Association of blood calcium concentration in the first 3 days after parturition and energy balance metabolites at day 3 in milk with disease and production outcomes in multiparous Jersey cows

Research exploring specific associations of markers of negative energy balance and Ca in postpartum Jersey cows with lactation performance is lacking. Our objectives were to evaluate the associations of total Ca concentration (tCa) measured at 1 through 3 d in milk (DIM) and free fatty acids (FFA), β-hydroxybutyrate (BHB), and glucose measured at 3 DIM with (1) the risk of multiparous Jersey cows being diagnosed with early-lactation diseases and culling, (2) milk production in the first 9 wk of lactation, and (3) the risk of pregnancy in the first 150 DIM. A cohort study was performed in 1 dairy herd in Texas. Multivariable Poisson regression models were built to evaluate the association of the analytes of interest with the risks of early-lactation diseases and culling in the first 60 DIM (i.e., binary outcomes). Linear mixed models were used to evaluate the association of the analytes of interest with milk production within the first 9 wk of lactation, and a Cox proportional hazard model was built to assess the risk of pregnancy within 150 DIM. A total of 380 cows were used in the final analyses. Total Ca measured at 1 through 3 DIM was not associated with the risk of metritis. Cows with increased FFA and BHB had an increased risk of being diagnosed with metritis and clinical mastitis, respectively. Increased concentrations of glucose and FFA and decreased tCa at 3 DIM were associated with an increased risk of culling. Reduced tCa concentrations at 1 DIM (≤1.84 mmol/L) and 2 DIM (≤2.04 mmol/L) were associated with increased milk production across the first 9 wk of lactation compared with tCa concentrations above those thresholds. Total Ca was not associated with milk production when assessed at 3 DIM, whereas increased FFA (≥0.37 mmol/L) and decreased glucose (≤2.96 mmol/L) at 3 DIM were associated with increased milk production. None of the metabolites measured were associated with the risk of pregnancy in the first 150 DIM. Our results demonstrate that tCa concentration assessed in the first 3 DIM show temporary associations with milk production and culling in multiparous Jersey cows. Although increased concentration of FFA assessed at 3 DIM was associated with greater milk yield, it was a detrimental factor for the risk of metritis. This study attempted to better elucidate the relationship of tCa, FFA, BHB, and glucose assessed in early postpartum with health and performance of Jersey cows. Based on this study, assessments performed at 3 DIM using tCa concentration ≤1.99 mmol/L for increased risk of early-lactation culling and FFA ≥0.43 mmol/L for increased risk of metritis could be used as starting points. More studies evaluating the dynamics of energy balance markers and tCa in postpartum Jersey cows using a greater number of herds are needed to better inform dairy consultants on critical levels for exacerbated postpartum negative energy balance and subclinical hypocalcemia for the Jersey breed.     

Predicting hyperketonemia by logistic and linear regression using test-day milk and performance variables in early-lactation Holstein and Jersey cows

Although cowside testing strategies for diagnosing hyperketonemia (HYK) are available, many are labor intensive and costly, and some lack sufficient accuracy. Predicting milk ketone bodies by Fourier transform infrared spectrometry during routine milk sampling may offer a more practical monitoring strategy. The objectives of this study were to (1) develop linear and logistic regression models using all available test-day milk and performance variables for predicting HYK and (2) compare prediction methods (Fourier transform infrared milk ketone bodies, linear regression models, and logistic regression models) to determine which is the most predictive of HYK. Given the data available, a secondary objective was to evaluate differences in test-day milk and performance variables (continuous measurements) between Holsteins and Jerseys and between cows with or without HYK within breed. Blood samples were collected on the same day as milk sampling from 658 Holstein and 468 Jersey cows between 5 and 20 d in milk (DIM). Diagnosis of HYK was at a serum β-hydroxybutyrate (BHB) concentration ≥1.2 mmol/L. Concentrations of milk BHB and acetone were predicted by Fourier transform infrared spectrometry (Foss Analytical, Hillerød, Denmark). Thresholds of milk BHB and acetone were tested for diagnostic accuracy, and logistic models were built from continuous variables to predict HYK in primiparous and multiparous cows within breed. Linear models were constructed from continuous variables for primiparous and multiparous cows within breed that were 5 to 11 DIM or 12 to 20 DIM. Milk ketone body thresholds diagnosed HYK with 64.0 to 92.9% accuracy in Holsteins and 59.1 to 86.6% accuracy in Jerseys. Logistic models predicted HYK with 82.6 to 97.3% accuracy. Internally cross-validated multiple linear regression models diagnosed HYK of Holstein cows with 97.8% accuracy for primiparous and 83.3% accuracy for multiparous cows. Accuracy of Jersey models was 81.3% in primiparous and 83.4% in multiparous cows. These results suggest that predicting serum BHB from continuous test-day milk and performance variables could serve as a valuable diagnostic tool for monitoring HYK in Holstein and Jersey herds.     

