Effect of 25-hydroxyvitamin D3 during prepartum transition and lactation on production,reproduction, and health of lactating dairy cows

We hypothesized that feeding 25-hydroxyvitamin D₃ [25-(OH)D₃] during lactation and prepartum in conjunction with negative dietary cation-anion difference diets would improve milk production, increase the probability of pregnancy, and reduce the incidence of postcalving diseases. Cows from 4 dairies with prepartum transition diets negative in dietary cation-anion difference were used in 2 randomized cohort experiments. In Experiment 1 (Exp. 1), cows were assigned to control [CON; n = 645; no 25-(OH)D₃] or treatment [TRT; n = 537; 2 mg/d of 25-(OH)D₃ from ～21 d prepartum to parturition and 1 mg/d in lactation] groups at ～21 d prepartum. Cows were monitored for weekly milk yield, milk composition every 60 d, and health and reproductive measures. In Experiment 2 (Exp. 2), cows (n = 2,064; median 147 d in milk) were assigned to 4 groups and monitored for the same measures as in Exp. 1 to the end of that lactation (L1), the subsequent transition (～21 d prepartum to parturition), and the next lactation (L2). Groups were as follows, with the amount of 25-(OH)D₃ fed (mg/d) indicated in parentheses for L1, transition, and L2, respectively: (A) control-control (CON-CON; 0–0–0), (B) treatment-treatment (TRT-TRT; 1–2–1), (C) control-treatment (CON-TRT; 0–2–1), and (D) treatment-control (TRT-CON; 1–0–0). For L1, a total of 1,032 cows entered the control groups A or C and a total of 1,032 cows in groups B or D. The number of cows in groups A to D that entered L2 was 521, 523, 273, and 248, respectively. Blood calcium, phosphorus, and 25-(OH)D₃ concentrations were measured from 17 cows/group at 5 times. In Exp. 1, TRT cows had 0.2 lower log somatic cell count than CON cows (4.21 ± 0.045 vs. 4.01 ± 0.050, respectively) and multiparous TRT cows had 41 ± 23% higher probability of pregnancy/day than multiparous CON cows, resulting in a 22-d median decrease in time to pregnancy. Primiparous TRT cows had 1.67 ± 0.40 times greater odds of mastitis/day than primiparous CON cows. In Exp. 2 TRT-TRT cows had between 16 and 29% lower probability to be bred/day than other groups. Multiparous CON-CON and TRT-CON cows had 20 ± 8% and 30 ± 17% greater probability of pregnancy, respectively, than multiparous TRT-TRT cows. Serum calcium concentrations were not affected by group, but phosphorus and 25-(OH)D₃ concentrations were highest in the TRT-TRT cows. The study provides further insights into the use of 25(OH)D₃ in transition and lactation.     

Effect of prepartum source and amount of vitamin D supplementation on lactation performance of dairy cows

The objectives of this experiment were to determine the effects of supplementing 25-hydroxyvitamin D₃ (calcidiol, CAL) compared with vitamin D₃ (cholecalciferol, CHOL) at 1 or 3 mg/d in late gestation on production outcomes of dairy cows. One hundred thirty-three parous and 44 nulliparous pregnant Holstein cows were enrolled in the experiment. Cows were blocked by parity and previous lactation milk yield (parous) or genetic merit (nulliparous) and assigned randomly to receive 1 or 3 mg/d of CAL or CHOL in a 2 × 2 factorial arrangement of treatments (CAL1, CAL3, CHOL1, and CHOL3). Treatments were provided to individual cows as a top-dress to the prepartum diet from 250 d in gestation until parturition. The prepartum diet had a dietary cation-anion difference of ?128 mEq/kg of dry matter. Production and disease were evaluated for the first 42 d in milk, and reproduction was evaluated to 300 d in milk. Incidence of postpartum diseases did not differ among treatments. Feeding CAL compared with CHOL increased yields of colostrum and colostrum fat, protein, and total solids, resulting in an increased amount of net energy for lactation secreted as colostrum (CHOL = 7.0 vs. CAL = 9.0 ± 0.7 Mcal). An interaction between source and amount was observed for milk yield: CAL3 increased milk yield compared with CHOL3 (CHOL3 = 34.1 vs. CAL3 = 38.7 ± 1.4 kg/d) but milk yield did not differ between CAL1 and CHOL1 (CHOL1 = 36.9 vs. CAL1 = 36.4 ± 1.4 kg/d). Concentrations of serum calcidiol on day of calving and average serum Ca from d 2 to 11 postpartum were positively associated with milk yield in the first 42 d in milk. Interactions between source and amount of vitamin D were also observed for pregnancy after first AI: the percentage of cows receiving CHOL1 and CAL3 that became pregnant was smaller than that of cows receiving CHOL3 and CAL1. However, pregnancy per AI and pregnancy by 300 d in milk did not differ among treatments. Overall, CAL3 increased milk yield compared with CHOL3, whereas in cows fed 1 mg/d (CAL1 and CHOL1), the source of vitamin D generally had no effect. The effect of CAL3 may be explained in part by serum CAL concentrations and postpartum serum Ca, which were associated with milk yield.     

