Association of subclinical hypocalcemia dynamics with dry matter intake, milk yield, and blood minerals during the periparturient period.

Subclinical hypocalcemia (SCH) affects many high-producing dairy cows in the postpartum period. Recent work has shown that cows experiencing prolonged or delayed SCH are at increased risk for disease and produce less milk than cows experiencing a transient reduction in or normal concentrations of plasma Ca following parturition. Our objective was to determine the association between different postpartum SCH dynamics with pre- and postpartum dry matter intake (DMI), milk yield, and blood mineral concentrations. Data were retrospectively collected from multiparous Holstein cows (n = 78), and cows were classified into 1 of 4 SCH groups based on mean blood total Ca (tCa) concentrations at 1 and 4 d in milk (DIM): normocalcemic (NC; [tCa] >1.95 mmol/L at 1 DIM and >2.2 mmol/L at 4 DIM, n = 28); transient SCH (tSCH; [tCa] ≤1.95 mmol/L at 1 DIM and >2.2 mmol/L at 4 DIM, n = 27); delayed SCH (dSCH; [tCa] >1.95 mmol/L at 1 DIM and ≤2.2 mmol/L at 4 DIM, n = 6); and persistent SCH (pSCH; [tCa] ≤1.95 mmol at 1 DIM and ≤2.2 mmol/L at 4 DIM, n = 17). Linear mixed models were created to analyze the change in pre- and postpartum DMI, milk yield, and blood mineral concentrations over time as well as differences between SCH groups. Prepartum intake was similar between groups, but the NC and tSCH cows consumed more feed than the pSCH or dSCH cows during the first 3 wk of lactation. The tSCH cows produced more milk than the other 3 groups during the first 6 wk of lactation. Postpartum blood tCa and Mg were different between SCH groups and were highest in the NC cows and lowest in the pSCH cows. Our results suggest that the high level of DMI consumed by the NC and tSCH cows in the postpartum period supported an appropriate homeostatic response to the increased Ca demands of lactation, allowing for higher milk yield compared with their counterparts experiencing delayed or prolonged episodes of SCH.

Acute-phase protein α-1-acid glycoprotein is negatively associated with feed intake in postpartum dairy cows.

α-1-Acid glycoprotein (AGP) is an acute-phase protein that may suppress dry matter intake (DMI), potentially by acting on the leptin receptor in the hypothalamus. Our objectives were to characterize plasma AGP concentration and associations with DMI during the transition period, and to determine the utility of AGP to identify or predict cows with low DMI. Plasma samples (n = 2,086) from 434 Holstein cows in 6 studies were analyzed on d -21, -13 ± 2, -3, 1, 3, 7 ± 1, 14 ± 1, and 21 ± 1 relative to parturition. A commercially available ELISA kit specific for bovine AGP was validated, and 2 internal controls were analyzed on each plate with interplate variation of 15.0 and 17.3%, respectively. Bivariate analysis was used to assess the relationship between AGP and DMI. For significant associations, treatment(study) was added to the model, and quadratic associations were included in the model if significant. Plasma AGP concentration (±SEM) increased from 213 ± 37.3 µg/mL on d -3 to 445 ± 60.0 µg/mL on d 14. On d 3, AGP was associated negatively with DMI in a quadratic manner for wk 1 and wk 2 and linearly for wk 3. Day 7 AGP was associated negatively with DMI in a quadratic manner for wk 2 and linearly for wk 3. Similarly, d 14 AGP was negatively associated with DMI for wk 3 and wk 4. As d 3 AGP concentration increased over the interquartile range, a calculated 1.4 (8.5%), 0.5 (2.7%), and 0.4 (1.9%) kg/d reduction in predicted DMI was detected during wk 1, 2, and 3, respectively. Using bivariate analysis, d 3 AGP explained 10% of the variation in DMI during wk 1. We explored the clinical utility of d 3 AGP to diagnose low DMI, defined as wk 1 DMI >1 standard deviation below the mean. Receiver operating characteristic analysis identified a threshold of 480.9 µg/mL, providing 76% specificity and 48% sensitivity (area under the curve = 0.60). Limited associations occurred between AGP and blood biomarkers; however, AGP was associated with plasma haptoglobin concentration postpartum and incidence of displaced abomasum, retained placenta, and metritis. These results demonstrate a negative association between plasma AGP concentration and DMI in early-postpartum dairy cows, although its diagnostic performance was marginal. Further investigation into whether AGP directly suppresses DMI in dairy cattle is warranted.