Assessment of milk yield and composition,early reproductive performance,and herd removal in multiparous dairy cattle based on the week of diagnosis of hyperketonemia in early lactation

The purpose of this retrospective cohort study was to evaluate the effects of the timing of hyperketonemia (HYK) diagnosis during early lactation on milk yield and composition, reproductive performance, and herd removal. Plasma β-hydroxybutyrate (BHB) was measured twice a week during the first 2 wk of lactation in 362 multiparous Holstein cows for the diagnosis of HYK. In each week, cows were diagnosed as HYK positive (HYK+) if the plasma BHB concentrations were ≥1.2 mmol/L in at least one of the tests for the week evaluated. Milk-related outcomes (first 10 monthly milk tests) included milk yield, milk fat and protein content, milk urea nitrogen (MUN), and linear score of somatic cell count. Other performance outcomes of interest included risk of pregnancy within 150 d in milk (DIM) and herd removal (i.e., culling or death) within 300 DIM. Statistical models were built separately for cows diagnosed with HYK during the first week of lactation (wk1) and for cows diagnosed during the second week of lactation (wk2). All models for wk2 were adjusted by HYK diagnosed in wk1, along with other potential confounder variables. The association between HYK in each week and milk-related outcomes was assessed using generalized estimated equation models that accounted for repeated measures. Time to pregnancy and time to herd removal were analyzed using Cox's proportional hazard regression models. Seventy-eight cows (21.5%) tested positive for HYK during wk1, 60 cows (16.6%) in wk2, and 29 cows (8.0%) in both weeks. Hyperketonemia during wk1 was associated with a milk yield reduction of 3.7 kg [95% confidence interval (CI): ?6.67 to ?0.76] per cow per day throughout the lactation. Meanwhile, we did not observe evidence of an association between HYK diagnosed during wk2 and milk yield. During the first 2 monthly milk tests, cows diagnosed as HYK+ in wk1 had greater fat (0.42%; 95% CI: 0.16 to 0.67) and MUN (0.75 mg/dL; 95% CI: 0.26 to 1.24) content in milk than HYK-negative (HYK?) cows. We did not detect any evidence of an association between HYK diagnosed in wk2 and these outcomes. The HYK+ cows in wk1 had a 30% [hazard ratio (HR) = 0.70; 95% CI: 0.48 to 1.01] lower risk of pregnancy within 150 DIM and 2.48 times (95% CI: 1.63 to 2.89) higher risk of herd removal within 300 DIM than HYK? cows. Conversely, no evidence of association was observed between HYK+ cows in wk2 and risk of pregnancy by 150 DIM (HR = 0.98; 95% CI: 0.64 to 1.51) or removal from the herd within 300 DIM (HR = 0.91; 95% CI: 0.52 to 1.60). Our findings indicate that HYK diagnosed during wk1 of lactation is associated with negative performance in terms of milk yield, reproduction, and herd removal. No evidence of association was found for the same outcomes when HYK was diagnosed in wk2. Our results suggest the need to consider the timing when HYK is diagnosed when investigating its association with performance 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.     

Relationships between body condition,body condition loss,and serum metabolites during the transition period in primiparous and multiparous cows

In the transition period from late gestation to early lactation, dairy cows undergo tremendous metabolic changes. Insulin is a relevant antilipolytic factor. Decreasing serum concentrations of insulin and glucose, increasing serum concentrations of nonesterified fatty acids (NEFA) and β-hydroxybutyrate (BHB), and changes in body condition score (BCS) reflect the negative energy balance around calving. This study investigated peripartum metabolic adaptation in 359 primiparous and 235 multiparous German Holstein cows from a commercial dairy herd under field conditions. Body condition score was recorded and blood samples were taken 10 to 1 d prepartum, 2 to 4 d postpartum, and 12 to 20 d postpartum. Generalized mixed models and generalized estimation equations were applied to assess associations between prepartum BCS; BCS changes during the transition period; insulin, glucose, NEFA, and BHB serum concentrations; and milk yield, which was taken from an electronic milk meter from d 6 of lactation. Serum insulin concentrations of multiparous postpartum cows were lower compared with prepartum, and compared with primiparous cows. In general, primiparous cows had lower postpartum NEFA and BHB concentrations than multiparous cows. In primiparous cows, we identified a positive association between prepartum BCS and prepartum serum insulin concentration. Prepartum obese multiparous cows, but not primiparous cows, were characterized by higher postpartum serum NEFA and BHB concentrations and lower milk yield than other cows in the same parity class. Primiparous cows with a smaller degree of BCS loss during the transition period had higher postpartum insulin and lower NEFA concentrations and lower milk yield than other primiparous cows. In conclusion, primiparous cows had less lipolysis and lower milk yield than multiparous cows, associated with higher insulin concentrations. Avoiding high body condition loss during the transition period is a main factor in preventing peripartal metabolic imbalances of glucose and fat metabolism.     