Effects of prepartum dietary cation-anion difference and source of vitamin D in dairy cows: Vitamin D,mineral, and bone metabolism

Pregnant Holstein cows, 28 nulliparous and 51 parous, were blocked by parity and milk yield and randomly allocated to receive diets that differed in dietary cation-anion difference (DCAD), +130 or ?130 mEq/kg, and supplemented with either calcidiol or cholecalciferol at 3 mg/11 kg of dry matter from 255 d of gestation until parturition. Blood was sampled thrice weekly prepartum, and on d 0, 1, 2, 3, 6, 9, 12, 15, 18, 21, 24, 27, and 30 postpartum to evaluate effects of the diets on vitamin D, mineral and bone metabolism, and acid-base status. Blood pH and concentrations of minerals, vitamin D metabolites, and bone-related hormones were determined, as were mineral concentrations and losses in urine and colostrum. Supplementing with calcidiol increased plasma concentrations of 25-hydroxyvitamin D₃, 3-epi 25-hydroxyvitamin D₃, 25-hydroxyvitamin D₂, 1,25-dihydroxyvitamin D₃, and 24,25-dihydroxyvitamin D₃ compared with supplementing with cholecalciferol. Cows fed the diet with negative DCAD had lesser concentrations of vitamin D metabolites before and after calving than cows fed the diet with positive DCAD, except for 25-hydroxyvitamin D₂. Feeding the diet with negative DCAD induced a compensated metabolic acidosis that attenuated the decline in blood ionized Ca (iCa) and serum total Ca (tCa) around calving, particularly in parous cows, whereas cows fed the diet with positive DCAD and supplemented with calcidiol had the greatest 1,25-dihydroxyvitamin D₃ concentrations and the lowest iCa and tCa concentrations on d 1 and 2 postpartum. The acidogenic diet or calcidiol markedly increased urinary losses of tCa and tMg, and feeding calcidiol tended to increase colostrum yield and increased losses of tCa and tMg in colostrum. Cows fed the diet with negative DCAD had increased concentrations of serotonin and C-terminal telopeptide of type 1 collagen prepartum compared with cows fed the diet with positive DCAD. Concentrations of undercarboxylated and carboxylated osteocalcin and those of adiponectin did not differ with treatment. These results provide evidence that dietary manipulations can induce metabolic adaptations that improve mineral homeostasis with the onset of lactation that might explain some of the improvements observed in health and production when cows are fed diets with negative DCAD or supplemented with calcidiol.     

Effect of source and amount of vitamin D on serum concentrations and retention of calcium,magnesium, and phosphorus in dairy cows

The objectives of the experiment were to determine the effects of supplementing 2 amounts of 25-hydroxyvitamin D₃ (calcidiol; CAL) compared with equal amounts of vitamin D₃ (cholecalciferol; CHOL) on serum concentrations, absorptions, and retentions of Ca, Mg, and P in periparturient dairy cows. One hundred seventy-seven (133 parous and 44 nulliparous) pregnant Holstein cows were enrolled in the experiment. Cows were blocked by parity and previous lactation milk yield (parous) or genetic merit for energy-corrected milk yield (nulliparous) and assigned randomly to receive 1 or 3 mg/d of CAL or CHOL in a 2 × 2 factorial arrangement of treatments. Treatments were provided to individual cows as a top-dress to the prepartum diet from 250 d gestation until parturition. The prepartum diet had a dietary cation-anion difference of ?128 mEq/kg of dry matter. All cows were fed a common postpartum diet containing 46 μg of vitamin D₃/kg of dry matter without further supplementation of treatments. Concentrations of vitamin D metabolites, Ca, Mg, and P in serum were measured pre- and postpartum, in addition to total-tract digestibility and urinary excretion of Ca, Mg, and P in the prepartum period. Feeding 3 mg compared with 1 mg of CAL increased serum 25-hydroxyvitamin D₃ (CAL1 = 94 vs. CAL3 = 173 ± 3 ng/mL). In comparison, the increment in serum 25-hydroxyvitamin D₃ from feeding 3 mg compared with 1 mg of CHOL was small (CHOL1 = 58 vs. CHOL3 = 64 ± 3 ng/mL). Feeding CAL increased prepartum concentration of P in serum compared with CHOL (CHOL = 1.87 vs. CAL = 2.01 ± 0.02 mM), regardless of the amount fed, but neither source nor amount affected prepartum Ca or Mg in serum. Feeding CAL increased serum Ca and P for the first 11 d postpartum compared with CHOL (CHOL = 2.12 vs. CAL = 2.16 ± 0.01 mM serum Ca; CHOL = 1.70 vs. CAL = 1.78 ± 0.02 mM serum P) but the amount of vitamin D did not affect postpartum concentrations of Ca, Mg, and P in serum. Feeding CAL increased prepartum apparent digestibility of Ca compared with CHOL (CHOL = 26.6 vs. CAL = 33.5 ± 2.8%) but treatments did not affect Ca retention prepartum. Neither source nor amount of vitamin D affected Mg and P apparent digestibility, but CAL decreased the concentration of P excreted in urine during the prepartum period (CHOL = 1.8 vs. CAL = 0.8 ± 0.3 g/d). Calcidiol tended to increase the amount of Ca secreted in colostrum (CHOL = 9.1 vs. CAL = 11.2 ± 0.9 g/d) and Ca excreted in urine postpartum (CHOL = 0.4 vs. CAL = 0.6 ± 0.1 g/d) compared with CHOL. Collectively, feeding CAL at 1 or 3 mg/d compared with CHOL in the last 24 d of gestation is an effective way to increase periparturient serum P concentration and postpartum serum Ca of dairy cows fed a prepartum diet with negative DCAD.     

