Cow-level factors associated with subclinical hypocalcemia at calving in multiparous Jersey cows

The objective of our study was to identify cow-level factors associated with subclinical hypocalcemia at calving (SCH) in multiparous Jersey cows. A total of 598 Jersey and 218 Jersey × Holstein crossbreed cows from 2 commercial dairy herds were enrolled in a retrospective cohort study. Blood samples to determine total Ca concentration were collected from the coccygeal vessels at 3 h 19 min (±2 h 33 min) after calving. We used 2 serum Ca concentration thresholds to define SCH: <2.00 mmol/L (SCH-2.00) and <2.12 mmol/L (SCH-2.12). We evaluated the association of cow-level factors with SCH with multivariable Poisson regression models. Variables evaluated for association with SCH were herd; parity (2, 3, and ≥4); breed; previous lactation length and 305-d mature-equivalent milk yield; previous lactation first test milk yield and last test somatic cell count; lengths of calving interval, gestation, dry, and close-up periods; body condition and locomotion scores at calving; calving ease; and calf sex for singletons. We categorized continuous variables into quartiles (≤25th percentile, interquartile range and ≥75th percentile). The prevalence of SCH among Jersey cows was 40 (SCH-2.00) and 64% (SCH-2.12). Jersey cows of higher parity had greater risk of SCH-2.00 and SCH-2.12. The risk of SCH-2.12 was higher after birthing male calves. We also found a tendency for previous lactation length and previous lactation 305-d mature-equivalent milk yield effect to affect risk of SCH-2.12. The risk of SCH-2.12 was lower for cows that had a previous lactation length shorter than the 25th percentile compared with cows that had a previous lactation length within the interquartile range. The risk of SCH-2.12 was higher for cows that had a previous lactation 305-d mature-equivalent milk yield below the 25th percentile compared with cows that had a previous lactation 305-d mature-equivalent milk yield above the 75th percentile. Also, Jersey × Holstein crossbreed was associated with increased risk of SCH-2.00. In the multivariable analysis, we observed no association between SCH and previous lactation first test milk yield; last test somatic cell count; lengths of calving interval, gestation, dry, and close-up periods; body condition and locomotion scores at calving; and calving ease. Our study identified parity, breed, calf sex, previous lactation length, and previous lactation 305-d mature-equivalent milk yield as cow-level factors associated with SCH in multiparous Jersey cows.     

Field trial of 2 calcium supplements on early lactation health and production in multiparous Holstein cows

Our objectives were to measure serum Ca concentrations in the first 48 h postpartum in cows supplemented with oral Ca or subcutaneous Ca and nonsupplemented cows and evaluate the effect of these treatments on the incidence of metritis, displaced abomasum, mastitis, and early lactation disease (any of the diseases milk fever, retained placenta, metritis, or displaced abomasum), removal from the herd, pregnancy to first insemination, and average daily milk yield for the first 10 wk of lactation. We conducted 2 experiments on 1 commercial herd in New York State. In experiment 1, multiparous Holstein cows (n = 30) were blocked by parity (2 and ≥3) and sequentially assigned at calving to nontreated control (CON, n = 10), subcutaneous administration of 500 mL 23% Ca gluconate at calving (SC, n = 10), or administration of an oral Ca bolus containing 43 g of calcium at calving and again 12 h later (OB, n = 10). Blood was collected before treatment and at 1, 2, 4, 8, 12, 24, and 48 h thereafter for measurement of serum total Ca concentration. In experiment 2, 1,478 multiparous Holstein cows were sequentially assigned by calving date to the same 3 treatments (CON, n = 523; SC, n = 480; OB, n = 475). In experiment 1, SC cows had greater Ca concentrations from 1 through 12 h post-treatment and OB cows had greater Ca concentrations at 1 and 24 h post-treatment compared with CON cows. We found no difference in risk of metritis, displaced abomasum, early lactation disease diagnosis, or pregnancy to first insemination among treatments. Treatment with SC or OB had no effect on average daily milk yield compared with CON cows (CON = 46.7 kg; SC = 47.1 kg; OB = 47.0 kg). Cows treated with SC or OB that had a high relative herd milk rank in the previous lactation were almost half as likely to be diagnosed with mastitis in the first 60 DIM compared with CON cows [risk ratio (RR)_SC = 0.57, RR_OB = 0.54]; however, we found no difference in risk of mastitis among treatments for cows with low relative herd milk rank. Second-parity cows fed a negative prepartum dietary cation-anion difference ration and treated with SC or OB were more likely to be removed from the herd than CON cows (RR_SC = 3.91, RR_OB = 4.72); this difference was not observed in second-parity cows fed a neutral prepartum dietary cation-anion difference ration or in parity ≥3 cows. Although Ca supplementation increased serum Ca, this effect did not greatly improve milk production or health and reproductive outcomes.     

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.     

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.     

