Relationship of plasma beta-carotene and vitamin A to luteal function in postpartum cattle

The influence of plasma concentrations of beta-carotene and vitamin A on in vivo progesterone production by bovine corpora lutea after gonadotropin-releasing hormone-induced LH release was assessed in 39 postpartum dairy cows. Thirty Holsteins and nine Jerseys were given 100 micrograms gonadotropin-releasing hormone on d 12 of an estrous cycle, which began from 30 to 49 d postpartum. Concentrations of beta-carotene and vitamin A in plasma and progesterone and LH in serum were determined prior to gonadotropin-releasing hormone injection (0 h); serum progesterone and LH concentrations were also determined 1, 2, and 3 h after injection of gonadotropin-releasing hormone. Serum concentrations of progesterone and LH were increased by gonadotropin-releasing hormone. Incremental progesterone production in an analysis of covariance was influenced by breed as well as the interactions of breed with vitamin A, of season with beta-carotene, and of season with vitamin A. The regression coefficients were positive for beta-carotene and negative for vitamin A in all cases. In conclusion, luteal function in the postpartum cow appears to be related to plasma concentrations of beta-carotene and vitamin A.     

Progesterone production in bovine luteal cells treated with drugs that modulate nitric oxide production

The aim of this study was to investigate the influence of nitric oxide (NO) donors (S-nitroso-L-acetyl penicillamine, spermine-NO complex and sodium nitroprusside) and NO synthase inhibitors (N(omega)-nitro-L-arginine methyl ester, N(omega)-nitro-l-arginine, and (+/-)-2-amino-5,6-dihydro-6-methyl-4H-1,3-thiazine) on progesterone production by dispersed bovine luteal cells cultured for 24 h. All NO donors inhibited progesterone production and increased nitrite or nitrate concentration in the medium in a dose-dependent manner. Secretion of progesterone was reduced to 75% (P < 0.01), 56% (P < 0.001) and 37% (P < 0.001) by S-nitroso-L-acetyl penicillamine; to 65% (P < 0.001), 45% (P < 0.001) and 33% (P < 0.001) by spermine-NO complex and to 77% (P < 0.05), 74% (P < 0.01) and 54% (P < 0.001) by sodium nitroprusside treatments at concentrations of 10(-5), 10(-4) and 10(-3) mol l(-1), respectively, compared with the concentration of this hormone measured in cells cultured in medium alone. NO synthase inhibitors decreased significantly (P < 0.05) nitrite or nitrate concentration and increased progesterone secretion with different potency at different doses. Significant increases in progesterone production were observed after N(omega)-nitro-L-arginine methyl ester treatment at a concentration of 10(-5) mol l(-1) and 10(-4) mol l(-1), and after N(omega)-nitro-l-arginine administration at a concentration of 10(-6) mol l(-1) (P < 0.01) and 10(-5) mol l(-1) (P < 0.05), compared with the concentration of this hormone measured in control cells. The results indicate that both NO donors and NO synthase inhibitors regulate steroidogenesis in cultured bovine luteal cells from days 10 to 14 of the oestrous cycle; however, the degree of progesterone inhibition by NO donors and stimulation by NO synthase inhibitors was dependent on the drug used.     

In vitro effects of beta-carotene and vitamin A on peripartum bovine peripheral blood mononuclear cell proliferation

The effects of beta-carotene, retinol, and retinoic acid on function of mononuclear cells during the peripartum period was assessed in vitro. Blood was collected from 14 Holstein cows on wk -4, -1, 0, 1, and 4 postpartum, and mononuclear cells were obtained by gradient centrifugation. Mononuclear cell proliferation induced by concanavalin A was measured in the presence of beta-carotene, retinol (1 x 10(-9) and 1 x 10(-8) M), and retinoic acid (1 x 10(-10) and 1 x 10(-9) M). Retinol and beta-carotene had no effect on spontaneous cell proliferation, whereas retinoic acid was suppressive. However, 1 x 10(-9) M beta-carotene enhanced concanavalin A-induced proliferation at wk -1, whereas 1 x 10(-8) M beta-carotene was suppressive at wk -4. Retinoic acid suppressed concanavalin A-induced proliferation at wk 0, but retinol had no effect. These results suggest a mechanism by which beta-carotene affords the mammary gland protection against infection immediately prepartum.     