Effect of dietary supplements of biotin,intramuscular injections of vitamin B12, or both on postpartum lactation performance in multiparous dairy cows

The current study was conducted to investigate the effects of dietary supplementation of biotin, intramuscular injections of vitamin B₁₂ (VB₁₂), or both beginning at the prepartum period on feed intake and lactation performance in postpartum dairy cows. Forty-eight dairy cows were allocated into 12 blocks, based on parity and milk yield of the previous lactation cycle, and randomly assigned to 1 of 4 treatments. Supplementation of VB₁₂ (weekly intramuscular injections of 0 or 10 mg) and biotin (dietary supplements of 0 or 30 mg/d) were used in a 2 × 2 factorial arrangement in a randomized complete block design of 12 blocks with repeated measures. The study started at 3 wk before the expected calving date and ended at 8 wk after calving. Feed intake and lactation performance (milk yield and composition) were recorded weekly after calving. Blood variables were measured on d ?10, 0, 8, 15, 29, 43, and 57 relative to calving. When VB₁₂ was given, the cows had greater feed intake, better lactation performance and lower body weight loss in the postpartum period compared with animals without injection of VB_12. The VB₁₂-injected cows had lower plasma nonesterified fatty acids and β-hydroxybutyrate concentrations but higher plasma superoxide dismutase activity compared with cows without VB₁₂. Cows fed a biotin supplement had higher milk protein yield (6 and 8 wk) and lactose yield (6–8 wk), compared with animals without biotin. However, under the present experimental conditions, we found no additive effect of a combined supplement of biotin and vitamin B₁₂ on lactation performance of dairy cows.     

Short communication: Effects of dietary 5,6-dimethylbenzimidazole supplementation on vitamin B12 supply,lactation performance,and energy balance in dairy cows during the transition period and early lactation

The current study was conducted to investigate the effects of 5,6-dimethylbenzimidazole (DMB) supplementation to the feed during the transition period and early lactation on the vitamin B₁₂ supply, lactation performance, and energy balance in postpartum cows. Twenty-four prepartum Holstein dairy cows were divided into 12 blocks based on their parity and milk yield at the last lactation and were then randomly allocated to 1 of 2 treatments: a basal diet without DMB (control) or a treatment diet that contained 1.5 g of DMB/d per cow. The study started at wk 3 before the expected calving day and ended at wk 8 postpartum. The feed intake and the lactation performance were measured weekly after calving. Blood parameters were measured on d ?10, 0, 8, 15, 29, 43, and 57 relative to the calving day. Body weight was measured on the calving day and on d 57 after calving. The yields of milk, protein, and lactose in cows fed DMB were higher than in the control throughout the whole postpartum stage. On wk 8 postpartum, the vitamin B₁₂ content in the milk and sera was greater in cows fed DMB than in the control. The overall body weight loss from wk 1 to 8 postpartum was less in cows fed DMB than in the control. The plasma content of nonesterified fatty acids and β-hydroxybutyric acid was significantly lower in cows fed DMB than in the control throughout the whole experimental stage. In conclusion, dietary DMB fed during the transition period and early lactation improved the vitamin B₁₂ supply, milk production, and energy balance of postpartum dairy cows.     

Associations between bone and energy metabolism in cows fed diets differing in level of dietary cation-anion difference and supplemented with cholecalciferol or calcidiol

Bone-derived hormones play an important role in metabolism. This study examined the hypothesis that interactions between bone and energy metabolism, particularly those involving osteocalcin, are present in dairy cattle and have feedback mechanisms over time. Associations between metabolites in blood were examined in 32 Holstein cows blocked by parity and milk yield and randomly allocated to diets containing either 0.27 mg/kg dry matter (DM) calcidiol or cholecalciferol for an anticipated intake of 3 mg/d (120,000 IU/d) at 11 kg of DM, and positive (+130 mEq/kg DM) or negative (?130 mEq/kg DM) dietary cation-anion difference (DCAD) from 252 d of gestation to calving. Blood was sampled every 3 d, from 9 d prepartum to 30 d postpartum, and plasma concentrations of vitamin D₃, 25-hydroxyvitamin D₃, adiponectin, C-telopeptide of type 1 collagen (CTX1), glucose, insulin-like growth factor 1 (IGF1), insulin, undercarboxylated osteocalcin (uOC), and carboxylated osteocalcin (cOC) were determined. Feeding calcidiol compared with cholecalciferol increased plasma concentrations of 25-hydroxyvitamin D₃ pre- (264.2 ± 8.0 vs. 61.3 ± 8.0 ng/mL) and postpartum (170.8 ± 6.2 vs. 51.3 ± 6.2 ng/mL) but decreased concentrations of vitamin D₃ pre- (1.2 ± 0.6 vs. 14.5 ± 0.6 ng/mL) and postpartum (1.9 ± 0.4 vs. 3.2 ± 0.6 ng/mL). Prepartum, cows fed the negative DCAD diet had reduced concentrations of vitamin D₃ and glucose compared with cows fed a positive DCAD. The combination of negative DCAD and cholecalciferol reduced IGF1 concentrations prepartum. The DCAD treatment had no effect on postpartum concentrations of metabolites. Nulliparous cows had increased concentrations of OC, CTX1, IGF1, glucose, and insulin compared with parous cows. Time series analysis identified associations between metabolites on the same day and over 3-d lags up to ±9 d that suggest feedback between 25-hydroxyvitamin D₃ and vitamin D₃ in the negative lags, indicating that 25-hydroxyvitamin D₃ may exert feedback on vitamin D₃ but not vice versa. We found evidence of a feedback mechanism between vitamin D₃ and IGF1, with positive effect size (ES) on the same day and 3 d later, and negative ES 9 d later, that was more evident in cholecalciferol-fed cows. This suggests an important role of IGF1 in integrating bone metabolism with energy and protein metabolic pathways. Evidence of feedback was found between uOC and particularly cOC with IGF1, with positive ES on the same day but negative ES 6 d before and 6 d after. An association between uOC or cOC and IGF1 has not been previously identified in cattle and suggests that both uOC and cOC may have marked biological activity. Associations between OC and insulin identified in mice were not observed herein, although associations between OC and glucose were similar to those between IGF1 and glucose, supporting associations between glucose, OC, and IGF1. We provide further statistical evidence of crosstalk between vitamin D compounds, bone hormones, and energy metabolism in cattle. In particular, associations between uOC or cOC and IGF1 may provide links between prepartum diets and observations of prolonged increases in milk production and allow better control of peripartum metabolism.     