Relationships between body condition score change,prior mid-lactation phenotypic residual feed intake,and hyperketonemia onset in transition dairy cows

Extensive efforts have been made to identify more feed-efficient dairy cows, yet it is unclear how selection for feed efficiency will influence metabolic health. The objectives of this research were to determine the relationships between residual feed intake (RFI), a measure of feed efficiency, body condition score (BCS) change, and hyperketonemia (HYK) incidence. Blood and milk samples were collected twice weekly from cows 5 to 18 d postcalving for a total of 4 samples. Hyperketonemia was diagnosed at a blood β-hydroxybutyrate (BHB) ≥1.2 mmol/L and cows were treated upon diagnosis. Dry period, calving, and final blood sampling BCS was recorded. Prior mid-lactation production, body weight, body weight change, and dry matter intake (DMI) data were used to determine RFI phenotype, calculated as the difference between observed DMI and predicted DMI. The maximum BHB concentration (BHB_max) for each cow was used to group cows into HYK or not hyperketonemic. Lactation number, BCS, and RFI data were analyzed with linear and quadratic orthogonal contrasts. Of the 570 cows sampled, 19.7% were diagnosed with HYK. The first positive HYK test occurred at 9 ± 0.9 d postpartum and the average BHB concentration at the first positive HYK test was 1.53 ± 0.14 mmol/L. In the first 30 d postpartum, HYK-positive cows had increased milk yield and fat concentration, decreased milk protein concentration, and decreased somatic cell count. Cows with a dry BCS ≥4.0, or that lost 1 or more BCS unit across the transition to lactation period, had greater BHB_max than cows with lower BCS. Prior-lactation RFI did not alter BHB_max. Avoiding over conditioning of dry cows and subsequent excessive fat mobilization during the transition period may decrease HYK incidence; however, RFI during a prior lactation does not appear to be associated with HYK onset.     

Transition cow nutrition and management strategies of dairy herds in the northeastern United States: Part II—Associations of metabolic- and inflammation-related analytes with health,milk yield,and reproduction

The objectives were as follows: (1) establish cow-level thresholds for prepartum nonesterified fatty acids (NEFA) and postpartum NEFA, β-hydroxybutyrate (BHB), and haptoglobin (Hp) concentrations associated with negative health events; (2) evaluate cow-level associations between biomarkers and 305-d mature equivalent milk at the fourth test day (ME305) and reproductive performance; and (3) identify herd-alarm levels (proportion of cows sampled above the critical threshold) for biomarkers that are associated with herd-level changes in disorder incidence (displaced abomasum and clinical ketosis), reproductive performance, and ME305. In a prospective cohort study, 1,473 cows from 72 farms were enrolled from the northeastern United States. Blood samples were collected from the same 11 to 24 cows per herd during the late-prepartum and early-postpartum periods. Whole blood was analyzed for postpartum BHB concentrations; plasma was analyzed for prepartum and postpartum NEFA and postpartum Hp concentrations. Critical thresholds for the biomarkers associated with health events for all cows were established using a receiver operating characteristic curve analysis. Poisson, linear mixed effects, and Cox proportional hazards models investigated the association of the biomarkers with health and performance. The prepartum NEFA and Hp threshold associated with culling was ≥0.17 mmol/L and 0.45 g/L, respectively. The postpartum NEFA and BHB thresholds associated with diagnosis of metritis, displaced abomasum, or clinical ketosis were ≥0.46 mmol/L and ≥0.9 mmol/L, respectively. Multiparous cows with prepartum NEFA concentration ≥0.17 mmol/L produced 479 kg less ME305. Multiparous and primiparous cows with postpartum NEFA concentration ≥0.46 mmol/L produced 280 kg less and 446 kg more ME305, respectively. Cows with BHB concentration ≥0.9 and ≥1.1 mmol/L produced 552 kg more ME305 and had a 20% decreased risk of pregnancy within 150 d in milk, respectively; however, multiparous cows with BHB concentration ≥1.5 mmol/L produced 376 kg less ME305. Cows with Hp concentration ≥0.45 g/L produced 492 kg less ME305 and had 28% decreased risk of pregnancy within 150 DIM. Cows with Hp concentration ≥0.45 g/L had 19% decreased pregnancy risk to first service (PRFS). Herds above the herd-alarm levels for prepartum NEFA had a 6.0-percentage unit increase in disorder incidence and a 6.0-percentage unit decrease in 21-d pregnancy rate (PR) for multiparous cows, a 3.9-percentage unit increase in PR and a 5.8-percentage unit increase in the probability of pregnancy for primiparous cows. Herds above the herd-alarm levels for postpartum NEFA had a 5.8- and 4.2-percentage unit increase in disorder incidence for multiparous and primiparous cows, respectively, a 789 kg decrease in ME305 for multiparous cows, and a 6.8- and 6.3-percentage unit decrease and increase in PR and PRFS for multiparous cows, respectively. Herds above the herd-alarm levels for BHB had an 8.5-percentage unit increase in disorder incidence, a 332 and 229 kg increase in ME305 for primiparous and multiparous cows, respectively, and a 3.2-, 5.2-, and 7.0-percentage unit decrease in PR, probability of pregnancy, and PRFS, respectively. Herds above the herd-alarm levels for postpartum Hp had a 5.3-percentage unit increase in disorder incidence. At the cow level and herd level, elevated biomarker concentrations were associated with an increased disorder risk and varied performance responses.     

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.     