Stimulation of luteal cell progesterone production by lipoproteins from cows fed control or fat-supplemented diets.

Plasma lipoproteins from lactating dairy cows fed 0 or 7% supplemental fat were examined for their composition and ability to stimulate luteal cell progesterone production in vitro. Ultracentrifugation was utilized to isolate blood lipoproteins, and heparin affinity chromatography allowed separation of lipoprotein fractions based on the presence (low density lipoproteins) or absence of apolipoprotein B (high density lipoproteins). A portion of high density lipoproteins was fractionated by size, utilizing gel filtration chromatography. Slaughterhouse corpora lutea were dissociated, and plasma lipoproteins were added to the luteal cells on d 3 of culture and incubated for 48 h. In Experiment 1, blood was collected from heifers fed a diet that was not supplemented with fat. The addition of cholesterol from large, high density lipoproteins with a high cholesterol to protein ratio to luteal cultures increased progesterone production by an average of 17% compared with the addition of cholesterol from small, high density lipoproteins with a low cholesterol to protein ratio. In Experiment 2, electrophoretic mobility, apolipoprotein composition, and size of lipoproteins from control and fat-supplemented cows were similar. Lipoproteins from cows assigned to either a control or fat-supplemented diet showed no difference in their ability to stimulate progesterone production. Increased plasma progesterone concentration in lactating dairy cows fed supplemental fat does not appear to be mediated by alterations in lipoprotein composition.     

Role of gap junctions in regulation of progesterone secretion by ovine luteal cells in vitro

To evaluate the role of gap junctions in the regulation of progesterone secretion, two experiments were conducted. In Experiment 1, luteal cells obtained on days 5, 10, and 15 were cultured overnight at densities of 50 x 10(3), 100 x 10(3), 300 x 10(3), and 600 x 10(3) cells/dish in medium containing: (1) no treatment (control), (2) LH, or (3) dbcAMP. In Experiment 2, luteal cells from days 5 and 10 of the estrous cycle were transfected with siRNA, which targeted the connexin (Cx) 43 gene. In Experiment 1, progesterone secretion, Cx43 mRNA expression, and the rates of gap junctional intercellular communication (GJIC), were affected by the day of the estrous cycle, cell density, and treatments (LH or dbcAMP). The changes in progesterone secretion were positively correlated with the changes in Cx43 mRNA expression and the rates of GJIC. Cx43 was detected on the luteal cell borders in every culture, and luteal cells expressed 3beta-hydroxysteroid dehydrogenase. In Experiment 2, two Cx43 gene-targeted sequences decreased Cx43 mRNA expression and progesterone production by luteal cells. The changes in Cx43 mRNA expression were positively correlated with changes in progesterone concentration in media. Thus, our data demonstrate a relationship between gap junctions and progesterone secretion that was supported by (1) the positive correlations between progesterone secretion and Cx43 mRNA expression and GJIC of luteal cells and (2) the inhibition of Cx43 mRNA expression by siRNA that resulted in decreased production of progesterone by luteal cells. This suggests that gap junctions may be involved in the regulation of steroidogenesis in the ovine corpus luteum.     

Bovine vitamin A and beta-carotene intake and lactational status. 1. Responsiveness of peripheral blood polymorphonuclear leukocytes to vitamin A and beta-carotene challenge in vitro

Dietary vitamin A and beta-carotene were assessed on their interaction with lactational status to influence neutrophil function in vitro. Cows were fed 1) 53,000 IU or 2) 213,000 IU vitamin A, or 3) 53,000 IU vitamin A plus 400 mg beta-carotene/cow per d from 6 wk before to 2 wk after dry off. Blood neutrophils were isolated the day of dry off and 2 wk after dry off and incubated with retinol, retinoic acid, or beta-carotene. Phagocytosis and kill of Staphylococcus aureus were measured. Across all treatments, kill was higher after dry off than before dry off. Phagocytosis tended to be lower after dry off than before in cows fed vitamin A only. In vitro, 10(-6) M beta-carotene stimulated phagocytosis after dry off and kill before dry off in cows fed vitamin A only. In general, retinol and retinoic acid suppressed phagocytosis but did not affect kill. Neutrophils from cows fed high amounts of vitamin A were more susceptible to in vitro suppression than those from cows fed adequate amounts of vitamin A. Therefore, vitamin A and beta-carotene supplementation interacts with lactational status to influence the responsiveness of bovine neutrophils to vitamin challenge in vitro.     