Association of low serum calcium concentration after calving with productive and reproductive performance in multiparous Jersey cows

Consequences of postpartum low blood calcium (Ca) concentration are still under study and literature describing this condition in Jersey cows is scarce. A prospective cohort study was conducted to evaluate the association of low serum Ca concentration shortly after calving with milk and energy-corrected milk yields, somatic cell count linear score, and pregnancy to first service and within 150 d in milk in multiparous Jersey cows from 2 commercial herds. Blood samples for serum Ca determination were collected on average at 3 h 10 min postpartum from 352 multiparous Jersey cows. Productive data up to the 10th monthly test were obtained from the Dairy Herd Improvement Association and reproductive data were obtained from herd records. Multiple linear, log-binomial, and Cox's proportional hazards regressions were used to evaluate the association of low serum Ca concentration with productive and reproductive outcomes. Serum Ca concentration ≤2.18 mmol/L was associated with 1.43 and 1.85 kg/d more milk and energy-corrected milk. However, lower serum Ca concentrations were associated with a 0.28-unit-higher somatic cell count linear score per monthly test (Ca ≤2.00 mmol/L), and decreased pregnancy risk at first service (risk ratio = 0.64; Ca ≤1.94 mmol/L) and hazard of pregnancy within 150 d in milk (hazard ratio = 0.40; Ca ≤1.90 mmol/L). The present study is based on a convenience sample of multiparous Jersey cows from 2 commercial herds; further research including more herds and additional blood Ca determinations is needed to describe postpartum blood Ca dynamics and its association with productive and reproductive outcomes for the Jersey breed.     

Timothy hay with a low dietary cation-anion difference improves calcium homeostasis in periparturient Holstein cows

The current study was undertaken to evaluate the effects of feeding timothy (Phleum pratense L.) hay differing in dietary cation-anion difference (DCAD) on the capability of cows to maintain calcium homeostasis around parturition. We hypothesized that feeding low-DCAD timothy hay during the prepartum period would induce a mild metabolic acidosis prepartum and improve calcium homeostasis postpartum with no effect on dry matter intake. Forty-one dry pregnant Holstein cows entering their second lactation or greater were used in a randomized complete block design. Timothy hay was obtained from an established timothy stand under a pivot irrigation system. Low-DCAD timothy hay was produced by fertilizing the area between the second and third pivot towers at a rate of 224 kg of CaCl₂/ha, and control timothy hay (high DCAD) was grown on the area between the fourth and fifth pivot towers of the same field. The chloride concentration was 1.07 and 0.15% on a dry matter (DM) basis, and the DCAD was 1.2 and 21.6 mEq/100 g of DM for the low- and high-DCAD timothy hay, respectively. Experimental diets, containing timothy hay at 63% of dietary DM, were fed ad libitum starting 30 d before the expected calving date. The DCAD values were 1.6 vs. 14.5 mEq/100 g of DM for the low- and high-DCAD timothy-based diets, respectively. At the beginning of the study, urine pH and blood bicarbonate concentration averaged 8.22 ± 0.06 and 28.5 ± 0.3 mM, respectively. The low-DCAD timothy diet decreased urine pH compared with the high-DCAD timothy diet on d 21 (7.75 vs. 8.31), d 14 (7.69 vs. 8.22), and d 7 (7.50 vs. 8.19) before calving, and it also decreased the prepartum blood bicarbonate concentration by 2 mM. In addition, cows fed the low-DCAD timothy diet had greater blood ionized calcium concentration prepartum (1.22 vs. 1.19 mM), greater blood ionized calcium concentration at 0 and 8 h after calving, and similar prepartum dry matter intake. These results indicate that timothy hay differing in DCAD affects the acid-base balance of periparturient dairy cows, and that low-DCAD timothy hay improves calcium homeostasis postpartum with no negative effect on dry matter intake.     