Effects of feeding synthetic zeolite A during the prepartum period on serum mineral concentration, oxidant status, and performance of multiparous Holstein cows.

The objective of this study was to determine the effects of feeding synthetic zeolite A for 3 wk before expected calving on peripartal serum mineral concentrations, hypocalcemia, oxidant status, and performance. Holstein cows (n = 55) entering their second or greater lactations were assigned randomly to 1 of 2 dietary treatments starting 21 d before expected calving: control (CON: 40% corn silage, 33% wheat straw, and 27% concentrate; n = 29) or experimental [EXP: CON plus zeolite A (X-Zelit, Protekta Inc., Lucknow, ON, Canada/Vilofoss, Graasten, Denmark; n = 26) at an inclusion rate of 3.3% of dry matter, targeting 500 g/d as-fed]. Cows were fed the same postpartum diet and housed in individual tiestalls through 28 d in milk. Cows fed EXP had higher serum Ca concentrations as parturition approached and during the immediate postpartum period. Serum P concentrations were lower for the EXP-fed cows during the prepartum period and the first 2 d of lactation, whereas serum Mg concentrations were lower than those of the CON-fed cows only during the immediate periparturient period. Cows fed EXP had decreased prevalence of subclinical hypocalcemia (SCH) from d -1 through 3 relative to day of parturition, with the largest difference occurring within the first day postpartum. Prepartum dry matter intake tended to be decreased and rumination was decreased in cows fed EXP; however; postpartum dry matter intake, rumination, milk yield, milk component yield, and colostrum measurements did not differ between treatments. Cows fed EXP tended to have increased hazard of pregnancy by 150 d in milk when controlling for parity compared with CON-fed cows; potential reproductive benefits merit further study. This study demonstrated that zeolite A supplementation during the prepartum period results in markedly improved serum Ca concentrations around parturition and similar postpartum performance compared with controls and is effective at decreasing hypocalcemia in multiparous Holstein cows.

Epidemiology of subclinical hypocalcemia in early-lactation Holstein dairy cows: The temporal associations of plasma calcium concentration in the first 4 days in milk with disease and milk production.

The objectives of this study were to characterize the epidemiology of subclinical hypocalcemia (SCH) in Holstein dairy cows by assessing the temporal associations of plasma Ca concentrations in the first 4 d in milk (DIM) with the risk of cows being diagnosed with metritis or displaced abomasum (or both), and milk production across the first 15 wk of lactation. A prospective cohort study was conducted in 2 dairy herds in New York State, in which cows had a blood sample collected daily for the first 4 DIM. A total of 396 Holstein cows (137 primiparous and 259 multiparous) were enrolled. Multivariable Poisson regression models were built to evaluate the associations of plasma Ca concentration at each of the 4 d following parturition with the risk of primiparous cows being diagnosed with metritis and multiparous cows being diagnosed with metritis, displaced abomasum, or both. Similarly, generalized linear mixed models were built to evaluate the associations of plasma Ca concentration with milk production across the first 15 wk of lactation. Plasma Ca concentration was assessed on a continuous scale in all models; dichotomization and SCH classification only occurred in the final models if the Ca concentration variable was meaningful by creating an optimized threshold based on receiver operating characteristic curve analysis. Plasma Ca concentration assessed at 1 DIM was not associated with the risk of metritis in primiparous cows, but an association was observed at 2, 3, and 4 DIM (critical thresholds were plasma Ca concentration ≤2.15, 2.10, and 2.15 mmol/L, respectively). Plasma Ca concentration was associated with the risk of metritis or displaced abomasum diagnosis (or both) for 2nd parity animals at 2 DIM (threshold ≤1.97 mmol/L), and at 4 DIM for 3rd and greater lactations (threshold ≤2.20 mmol/L). Reduced plasma Ca concentration was associated with higher milk production when assessed at 1 DIM in primiparous and multiparous cows, and lower milk production when assessed at 4 DIM in multiparous cows only. For primiparous cows, plasma Ca concentration was not associated with lower milk production at any of the DIM assessed. In conclusion, assessments of SCH at the individual cow level must take into account the DIM of Ca concentration measurement and parity of the cow, as the epidemiology of the disorder was demonstrated to be highly dependent on these variables. This study advances the knowledge of the epidemiology of SCH and better establishes thresholds for optimizing SCH diagnosis.