Bovine vitamin A and beta-carotene intake and lactational status. 2. Responsiveness of mitogen-stimulated peripheral blood lymphocytes to vitamin A and beta-carotene challenge in vitro

The interaction of dietary vitamin A and beta-carotene with lactational status on the in vitro proliferation of mitogen-induced peripheral blood lymphocytes was studied. Cows were fed (IU/cow per d) 1) 53,000 IU vitamin A, 2) 213,000 IU vitamin A, or 3) 53,000 IU vitamin A plus 400 mg beta-carotene from 6 wk before to 2 wk after dry off. Lymphocytes were incubated with retinol, retinoic acid, or beta-carotene. Concanavalin A-induced blastogenesis was inhibited by 10(-6) M retinol and 10(-8) M retinoic acid in cows fed 53,000 IU vitamin A before dry off. In contrast, 10(-7) M retinol and 10(-7) M retinoic acid stimulated Concanavalin A-induced blastogenesis for cows fed vitamin A plus beta-carotene before dry off. After dry off, retinol and retinoic acid did not affect Concanavalin A-induced blastogenesis in all treatment groups. In vitro, 10(-5) M beta-carotene inhibited Concanavalin A-induced blastogenesis before and after dry off in all treatment groups. Blastogenesis in the absence of mitogen stimulation or induced by lipopolysaccharide was inhibited by all vitamins before and after dry off in all treatment groups. These data indicate that vitamin A and beta-carotene supplementation interact with lactational status to influence the responsiveness of bovine blood lymphocytes to vitamin challenge in vitro.     

Modulation of fat-soluble vitamin concentrations and blood mononuclear leukocyte populations in milk replacer-fed calves by dietary vitamin A and beta-carotene

Dairy calves (n = 18), separated from dams at birth, were fed 1 L of pooled-colostrum. For the remaining 7 wk of the study, they were fed one of three diets consisting of either a custom-formulated milk replacer without vitamin A (controls), or supplemented with retinyl palmitate (equivalent to 32,000 IU of vitamin A/d) or with beta-carotene (equivalent to 20,000 IU of vitamin A/d). Plasma retinol, beta-carotene, and RRR-alpha-tocopherol concentrations were lowest at birth, and increased substantially from birth to 1 wk postpartum in all groups, a probable consequence of ingestion of colostrum. From 1 to 7 wk of age, retinol concentrations were greatest in retinyl palmitate-supplemented calves, intermediate in beta-carotene-supplemented calves and lowest in control calves. At 2, 3, 5, 6, and 7 wk, RRR-alpha-tocopherol concentrations were lower in retinyl palmitate-supplemented calves than in control calves. A negative correlation between plasma retinol and vitamin E concentrations existed from wk 2 to 7, suggesting vitamin A influences the absorption and distribution of RRR-alpha-tocopherol. Supplemental retinyl palmitate, but not beta-carotene, was associated with a reduction in the percentage of blood mononuclear leukocytes expressing CD2, CD4, and CD8-T cell antigens and interleukin-2 receptors. By wk 7, leukocyte populations from retinyl palmitate-supplemented calves were more similar to those from adult cattle than those from control calves, suggesting that supplemental vitamin A, as retinyl palmitate, affects the maturation of the neonatal immune system. Differences in the composition of blood mononuclear leukocyte populations may represent changes in immune competency.     