Increased serum serotonin improves parturient calcium homeostasis in dairy cows

Hypocalcemia in dairy cows is caused by the sudden increase in calcium demand by the mammary gland for milk production at the onset of lactation. Serotonin (5-HT) is a key factor for calcium homeostasis, modulating calcium concentration in blood. Therefore, it is hypothesized that administration of 5-hydroxy-l-tryptophan (5-HTP), a 5-HT precursor, can increase 5-HT concentrations in blood and, in turn, induce an increase in blood calcium concentration. In this study, 20 Holstein dairy cows were randomly assigned to 2 experimental groups. Both groups received a daily i.v. infusion of 1 L of either 0.9% NaCl (C group; n = 10) or 0.9% NaCl containing 1 mg of 5-HTP/kg of BW (5-HTP group, n = 10). Infusions started d 10 before the estimated parturition and ceased the day of parturition, resulting in at least 4 d of infusion (8.37 ± 0.74 d of infusion). Until parturition, blood samples were collected every morning before the infusions, after parturition samples were taken daily until d 7, and a final sample was collected on d 30. Milk yield was recorded during this period. No differences between groups were observed for blood glucose, magnesium, and β-hydroxybutyrate. Cows receiving the 5-HTP infusion showed an increase in fatty acid concentrations from d ?3 to ?1 before parturition. Serum 5-HT concentrations were increased at d ?4 related to parturition until d 5 postpartum in the 5-HTP group compared with the C group. In addition, cows from the 5-HTP group had increased 5-HT concentrations in colostrum, but not in mature milk, on d 7 postpartum. Serum calcium concentrations decreased in both groups around parturition; however, calcium remained higher in the 5-HTP group than in controls, with a significant difference between groups on d 1 (1.62 ± 0.08 vs. 1.93 ± 0.09 mmol/L in control and 5-HTP groups, respectively) and d 2 (1.83 ± 0.06 vs. 2.07 ± 0.07 mmol/L in control and 5-HTP groups, respectively). Additionally, colostrum yield (first milking) was lower in the 5-HTP group compared with the C group, but without consequences on colostrum IgG concentrations. Milk yield did not differ between groups during the rest of the experiment. The study data were consistent with the concept that infusion of 5-HTP to dairy cows increases blood 5-HT concentrations, which in turn is a significant regulatory component in the chain of effectors that affect calcium status around parturition, hence the occurrence of clinical or subclinical hypocalcemia.     

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 β-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.     

Effect of oral calcium bolus supplementation on early-lactation health and milk yield in commercial dairy herds

The objective of this study was to evaluate the effect of supplementation with oral Ca boluses after calving on early-lactation health and milk yield. Cows in their second lactation or greater (n = 927) from 2 large dairies in Wisconsin were enrolled during the summer of 2010. Both herds were fed supplemental anions during the prefresh period and less than 1% of fresh cows were treated for clinical milk fever. Cows were scored before calving for lameness and body condition, and then randomly assigned to either a control group or an oral Ca bolus-supplemented group. Control cows received no oral Ca boluses around calving. Cows in the oral Ca bolus group received 2 oral Ca boluses (Bovikalc, Boehringer Ingelheim, St. Joseph, MO), one bolus 0 to 2 h after calving and the second 8 to 35 h after calving. The oral Ca bolus administration schedule allowed fresh cows to be restrained in headlocks only once daily. Whole-blood samples were collected immediately before the second oral Ca bolus was given and were analyzed for ionized Ca (Ca²⁺) concentration. Early-lactation health events were recorded and summed for each cow. Only 6 cases (0.6% of calvings) of clinical milk fever occurred during the trial, and only 14% of cows tested were hypocalcemic (Ca²⁺ less than 1.0 mmol/L) at 8 to 35 h after calving. Mean Ca²⁺ concentrations were not different between the control and oral Ca bolus-supplemented groups. Blood samples from the cows given oral Ca boluses were collected an average of 20.6 h after administration of the first bolus. Subpopulations of cows with significant responses to oral Ca bolus supplementation were identified based on significant interactions between oral Ca bolus supplementation and covariates in mixed multiple regression models. Lame cows supplemented with oral Ca boluses averaged 0.34 fewer health events in the first 30 d in milk compared with lame cows that were not supplemented with oral Ca boluses. Cows with a higher previous lactation mature-equivalent milk production (greater than 105% of herd rank) and supplemented with oral Ca boluses produced 2.9 kg more milk at their first test after calving compared with cows with higher previous lactation milk yields that were not supplemented. Results of this study indicate that lame cows and higher producing cows responded favorably to supplementation with oral Ca boluses. Supplementing targeted subpopulations of cows with oral Ca boluses was beneficial even for dairies with a very low incidence of hypocalcemia.     

Serum calcium dynamics within the first 3 days in milk and the associated risk of acute puerperal metritis

The objectives of this study were to evaluate postpartum serum calcium dynamics for different parity groups of dairy cows and to assess whether serum calcium concentration on d 0, 1, and 3 postpartum was associated with the risk of developing acute puerperal metritis (APM). The study took place on a commercial dairy farm in northern Germany and included 4,043 Holstein dairy cows. Calving difficulties, such as dystocia, twins, or stillbirth, were recorded. Blood samples were obtained on d 0, 1, and 3 after calving for analysis of serum calcium concentration. Animals were examined daily for clinical symptoms of retained placenta, APM, mastitis, and displaced abomasum until 10 d in milk. To determine serum calcium dynamics postpartum, we performed repeated-measures ANOVA with first-order autoregressive covariance. A logistic regression model was used to evaluate the association of serum calcium concentration with the risk of developing metritis. Serum calcium concentration was affected by time relative to calving, parity, and APM. Increasing parity negatively affected serum calcium concentration on d 0 and 1. Serum calcium concentration reached its lowest level on d 1 and 3 in multiparous and primiparous cows, respectively. The concentration increased from d 1 to 3 in multiparous cows and decreased from d 0 to 3 in primiparous cows. The association of APM and serum calcium dynamics varied by parity. On d 3, serum calcium concentration was significantly lower in animals with subsequent APM than in those without APM. The overall incidence of APM was 12.0% (primiparous cows, 20.4%; multiparous cows, 8.6%). An association existed between serum calcium concentration on d 3 after calving and APM. Primiparous cows had an odds ratio of 0.12 for serum calcium concentration on d 3, indicating that a primiparous cow with serum calcium concentration of 2.5 mmol/L had a 88% lower chance of developing APM compared with a cow with a concentration of 1.5 mmol/L. Multiparous cows had an odds ratio of 0.34 for serum calcium concentration on d 3, indicating that a multiparous cow with serum calcium concentration of 2.5 mmol/L had a 66% lower chance of developing APM compared with a cow with a concentration of 1.5 mmol/L. Primiparous cows with low serum calcium concentration had the highest predicted probability of developing APM. Our results reveal a dynamic in serum calcium concentration in the first 3 d in milk. Consequently, the day of sampling and the observed risk period for hypocalcemia are important when conducting epidemiological studies to evaluate associations between hypocalcemia and clinical diseases.     

