Assessment of systemic inflammation following oral calcium supplementation in healthy postpartum multiparous dairy cows-A randomized controlled trial.

Around parturition, dairy cows inevitably and perhaps necessarily experience some degree of systemic inflammation, but when excessive or dysregulated, it may contribute to health disorders. As immune activation decreases blood Ca, greater extracellular Ca availability may potentiate or sustain inflammation. We hypothesized that in clinically healthy multiparous cows, postpartum administration of supplemental Ca would increase serum concentrations of inflammatory markers. The objective of this randomized controlled trial was to investigate a possible effect of supplementing calcium (Ca) on postpartum systemic inflammation in dairy cows. Healthy cows (n = 101) from 2 commercial dairy farms in Ontario calving into parity 2, 3, or 4 were enrolled. Cows were blocked by parity and randomly assigned to receive an oral bolus of Ca (42 g of Ca) within 12 h after calving and a second bolus 12 h later (TRT; n = 51), or no Ca supplementation (CON; n = 49). Concentrations in serum of total Ca (tCa), haptoglobin (Hp), and albumin (ALB) were assessed at d 0 (within 12 h postpartum), 0.5 (12 h later), 1, 2, 4, 6, and 8 postpartum; ionized calcium was assessed at d 0, 0.5, 2, and 4, and lipopolysaccharide-binding protein (LBP) and serum amyloid A (SAA) were assessed at d 0, 2, and 4. Multivariable linear regression models of each outcome accounting for repeated measures included treatment, parity (2 vs. 3 or 4), farm, sampling day, baseline concentration (d 0), and interactions of treatment with farm, parity, and day. Results are presented as least squares means and 95% confidence intervals. Concentration of tCa tended to be greater at d 0.5 (TRT 2.07 mmol/L [2.03-2.12]; CON 2.01 [1.96-2.06]) but was lesser at d 2 (TRT 2.18 [2.13-2.23]; CON 2.27 [2.23-2.32]) in TRT than CON cows. Concentrations of LBP were greater in TRT (2.28 ng/mL [2.06-2.50]) than CON (1.99 [1.77-2.21]) in parity 2, but not different in older cows (TRT 2.28 ng/mL [2.06-2.50]; CON 1.99 [1.77-2.21]). Concentrations of SAA were greater in TRT than CON cows at d 2 (TRT 135 ug/mL [124-146]; CON 114 [75-106]). Treatment had no effect on ALB or Hp. In clinically healthy cows, oral Ca supplementation had a small transient effect on blood tCa and little indication of increasing inflammation based on the analytes evaluated.

Postpartum health is associated with detection of estrus by activity monitors and reproductive performance in dairy cows.