Vitamin D3 synthesis in the entire skin surface of dairy cows despite hair coverage

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.     

Reduced serum vitamin D concentrations in healthy early-lactation dairy cattle

Cattle obtain vitamin D by ingestion or cutaneous exposure to UV light. Dairy cattle diets are frequently supplemented with vitamin D to compensate for limited sun exposure or during times of increased metabolic demands, such as the periparturient period, to maintain calcium homeostasis. Whether housing and supplemental vitamin D practices supply adequate amounts of vitamin D to optimally support the transition from gestation to lactation in dairy cattle is unknown. Our objective was to determine how serum vitamin D concentrations of dairy cows change with season, age, parity, and stage of lactation. Clinically healthy cows (n = 183) from 5 commercial dairies were enrolled in the study. Serum samples were collected at dry off, within 7 d of entering the close-up group, and within 7 d after calving (calving+7). Vitamin D status was determined by measuring serum 25-hydroxyvitamin D [25(OH)D] by radioimmunoassay. We performed repeated-measures mixed-effects linear regression to determine the effects of season, age, parity, and lactation stage (dry off, close-up, and calving+7) on 25(OH)D concentrations in serum. Bivariable analysis indicated that parity, age, and season were not associated with serum 25(OH)D concentrations. Sample period affected 25(OH)D concentrations, with the highest 25(OH)D levels at dry off (99.7 ± 1.9 ng/mL) followed by close up (93.8 ± 2.1 ng/mL), with the lowest levels at calving+7 (82.6 ± 1.7 ng/mL). These data showed a large depletion of 25(OH)D in dairy cattle postpartum compared with late prepartum, although the biological significance of this change in these healthy cattle is unclear. Consumption of serum 25(OH)D by immune system functions and calcium homeostasis in early lactation likely caused the reduction in serum 25(OH)D concentrations after calving. These results suggest that determining whether serum 25(OH)D concentrations are associated with the incidence of transition period disease is an appropriate next step. Assessing the effects of enhanced vitamin D supplementation of cows in early lactation on postpartum diseases may be warranted.     

Effect of supplementing rumen-protected methionine,lysine, and histidine to low-protein diets on the performance and nitrogen balance of dairy cows

Lowering the dietary protein content can reduce N excretions and NH₃ emissions from manure and increase milk N efficiency of dairy cows. However, milk yield (MY) and composition can be compromised due to AA deficiency. Methionine and Lys are known as first limiting EAA for dairy cows, and recently His is also mentioned as limiting, especially in grass-based or low-protein diets. To examine this, a trial was conducted with a 3-wk pre-experimental adaptation period (diet 16.5% crude protein), followed by a depletion period of 4 wk, in which 39 cows (average ± standard deviation: 116 ± 29.3 d in milk, 1.8 ± 1.2 lactations, 638 ± 73.2 kg of body weight, and 32.7 ± 5.75 kg MY/d) received a low-protein diet (CTRL) (14.5% crude protein). Then, taking into account parity, His plasma concentration, and MY, cows were randomly assigned to 1 of 3 treatment groups during the rumen-protected (RP) AA period of 7 wk; (1) CTRL; (2) CTRL + RP-Met + RP-Lys (MetLys); (3) CTRL + RP-Met + RP-Lys + RP-His (MetLysHis). Products were dosed, assuming requirements for digestible (d) Met, dLys, and dHis being, respectively, 2.4%, 7.0%, and 2.4% of intestinal digestible protein. In the cross-back period of 5 wk, all cows received the CTRL diet. During the last week of each period, a N balance was conducted by collecting total urine and spot samples of feces. Total feces production was calculated using the inert marker TiO₂. Statistical analysis was performed with a linear mixed model with cow as random effect and data of the last week of the pre-experimental period used as covariate for the animal performance variables. No effect of supplementing RP-Met and RP-Lys nor RP-Met, RP-Lys, and RP-His on feed intake, milk performance, or milk N efficiency was observed. However, the plasma AA profile indicated additional supply of dMet, dLys, and dHis. Nevertheless, evaluation of the AA uptake relative to the cow's requirements showed that most EAA (exclusive Arg and Thr) were limiting over the whole experiment. Only dHis was sufficiently supplemented during the RP-AA period due to an overestimation of the diet's dMet and dLys supply in the beginning of the trial. The numerically increased milk urea N and urinary N excretion when RP-Met, RP-Lys, and RP-His were added to the low-protein diet suggest an increased catabolism of the excess His.     

Blood mineral, hormone, and osteocalcin responses of multiparous Jersey cows to an oral dose of 25-hydroxyvitamin D3 or vitamin D3 before parturition