Evaluation of peripartal calcium status,energetic profile,and neutrophil function in dairy cows at low or high risk of developing uterine disease

In this prospective cohort study, Holstein cows considered to be at high risk of developing metritis (dystocia, twins, stillbirth, retained placenta, or their combination) were matched with herdmates at low risk of developing metritis (normal calving) and monitored daily for rectal temperature and uterine discharge during the first 12 d in milk (DIM). Blood was sampled on d 0, 1, and 3 postpartum for assessment of neutrophil phagocytic and oxidative burst activities. Blood was also sampled at 0, 1, 2, 3, 4, 7, and 12 DIM for determination of serum concentrations of Ca, K, Mg, nonesterified fatty acids, β-hydroxybutyrate, and glucose. On the basis of receiver operator characteristic curves, subclinical hypocalcemia (SCH) was defined as a serum Ca concentration ≤8.59 mg/dL in at least 1 sample in the first 3 DIM. The overall incidences of metritis and puerperal metritis were 47.3 and 30%, respectively. Concentration in blood and percentages of neutrophils undergoing phagocytosis and oxidative burst were all reduced in cows with SCH compared with normocalcemic cows. Cows with SCH were at a greater risk of developing fever, metritis, and puerperal metritis compared with normocalcemic cows. Among cows at low risk of developing metritis, those with SCH had a greater incidence of metritis (40.7%) compared with normocalcemic cows (14.3%). Similarly, among cows at high risk of developing metritis, cows with SCH had a greater incidence of metritis (77.8%) compared with normocalcemic cows (20.0%). Cows with SCH had elevated concentrations of nonesterified fatty acids and β-hydroxybutyrate compared with normocalcemic cows. The relative risk of developing metritis decreased by 22% for every 1 mg/dL increase in serum Ca. Neither metritis nor SCH influenced the resumption of estrous cyclicity by 38 DIM, but cows with SCH had a reduced pregnancy rate and a longer interval to pregnancy compared with normocalcemic cows. Finally, the population risk to develop uterine diseases attributable to SCH was 66.6% for metritis and 91.3% for puerperal metritis in the present study.     

Associations of subclinical hypocalcemia with fertility in a herd of grazing dairy cows