Differential effects of a single dose of oral calcium based on postpartum plasma calcium concentration in Holstein cows.

Our objectives were to determine (1) the effect of a single dose of an oral Ca bolus within 24 h after parturition on plasma Ca concentration, (2) the response of primiparous (PP) and multiparous (MP) cows to this supplementation strategy, and (3) differential responses based on plasma Ca at enrollment. For objective 1, cows from 1 commercial dairy in New York State were enrolled within 19 h after parturition (mean ± standard deviation = 8.3 ± 5.3 h) and randomized within parity group (first, second, and ≥third) to control [CON (n = 25); no placebo] or a single dose bolus treatment [BOL (n = 25); 3 oral Ca boluses supplying 54 to 64 g of Ca]. Plasma Ca was measured repeatedly between 1 and 24 h following treatment. For objectives 2 and 3, cows on 6 commercial farms in New York State were assigned to treatment as for objective 1 (CON, n = 1,973; BOL, n = 1,976). Herd records for health, reproduction, and Dairy Herd Improvement Association test day milk production were collected. Mixed effect multivariable models were developed using repeated measures ANOVA, Poisson regression, or proportional hazard models. Objective 2 analyses considered treatment with periparturient risk factors, whereas objective 3 analyses also considered Ca status. No difference was observed for plasma Ca between 1 and 24 h after treatment. Primiparous cows assigned to BOL calving at >712 d old had decreased risk of one or more health disorders [≤30 d in milk; risk ratio (RR) = 0.65, 95% confidence interval (CI) = 0.51 to 0.84] and those with body condition score >3.5 responded to BOL with increased milk production (CON = 31.7 ± 1.1, BOL = 35.1 ± 1.1 kg/d), as did those with days carried calf >277 (CON = 31.9 ± 1.0, BOL = 34.7 ± 1.0 kg/d). Reduced risk of one or more health disorders was observed in parity ≥3 (RR = 0.85, 95% CI = 0.81 to 0.89) and MP cows with body condition score >3.5 (retained placenta; RR = 0.70, 95% CI = 0.58 to 0.84) or that were lame (displaced abomasum; RR = 0.49, 95% CI = 0.32 to 0.75). Differential responses for PP cows by Ca status were minimal. For MP cows with low plasma Ca, BOL decreased risk of additional Ca treatment (≤1.8 mmol/L; RR = 0.57, 95% CI = 0.40 to 0.80) as well as risk of one or more health disorders (≤2.15 mmol/L; RR = 0.90, 95% CI = 0.85 to 0.95). Supplementation with a single oral dose of Ca could be targeted to periparturient risk groups for improved health. Calcium status did not differentiate responses of PP cows, but MP cows with low Ca at parturition had improved health status when supplemented.

Association of immediate postpartum plasma calcium concentration with early-lactation clinical diseases, culling, reproduction, and milk production in Holstein cows.