Molecular control of luteal secretion of progesterone

Cholesterol provided by low- or high-density lipoprotein is the precursor for biosynthesis of progesterone. Once inside the cell, cholesterol can be used for steroidogenesis or esterified with long-chain fatty acids and stored as cholesterol esters in lipid droplets. When it is needed for steroidogenesis, free cholesterol is transported to the mitochondrion via a mechanism that involves cytoskeletal elements and sterol carrier proteins. Cytochrome P450 cholesterol side chain cleavage enzyme complex converts the cholesterol to pregnenolone, which is then converted to progesterone by 3beta-hydroxysteroid dehydrogenase/delta5,delta4 isomerase in the smooth endoplasmic reticulum. Transport of cholesterol from the cytoplasm to the inner mitochondrial membrane is both the rate-limiting step in progesterone biosynthesis and the step most acutely influenced by second messengers. Steroidogenic acute regulatory protein (StAR) and peripheral-type benzodiazepine receptors (PBR) are involved in this transport. StAR may bind cholesterol in the cytosol and transport it to the mitochondrial membrane where PBR is involved in transport from the outer to the inner mitochondrial membrane. Phosphorylation of StAR by protein kinase A (PKA) stimulates cholesterol transport, whereas phosphorylation by PKC may inhibit this process. Endozepine, the natural ligand for PBR, also appears to be involved in regulation of the rate of cholesterol transport to the inner mitochondrial membrane and to play a role in the stimulatory effects of PKA on steroidogenesis. Increased concentrations of endozepine were detected in large luteal cells, and may explain the increased progesterone secretion from this type of cell. Fluorescence energy transfer procedures indicate that StAR associates with PBR in mitochondrial membranes. A model is presented for the proposed interactions of StAR, PBR and endozepine in the transport of cholesterol from the outer to the inner mitochondrial membrane.     

Effect of dietary vitamin A and beta-carotene on polymorphonuclear leukocyte and lymphocyte function in dairy cows during the early dry period

Vitamin A and beta-carotene improved mammary health in dairy cows around dry off. To define possible mechanisms, cows were fed 1) 53,000 IU vitamin A, 2) 213,000 IU vitamin A, or 3) 53,000 IU vitamin A plus 400 mg beta-carotene/cow per d (n = 10/treatment) from 6 wk before to 2 wk after dry off. Blood polymorphonuclear neutrophil function (phagocytosis, kill, and chemotaxis) and lymphocyte proliferation were measured at wk -6, 0 (dry off), and 2. Concentrations of vitamin A in serum did not differ across vitamin treatments. beta-Carotene in serum was elevated in cows fed beta-carotene. Treatment did not influence phagocytosis or kill. Kill ability increased after dry off in all treatment groups, but phagocytosis tended to decrease after dry off in cows fed vitamin A only. Lymphocyte blastogenesis stimulated by concanavalin A on wk 2 for cows fed 53,000 IU vitamin A but did not vary in the other two groups. Lipopolysaccharide-stimulated blastogenesis peaked at wk 0 and then decreased to pretreatment values by wk 2 in cows fed 213,000 IU vitamin A. These data indicate lymphocyte function is influenced by vitamin A supplementation and that beta-carotene supplementation seems to exert a stabilizing effect on neutrophil and lymphocyte function during the period around dry off.     

Effect of intrauterine and intramuscular administration of recombinant bovine interferon alpha 1 on luteal lifespan in cattle

Intrauterine and intramuscular administration of interferon was tested for effectiveness in extending luteal lifespan in cattle. Intrauterine infusion of 1 mg of recombinant bovine interferon-alpha I1, twice daily, to lactating dairy cows from d 14 to 21 after estrus extended interestrous interval (30.4 +/- 1.91 d versus 24.8 +/- .58 d) and functional lifespan of the corpus luteum (28.4 +/- 2.01 d versus 23.6 +/- .75 d). In another experiment, twice daily intramuscular injection of 20 mg interferon to Simmental heifers from d 15 to 19 extended interestrous intervals (24.6 +/- 1.36 d versus 20.6 +/- .49 d) and functional lifespan of the corpus luteum (23.2 +/- .37 d versus 20.2 +/- .73 d). In a third experiment, pubertal dairy heifers received twice daily intramuscular injections of 0, 2.5, 5.0, or 10.0 mg/injection of interferon from d 14 to 21 after estrus. The three interferon-treated groups had longer interestrous intervals and functional luteal lifespans than the control group. Interestrous intervals were 22.0 +/- .68, 24.0 +/- 1.14, 24.6 +/- 1.17, and 25.4 +/- .97 d, respectively. The present data strengthen the theory that an interferon-alpha-like molecule can regulate luteal function in cattle. Such a regulatory compound might prove useful in schemes to reduce embryonic mortality caused by aberrant secretion of embryonic interferon.     