The main objective was to assess the associations of subclinical hypocalcemia (SCH), diagnosed at parturition (SCH-0) and 7 d in milk (SCH-7), with fertility in a herd of grazing dairy cows. Additional objectives were to characterize Ca concentration on 0 and 7 d in milk (DIM), assessing the risk factors for SCH-0 and SCH-7 and also the relationship with health status (metritis, endometritis, subclinical ketosis, and culling). A prospective observational study was carried out in a dairy farm in Argentina. Holstein cows (n = 126) were body condition scored (BCS, 1–5) on ?21 ± 3, 0, 7 ± 3, and 28 ± 7 DIM and blood was collected on 0 and 7 ± 3 DIM to determine Ca and β-hydroxybutyrate concentrations. Calcium concentrations <2.0 and <2.14 mmol/L were used to define SCH-0 and SCH-7, respectively. The associations of SCH with (1) the odds for pregnancy to first service (P1AI) and pregnancy by 100 DIM (P100) were evaluated by logistic models, (2) the services per pregnancy was evaluated by a Poisson regression model, and (3) the hazards of insemination and pregnancy were evaluated with proportional hazards regression models whereas median days from calving to first insemination and to pregnancy were estimated by Kaplan-Meier method. Additionally, Ca concentration was assessed by linear regression models, and the associations of SCH-0 and SCH-7 with the odds for metritis, endometritis, subclinical ketosis, and culling were evaluated by logistic models. Calcium concentrations were similar at 0 and 7 DIM (2.40 vs. 2.41 mmol/L, respectively); they were higher in cows calving in fall than in summer (2.58 vs. 2.24 mmol/L), and they also were higher in primiparous than in multiparous cows (2.53 vs. 2.28 mmol/L, respectively). The proportion of cows having SCH-0 and SCH-7 was 27.3 and 39.3%, respectively. Fall-calving cows had lower odds for SCH-0 [odds ratio (OR) = 0.31, 95% confidence interval (CI) = 0.12–0.86] than summer-calving cows, multiparous cows had higher odds for SCH-0 (OR = 3.96, 95% CI = 1.09–14.39) than primiparous cows, and cows with prepartum BCS ≥3.00 had higher odds for SCH-0 (OR = 4.03, 95% CI = 1.17–13.89) than in cows with BCS <3.00. Conversely, parity and prepartum BCS were not important predictors for SCH-7. Surprisingly, SCH-0 was not a risk factor for SCH-7. Cows with SCH-0 had lower odds for P1AI (OR = 0.26, 95% CI = 0.07–0.99) than normocalcemic cows, given that P1AI was 14 versus 38%, respectively. The hazard of first service was not associated with SCH-0 (hazard ratio = 1.03, 95% CI = 0.63–1.70) but cows with SCH-0 had lower hazard of pregnancy (hazard ratio = 0.39, 95% CI = 0.16–0.98) and took 32 d longer to get pregnant (105 vs. 73) than normocalcemic cows. Conversely, SCH-7 was not associated with fertility. Finally, SCH-0 and SCH-7 were associated with the odds for subclinical ketosis and metritis, respectively. In conclusion, SCH-0 but not SCH-7 is associated with reduced fertility in a herd of grazing dairy cows, but both were associated with health status.     

Effect of induced subclinical hypocalcemia on physiological responses and neutrophil function in dairy cows

The objectives were to study the effects of induced subclinical hypocalcemia [SCH, blood ionized Ca (iCa²⁺) <1.0 mM, without recumbency] on physiological responses and function of immune cells in dairy cows. Ten nonpregnant, nonlactating Holstein cows were blocked by lactation and assigned randomly to a normocalcemic (NC; intravenous infusion of 0.9% NaCl i.v. plus 43 g of oral Ca, as Ca sulfate and Ca chloride, at −1 and 11 h) or an induced SCH [SCHI, 5% ethylene glycol tetraacetic acid (EGTA), a selective iCa²⁺ chelator, intravenous infusion] treatment for 24 h, using a crossover design. The sequence of treatments was either NC–SCHI or SCHI–NC, with a 6-d washout period. Ionized Ca was evaluated before, hourly during the infusion period, and at 48 and 72 h, to monitor concentrations and adjust the rate of infusion, maintaining blood iCa²⁺ <1.0 mM in SCHI throughout the 24-h infusion period. Additional measurements included heart and respiratory rates, rectal temperature, dry matter intake, rumen contractions, whole-blood pH, concentrations of glucose and K in whole blood, concentrations of total Ca, Mg, nonesterified fatty acids, β-hydroxybutyrate, and insulin in plasma, and urinary excretion of Ca. Total and differential leukocyte count in blood was also performed. The concentration of cytosolic iCa²⁺ in neutrophils and lymphocytes was quantified and neutrophil function was assayed in vitro. Infusion of a 5% EGTA solution successfully induced SCH in all SCHI cows, resulting in decreased blood iCa²⁺ concentrations throughout the 24-h treatment period (0.77 ± 0.01 vs. 1.26 ± 0.01 mM iCa²⁺). Induction of SCH reduced dry matter intake on the day of infusion (5.3 ± 0.8 vs. 9.1 ± 0.8 kg/d) and rumen contractions (1.9 ± 0.2 vs. 2.7 ± 0.2 contractions/2 min) for the last 12 h of infusion. Cows in SCHI had decreased plasma insulin concentration (1.44 ± 0.23 vs. 2.32 ± 0.23 ng/mL) evident between 6 and 18 h after the beginning of the infusion, accompanied by increased concentrations of glucose (4.40 ± 0.04 vs. 4.17 ± 0.04 mM). Plasma nonesterified fatty acids concentration was greater for SCHI than NC cows (0.110 ± 0.019 vs. 0.061 ± 0.014 mM). Neutrophils of cows in SCHI had a faster decrease in cytosolic iCa²⁺ after stimulation with ionomycin (9.9 ± 1.0 vs. 13.6 ± 1.4 Fluo-4:Fura Red post-end ratio) in vitro. Furthermore, induction of SCH reduced the percentage of neutrophils undergoing phagocytosis (22.1 ± 2.1 vs. 29.3 ± 2.1%) and reduced the oxidative burst response after incubation of pathogenic bacteria (16.1 ± 1.7 vs. 24.2 ± 1.7%). Subclinical hypocalcemia compromised appetite, altered metabolism, and impaired function of immune cells in dairy cows.     