Twenty-seven multiparous Jersey cows were randomly assigned to receive an oral bolus containing corn starch (control, CON), corn starch plus 15 mg of 25-hydroxyvitamin D₃ (25-OH), or 15 mg of cholecalciferol (D₃) at 6 d before expected parturition. Cows were maintained in individual box stalls from 20 d before expected parturition and fed a common diet. Jugular blood samples were collected at −14, −13, −5, −4, −3, −2, −1 d before expected calving, at calving, and at 1, 3, 5, 7, 9, 11, 13, 28, 56, and 84 d postcalving. After calving, cows were housed in 1 pen in a free-stall barn and consumed a common diet. Colorimetric assays were used to analyze Ca, P, and Mg concentrations in serum. Serum concentrations of osteocalcin (OC), an indicator of bone formation, serum 25-hydroxyvitamin D₃, and parathyroid hormone (PTH) were determined in samples obtained from d −5 through d 13. The 9 control multiparous cows and 5 untreated primiparous cows were used to evaluate the effect of parity on the variables that were measured. There was no effect of parity on Ca, PTH, or 25-OH concentration. Compared with second-lactation cows and older cows (>2 lactations), first-lactation cows had greater serum OC (22.3, 32.0, and 48.3 ng/mL, respectively), indicating that younger animals were forming more bone. Blood Ca, P, and Mg decreased near the time of calving and then increased over time. Serum 25-hydroxyvitamin D₃ was greater for cows dosed with 25-OH (119.0 ng/mL) compared with those dosed with D₃ (77.5 ng/mL) or CON (69.3 ng/mL). Cows dosed with 25-OH tended to have lower serum PTH concentration, but treatments did not affect serum Ca, P, or Mg. Serum OC was greater in second-lactation cows compared with cows entering their third or fourth lactation but OC was unaffected by treatment. Although results indicated a 60% increase in serum 25-hydroxyvitamin D₃ due to a single oral dose of 25-OH before calving, the amount administered in this study apparently was not sufficient for initiation of any improvement in Ca homeostasis at parturition.     

Methyl donor supplementation suppresses the progression of liver lipid accumulation while modifying the plasma triacylglycerol lipidome in periparturient Holstein dairy cows

Co-supplementation of methyl donors may lower hepatic lipid content in transition cows. To define the ability of methyl donor supplementation (MDS) to reduce hepatic lipid content and modify the plasma lipidome, 30 multiparous Holstein cows (2.04 ± 0.69 lactations; 689 ± 58 kg of body weight; 3.48 ± 0.10 units of body condition score) were fed a ration with or without rumen-protected methyl donors (22 g/d of Met, 10 g/d of choline chloride, 3 g/d of betaine, 96 mg/d of riboflavin, and 1.4 mg/d of vitamin B₁₂) from d ?28 before expected calving through d 14 postpartum. Cows were randomly enrolled based on predefined selection criteria (body condition score and parity). Base diets without MDS were formulated for gestation (15.4% crude protein with a predicted Lys-to-Met ratio of 3.25; 1.44 Mcal of net energy for lactation/kg of dry matter) and lactation (16.6% crude protein with a predicted Lys-to-Met ratio of 3.36; 1.64 Mcal of net energy for lactation/kg of dry matter). Blood sampling occurred from d ?28 relative to expected calving through d 14 postpartum. Liver tissue was biopsied at d ?28 relative to expected calving and on d 5 and 14 postpartum. In addition to routine analyses, serum AA concentrations on d 10 and 12 were quantified using mass spectrometry. Plasma triacylglycerol (TAG) and cholesteryl esters (CE) were qualitatively measured using time-of-flight mass spectrometry. Data were analyzed using a mixed model with repeated measures. Dry matter intake and milk yield were not modified by MDS. The transition from d ?28 relative to expected parturition to d 14 postpartum was characterized by increased plasma fatty acid (0.15 to 0.71 mmol/L) and β-hydroxybutyrate (0.34 to 0.43 mmol/L) levels and liver lipid content (3.91 to 9.16%). Methyl donor supplementation increased the serum Met level by 26% and decreased the serum Lys-to-Met ratio by 21% on d 10 and 12, respectively. Moreover, the increase in hepatic lipid content from d 5 through 14 postpartum was suppressed with MDS relative to control (3.57 vs. ?0.29%). Dietary MDS modified the TAG and CE lipidome. For example, MDS increased plasma TAG 46:3 (carbon number:double bond) by 116% relative to control cows on d 5 postpartum. Moreover, MDS tended to increase plasma CE 34:6. In contrast, MDS lowered plasma TAG 54:8 by 39% relative to control cows on d 5 postpartum. We concluded that in the absence of gains in dry matter intake and milk and milk protein yields, dietary MDS slows the progression of hepatic lipid accumulation and modifies the plasma TAG lipidome in transition cows.     

Periparturient responses of multiparous Holstein cows fed different dietary phosphorus concentrations prepartum

Our objective was to compare the effects of different prepartum dietary phosphorus concentrations on periparturient metabolism and performance. Forty-two late pregnant multiparous Holstein cows were fed 0.21, 0.31, or 0.44% P (dry basis) for 4 wk before expected calving. After parturition, all cows were fed a common lactation diet (0.40% P). In the prepartum period, cows fed 0.21% P had lower blood serum P concentrations compared with cows fed 0.31 or 0.44% P. However, serum P concentrations of all cows were within the normal range (4 to 8 mg/dL) until the day of calving when average concentrations dropped below 4 mg/dL. From 3 to 14 d postpartum, serum P of cows fed 0.21% P was greater than that of cows fed 0.31 or 0.44% P. No cows presented with or were treated for clinical hypophosphatemia in the periparturient period. Total serum Ca was lower before calving through 2 d postpartum for cows fed 0.44% P compared with those fed 0.21 or 0.31%. Prepartum dietary P treatments did not alter blood osteocalcin, hydroxyproline, and deoxypyridinoline, indicators of bone metabolism, or concentrations of parathyroid hormone or 1,25-dihydroxyvitamin D₃. Energy-corrected milk yield and milk composition (first 28 d of lactation) were not affected by prepartum dietary P concentrations. It is concluded that feeding 0.21% P (34 g of P/cow daily) prepartum is adequate for periparturient multiparous Holstein cows with high metabolic demands and genetic potential for milk production. No adverse effects on periparturient health, dry matter intake, or 28-d lactation performance resulted.     