The objective of this study was to evaluate the association of postpartum plasma Ca concentration with early-lactation disease outcomes, culling within 60 d in milk, pregnancy to first service, and milk production. A total of 1,453 cows from 5 commercial dairy farms in New York State were enrolled in a prospective cohort study from February to November 2015. Blood samples were collected within 12 h of parturition, and plasma was submitted to a diagnostic laboratory for total Ca measurement. Early-lactation disease, reproductive performance, and milk production from Dairy Herd Improvement Association (DHIA) test-day data were compiled from each farm's management software. Multivariable Poisson regression models were built to evaluate the association of plasma Ca with the risks of retained placenta (RP), metritis, displaced abomasum (DA), clinical mastitis, culling within 60 d in milk, and pregnancy to first service. Repeated-measures ANOVA were used to evaluate the association of Ca at parturition with milk production across the first 9 DHIA tests. Herd was considered a random effect in all models. Primiparous cows were modeled separately from multiparous cows if differential responses were observed. Calcium was not associated with the risk of RP, metritis, clinical mastitis, or pregnancy to first service in primiparous or multiparous cows. For multiparous cows only, higher Ca concentration tended to be associated with increased culling within the first 60 d in milk. Multiparous cows with Ca ≤1.85 mmol/L had an increased risk of being diagnosed with a DA compared with cows with Ca >1.85 mmol/L. For the milk production models, Ca was not associated with the amount of milk produced within the first 9 DHIA tests in primiparous cows; however, multiparous cows with Ca ≤1.95 mmol/L produced, on average, 1.1 kg more milk per day across the 9 DHIA tests than their multiparous counterparts with Ca >1.95 mmol/L. Our results indicate that plasma Ca concentration measured within 12 h of parturition is a poor predictor of early-lactation health outcomes. Reduced Ca concentration in the immediate postpartum period was associated with higher milk production in multiparous cows. From these results, we caution that studies attempting to categorize subclinical hypocalcemia based on a single sample in the immediate postpartum period could misclassify the disorder.

Short communication: Relationship between methods for measurement of serum electrolytes and the relationship between ionized and total calcium and neutrophil oxidative burst activity in early postpartum dairy cows.

The objectives of this study were to (1) compare a test for serum measurement of total Ca (tCa), Mg, and P (VetTest Chemistry Analyzer, IDEXX Laboratories Inc., Westbrook, ME) to reference methods (spectrophotometric assays on a Beckman Coulter 640e automated clinical chemistry analyzer; Beckman Coulter, Brea, CA), (2) determine the relationship between ionized Ca (iCa) and reference method tCa in the immediate postpartum period, and (3) assess the relative value of these blood Ca indices as predictors of neutrophil oxidative burst activity. Samples were collected from multiparous Holstein cows (n = 33) over the first 5 d in milk. A total of 183 samples for objective 1 and 181 samples for objective 2 were available. Neutrophil oxidative burst activity was assessed once between 2 and 5 d in milk (n = 29). Linear regression demonstrated strong relationships between serum tCa, Mg, and P concentrations measured by the VetTest compared with the reference method. Bland Altman analysis indicated that the VetTest values were higher than the reference method by 0.22 mmol/L for tCa, 0.12 mmol/L for Mg, and 0.16 mmol/L for P. Compared with hypocalcemia categorized at ≤2.0 or ≤2.125 mmol/L with the reference method tCa, thresholds for the VetTest measured tCa of ≤2.23 mmol/L (sensitivity = 87%, specificity = 89%) or ≤2.30 mmol/L (sensitivity = 86%, specificity = 96%) could be used. The relationship between whole-blood iCa and reference method serum tCa differed by sampling time point after calving. Compared with identification of hypocalcemia with serum tCa measurements from the reference method (thresholds of ≤2.0 and 2.125 mmol/L), a whole-blood iCa threshold of ≤1.17 mmol/L resulted in the highest combined sensitivities (94 and 82%) and specificities (80 and 94%) at either threshold. Ionized Ca measurements were more consistently related to outcomes of neutrophil oxidative burst activity measured in vitro. The VetTest measurements of serum tCa reliably identified hypocalcemia when thresholds were adjusted to account for the bias of the test. The variation in the relationship between iCa and reference method tCa in the days following parturition suggest that these measures cannot be used interchangeably as indicators of Ca status. The more consistent associations between iCa and in vitro measures of neutrophil function, compared with tCa, indicated that this may be a more sensitive predictor of functional outcomes associated with postpartum Ca status.

