D2 or D3 at the end of the study. However, milk yield, as well as protein and fat content of the milk, was not influenced by vitamin D treatments. There was no difference obtained in the measured immune parameters: Leucocyte populations, somatic cell count, immunoglobulin concentrations in plasma and milk, and antigen-stimulated cytokine productions did not change in response to vitamin D repletion or difference in vitamin D sources, and no relations to plasma 25(OH)D levels were identified. Despite the fact that plasma 25(OH)D increased from a very low level to normal range, the present study did not show any effect of vitamin D repletion on the tested immune parameters of healthy dairy cows. Therefore, in this study, it was concluded that repletion to physiologically normal plasma 25-hydroxyvitamin D levels of vitamin D-depleted healthy dairy cows had no influence on immune parameters.

The effect of heat treatment on biohydrogenation of linoleic acid (LA) and linolenic acid (LNA) and formation of stearic acid (SA), cis-9, trans-11 conjugated LA (CLA), trans-10, cis-12 CLA and trans-vaccenic acid (VA) was studied in in vitro incubations with diluted rumen fluid as inoculum and partly defatted flaxseed (DF) and partly defatted sunflower (DS) as test feeds. Feeds were heated in a laboratory oven at 110 °C for 0 (unheated), 45, or 90 min. Michaelis-Menten kinetics was applied for quantifying biohydrogenation rate. The DF heated for 90 min showed the lowest biohydrogenation rate of LNA and LA, indicated by the lowest V max value (P < 0.04 and P < 0.03, respectively). The DS heated for 45 min had the lowest biohydrogenation rate of LNA, indicated by the lowest V max value (P < 0.04). In conclusion, heat treatment decreased biohydrogenation of LA and LNA in DF and LNA in DS.

Vitamin D has been found have various biological effects that may be potent in preventing bovine mastitis. Two forms of vitamin D, vitamin D 2 (D 2 ) and vitamin D 3 (D 3 ), can be hydroxylated to functional metabolites in cattle. The objectives of the present study were to investigate the potential of vitamin D compounds for controlling bovine mastitis using in vitro cell models, and to compare the differences between D 2 and D 3 compounds. Results showed that D 2 compounds have comparable effects to their D 3 analogues on inhibiting MAC-T cell viability in vitro. S. aureus growth was inhibited by high concentrations of D 2 , D 3 , 25(OH)D 2 and 25(OH)D 3 . 25(OH)D 2 and 25(OH)D 3 induced CYP24A1 expression but reduced VDR mRNA expression, whereas the expression of CYP27B1, occludin, and E-cadherin did not change. Additionally, the induction of CYP24A1 expression by 25(OH)D 3 was higher than that of 25(OH)D 2 , which may contribute to their differences in inhibiting cell viability. S. aureus invaded into MAC-T cells and universally inhibited gene expressions. Pre-treat MAC-T cells with 25(OH)D 2 reduced S. aureus adhesion while pre-treatment with 25(OH)D 3 inhibited S. aureus invasion, but neither of the compounds attenuated the S. aureus-induced gene expression reduction. In conclusion, the present study shows that D 2 compounds have comparable effects on inhibiting cell viability and S. aureus invasion to their D 3 analogues in vitro, suggesting that D 2 and its metabolites have potential in controlling bovine mastitis., (Copyright © 2017 Elsevier B.V. All rights reserved.)

A sufficient but balanced vitamin supplementation is a prerequisite for a satisfactory growth pattern and an effective immune system in mink and all other species. The fat-soluble vitamins are very sensitive to over- or under-supply because they interact with each other with respect to dose-response and chemical form. The purpose of the present study was to investigate the effect of increasing the amount of retinol in combination with RRR-α-tocopherol or all-rac-α-tocopherol in the feed given to growing mink on their retinol, cholecalciferol and α-tocopherol concentrations in plasma and selected organs. The results showed that the mink met their retinol requirements from the basal diet, but there were no negative effects of supplying various amounts of retinol on their plasma α-tocopherol concentrations. On the other hand, the study showed that the cholecalciferol status in plasma, assessed as the 25-hydroxycholecalciferol concentration, was low when retinol was supplemented in the feed at high levels. In addition, supplementation with RRR-α-tocopherol in the feed negatively affected the plasma concentration of 25-hydroxycholecalciferol compared with supplementation with all-rac-α-tocopherol. In general, female mink had higher concentrations of fat-soluble vitamins in plasma than male mink.

Vitamin D has become one of the most discussed nutrients in human nutrition, which has led to an increased interest in milk as a vitamin D source. Problems related to fortifying milk with synthetic vitamin D can be avoided by securing a high content of natural vitamin D in the milk by supplying dairy cows with sufficient vitamin D. However, choosing the most efficient route and form of supplementation requires insight into how different vitamin D metabolites are transported in the body of cattle. There are two forms of vitamin D: vitamin D2 (D2) and vitamin D3 (D3). Vitamin D2 originates from fungi on roughage. Vitamin D3 originates either from endogenous synthesis in the skin or from feed supplements. Vitamin D2 is chemically different from, and less physiologically active than, D3. Endogenous and dietary D3 is chemically similar but dietary D3 is toxic, whereas endogenous D3 appears well regulated in the body.