Effects of supplementing a Saccharomyces cerevisiae fermentation product during the periparturient period on performance of dairy cows fed fresh diets differing in starch content

The objective of this study was to evaluate the effects of supplementing a Saccharomyces cerevisiae fermentation product (SCFP; NutriTek, Diamond V, Cedar Rapids, IA) during the periparturient period (d ?28 ± 3 to 44 ± 3 relative to calving) on dry matter intake (DMI), milk production, apparent total-tract nutrient digestibility, and postpartum ovarian activity of dairy cows fed fresh diets varying in starch content. From d 28 ± 3 before the expected calving date until d 44 ± 3 after calving, 117 Holstein cows were fed diets with SCFP (SCFP; n = 59) or without (control, CON; n = 58). A common, basal, controlled-energy close-up diet (net energy for lactation: 1.43 Mcal/kg; 13.8% starch) was fed before calving. Cows within each treatment (CON or SCFP) were fed either a low- (LS; 22.1% starch) or high-starch (HS; 28.3% starch) diet from d 1 to 23 ± 3 after calving (fresh period), resulting in 4 treatment groups: LS-CON (n = 30), LS-SCFP (n = 29), HS-CON (n = 28), and HS-SCFP (n = 30). All cows were fed the HS diets from d 24 ± 3 to 44 ± 3 after calving (post-fresh period). Cows were assigned to treatment balanced for parity, body condition score, body weight, and expected calving date. Milk yield was higher for cows fed the LS diets compared with those fed the HS diets during the fresh period (34.1 vs. 32.1 kg/d), whereas DMI and 3.5% fat-corrected milk yield (FCM) were not affected by dietary starch content, and LS cows tended to lose more body condition than HS cows (?0.42 vs. ?0.35 per 21 d) during the fresh period. Overall DMI during the close-up and fresh periods did not differ between SCFP and CON cows. However, SCFP supplementation transiently increased DMI on d 1 (13.0 vs. 11.9 kg/d) and 5 (15.5 vs. 14.1 kg/d) after calving compared with CON. During the post-fresh period, SCFP cows tended to eat less than CON cows (19.8 vs. 20.6 kg/d) but had similar 3.5% FCM (44.9 vs. 43.6 kg/d), resulting in greater feed efficiency for SCFP cows (FCM/DMI; 2.27 vs. 2.13). Neither starch content of fresh diets nor SCFP supplementation affected the interval from calving to first ovulation or the incidence of double ovulation. These findings suggest that feeding low-starch diets during the fresh period can increase milk production of dairy cows during the fresh period, and that supplementation of SCFP may increase feed intake around calving and feed efficiency in the post-fresh period.     

Effects of rumen-protected choline and monensin on milk production and metabolism of periparturient dairy cows

Choline and monensin may be supplemented during the transition period with the objectives of aiding in fat metabolism and improving energy balance, respectively. The objectives of this study were to determine the effects of supplementing rumen-protected choline (RPC) and monensin in a controlled-release capsule (CRC) on metabolism, dry matter intake, milk production, and liver function in transition dairy cattle. Three weeks before expected calving, 182 Holsteins were randomly assigned to receive one of the following: a monensin CRC, 56 g/d of RPC until 28 d in milk, CRC + RPC, or neither supplement (control). Blood samples were collected at enrollment, 1 wk before calving, and in the first and second weeks after calving. Liver biopsies were obtained from multiparous cows randomly selected from each treatment group within 24 h and again 3 wk postpartum. Daily milk production was recorded through 60 d in milk. There were no interactions of the effects of RPC and CRC on any of the outcomes measured. Overall, cows that received RPC produced 1.2 kg/d more milk in the first 60 d of lactation, but this effect was attributable to an increase in milk production of 4.4 kg/d among cows with a body condition score ≥4 at 3 wk before calving; fat cows that received RPC ate 1.1 kg of DM/d more from wk 3 before calving through wk 4 after calving. Monensin supplementation significantly increased serum concentrations of glucose and urea, lowered concentrations of β-hydroxybutyric acid and aspartate aminotransferase in the peripartum period, and increased liver glycogen content at 3 wk into lactation. The metabolic effects of CRC are consistent with previous studies, and the effects on liver are novel. The mechanism by which RPC increased milk production was not revealed in this study and merits further research.     

Effects of supplements of folic acid, vitamin B12, and rumen-protected methionine on whole body metabolism of methionine and glucose in lactating dairy cows

The present experiment was undertaken to determine the effects of dietary supplements of rumen-protected methionine and intramuscular injections of folic acid and vitamin B₁₂, given 3 wk before to 16 wk after calving, on glucose and methionine metabolism of lactating dairy cows. Twenty-four multiparous Holstein cows were assigned to 6 blocks of 4 cows each according to their previous milk production. Within each block, 2 cows were fed a diet estimated to supply methionine as 1.83% metabolizable protein, equivalent to 76% of methionine requirement, whereas the 2 other cows were fed the same diet supplemented daily with 18 g of rumen-protected methionine. Within each diet, the cows were administrated either no vitamin supplement or weekly intramuscular injections of 160 mg of folic acid plus 10 mg of vitamin B_12. To investigate metabolic changes at 12 wk of lactation, glucose and methionine kinetics were measured by isotope dilution using infusions of 3[U-¹³C]glucose, [¹³C]NaHCO₃ and 3[1-¹³C,²H₃] methionine. Milk and plasma concentrations of folic acid and vitamin B₁₂ increased with vitamin injections. Supplementary B-vitamins increased milk production from 34.7 to 38.9 ± 1.0 kg/d and increased milk lactose, protein, and total solids yields. Whole-body glucose flux tended to increase with vitamin supplementation with a similar quantitative magnitude as the milk lactose yield increase. Vitamin supplementation increased methionine utilization for protein synthesis through increased protein turnover when methionine was deficient and through decreased methionine oxidation when rumen-protected methionine was fed. Vitamin supplementation decreased plasma concentrations of homocysteine independently of rumen-protected methionine feeding, although no effect of vitamin supplementation was measured on methionine remethylation, but this could be due to the limitation of the technique used. Therefore, the effects of these B-vitamins on lactation performance were not mainly explained by methionine economy because of a more efficient methylneogenesis but were rather related to increased glucose availability and changes in methionine metabolism.     