The objective of this prospective observational study was to investigate associations of postpartum health with estrus detection (ED) by activity monitors and pregnancy outcomes in dairy cows. A total of 1,743 Holstein cows from 2 commercial dairy herds in Ontario, Canada were enrolled 3 wk before expected parturition and examined for health variables until 9 wk postpartum. Body condition score (BCS) and lameness were measured at 3 wk prepartum, and serum concentrations of total Ca, haptoglobin (Hp), and nonesterified fatty acids were measured at 2 and 6 ± 2 d in milk (DIM), and blood ß-hydroxybutyrate (BHB) and metritis were assessed at 4, 8, 11, and 15 ± 2 DIM. Cows were examined for purulent vaginal discharge (PVD) and endometritis (ENDO) by endometrial cytology at wk 5, for lameness at wk 3 and 7, for BCS at wk 9 postpartum, and for time to onset of cyclicity by biweekly serum progesterone (P4) measurements. Additional disease data were obtained from farm records. Reproductive management for first AI was primarily based on ED by activity monitors until at least 75 DIM, and cows not detected in estrus were synchronized. Data were analyzed in multivariable logistic or Cox proportional hazards regression models including blood markers, health variables, potential covariates, and herd as a random effect. Estrus was detected in 77% of primiparous and 66% of multiparous cows between 50 or 55 DIM and 75 DIM. In 1,246 cows, the model-predicted probability of ED (percentage point difference) was lower in cows that had retained placenta (-14%), ENDO (-7%), PVD (-8%), delayed cyclicity (no P4 > 1 ng/mL by wk 9; -12%), or ≥0.5-point BCS loss (-14%) compared with cows without each of these risk factors, and it was negatively associated with blood BHB at 15 DIM. Considering only variables measured on farm (not requiring laboratory analysis), the probability of ED was lower (56 vs. 81%) in cows with >1 risk factor compared with cows without risk factors. The predicted probability of pregnancy at first artificial insemination (percentage point difference) was lower in cows that had ENDO (-7%) or PVD (-7%), and negatively associated with serum Hp at 6 ± 2 DIM. In cows detected in estrus by 75 DIM (n = 888), risk factors for reduced pregnancy rate by 250 DIM (adjusted hazard ratio (AHR); 95% confidence intervals) included difficult calving (AHR: 0.67; 0.45 to 1.00), metritis (AHR: 0.79; 0.61 to 1.01), PVD (AHR: 0.79; 0.65 to 0.97), or lameness (AHR: 0.79; 0.62 to 1.01), and it was negatively associated with serum Hp at 6 ± 2 DIM. Monitoring postpartum health may be used to identify cows that are more or less likely to be detected in estrus by activity monitors and to become pregnant in a timely manner. This would support a selective reproductive management program with targeted interventions.

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 ß-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.

Association of metabolic markers with neutrophil function in healthy postpartum dairy cows.

The objective of this study was to evaluate the associations of serum markers for systemic inflammation, liver, mineral, and energy status, and blood neutrophil counts with the function of circulating neutrophils in postpartum dairy cows. Blood samples were collected from 21 healthy Holstein cows at 5, 10, 14, and 21 d postpartum. Serum samples were used to measure concentrations of total calcium, phosphorus, magnesium, total protein, albumin, globulin, cholesterol, urea, glucose, gamma-glutamyl transferase, aspartate aminotransferase, glutamate dehydrogenase, haptoglobin (Hp), ß-hydroxybutyrate, non-esterified fatty acids (NEFA), and insulin-like growth factor-1. The shift of percentage of activated neutrophils for phagocytosis (PPC) and oxidative burst (POB) and the median fluorescence intensity (MFI) for PC (MFIPC), OB (MFIOB), and endocytic and proteolytic degradation measured via DQ-ovalbumin (MFIDQ) were evaluated using flow cytometry. Mixed linear regression models were used to assess the associations of serum concentrations of metabolites and blood neutrophil counts with each neutrophil function assay outcome accounting for d postpartum, parity, and body condition score. Pearson correlation tests (r) were used to describe the relationships of metabolites and neutrophil counts with neutrophil function assays that were significant in the regression models. Greater serum concentrations of Hp were associated with lesser OB function (POB and MFIOB; r = -0.2 for both), but greater PPC and MFIDQ (r = 0.3 for both). Greater serum NEFA and cholesterol were associated with lesser (r = -0.3) and greater (r = 0.2) POB function, respectively. Blood neutrophil counts were associated with PPC (r = -0.2) and MFIDQ (r = -0.3). At 5 d postpartum, greater serum total protein was associated with greater OB function (POB and MFIOB; r = 0.4 for both). Other markers were not associated with neutrophil function. Some metabolic and inflammatory markers in healthy postpartum dairy cows were associated with circulating neutrophil function. However, these associations only explained a small proportion of the variance in neutrophil function. Serum Hp concentration was most associated with neutrophil function changes but had opposite directions of association with OB- and PC-related functions. Future studies should focus on understanding the mechanisms by which Hp and other metabolic indicators affect neutrophil function in healthy and diseased postpartum dairy cows.