The effect of source of supplemental dietary calcium and magnesium in the peripartum period, and level of dietary magnesium postpartum, on mineral status, performance, and energy metabolites in multiparous Holstein cows.

The objective of this study was to determine the effects of feeding different supplemental sources of Ca and Mg in the peripartum period, and different dietary levels of Mg postpartum, on plasma mineral status, performance, and aspects of energy metabolism in transition dairy cows. Multiparous Holstein cows (n = 41) were used in a completely randomized design with a 2 × 2 factorial arrangement of treatments starting at 28 d before expected parturition. Main effects were source assignments (CS = common sources of supplemental Ca and Mg, or MA = a blend of common and commercial mineral sources with supplemental minerals primarily from a commercial Ca-Mg dolomite source; MIN-AD, Papillon Agricultural Company Inc., Easton, MD) beginning at 21 d before due date; cows were further randomized within source treatments to 1 of 2 levels of Mg supplementation (LM = formulated postpartum diet Mg at 0.30% of dry matter (DM), or HM = formulated postpartum diet Mg at 0.45% of DM) beginning within 1 d after parturition. Final treatment groups included the following: common source, low Mg (CS-LM, n = 11); common source, high Mg (CS-HM, n = 11); MIN-AD, low Mg (MA-LM, n = 10); and MIN-AD, high Mg (MA-HM, n = 9). Treatment diets were fed and data collected through 42 d in milk. Postpartum plasma Mg concentrations tended to be higher for cows fed HM and cows fed CS, but no effects were observed on peripartum plasma Ca concentrations. Peripartum plasma P concentrations were higher for cows fed MA. Dry matter intake (DMI) in the prepartum period was higher for cows fed MA (CS = 15.9 vs. MA = 16.8 kg/d) and postpartum DMI was higher in some groups depending on week. Plasma nonesterified fatty acid concentrations were lower for cows fed MA during both the prepartum and postpartum periods. A source by level interaction was observed for postpartum plasma ß-hydroxybutyrate (BHB) concentrations such that cows fed CS-LM had numerically higher BHB and cows fed MA-LM had numerically lower BHB (geometric means; CS-LM = 7.9, CS-HM = 6.9, MA-LM = 6.3, and MA-HM = 7.3 mg/dL) than cows fed the other 2 treatments. Higher milk fat yield, milk fat content, and fat- and energy-corrected yield during wk 1 for cows fed MA resulted in source by week interactions for these outcomes. This study demonstrated that varying supplemental Ca and Mg sources and feeding rates had minimal effect on plasma Ca status despite differences in plasma Mg and P concentrations. Effects on DMI and plasma energy metabolites suggest an opportunity for strategic use of mineral sources in the transition period to promote metabolic health.

Effects of prepartum dietary cation-anion difference on aspects of peripartum mineral and energy metabolism and performance of multiparous Holstein cows.