Intramammary 25-hydroxyvitamin D3 treatment modulates innate immune responses to endotoxin-induced mastitis

Vitamin D signaling in response to pathogen-associated molecules contributes to activation of innate immune responses of bovine monocytes. We hypothesized that lipopolysaccharide (LPS) of bacteria associated with mastitis in dairy cows activates the vitamin D pathway in innate immune cells of the udder and that increasing availability of 25-hydroxyvitamin D₃ [25(OH)D₃] would augment expression of vitamin D-associated genes. The objective of this experiment was to determine the effects of intramammary LPS and 25(OH)D₃ treatments on activation of the vitamin D pathway and innate immune responses of mammary immune cells. Individual mammary quarters of 5 lactating cows were treated with placebo control, 100 μg of 25(OH)D₃, 5 μg of LPS, or a combination of 100 μg of 25(OH)D₃ and 5 μg of LPS. Somatic cells from milk were evaluated for percentage of neutrophil and macrophage populations and expression of genes associated with vitamin D metabolism and innate immunity. Data from samples collected from 4 to 12 h after challenge were analyzed for main effects of LPS and 25(OH)D₃ treatments, treatment interactions, and simple effects of 25(OH)D₃ treatment. Data from samples collected at the time of challenge were used as covariates. The percentages of neutrophils in milk at 8 h postchallenge were 58 ± 10, 82 ± 11, 89 ± 10, and 63 ± 10% of total cells in milk from control, 25(OH)D₃, LPS, and LPS plus 25(OH)D₃ glands, respectively, such that the interaction of LPS and 25(OH)D₃ was significant. Expression of the vitamin D 1α-hydroxylase (CYP27B1) and vitamin D receptor genes was upregulated by LPS treatment in total cells, macrophages, and neutrophils in milk. In addition, expression of the vitamin D 24-hydroxylase (CYP24A1) gene in milk somatic cells was upregulated by 25(OH)D₃ and LPS treatments. The inducible nitric oxide synthase (iNOS), chemokine (C-C-motif) ligand 5 (CCL5), β-defensin 3 (DEFB3), DEFB7, and DEFB10 genes were upregulated by LPS treatment in total cells and neutrophils from milk. Expression of iNOS in milk somatic cells tended to be affected by the interaction between LPS and 25(OH)D₃, such that 25(OH)D₃ tended to increase iNOS in the absence of LPS but not in the presence of LPS. Furthermore, expression of CCL5 in macrophages was downregulated by 25(OH)D₃. In conclusion, intramammary endotoxin challenge activates the vitamin D pathway in mammary macrophages and neutrophils, and intramammary 25(OH)D₃ treatment alters the percentage of neutrophils and expression of immune genes in milk somatic cells.     

Effect of source and amount of vitamin D on function and mRNA expression in immune cells in dairy cows

Objectives were to determine the effect of supplementing 2 sources of vitamin D, cholecalciferol (CH) or calcidiol (CA), at 1 (1mg) or 3 mg/d (3mg) prepartum on concentrations of vitamin D metabolites in plasma, measures of innate immune function, and leukocyte mRNA expression. Parous Holstein cows (n = 99) were assigned to a daily treatment administered as top-dress containing either 1 or 3 mg of CH (CH1 or CH3) or of CA (CA1 or CA3) from 250 d of gestation until calving. Plasma concentrations of vitamin D, immune cell population in blood, cell adhesion markers, and granulocyte phagocytosis and oxidative burst were evaluated pre- and postpartum. The mRNA expression in leukocytes was determined at 270 d of gestation and 3 d postpartum for genes involved in cell migration, pathogen recognition receptors, cell signaling, cytokines, antimicrobial mechanisms, oxidative burst, and Ca and vitamin D metabolism. Concentrations of vitamin D₃ increased in cows fed CH, whereas those of 25-hydroxyvitamin D₃ increased in cows fed CA. Percentage of granulocytes from total leukocytes differed with amount of vitamin D pre- (1mg = 24.5 vs. 3mg = 37.9%) and postpartum (1mg = 22.0 vs. 3mg = 31.0%), thus shifting mononuclear cells in the opposite direction pre- (1mg = 75.5 vs. 3mg = 62.1%) and postpartum (1mg = 78.0 vs. 3mg = 69.0%). Granulocytes displaying phagocytosis (1mg = 69.0 vs. 3mg = 62.9%) and intensity of phagocytosis prepartum (1mg = 7.46 vs. 3mg = 7.28) tended to be less in cows fed 3mg compared with 1mg. During prepartum, CA increased mRNA expression of genes related to cell adhesion and migration (CD44, ICAM1, ITGAL, ITGB1, LGALS8, SELL), pathogen recognition receptor (NOD2, TLR2, TLR6), cell signaling (FOS, JUN, NFKB2), cytokine signaling (IL1B, IL1R1, IL1RN), antimicrobial mechanisms (CTSB, LYZ), and Ca metabolism (ATP2B1, STIM1, TRPV5) compared with CH. Similarly, postpartum, CA increased mRNA expression of genes related to cell adhesion and migration (CXCR2, SELL, TLN1), cell signaling (AKT2), cytokines (CCL2, IL1R1, ILRN), antimicrobial mechanisms (DEFB3), oxidative burst (RAC2), and calcium metabolism (CALM3) compared with CH. Feeding additional vitamin D in the last 3 wk of gestation changed the profile of blood leukocytes and attenuated granulocyte phagocytosis during the transition period, whereas supplementing CA prepartum increased mRNA expression of genes involved in immune cell function, including genes related to pathogen recognition and antimicrobial effects of leukocytes.     