The effect of zeolite A supplementation in the dry period on periparturient calcium,phosphorus, and magnesium homeostasis

One potential way of preventing parturient hypocalcemia in the dairy cow is to feed dry cow rations very low in calcium (<20 g/d); but, because it is difficult to formulate rations sufficiently low in calcium, this principle has been almost abandoned. Recent studies have shown, however, that it is possible to prevent milk fever, as well as subclinical hypocalcemia, by supplementing the dry cow ration with sodium aluminium silicate (zeolite A), which has the capacity to bind calcium. The aim of this study was to further evaluate the effect, if any, of such supplementation on other blood constituents, feed intake, and milk production in the subsequent lactation. A total of 31 pregnant dry cows about to enter their third or later lactation were assigned as experimental or control cows according to parity and expected date of calving. The experimental cows received 1.4 kg of zeolite pellets per d (0.7 kg of pure zeolite A) for the last 2 wk of pregnancy. Blood samples were drawn from all cows 1 wk before the expected date of calving, at calving, at d 1 and 2 after calving, and 1 wk after calving. Additionally, a urine sample was drawn 1 wk before the expected date of calving. Zeolite supplementation significantly increased the plasma calcium level on the day of calving, whereas plasma magnesium as well as inorganic phosphate was suppressed. Serum 1,25(OH)2D was significantly increased 1 wk before the expected date of calving among the experimental cows, whereas there was no difference in the urinary excretion of the bone metabolite deoxypyridinoline between the two groups. Feed intake was decreased among the zeolite-treated cows during the last 2 wk of pregnancy. No effect was observed on milk yield, milk fat, and milk protein in the subsequent lactation. The mechanisms and interactions involved in zeolite supplementation are discussed in relation to the observed improvement in parturient calcium homeostasis and to the observed depression in blood magnesium and inorganic phosphate.     

Blood calcium concentration and performance in periparturient and early lactating dairy cows is influenced by plant bioactive lipid compounds

Previous studies ex vivo suggested that plant bioactive lipid compounds (PBLC) can increase ruminal calcium absorption. Therefore, we hypothesized that PBLC feeding around calving may potentially counteract hypocalcemia and support performance in postpartum dairy cows. The corresponding aim of the study was to investigate the effect of PBLC feeding on blood minerals in Brown Swiss (BS) and hypocalcemia-susceptible Holstein Friesian (HF) cows during the period from d −2 to 28 relative to calving and on milk performance until d 80 of lactation. A total of 29 BS cows and 41 HF cows were divided each into a control (CON) and PBLC treatment group. The latter was supplemented with 1.7 g/d menthol-rich PBLC from 8 d before expected calving to 80 d postpartum. Milk yield and composition, body condition score and blood minerals were measured. Feeding PBLC induced a significant breed × treatment interaction for iCa, supporting that PBLC increased iCa exclusively in HF cows; the increase was 0.03 mM over the whole period and 0.05 mM from d 1 to 3 after calving. Subclinical hypocalcemia was seen in one BS-CON and 8 HF-CON cows and 2 BS-PBLC and 4 HF-PBLC cows. Clinical milk fever was detected only in HF cows (2 HF-CON and one HF-PBLC). Other tested blood minerals, such as sodium, chloride, and potassium, as well as blood glucose, were neither affected by PBLC feeding nor breed, nor were their 2-way interactions, except for higher sodium levels in PBLC cows on d 21. Body condition score showed no effect of treatment, except for a lower body condition score in BS-PBLC compared with BS-CON at d 14. Dietary PBLC increased milk yield, milk fat yield, and milk protein yield at 2 consecutive dairy herd improvement test days. As indicated by treatment × day interactions, energy-corrected milk yield and milk lactose yield were increased by PBLC on the first test day only, and milk protein concentration decreased from test d 1 to test d 2 in CON only. The concentrations of fat, lactose, and urea, as well as somatic cell count, were not affected by treatment. The weekly milk yield over the first 11 wk of lactation was 29.5 kg/wk higher for PBLC versus CON across breeds. It is concluded that the applied PBLC induced a small but measurable improvement of calcium status in HF cows in the study period and had additional positive effects on milk performance in both breeds.     

Blood calcium dynamics after prophylactic treatment of subclinical hypocalcemia with oral or intravenous calcium