The effect of human chorionic gonadotrophin on the expression of progesterone receptors in human luteal cells in vivo and in vitro

The human corpus luteum expresses genomic progesterone receptors (PRs) suggesting that progesterone may have an autocrine or paracrine role in luteal function. We hypothesised that the reduction in luteal PR reported in the late-luteal phase augmented progesterone withdrawal and had a role in luteolysis. We therefore tested the hypothesis that luteal rescue with human chorionic gonadotrophin (hCG) would maintain PR expression. PR was immunolocalised to different cell types in human corpora lutea (n = 35) from different stages of the luteal phase and after luteal rescue with exogenous hCG. There was no change in the staining intensity of theca-lutein cell or stromal cell PR throughout the luteal phase or after luteal rescue. In the late-luteal phase, granulosa-lutein cell PR immunostaining was reduced (P < 0.05) but the trend to reduction was also seen after luteal rescue with hCG (P = 0.055). To further investigate the effect of hCG on granulosa-lutein cell PR expression, an in vitro model system of cultured human luteinised granulosa cells was studied. Cells were cultured for 12-13 days exposed to different patterns of hCG and aminoglutethamide to manipulate progesterone secretion (P < 0.0001). Expression of PR A/B and PR B isoforms was examined by quantitative real-time RT-PCR. PR A/B mRNA was lower (P < 0.05) after 11-13 days of culture than after 7 days of culture. This reduction could not be prevented by hCG in the presence (P < 0.05) or absence (P < 0.05) of stimulated progesterone secretion. The expression of PR B mRNA showed a similar pattern (P = 0.054). Simulated early pregnancy in vivo and hCG treatment of luteinised granulosa cells in vitro did not appear to prevent the down-regulation of PR seen during luteolysis.     

Effects of supplemental vitamin A or beta-carotene during the dry period and early lactation on udder health.

Effects of vitamin A or beta-carotene supplementation during the dry period and early lactation on the frequency of new intramammary infection and clinical mastitis and on SCC and milk yield were examined. Eighty-two Holstein cows were randomly assigned to one of three groups: 1) 50,000 IU/d of vitamin A per cow (approximately equivalent to 1978 NRC recommended daily intake for dairy cows); 2) 170,000 IU/d of vitamin A per cow; or 3) 50,000 IU/d of vitamin A plus 300 mg of beta-carotene per cow. Cows were supplemented during the 2 wk before drying off, throughout the dry period, and for the first 6 wk of lactation. Concentrations of serum vitamin A did not differ among treatment groups but tended to decrease for all treatment groups from 14 d before drying off to calving. After calving, serum vitamin A tended to increase in all groups through wk 6 of lactation. Serum beta-carotene tended to be higher in beta-carotene-supplemented cows at dry-off, in the early dry period, and again during lactation. Serum beta-carotene decreased sharply in all groups during the prepartum period. The frequency of clinical mastitis and of new intramammary infection during the dry period, near parturition, and for the first 6 wk of lactation did not differ among treatment groups. The percentage of quarters newly infected over the entire trial was 26.8 in the control, 25.0 in the high vitamin A, and 30.6 in the beta-carotene group. Pathogens isolated most frequently were coagulase-negative staphylococci, streptococci other than Streptococcus agalactiae, and coliforms.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of bovine somatotropin on production and reproduction in prepubertal Friesian heifers

This experiment was conducted to evaluate the effect of bST on average daily gain, onset of puberty, first lactation milk yield, and reproductive efficiency in Friesian heifers. Heifers (n = 16 per treatment) were allocated to either: 1) control (1.5 ml of vehicle) or 2) bST (15 mg of bST in 1.5 ml of vehicle) using a randomized complete block design. Subcutaneous injections of bST were administered once daily from 7 mo of age for 120 d or until heifers reached puberty. Heifers were weighed every 2 wk, and blood samples were collected twice weekly after heifers reached 200 kg. Progesterone concentrations were used to determine onset of puberty. Heifers were bred between 16 and 18 mo of age and, following parturition, milk yield and composition were recorded twice weekly and once every 2 wk, respectively. Heifers assigned to bST treatment had an average daily gain (kg) of .8 compared with .7 in control heifers. Number of days from birth to onset of puberty for bST-treated heifers was 401 compared with 381 for control heifers. Treatment with bST had no effect on milk yield, milk composition, or reproductive efficiency during the first lactation. These data demonstrate that daily administration of bST to Friesian heifers from 7 to 11 mo of age does not affect average daily gain, onset of puberty, reproductive efficiency, or first lactation milk yield of heifers.     