The number of dairy cows without access to pasture or sunlight is increasing; therefore, the content of vitamin D in dairy products is decreasing. Ultimately, declining vitamin D levels in dairy products will mean that dairy products are a negligible source of natural vitamin D for humans. We tested the ability of a specially designed UVB lamp to enhance the vitamin D3 content in milk from dairy cows housed indoors. This study included 16 cows divided into 4 groups. Each group was exposed daily to artificial UVB light simulating 1, 2, 3, or 4 h of summer sun at 56°N for 24 d, and the group with simulated exposure to 2 h of summer sun daily continued to be monitored for 73 d. We found a significant increase in 25-hydroxyvitamin D3 (25OHD3) levels in plasma as well as vitamin D3 and 25OHD3 levels in milk after daily exposure for 24 d in all treatment groups. Extending daily exposure to artificial UVB light to 73 d did not lead to an increase of vitamin D3 or 25OHD3 level in the milk. In conclusion, the change in production facilities for dairy cows providing cows with no access to pasture and sunlight causes a decrease of vitamin D levels in dairy products. This decrease may be prevented by exposing cows to artificial UVB light in the stable., (Copyright © 2015 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

In vitro studies with skin samples or pure precursors of cholecalciferol indicated that cholecalciferol synthesis during UV light exposure is a non-linear process. However, in vitro studies indicate nothing about the relationship between sunlight exposure and physiological cholecalciferol status of living organisms. Due to the lack of cholecalciferol in plant material, this relationship is important for herbivores including domestic cattle, particularly in organic agriculture, because the use of synthetic additives, like cholecalciferol, is restricted in order to fulfil the principles of sustainable organic production. The major physiological metabolite of cholecalciferol is the liver-derived 25-hydroxycholecalciferol (25(OH)D₃). The purpose of the present study was to determine the relationship between sunlight exposure and 25(OH)D₃ status in vivo in large herbivores during mid-summer at 56°N. Five groups of cows were given access to pasture during 15, 30, 75, 150 or 300 min daily for 28 d in June and plasma analysed for 25(OH)D₃. Animals allowed 15, 30 or 75 min of daily access to pasture showed a declining linear relationship between plasma 25(OH)D₃ and sampling day in contrast to animals allowed 150 or 300 min of pasture access which showed linear increasing plasma 25(OH)D₃ status. Determined from the slopes of 25(OH)D₃ concentration curves within treatments, breakeven for maintaining the initial 25(OH)D₃ status of 45 nmol/l was 90 min pasture access per d during summer at 56°N.

Two physiologically important forms of vitamin D exist: vitamin D(2) and vitamin D(3), which by liver based hydroxylase enzymes are converted to 25-hydroxyvitamin D(2) and 25-hydroxyvitamin D(3), respectively. These hydroxylated metabolites of vitamin D are measured in plasma to assess the vitamin D status of animals and humans. Therefore cheap and reliable analytical methods are very much in demand in nutritional and physiological research. After saponification and extraction of plasma or serum samples the current method uses reverse phase high performance liquid chromatography on a C(30) column and with UV detection at 265nm for quantifying vitamin D(2), vitamin D(3), 25-hydroxyvitamin D(2), and 25-hydroxyvitamin D(3). The method proved versatile with respect to plasma lipid content, sample amount, and plasma concentration of the vitamin D metabolites as it was tested using plasma from six different species: cattle, pigs, poultry, mink, horses, and humans. In cattle plasma recoveries were between 86.6 and 101.0%, within day error between 0.9 and 5.9%, and between day error between 0.2 and 1.7%. However, depending on species and sample amount error percentages varied. When running the method on standard reference material® 972 "Vitamin D in human serum" from the National Institute of Standards and Technology (NIST) (Gaithersburg, USA) the results for 25-hydroxyvitamin D(2) and 25-hydroxyvitamin D(3) concentrations were within the boundaries provided by NIST, reflected by Z-scores between 0.1 and 0.9., (Copyright © 2011 Elsevier B.V. All rights reserved.)

Use of synthetic feed additives, including synthetic vitamin D3 (D3) in the feed for cows and other ruminants, is not consistent with the international principles of organic farming. If dairy farmers wish to produce in accordance with the organic principles, production animals would be left with only their endogenous production of D3 from summer sunlight as a source of D3. To examine the impact of supplemental synthetic D3 from the feed on the D3 status of dairy cattle in organic production in Nordic countries, 20 high-yielding dairy cows and 30 dairy steers were divided into two groups: one supplemented with synthetic D3 in the feed and one not supplemented with synthetic D3. Vitamin D3 status of the animals was assessed by measuring the concentration of the liver-derived 25-hydroxyvitamin D3 (25OHD3) in plasma. Results showed that 25OHD3 concentration in plasma from dairy cattle as well as from steers decreased during winter for both supplemented and unsupplemented groups. Unsupplemented cows and steers had approximately 2 ng 25OHD3 per ml plasma during winter, whereas supplemented animals had between 10 (cows) and 30 (steers) ng/ml. During summer and autumn there was no additive effect of supplementing with synthetic D3 since unsupplemented and supplemented animals had the same D3 status at this time of year. In all cows summer concentrations of 25OHD3 were 20-25 ng/ml and in all steers 40-50 ng/ml plasma. The decrease in vitamin D3 status during winter indicates that cows and steers are able to store D3 only to a limited extent. The results also show that cows or steers fed supplemental D3 according to Swedish recommendation throughout the year are not able to maintain their summer value of 25OHD3 during winter.