A randomized controlled trial examining the effects of treatment with propylene glycol and injectable cyanocobalamin on naturally occurring disease,milk production,and reproductive outcomes of dairy cows diagnosed with concurrent hyperketonemia and hypoglycemia

The objective of this study was to assess the effects of treatment with propylene glycol (PG) and cyanocobalamin (B12) on health, milk production, and reproductive outcomes of cows diagnosed with hyperketonemia (HK), hypoglycemia (HG), or concurrent HKHG. Glucose and β-hydroxybutyric acid (BHBA) concentrations were assessed in whole blood using a handheld device in lactating dairy cows (n = 2,418) between 3 and 9 d postpartum. Cows categorized as HK (n = 232, BHBA ≥1.2 mmol/L), HG (n = 161, glucose ≤2.2 mmol/L), and concurrent HKHG (n = 204, BHBA ≥1.2 mmol/L, and glucose ≤2.2 mmol/L) were randomized to receive treatment or to remain untreated (control). Treatment consisted of a single dose of B12 (10 mg, intramuscularly) and 300 mL of PG orally for 5 d, starting on the day of cow-side testing. Milk production, health, and reproductive outcomes were analyzed according to groups. Statistical analysis was carried out using SAS version 9.4 (SAS/STAT, SAS Institute Inc.). Treatment in HG cows decreased clinical ketosis, increased milk production in the fifth week of lactation for multiparous cows, and tended to increase 305-d mature-equivalent milk yield (305ME) for primiparous cows compared with untreated cows with the same metabolic profile. For cows with HKHG, treatment increased 305ME in multiparous cows and tended to increase 305ME in primiparous cows. No differences were found for treatment among any of the metabolic groups regarding reproductive outcomes, nor were any treatment effects found among HK cows. Glycemic status may help identify metabolically challenged early postpartum dairy cows, which may have differential response to PG and B12 treatment.     

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.     

Association of postpartum hypocalcemia with early-lactation milk yield,reproductive performance,and culling in dairy cows

Periparturient hypocalcemia is frequently observed and considered as a gateway disease that is associated with various health issues. The objective of this study was to evaluate the association of hypocalcemia with early-lactation milk yield, reproductive performance, and culling across a large number of different managerial systems. A prospective cohort study was conducted based on a convenience sample of 125 dairy herds from 8 federal states of Germany between February 2015 and August 2016. A blood sample was drawn from 1,709 animals within 48 h after parturition and analyzed for serum calcium concentration. After discarding cows (n = 283) with missing data, a total of 1,426 cows were considered for final analyses. The median time from calving to sampling was 14.0 h (interquartile range = 5.0–24.9 h). For each herd, a record of the herd management software was requested 150 d after the last cow was sampled. Serum calcium concentration of each cow was associated with early-lactation milk yield (Dairy Herd Improvement Association equivalent test 1 to 3), reproductive performance [days in milk (DIM) at first artificial insemination (AI), pregnancy at first AI, time to pregnancy within 150 DIM], and culling (until 60 DIM) data. Generalized linear mixed models were used to analyze continuous or categorical data. Shared frailty models were used for time to event data. Five different thresholds were used to define hypocalcemia. Thresholds ranged from 1.8 to 2.2 mmol/L using 0.1-mmol/L increments. Clinical hypocalcemia was defined as serum calcium concentration <2.0 mmol/L in combination with clinical signs (e.g., recumbency). The effect of hypocalcemia on milk yield was conditional on parity. In primiparous cows a serum calcium concentration <2.0 mmol/L (6.4% of cows were below this threshold) had no effect on milk production, whereas there was a tendency for multiparous cows with a serum calcium concentration <2.1 mmol/L (63.2% of cows were below this threshold) to produce 0.80 kg/d more milk compared with multiparous cows at or above the threshold. Multiparous cows suffering from clinical hypocalcemia produced 2.19 kg/d less milk compared with normocalcemic cows in early lactation. Calcium status was not associated with days to first insemination. Cows with a serum calcium concentration <1.9 mmol/L (34.6% of cows below this threshold) had decreased odds (odds ratio = 0.56) of pregnancy at first AI. A serum calcium concentration <1.8 mmol/L (24.1% of cows below this threshold) had a significant effect on time to pregnancy. Compared with animals with a serum calcium concentration ≥1.8 mmol/L, the hazard of becoming pregnant within 150 DIM was reduced when cows had a serum calcium concentration <1.8 mmol/L (hazard ratio = 0.68). Cows with a serum calcium concentration <2.0 mmol/L (44.3% of cows were below this threshold) had a 1.69 times greater hazard of being culled within the first 60 DIM compared with normocalcemic animals. The present study shows that the association of hypocalcemia with milk yield was conditional on parity and serum calcium concentration measured once within 48 h after calving. Considering reproductive performance and culling in early lactation, a negative effect of postpartum hypocalcemia was demonstrated.     

