The determination of the vitamin D metabolites on a single plasma sample: changes during parturition in dairy cows.

Methods have been developed for the precise measurement of the major known vitamin D metabolites in a single sample of cow plasma (~5 ml). The procedure involves initial extraction with methylene chloride-methanol followed by chromatography on Sephadex LH-20. 25-Hydroxyvitamin D₂ and 25-hydroxyvitamin D₃ were determined using high-pressure liquid chromatography and comparing ultraviolet absorption peak height with absorption peak heights of standards. The dihydroxylated metabolites were further purified and resolved by high-pressure liquid chromatography and determined by radioligand binding assays. The assays were employed to measure the total vitamin D metabolite levels in the plasma of paretic and normal dairy cows at parturition. Parturition had no effect on 25-hydroxyvitamin D levels in either group of cows (paretic, 37–44 ng/ml; normal, 35–38 ng/ml). However, normal cows did show lower mean 25-hydroxyvitamin D levels at every sampling period with the lowest levels in both groups occurring at 7 days postpartum. Plasma 25, 26-dihydroxyvitamin D was higher in paretic animals prepartum and at parturition (0.7–1.0 ng/ml) when compared to nonparetic animals (0.4–0.45 ng/ml). Similar levels (0.6 ng/ml) were observed in both groups postpartum. Cows developing parturient paresis showed a significant (P < 0.05) elevation of 1,25-dihydroxyvitamin D at parturition with a maximum level of 350 pg/ml attained at 1 day postpartum compared to prepartum levels of 60 pg/ml. Normal animals also showed a rise in plasma levels of 1,25-dihydroxyvitamin D with a maximum level of 185 pg/ml observed at 1 day postpartum. Plasma 24,25-dihydroxyvitamin D was initially higher in paretic cows (1.9 ng/ml) with a significant (P < 0.05) drop to 1.05 ng/ml occurring at parturition. This level was maintained for 7 days postpartum. The levels of this steroid were maintained at 1.3–1.4 ng/ml in the normal cows throughout the entire sampling period.     

Effect of dietary beta-carotene on the accumulation of beta-carotene and vitamin A in plasma and tissues of gilts

The absorption of beta-carotene in pigs is limited. Nevertheless beta-carotene might positively affect reproduction. In this study the absorption and tissue distribution of beta-carotene as well as its function as precursor of vitamin A was investigated in gilts that were fed according to one of three dietary treatments: VA (4000 IU vitamin A), VA + VA (4000 IU + 8300 IU) and VA + BC (4000 IU + 100 mg beta-carotene per kg diet) for 14 weeks. Only in the VA + BC group was beta-carotene detected in plasma (1-8 ng x mL(-1)), liver, adrenals and corpora lutea, indicating that pigs absorb intact beta-carotene at low rates. Liver levels of vitamin A were higher (P < 0.01) at comparable levels in the VA + VA and VA + BC group than in the VA group, indicating a conversion rate of beta-carotene to vitamin A of 40 to 1 on the basis of weight for beta-carotene at this level (100 mg x kg(-1)) in the diet. Higher levels of vitamin A in the uterus of the VA + BC group (P < 0.01) as well as the accumulation of beta-carotene in adrenals and corpora lutea might reflect some influence of beta-carotene on local vitamin A metabolism which might be of importance for reproductive performance in gilts.     

Effect of injection of beta-carotene or vitamin E and selenium on fertility of lactating dairy cows

Experiments tested whether supplemental antioxidants improved fertility. To test effects of beta-carotene, cows in a hot environment were injected with prostaglandin F2 alpha (PGF2 alpha) and were given 3 injections, i.m., of 800 mg beta-carotene or saline at Days -6 and -3 before the anticipated date of insemination and at insemination (n = 37-41 inseminated cows/group). There was no effect of beta-carotene on the proportion of cows detected in estrus following PGF2 alpha, timing of estrus after PGF2 alpha injection or pregnancy rate in inseminated cows. In a second trial, cows in a temperate climate received intramuscular injections of vitamin E (500 mg) and selenium (50 mg) at 30 d post partum (n = 97) or were untreated controls (n = 89). Treatment did not affect interval from calving to first insemination or the proportion of cows pregnant at first service, but it increased the pregnancy rate at second service (69.8 vs 52.1%; P = 0.07) and reduced services per conception (1.7 vs 2.0; P < 0.05) and interval from calving to conception (84.6 vs 98.1; P < 0.05). Thus, injection of vitamin E and selenium increased fertility in cattle that did not become pregnant at first service.     

Concentrations of vitamin A, beta-carotene and vitamin E in individual bovine follicles of different quality

The degree of atresia of the follicle had no influence on the intrafollicular concentrations of beta-carotene, vitamin E and cholesterol. This might result from the passive transfer of these substances from blood to follicular fluid bound to high density lipoproteins. However, concentrations of vitamin A in follicular fluid were significantly (P less than 0.001) influenced by follicle quality, with highest concentrations (0.32 microgram/ml) in non-atretic follicles and lowest values (0.15 microgram/ml) in greatly atretic follicles. The higher concentrations of vitamin A in healthy follicles might be due to a local conversion of beta-carotene into vitamin A in follicular structures. By influencing hormone and protein synthesis, vitamin A may have a potential for local modulation of follicular development and therefore be one of the factors controlling recruitment, selection and growth of the dominant follicle in cattle.