Induction of nitric oxide production by bovine mammary epithelial cells and blood leukocytes.

A recent study from our laboratory has shown that significant amounts of nitric oxide are released by somatic cells recovered during endotoxin-induced mastitis. The present study was undertaken to investigate which cell type(s) among milk somatic cell population can produce nitric oxide under inflammatory conditions. Nitric oxide release from mammary epithelial cell lines and from bovine neutrophils and monocytes extracted from blood was measured in response to cytokines and Escherichia coli lipopolysaccharides. An epithelial cell line isolated from bovine mammary gland, FbE cells, was found to release nitric oxide after exposure to interleukin-1beta. This nitric oxide production was completely abolished by addition of L-N6-(1-iminoethyl) lysine, a potent inducible nitric oxide synthase inhibitor. Bovine monocytes produced nitric oxide in response to recombinant bovine interferon-gamma alone or in combination with E. coli lipopolysaccharides. In these cells, nitric oxide release was reduced by the addition of inducible nitric oxide synthase inhibitors L-N6-(1-iminoethyl) lysine and aminoguanidine. Lipopolysaccharides and recombinant bovine interferon-gamma increased nitric oxide synthase mRNA in neutrophils, but nitric oxide release could not be detected under any of the experimental conditions used. These results show that bovine epithelial cells and mononuclear phagocytes produce nitric oxide under inflammatory conditions and suggest that these cell populations are responsible for nitric oxide release observed during mastitis.     

Effect of bovine interferon on acute changes in body temperature and serum progesterone concentrations in heifers

Bovine interferon-alpha I1 has extensive sequence and functional homology with the antiluteolytic protein, bovine trophoblast protein-1. Because of the possible use of interferon-alpha I1 as a drug that supplements embryonic secretion of bovine trophoblast protein-1, interferon-alpha I1 was tested for other biological actions that might affect its usefulness as a fertility-enhancing treatment. Experiments were performed to evaluate whether interferon-alpha I1 causes hyperthermia and an acute depression in circulating concentrations of progesterone. In four experiments, intramuscular administration of interferon-alpha I1 (range 1.25 to 20 mg) caused hyperthermia; average peak body temperatures of 40 to 40.4 degrees C occurred 2.5 to 6 h after injection. Temperatures returned to baseline 12 to 16 h later. The rise in rectal temperature could be reduced, but not totally alleviated, with concomitant administration of an inhibitor of prostaglandin synthesis. The maximal hyperthermic response was similar when interferon-alpha I1 was delivered via osmotic minipumps or through a series of intramuscular injections. The hyperthermic response decreased with repeated daily exposure to interferon-alpha I1. The increase in rectal temperatures was associated temporally with a decrease in serum progesterone. Effects of interferon-alpha I1 on body temperature and circulating progesterone could possibly limit its effectiveness in enhancing fertility.     

Characteristics of prolonged luteal phase identified by milk progesterone concentrations and its effects on reproductive performance in Holstein cows