Prediction of blood β-hydroxybutyrate content and occurrence of hyperketonemia in early-lactation,pasture-grazed dairy cows using milk infrared spectra

The objective of this study was to evaluate the ability of milk infrared spectra to predict blood β-hydroxybutyrate (BHB) concentration for use as a management tool for cow metabolic health on pasture-grazed dairy farms and for large-scale phenotyping for genetic evaluation purposes. The study involved 542 cows (Holstein-Friesian and Holstein-Friesian × Jersey crossbreds), from 2 farms located in the Waikato and Taranaki regions of New Zealand that operated under a seasonal-calving, pasture-based dairy system. Milk infrared spectra were collected once a week during the first 5 wk of lactation. A blood “prick” sample was taken from the ventral labial vein of each cow 3 times a week for the first 5 wk of lactation. The content of BHB in blood was measured immediately using a handheld device. After outlier elimination, 1,910 spectra records and corresponding BHB measures were used for prediction model development. Partial least square regression and partial least squares discriminant analysis were used to develop prediction models for quantitative determination of blood BHB content and for identifying cows with hyperketonemia (HYK). Both quantitative and discriminant predictions were developed using the phenotypes and infrared spectra from two-thirds of the cows (randomly assigned to the calibration set) and tested using the remaining one-third (validation set). A moderate accuracy was obtained for prediction of blood BHB. The coefficient of determination (R²) of the prediction model in calibration was 0.56, with a root mean squared error of prediction of 0.28 mmol/L and a ratio of performance to deviation, calculated as the ratio of the standard deviation of the partial least squares model calibration set to the standard error of prediction, of 1.50. In the validation set, the R² was 0.50, with root mean squared error of prediction values of 0.32 mmol/L, which resulted in a ratio of performance to deviation of 1.39. When the reference test for HYK was defined as blood concentration of BHB ≥1.2 mmol/L, discriminant models indicated that milk infrared spectra correctly classified 76% of the HYK-positive cows and 82% of the HYK-negative cows. The quantitative models were not able to provide accurate estimates, but they could differentiate between high and low BHB concentrations. Furthermore, the discriminant models allowed the classification of cows with reasonable accuracy. This study indicates that the prediction of blood BHB content or occurrence of HYK from milk spectra is possible with moderate accuracy in pasture-grazed cows and could be used during routine milk testing. Applicability of infrared spectroscopy is not likely suited for obtaining accurate BHB measurements at an individual cow level, but discriminant models might be used in the future as herd-level management tools for classification of cows that are at risk of HYK, whereas quantitative models might provide large-scale phenotypes to be used as an indicator trait for breeding cows with improved metabolic health.     

Associations of pre- and postpartum lying time with metabolic,inflammation, and health status of lactating dairy cows

The objective was to evaluate the associations of pre- and postpartum lying time (LT) with serum total calcium (Ca), nonesterified fatty acids (NEFA), β-hydroxybutyrate (BHB), and haptoglobin concentrations, hemogram, and health status of dairy cows. A total of 1,052 Holstein cattle (401 nulliparous heifers and 651 parous cows) from 3 commercial dairy farms were fitted with electronic data loggers (IceQube, IceRobotics, Edinburgh, UK) on a hind leg 14 ± 3 d before parturition (dpp) and removed at 14 ± 3 d in milk (DIM) to assess their LT. Lying time data were summarized and reported daily (min/d or h/d). Serum concentrations of NEFA (at 14 ± 3 and 7 ± 3 dpp), total serum calcium within 48 h after calving, and BHB (at 7 ± 3 and 14 ± 3 DIM) were determined. Serum concentration of haptoglobin was determined and a hemogram was performed on a subsample of 577 cows (237 primiparous and 340 multiparous) at 7 ± 3 DIM. Cases of milk fever, retained placenta, metritis, mastitis, pneumonia, and digestive disorders within 30 DIM were recorded and cows were categorized into 1 of 4 groups: (1) nondiseased (ND, n = 613; cows without ketosis and any other health conditions); (2) cows with only ketosis (KET, n = 152); (3) sick cows experiencing ≥1 health conditions, but without ketosis (SICK, n = 198); or (4) cows with ketosis plus at least one other health condition (KET+, n = 61). Data were analyzed using mixed linear regression models or logistic regression (MIXED or GLIMMIX procedures). Lying time within 14 dpp had a significant positive quadratic association with serum NEFA concentrations at 14 ± 3 and 7 ± 3 dpp but was not significantly associated with serum Ca concentration within 48 h after calving. Lying time during the first 14 DIM after parturition had a significant linear association with the risk of ketosis within 14 DIM. For every 1-h increment in mean LT (from 8 to 15 h/d) within the first 14 DIM after calving, the risk of diagnosis with ketosis within 14 DIM increased by 3.7 percentage points. Regardless of parity, a greater proportion of KET and KET+ groups had increased serum prepartum NEFA concentration (≥400 µEq/L) and increased body condition loss from 14 dpp to 28 DIM compared with SICK and ND cows. A greater proportion of multiparous KET and KET+ cows had hypocalcemia within 48 h after calving compared with ND and SICK cows, but we did not detect a significant association between hypocalcemia and health status on primiparous cows. Multiparous KET+ cows had significantly reduced neutrophils and white blood cell count compared with ND cows, but lymphocytes did not differ. Regardless of parity, KET+ and SICK cows had significantly higher concentrations of serum haptoglobin compared with ND cows. These results suggest that LT along with energy and Ca balance are critical for transition cow health.     