Influence of methionine supply on the response of lactational performance of dairy cows to supplementary folic acid and vitamin B12

The present experiment was undertaken to determine if the effects of supplementary folic acid on lactational performance were caused by improved methylneogenesis and if the supply in vitamin B₁₂ could affect this metabolic pathway. In this eventuality, supplementary Met, a major source of preformed methyl groups, should reduce the requirements for these vitamins. Sixty multiparous Holstein cows were assigned to 10 blocks of 6 cows each according to their previous milk production. Within each block, 3 cows were fed a diet estimated to supply Met as 1.83% metabolizable protein and 3 cows were fed the same diet supplemented with 18 g of rumen-protected methionine (RPM) to supply Met as 2.23% of metabolizable protein. Within each level of Met, cows received no vitamin supplement or weekly intramuscular injections of 160 mg of folic acid alone or combined with 10 mg of vitamin B₁₂ from 3 wk before to 16 wk after calving. There was no treatment effect on dry matter intake during pre- and postcalving periods: 13.4 ± 0.4 and 21.8 ± 0.4 kg/d, respectively. Milk production was not affected by RPM supplementation. Folic acid and vitamin B₁₂ given together tended to increase milk production during the 16 wk of lactation. This effect was more pronounced during the first 4 wk of lactation: 37.5, 37.7, and 40.3 ± 0.9 kg/d for cows receiving no vitamin supplement, folic acid alone, or folic acid combined with vitamin B₁₂, respectively. Milk fat yield was not affected by treatments. Lactose, crude protein, and total solid yields were greater, in early lactation, in cows injected with folic acid and vitamin B₁₂ together but this effect diminished as lactation progressed. Intramuscular injections of folic acid alone or combined with vitamin B₁₂ tended to decrease plasma concentrations of homocysteine from 5.51 μM with no vitamin supplement to 4.54 and 4.77 ± 0.37 μM, respectively. Results of the present experiment suggest that the effects of the combined supplement of folic acid and vitamin B₁₂ on lactational performance of dairy cows were not due to an improvement in methyl groups supply, because RPM supplement, a source of preformed methyl groups, did not alter the cow responsiveness to vitamin supplements.     

Replacing soybean meal with okara meal: Effects on production,milk fatty acid and plasma amino acid profile,and nutrient utilization in dairy cows

Okara meal is a byproduct from the production of soymilk and tofu and can potentially replace soybean meal (SBM) in dairy diets due to its high crude protein (CP) concentration and residual fat. The objective of this study was to investigate the effects of replacing SBM with okara meal on feed intake, yields of milk and milk components, milk fatty acid (FA) profile, nutrient utilization, and plasma AA concentration in lactating dairy cows. Twelve multiparous (65 ± 33 d in milk) and 8 primiparous (100 ± 35 d in milk) organically certified Jersey cows were paired by parity or days in milk, and within pair, randomly assigned to treatments in a crossover design with 21-d periods (14 d for diet adaptation and 7 d for data and sample collection). Diets were fed as total mixed ration formulated to be isonitrogenous and isofibrous and contained (dry matter basis) 50% mixed, mostly grass baleage, 2% sugarcane liquid molasses, 2% minerals-vitamins premix, and either (1) 8.1% SBM, 10% soyhulls, and 27.9% ground corn (CTRL); or (2) 15% okara meal, 8% soyhulls, and 23% ground corn (OKR). Dietary CP, ash-free neutral detergent fiber, and total FA averaged 15.4, 35.3, and 3.08% for CTRL and 15.9%, 36.3%, and 3.74% for OKR, respectively. Substitution of SBM with okara meal did not alter dry matter intake but increased intakes of CP and ash-free neutral detergent fiber. Additionally, no significant differences between treatments were observed for yields of milk and milk components, and concentrations of milk fat, lactose, and total solids. However, milk true protein concentration was lower in cows fed OKR (3.76%) versus CTRL (3.81%). Both milk urea N (8.51 vs. 9.47 mg/dL) and plasma urea N (16.9 vs. 17.8 mg/dL) concentrations decreased with OKR relative to the CTRL diet, respectively. Compared with CTRL, feeding OKR lowered the milk proportions of total odd-chain FA, de novo FA, and mixed FA and increased those of preformed FA, total n-6 FA, and total n-3 FA. The milk proportions of trans-10 18:1, trans-11 18:1, and cis-9,trans-11 18:2 were greater with feeding OKR versus the CTRL diet. The apparent total-tract digestibility of nutrients, urinary excretion of total purine derivatives (uric acid plus allantoin), and total N were not affected by treatments. Except for plasma Leu, which was lower in OKR compared with the CTRL diet, no other significant changes in the plasma concentrations of AA were observed. The plasma concentration of carnosine was lowest in cows receiving the OKR diet. Overall, our results revealed that okara meal can completely replace SBM without negatively affecting production and nutrient digestibility in early- to mid-lactation Jersey cows. Further research is needed to assess the economic feasibility of including okara meal in dairy diets, as well as the amount of okara meal that maximizes yields of milk and milk components in dairy cows in different stages of lactation.