A database of milk progesterone profiles consisting 497 lactations in 3 dairy herds from northern and western regions of Japan was used to identify the characteristics and associated risk factors for prolonged luteal phase (PLP) and its effects on subsequent reproductive performance in high-producing Holstein cows. Milk samples were collected twice weekly and progesterone concentrations in whole milk were determined by ELISA. Herds were visited monthly and examined by vaginoscopy and transrectal palpation. Resumption of ovarian cyclicity within 35 d postpartum followed by regular cycles was considered normal. Prolonged luteal phase was defined when progesterone concentrations were ≥5 ng/mL for ≥20 d of duration in any cycle postpartum. Delay of first ovulation to 35 to 60 d (delayed first ovulation type I), >60 d (delayed first ovulation type II), a luteal phase of <14 d except in the first cycle (short luteal phase), and the absence of luteal activity >14 d between 2 cycles (cessation of cyclicity) were the other categories of abnormal ovarian resumptions considered. The overall incidence of PLP in the 3 herds was 11.9% and a significantly higher proportion of PLP was observed in the first cycle postpartum compared with the second and third cycles. Approximately 83% of the PLP were 20 to 28 d in duration, and maximum progesterone concentration was significantly higher when PLP lasted >35 d compared with PLP of 20 to 35 d in duration. Higher parity, commencement of luteal activity ≤28 d postpartum, and postpartum complications significantly increased the occurrence of PLP within 90 d postpartum. Cows with PLP showed reduced conception rate to first artificial insemination (AI) and reduced pregnancy proportions within 100, 150, and 210 d postpartum. Based on survival analysis, PLP was associated with a 56% reduction in relative pregnancy rate and a 36% reduction in AI submission rate. Cows that experienced PLP had a longer interval from calving to first AI (74 d) and from calving to pregnancy (141 d) than cows without PLP (53 and 80 d), respectively. In conclusion, 11.9% of lactations had PLP, of which approximately two-thirds were seen in the first cycle postpartum. Most of the PLP were 20 to 28 d in duration. Higher parity, postpartum complications, and early commencement of luteal activity postpartum increased the risk for PLP. Occurrence of PLP adversely affected fertility by reducing pregnancy proportions and extending calving to conception interval.     

Peripartum changes of plasma and milk vitamin A and beta-carotene among dairy cows with or without mastitis

Over 12 mo we studied the relationship between peripartum concentrations of vitamin A and beta-carotene in blood plasma and milk of 93 Holsteins with or without subsequent mastitis. Blood was sampled daily from 7 days prepartum through 7 days postpartum and on alternate weeks through wk 10 of lactation. Milk samples were collected daily for 7 days postpartum and then biweekly for 10 wk. Somatic cell counts were on biweekly milk samples. Vitamin A and beta-carotene of blood plasma decreased rapidly prepartum to reach minimum concentrations at calving (vitamin A) or on day 4 to 6 postpartum (beta-carotene). Thereafter, both vitamin A and beta-carotene increased rapidly through 10 wk postpartum. Concentrations of vitamin A and beta-carotene in colostrum were higher than concentrations in milk. Cows with mastitis (somatic cells greater than 500,000 cells/ml milk) had lower vitamin A in blood plasma during days 0 to 7 and wk 2 and 4 postpartum than cows without mastitis. When data were analyzed with loge of somatic cell count as an independent regression variable, results were similar. In contrast to vitamin A, peripartum beta-carotene in blood plasma was higher among mastitic cows and was related to higher loge of somatic cell count. No significant difference was observed between mastitic and non-mastitic cows for vitamin A and beta-carotene in milk. Lower concentrations of plasma vitamin A and higher concentrations of beta-carotene during the immediate postpartum period were associated with higher milk somatic cell counts among dairy cows during lactation.     

Short communication: effect of vitamins E and C on cortisol production by bovine adrenocortical cells in vitro

The aim was to determine if vitamins E and C inhibit the release of cortisol from bovine adrenocortical cells when stimulated with ACTH in vitro. A factorial arrangement of treatments was used to culture bovine adrenocortical cells with different concentrations of vitamins E and C [(+)-α-tocopherol at 0, 2.3, and 16 μM and l-ascorbic acid at 0, 15, and 50 μM]. After 3 and 7 d of vitamin treatments, cell cultures were stimulated with ACTH (1 nM) for 24 h and the culture medium extracted to measure cortisol released by the cells using HPLC with UV detection. Vitamin E, vitamin C, or their combination did not affect the amount of cortisol released by the adrenal cultures to the media. Cortisol released by the adrenal cultures ranged from 33.6 ± 6.85 to 49.7 ± 8.01 nmol per 10⁷ cells. The modulation effect of vitamins E and C on the stress response does not take place at the cortex of the adrenal gland.     

Preservation of beta-carotene during production and storage of sterilized milk]

Safety of beta-carotene in sterilized dairy products under different methods of sterilization, keeping conditions, packing and food form of carotene has been investigated. The standard documentation on sterilized milk "Provita" with different level of fat and content of beta-carotene (0.25 mg/100 g) has been developed.