Effect of monensin on milk fatty acid profile in dairy cows and on the use of fatty acids for early diagnosis of elevated blood plasma concentrations of nonesterified fatty acids and hyperketonemia

This work examined the effects of precalving administration of continuous-release monensin capsule on postcalving milk fatty acid (FA) profile and on the accuracy of FA as a biomarker in the early identification of cows with elevated blood plasma nonesterified fatty acids (NEFA) and β-hydroxybutyrate (BHB) concentrations. Approximately 3 wk before expected calving, 203 multiparous Estonian Holstein cows were randomly divided into control (CO; n = 116) and experimental (MO; n = 87) groups, and a continuous-release capsule of monensin was administered to the MO cows. Blood samples were taken daily in the first 4 d postpartum, then on the sixth or seventh day in milk, twice in the second week, and thenceforth once per week until the end of the sixth week. Milk samples were taken once from 4 to 7 d in milk, twice in the second week, and thenceforth once per week. Blood samples were analyzed for NEFA and BHB, and milk was analyzed for FA concentrations. Cows with postpartum BHB concentrations ≥1.2 mmol/L at least once during the 6 wk were classified as hyperketonemic (HYK), and cows with NEFA concentrations ≥1.0 mmol/L as having elevated concentration of NEFA (NEFA_H). The ability of FA to predict NEFA_H and HYK cows was studied with logistic regression and receiver operating characteristic curve analysis and the identification accuracy was estimated by area under the receiver operating characteristic curve. For these analyses, we used FA measured on the ninth day after calving. Monensin administration affected FA mobilization and metabolism of the animals as blood NEFA were lower in the MO group on wk 1 and wk 3, and BHB values were considerably lower from wk 1 to wk 4 compared with the CO group. The FA dynamics were generally similar for MO and CO groups. Monensin administration resulted in higher concentrations of C15:0, C16:0, iso C17:0, anteiso C15:0, anteiso C17:0, total trans monounsaturated FA, and C18:2 cis-9,trans-11, and lower proportions of C18:0, C18:1 cis-9, and most of the iso FA. The identification accuracy of NEFA_H and HYK cows was higher in the CO compared with the MO group and for the identification of HYK compared with NEFA_H cows (0.75–0.77 vs. 0.78–0.80 in the CO group, and 0.61–0.66 vs. 0.68–0.75 in the MO group for NEFA_H vs. HYK, respectively). For all FA, the threshold values to identify NEFA_H and HYK cows were different in the CO and MO groups. Results suggest that specific threshold values for the identification of NEFA_H and HYK cows could be applicable only within similar feeding conditions and rumen environment.     

Reducing milking frequency from twice to once daily as an adjunct treatment for ketosis in lactating dairy cows—A randomized controlled trial

This randomized controlled trial investigated the effects of temporarily reducing milking frequency (MF) on the resolution of ketosis and milk production in dairy cows in early lactation. To detect ketosis [blood β-hydroxybutyrate (BHB) ≥1.2 mmol/L], Holstein cows were screened daily from 3 to 16 d in milk using a cow-side meter. Cows diagnosed with ketosis (n = 104) were randomly assigned to twice-daily milking (TDM) or reduced to once-daily milking (ODM) for 2 wk, then returned to twice-daily milking. Both treatment groups received a 5-d treatment of an oral propylene glycol drench (PG; 300 g) beginning on the afternoon of the diagnosis; cows received additional 5-d PG treatments if they had a ketotic test result (blood BHB ≥1.2 mmol/L) at least 4 d after finishing the first PG treatment. Blood BHB tests were conducted for the first 3 d after ketosis diagnosis, and then once every 3 d for 21 d of trial (DOT). Milk and milk component data were collected weekly for 15 wk following trial enrollment. The ODM group showed rapidly and markedly decreased blood BHB concentrations (primiparous cows: 1 DOT, 0.92 ODM vs. 1.22 TDM, 15 DOT, 0.55 vs. 0.81 mmol/L; multiparous cows: 1 DOT, 1.01 vs. 1.40, 15 DOT, 0.78 vs. 1.65 mmol/L). In addition, a logistic regression model indicated that ODM cows were less likely to have blood BHB concentrations ≥1.2 mmol/L [primiparous cows: 3 DOT: ODM 1% (95% confidence interval: 0–10%) vs. TDM 43% (30–58%), 15 DOT ODM 0% (0–0.2%) vs. TDM 22% (13–36%); multiparous cows: 3 DOT: ODM 33% (24–44%) vs. TDM 59% (48–69%), 15 DOT ODM 20.9% (13–31%) vs. TDM 64% (53–74%)]. The proportion of ODM cows that required additional treatments of PG were substantially lower than the TDM group (ODM: 39%; TDM: 64%) than the TDM cows during the initial 21-d period. However, during the 2-wk treatment period, cows in the ODM group produced 26% less milk and 25% less energy-corrected milk than the TDM cows. During wk 3 to 15, when all cows were milked twice daily, ODM cows produced less milk (?14%) and energy-corrected milk (?12%) compared with the TDM group. Milk protein percentage was greater, and milk fat percentage and linear score tended to be greater in the ODM group over 15 wk. In conclusion, a 2-wk reduction of MF in ketotic cows from twice to once daily with treatment with PG resolved ketosis and decreased blood BHB concentrations more effectively than treating TDM cows with PG alone. However, the 2-wk MF reduction had immediate and long-term (up to 13 wk after cessation of MF reduction) negative effects on milk production.     