Total serum Ca dynamics and urine pH levels were evaluated after prophylactic treatment of subclinical hypocalcemia after parturition in 33 multiparous Jersey × Holstein crossbreed cows. Cows were blocked according to their calcemic status at the time of treatment [normocalcemic (8.0–9.9 mg/dL; n = 15) or hypocalcemic (5.0–7.9 mg/dL; n = 18)] and randomly assigned to 1 of 3 treatments: control [no Ca supplementation (n = 11)]; intravenous Ca [Ca-IV (n = 11), 500 mL of 23% calcium gluconate (10.7 g of Ca and 17.5 g of boric acid as a solubilizing agent; Durvet, Blue Springs, MO)]; or oral Ca [Ca-Oral (n = 11), 1 oral bolus (Bovikalc bolus, Boehringer Ingelheim, St. Joseph, MO) containing CaCl₂ and CaSO₄ (43 g of Ca) 2 times 12 h apart]. Total serum Ca levels were evaluated at 0, 1, 2, 4, 8, 12, 16, 20, 24, 36, and 48 h, and urine pH was evaluated at 0, 1, 12, 24, 36, and 48 h after treatment initiation. Total serum Ca levels were higher for Ca-IV than for control and Ca-Oral cows at 1, 2, and 4 h after treatment initiation, but lower than Ca-Oral cows at 20, 24, and 36 h and lower than control cows at 36 and 48 h. At 1 h after treatment initiation, when serum Ca levels for Ca-IV cows peaked (11.4 mg/dL), a greater proportion of Ca-IV (n = 8) cows had total serum Ca levels >10 mg/dL than control (n = 0) and Ca-Oral (n = 1) cows. At 24 h after treatment initiation, when Ca-IV cows reached the total serum Ca nadir (6.4 mg/dL), a greater proportion of Ca-IV (n = 10) cows had serum Ca levels <8 mg/dL than control (n = 5) and Ca-Oral (n = 2) cows. Treatment, time, and treatment × time interaction were significant for urine pH. Mean urine pH was lower for Ca-Oral cows (6.69) than for control (7.52) and Ca-IV (7.19) cows. Urine pH levels at 1 h after treatment were lower for Ca-IV cows compared with both control and Ca-Oral cows, a finding likely associated with the iatrogenic administration of boric acid added as a solubilizing agent of the intravenous Ca solution used. At 12, 24, and 36 h, urine pH levels were lower for Ca-Oral cows compared with both control and Ca-IV cows. This was expected because the oral Ca supplementation used (Bovikalc) is designed as an acidifying agent. Wide fluctuations in blood Ca were observed after prophylactic intravenous Ca supplementation. The implications for milk production and animal health, if any, of these transient changes in total serum Ca have yet to be evaluated.     

Method comparison and validation of a prototype device for measurement of ionized calcium concentrations cow-side against a point-of-care instrument and a benchtop blood-gas analyzer reference method

The objective of this study was to assess an optimized ion-selective electrode Ca-module prototype as a potential cow-side device for ionized Ca (iCa) measurements in bovine blood. A linearity experiment showed no deviation from linearity over a range of iCa concentrations compared with a commercial point-of-care (POC) device commonly used in the field (POC_VS; VetScan i-STAT, Abaxis North America, Union City, CA) and a laboratory gold standard benchtop blood-gas analyzer [reference analyzer (RA); ABL-800 FLEX, Radiometer Medical, Copenhagen, Denmark]. Coefficient of variation on 3 samples with high, within-range, and low iCa concentrations ranged from 1.0 to 3.9% for the prototype. A follow-up validation experiment was performed, in which our objectives were to (1) assess the performance of the prototype cow-side against the POC_VS (farm gold-standard) using fresh non-anticoagulated whole-blood samples; (2) assess the performance of the prototype and the POC_VS against the RA in a diagnostic laboratory using blood collected in a heparin-balanced syringe; and (3) assess the agreement of the prototype and POC_VS on-farm (fresh non-anticoagulated whole blood) against the RA on heparin-balanced blood. Finally, sensitivity and specificity of the results obtained by the prototype and the POC_VS cow-side compared with the results obtained by the laboratory RA using 3 different iCa cut points for classification of subclinical hypocalcemia were calculated. A total of 101 periparturient Holstein cows from 3 dairy farms in New York State were used for the second experiment. Ionized Ca results from the prototype cow-side were, on average, 0.06 mmol/L higher than the POC_VS. With heparin-balanced samples under laboratory conditions, the prototype and POC_VS measured an average 0.04 mmol/L higher and lower, respectively, compared with the RA. Results from the prototype and POC_VS cow-side were 0.01 mmol/L higher and 0.05 mmol/L lower, respectively, compared with results from the laboratory RA on heparinized blood. Sensitivity and specificity for the prototype and the POC_VS under farm conditions at 3 potential subclinical hypocalcemia cut points were 100 and ≥93.5%, respectively. This novel ion-selective electrode Ca-module could become a rapid low-cost tool for assessing iCa cow-side, while qualitatively allowing classification of subclinical hypocalcemia on-farm.     

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.     