The objectives of this study were to determine the effect of decreasing dietary cation-anion difference [DCAD; (Na+ + K+) - (Cl- + S2-)] of the prepartum diet on aspects of mineral metabolism, energy metabolism, and performance of peripartum dairy cows. Multiparous Holstein cows (n = 89) were enrolled between 38 and 31 d before expected parturition and randomized to treatments in a completely randomized design (restricted to balance for previous 305-d mature equivalent milk production, parity, and body condition score) at 24 d before expected parturition. Treatments consisted of a low-K ration without anion supplementation [CON; n = 30, DCAD = +18.3 mEq/100 g of dry matter (DM)]; partial anion supplementation to a low-K ration (MED; n = 30, DCAD = +5.9 mEq/100 g of DM); and anion supplementation to a low-K ration to reach a targeted average urine pH between 5.5 and 6.0 (LOW; n = 29, DCAD = -7.4 mEq/100 g of DM). Cows were fed a common postpartum diet and data collected through 63 d in milk. Urine pH (CON = 8.22, MED = 7.89, and LOW = 5.96) was affected quadratically by decreasing prepartum DCAD. A linear relationship between urine pH and urine Ca:creatinine ratio was observed (r = -0.81). Plasma Ca concentrations in the postpartum period (d 0 to 14; CON = 2.16, MED = 2.19, and LOW = 2.27 mmol/L) were increased linearly with decreasing prepartum DCAD. A treatment by parity (second vs. third and greater) interaction for postpartum plasma Ca concentration suggested that older cows had the greatest response to the low DCAD diet and older cows fed LOW had decreased prevalence of hypocalcemia after calving. A quadratic effect of decreasing DCAD on prepartum DMI was observed (CON = 13.6, MED = 14.0, and LOW = 13.2 kg/d). Milk production in the first 3 wk postpartum was increased linearly with decreasing DCAD (CON = 40.8, MED = 42.4, and LOW = 43.9 kg/d) and DMI in this period also tended to linearly increase (CON = 20.2, MED = 20.9, and LOW = 21.3 kg/d). Overall, effects on intake and milk yield analyzed over wk 1 to 9 postpartum were not significant. This study demonstrates that feeding lower DCAD diets prepartum improves plasma Ca status in the immediate postpartum period and results in increased DMI and milk production in the 3 wk after parturition. Compared with no anion supplementation or lower levels of anion supplementation, greater improvements were observed with the lower DCAD feeding strategy, in which an average urine pH of 5.5 to 6.0 was targeted.

Risk factors associated with postpartum subclinical hypocalcemia in dairy cows.

Our study objectives were to evaluate the association of prepartum plasma Mg concentrations with subclinical hypocalcemia (SCH) classification at parturition and to evaluate the association of other cow-level risk factors with SCH classification at calving or at 2 d in milk (DIM). A total of 301 animals from 2 dairy herds located in New York were enrolled in a cohort study. Blood samples were collected at approximately 1 wk before the expected calving date, within 4 h of calving, and at 2 DIM. Prepartum samples had plasma macromineral concentrations (Ca, K, Mg, P), albumin, and ß-hydroxybutyrate analyzed. Samples collected at calving were analyzed for Ca only, and samples from 2 DIM had macromineral and albumin concentrations determined. Postpartum SCH was defined as Ca concentrations ≤2.1 mmol/L. The prevalence of SCH at calving was 2, 40, and 66% for first, second, and third or greater parities, respectively. Only 4% of cows could be classified with prepartum subclinical hypomagnesemia (Mg concentrations <0.8 mmol/L), which did not provide enough power to appropriately determine the association of plasma Mg with postpartum Ca concentrations and its effect on SCH classification. Multiparous cows with Ca concentrations ≤2.4 mmol/L in the prepartum period and third or greater parity cows had a higher risk of being categorized as SCH at calving [relative risk (RR) = 1.4 and 1.7, respectively]. The risk of SCH at 2 DIM was associated with the interaction of Ca status at calving and lameness score. Nonlame cows with Ca concentrations ≤2.1 mmol/L (RR = 3.2) and normocalcemic lame cows at parturition (RR = 3.4) were more likely to be SCH at 2 DIM compared with nonlame normocalcemic cows. In conclusion, we identified a prepartum Ca cut-point for identification of cows that are more likely to be classified as SCH at calving. Different risk factors were associated with SCH depending on the timing of diagnosis relative to parturition.