Effect of hyperketonemia on the diurnal patterns of energy-related blood metabolites in early-lactation dairy cows

Most dairy cows experience a period of energy deficit in early lactation, resulting in increased plasma concentrations of nonesterified fatty acids (NEFA) and β-hydroxybutyrate (BHB). Our objectives were to determine (1) the diurnal variation in plasma BHB and NEFA, (2) the correlation between plasma NEFA and BHB when accounting for diurnal changes, and (3) the effect of hyperketonemia (HYK) on the diurnal pattern of blood metabolites. Jugular catheters were placed in 28 multiparous Holstein cows between 3 and 9 days in milk, and blood samples were collected every 2 h for 96 h. Cows were retrospectively classified as HYK positive (HYK; n = 13) if they had plasma BHB concentrations ≥1.2 mmol/L for ≥3 study days, or HYK negative (non-HYK; n = 15) if they had plasma BHB concentrations ≥1.2 mmol/L for ≤2 study days. Generalized linear mixed models were used to analyze concentrations of analytes over time and differences in metabolites between HYK groups. The correlation between total plasma NEFA and BHB was analyzed by calculating the area under the curve for plasma NEFA and BHB for all cows. Plasma NEFA reached a peak approximately 2 h before morning feed delivery, falling to a nadir in the late evening. Plasma BHB was at a nadir at the time of morning feed delivery, peaking 4 h later. We observed a strong positive correlation between daily plasma NEFA and BHB. Additionally, HYK cows had greater concentrations of plasma NEFA and BHB than non-HYK cows. The HYK cows also experienced a greater magnitude of change in BHB throughout the day than the non-HYK cows. Our results suggest that the time relative to feeding should be considered when analyzing plasma metabolites, as classification of energy status may change throughout a day.     

An investigation of blood,milk, and urine test patterns for the diagnosis of ketosis in dairy cows in early lactation

Ketosis in dairy cattle is primarily diagnosed based on the concentrations of ketone bodies in the blood, milk, or urine. Cow-side tests using these fluids are available for rapid detection of elevated concentrations of ketone bodies. Although these tests have been extensively validated, the performance of different tests has not been compared over time. Our objectives were to investigate the relationship between point-of-care diagnostic tests measuring the concentrations of β-hydroxybutyrate (BHB) in blood (BT; Precision Xtra, Abbott Laboratories), BHB in milk (MT; Keto-Test, Elanco), and acetoacetate (AcAc) in urine (UT; Ketostix, Bayer Corporation) through cases of ketosis. Holstein cows (n = 148) were screened daily for hyperketonemia (HYK; blood BHB ≥1.2 mmol/L) from 3 to 16 d in milk (DIM); moreover, milk and urine samples were collected concomitantly and tested for ketones (ketosis thresholds: 100 µmol/L milk BHB and 5 mg/dL urine AcAc). Of the animals screened (n = 148), 74% were diagnosed with HYK. When diagnosed with HYK, cows were treated with propylene glycol orally once daily for 5 d. After the day of diagnosis (d 0), hyperketonemic cows were retested with BT, MT, and UT for 3 d (d 1, 2, and 3). We assessed the diagnostic test performance and time to ketosis (survival analyses and Cox proportional hazards models) of MT and UT compared with BT. Considering all paired samples (before and after diagnosis of HYK), MT had 61% sensitivity and 91% specificity, whereas the UT had 77% sensitivity and 94% specificity compared with BT. The specificity of MT and UT increased from d 0 to d 1, decreased on d 2, and increased on d 3. The median time to diagnosis of ketosis in blood was 5 DIM (95% CI 5 to 7 DIM); moreover, MT and UT had 2 d greater median time to diagnosis of ketosis compared with the BT [7 DIM (6 to 11 d); and 7 DIM (6 to 13 d), respectively]. We concluded that the UT is a more sensitive predictor of blood BHB concentration than the MT. The UT and MT tests diagnosed ketotic cows approximately 2 d later than the BT. The possible consequences of delay in detection of ketosis in milk and urine should be investigated.     

Effects of postpartum milking strategy on plasma mineral concentrations and colostrum,transition milk,and milk yield and composition in multiparous dairy cows