Short communication: Assessing the accuracy of inline milk fat-to-protein ratio data as an indicator of hyperketonemia in dairy cows in herds with automated milking systems

The objective of this study was to evaluate the accuracy of inline milk fat-to-protein (F:P) data to detect hyperketonemia (HYK) in herds with automated milking systems (AMS). The F:P ratio has been investigated as a tool for detecting HYK with moderate accuracy in past studies, but inline F:P data in AMS may also be useful for HYK screening. To assess the accuracy of these data in commercial settings, we monitored 484 cows from 9 AMS herds for their first 3 wk of lactation, taking blood samples once per week (n = 1,427). Positive cases of HYK were defined by whole-blood β-hydroxybutyrate (BHB) concentrations ≥1.2 or ≥1.4 mmol/L. Milk data were collected from the AMS software on each farm for each cow and converted into 4 different F:P values: (1) value from the same day as the BHB test; (2) 5-d centered-moving average (CMA); (3) 5-d backward-moving average (BMA); (4) 5-d forward-moving average (FMA). In linear regression models, all 4 values were associated with BHB, but slope estimates varied and R² were low: same day (slope = 0.95, R² = 0.07), CMA (slope = 1.05, R² = 0.07), BMA (slope = 0.65, R² = 0.04), and FMA (slope = 1.23, R² = 0.09). In logistic regression models, the odds of having HYK (BHB ≥1.2 mmol/L) increased with every 0.1-unit increase from the mean F:P ratio (1.16) using same-day values (odds ratio = 1.35, 95% confidence interval = 1.25–1.47) and CMA (odds ratio = 1.39, 95% confidence interval = 1.27–1.51). The same increase in F:P from mean BMA (1.14) and FMA (1.17) was associated with 1.22 and 1.49 times the odds of HYK, respectively. For all 4 F:P variations, we evaluated the sensitivity, specificity, positive predictive value, and negative predictive value of different F:P thresholds with HYK status. As the F:P threshold increased from 1.17 to 1.50, sensitivity decreased (range: 77 to 9%) but specificity increased (range: 58 to 96%). Same-day and CMA F:P cutoffs at which a balance was reached between sensitivity and specificity ranged from 1.18 to 1.22; however, even at these values we found high rates of false positives and negatives (range: 31–39%). These results suggest that inline milk F:P data from inconsistently calibrated sensors should not be used alone to detect HYK in AMS herds.     

Use of calcitriol to maintain postpartum blood calcium and improve immune function in dairy cows

Our objectives were to determine the effects of an injectable formulation of calcitriol on mineral metabolism and immune function in postpartum Holstein cows that received an acidogenic diet prepartum to minimize hypocalcemia. In experiment 1, cows within 6 h of calving received calcitriol (0, 200, or 300 μg) to determine the dose needed to increase plasma concentrations of Ca; 300 μg was sufficient to sustain Ca for at least 3 d. In experiment 2, multiparous cows were assigned randomly to receive only vehicle (control, n = 25) or 300 μg of calcitriol (n = 25) subcutaneously within the first 6 h after calving. Blood was sampled before treatment and 12 h later, then daily until 15 d in milk (DIM), and analyzed for concentrations of ionized Ca (iCa), total Ca (tCa), total Mg (tMg), and total P (tP), metabolites, and hormones. Urine was sampled in the first 7 DIM and analyzed for concentrations of tCa, tMg, and creatinine. Neutrophil function was evaluated in the first week postpartum. Dry matter intake and production performance were evaluated for the first 36 DIM. Calcitriol administration increased concentrations of calcitriol in plasma within 12 h of application from 51 to 427 pg/mL, which returned to baseline within 5 d. Concentrations of iCa and tCa increased 24 h after treatment with calcitriol. Concentrations of iCa (control = 1.08 vs. calcitriol = 1.20 mM), tCa (control = 2.23 vs. calcitriol = 2.33 mM), and tP (control = 1.47 vs. calcitriol = 1.81 mM) remained elevated in cows treated with calcitriol until 3, 5, and 7 DIM, respectively, whereas concentration of tMg (control = 0.76 vs. calcitriol = 0.67 mM) was less in calcitriol cows than control cows until 3 DIM. Concentrations of parathyroid hormone decreased in calcitriol cows compared with control cows (control = 441 vs. calcitriol = 336 pg/mL). Calcitriol tended to increase plasma concentrations of β-hydroxybutyrate and serotonin, but concentrations of glucose, nonesterified fatty acids, and C-telopeptide of type I collagen in plasma did not differ between treatments. Cows treated with calcitriol excreted more urinary tCa (control = 0.5 vs. calcitriol = 2.1 g/d) and tMg (control = 4.5 vs. calcitriol = 5.0 g/d) in the first 7 and 2 DIM, respectively, than control cows. Compared with control, calcitriol improved the proportion of neutrophils with oxidative burst (control = 31.9 vs. calcitriol = 40.6%), mean fluorescence intensity for oxidative burst (control = 90,900 vs. calcitriol = 99,746), and mean fluorescence intensity for phagocytosis (control = 23,887 vs. calcitriol = 28,080). Dry matter intake, yields of milk, and milk components did not differ between treatments. Administration of 300 μg of calcitriol at calving was safe and effective in increasing blood concentration of iCa and plasma concentrations of calcitriol, tCa, and tP for the first 6 d after treatment, and improved measures of innate immune function in early-lactation Holstein cows.