The effects of postpartum milking strategy on plasma mineral concentrations, blood β-hydroxybutyrate (BHB) concentration, and colostrum, transition milk, and first monthly test milk yield and composition were evaluated in 90 multiparous Jersey and Jersey × Holstein crossbreed cows from a commercial farm. Before first postpartum milking, cows were randomly assigned to the following milking strategies, implemented during the first 2 d postpartum: twice-a-day milking (M2, standard industry practice, milking every 12 h; n = 22), once-a-day milking (M1, milking every 24 h; n = 24), restricted milking (MR, 3-L milking every 12 h; n = 21), and delayed milking (MD, no milking for the first 24 h, and milking every 12 h afterward; n = 23). Blood samples for total plasma Ca, P, and Mg determination were collected from enrollment every 4 h up to 48 h, and at 3 d in milk. Blood BHB concentration was determined at 3 and 11 d in milk. Colostrum and transition milk yields were recorded, and samples were collected at each study milking for IgG and somatic cell count (SCC) determinations. Information for first monthly test milk yield and composition was obtained from the Dairy Herd Improvement Association. Statistical analyses were conducted using generalized multiple linear and Poisson regressions with Dunnett adjustment and M2 as reference group for mean comparisons. Overall, plasma Ca concentration within 48 h after enrollment was higher for MD (2.17 mmol/L), tended to be higher for MR (2.15 mmol/L), and was similar for M1 (2.09 mmol/L) compared with M2 cows (2.06 mmol/L). No statistically significant differences compared with M2 cows were observed for plasma P and Mg concentrations. Colostrum and transition milk and total Ca harvested within 48 h after enrollment were lower for M1, MR, and MD compared with M2 cows. The MD strategy prevented harvesting colostrum with >50 g of IgG/L. No statistically significant effects were detected on plasma mineral concentrations at 3 DIM, blood BHB concentration, colostrum and transition milk SCC within 48 h after enrollment, or milk yield, energy-corrected milk yield, and SCC at first monthly test. Our results suggest that postpartum plasma Ca concentration may be influenced by postpartum milking strategy, without interfering with future milk yield and udder health. Further studies should evaluate whether the proposed milking strategies in early postpartum affect production, reproduction, or health.     

Effects of folic acid and vitamin B12 supplementation on culling rate,diseases, and reproduction in commercial dairy herds

This study was undertaken to determine the effect of a combined folic acid and vitamin B₁₂ supplement given in early lactation on culling rate, metabolic disorders and other diseases, and reproduction in commercial dairy herds. A total of 805 cows (271 primiparous and 534 multiparous cows) in 15 commercial dairy herds were involved. Every 2 mo from February to December 2010 and within each herd, cows were assigned according to parity, previous 305-d milk production, and calving interval to 5 mL of either (1) saline 0.9% NaCl (control group) or (2) 320 mg of folic acid + 10 mg of vitamin B₁₂ (vitamin group). Treatments were administered weekly by intramuscular injections starting 3 wk before the expected calving date until 8 wk after parturition. A total of 221 cows were culled before the next dry period. Culling rate was not affected by treatment and was 27.5%; culling rate was greater for multiparous (32.2%) than for primiparous cows (18.8%). Within the first 60 d in milk (DIM), 47 cows were culled, representing 21.3% of total culling, and no treatment effect was noted. Ketosis incidence based on a threshold ≥100 µmol/L of β-hydroxybutyrate in milk was 38.3 ± 2.9% for the vitamin group and 41.8 ± 3.0% for the control group and was not affected by treatment. The combined supplement of folic acid and vitamin B₁₂ did not decrease incidence of retained placenta, displaced abomasum, milk fever, metritis, or mastitis. However, the incidence of dystocia decreased by 50% in multiparous cows receiving the vitamin supplement, although no effect was observed in primiparous cows. The first breeding postpartum for multiparous cows occurred 3.8 d earlier with the vitamin supplement compared with controls, whereas no treatment effect was seen for primiparous cows. Days open, first- and second-breeding conception rates, number of breedings per conception, and percentage of cows pregnant at 150 DIM were not affected by treatment. The reduced percentage of dystocia combined with the earlier DIM at first breeding for multiparous cows receiving the combined supplementation in folic acid and vitamin B₁₂ indicates that the vitamin supplement had a positive effect in older cows.     

Effects of short dry periods on performance and metabolic status in Holstein dairy cows

To evaluate effects of different dry period lengths on milk yield, milk composition, and energy balance of dairy cows, 122 multiparous and primiparous Holstein dairy cows were used in a completely randomized experimental design with 56-, 42-, and 35-d dry period lengths. Actual dry period lengths for respective treatments (TRT) were 56 ± 5.1 d, 42 ± 2.1 d, and 35 ± 2.7 d. Overall, cows in the 42- and 56-d TRT gained more body condition than those in 35-d TRT during the dry period; however, postpartum body condition score did not change substantially among the TRT. Although from 3 to 210 DIM, differences were not detected in the milk yield of multiparous cows between the 35- and 56-d TRT, primiparous cows in the 35-d TRT produced less milk than those in 56-d TRT. In primiparous cows, the milk production at wk 9, 10, and 11 of lactation was lower in the 35-d compared with the 56-d TRT. Primiparous cows in the 35-d compared with the 56-d TRT produced less milk protein. In the 35-d TRT, serum triglyceride concentration was greater in primiparous cows than in multiparous cows during the peripartum period. Among primiparous cows, those in the 56-d TRT had greater concentrations of nonesterified fatty acids than those in the 35-d TRT during the peripartum period. No significant differences were observed in concentrations of serum glucose, insulin, and insulin-like growth factor-I during early lactation among TRT. There was also no difference among TRT for incidence of metabolic disorders. Thus, this study indicates that shortening the dry period to 35 d may be beneficial in multiparous and overconditioned cows, but not in primiparous cows.