Impact of hyperketonemia in early lactation dairy cows on health and production

Data from 1,010 lactating lactating, predominately component-fed Holstein cattle from 25 predominately tie-stall dairy farms in southwest Ontario were used to identify objective thresholds for defining hyperketonemia in lactating dairy cattle based on negative impacts on cow health, milk production, or both. Serum samples obtained during wk 1 and 2 postpartum and analyzed for β-hydroxybutyrate (BHBA) concentrations that were used in analysis. Data were time-ordered so that the serum samples were obtained at least 1 d before the disease or milk recording events. Serum BHBA cutpoints were constructed at 200 μmol/L intervals between 600 and 2,000 μmol/L. Critical cutpoints for the health analysis were determined based on the threshold having the greatest sum of sensitivity and specificity for predicting the disease occurrence. For the production outcomes, models for first test day milk yield, milk fat, and milk protein percentage were constructed including covariates of parity, precalving body condition score, season of calving, test day linear score, and the random effect of herd. Each cutpoint was tested in these models to determine the threshold with the greatest impact and least risk of a type 1 error. Serum BHBA concentrations at or above 1,200 μmol/L in the first week following calving were associated with increased risks of subsequent displaced abomasum [odds ratio (OR) = 2.60] and metritis (OR = 3.35), whereas the critical threshold of BHBA in wk 2 postpartum on the risk of abomasal displacement was ≥1,800 μmol/L (OR = 6.22). The best threshold for predicting subsequent risk of clinical ketosis from serum obtained during wk 1 and wk 2 postpartum was 1,400 μmol/L of BHBA (OR = 4.25 and 5.98, respectively). There was no association between clinical mastitis and elevated serum BHBA in wk 1 or 2 postpartum, and there was no association between wk 2 BHBA and risk of metritis. Greater serum BHBA measured during the first and second week postcalving were associated with less milk yield, greater milk fat percentage, and less milk protein percentage on the first Dairy Herd Improvement test day of lactation. Impacts on first Dairy Herd Improvement test milk yield began at BHBA ≥1,200 μmol/L for wk 1 samples and ≥1,400 μmol/L for wk 2 samples. The greatest impact on yield occurred at 1,400 μmol/L (−1.88 kg/d) and 2,000 μmol/L (−3.3 kg/d) for sera from the first and second week postcalving, respectively. Hyperketonemia can be defined at 1,400 μmol/L of BHBA and in the first 2 wk postpartum increases disease risk and results in substantial loss of milk yield in early lactation.     

Effects of rumen-protected Capsicum oleoresin on immune responses in dairy cows intravenously challenged with lipopolysaccharide

The objective of this experiment was to investigate the effects of rumen-protected Capsicum oleoresin (RPC) on productivity and immune responses including feed intake, milk yield and composition, white and red blood cells, lipid peroxidation, and blood concentration of cortisol, haptoglobin, glucose, and insulin in lactating dairy cows experimentally challenged with lipopolysaccharide (LPS). The experiment was a replicated 3 × 3 Latin square design with 9 multiparous Holstein cows in three 28-d periods. Treatments were 0 (control), 100, and 200 mg of RPC/cow per day, mixed with small portions of the total mixed ration and top-dressed. Bacterial LPS was intravenously administered at 1.0 μg/kg of body weight in the last week of each experimental period, and blood samples were collected at 0, 2, 4, 8, and 24 h after administration. Dry matter intake, milk yield, and white blood cells including neutrophils, lymphocytes, monocytes, and eosinophils were decreased, and rectal temperature, hemoglobin, and serum concentrations of cortisol and haptoglobin were increased by LPS. Red blood cells, platelets, and plasma concentration of thiobarbituric acid reactive substances were not affected by LPS. Dry matter intake, milk yield, and milk composition in the 5 d post-LPS challenge were not affected by RPC. Rectal temperature, white blood cells, red blood cells, hemoglobin, and platelets were also not affected by RPC. Compared with the control, RPC tended to decrease cortisol at 2 h following LPS challenge and decreased haptoglobin concentration in serum across sampling points. Concentration of thiobarbituric acid reactive substances in plasma was decreased by RPC at 24 h post-LPS challenge. Glucose and insulin were not affected by RPC, but serum insulin concentration at 8 h was lowered by RPC compared to the control. Collectively, RPC had no or subtle effects on feed intake, milk yield and composition, rectal temperature, white and red blood cells, and serum glucose and insulin concentration in dairy cows challenged by LPS. However, RPC tended to decrease cortisol and decreased concentrations of haptoglobin and thiobarbituric acid reactive substances in blood following LPS challenge. Data suggest that dietary supplementation of RPC may modulate acute phase responses induced by bacterial infection in lactating dairy cows.     

Effects of weekly regrouping of prepartum dairy cows on innate immune response and antibody concentration

Objectives were to evaluate the effects of a stable prepartum grouping strategy on innate immune parameters, antibody concentration, and cortisol and haptoglobin concentrations of Jersey cows. Cows (253 ± 3 d of gestation) were paired by gestation length and assigned randomly to the stable (all-in-all-out; AIAO) or traditional (TRD) treatment. In the AIAO treatment, groups of 44 cows were moved into a pen where they remained for 5 wk, whereas in the TRD treatment, approximately 10 cows were moved into a pen weekly to maintain stocking density (44 cows for 48 headlocks). Pens were identical in size and design and each pen received each treatment a total of 3 times (6 replicates; AIAO, n = 259; TRD, n = 308). A subgroup of cows (n = 34/treatment) was selected on wk 1 of each replicate from which blood was sampled weekly from d −14 to 14 (d 0 = calving) to determine polymorphonuclear leukocyte (PMNL) phagocytosis, oxidative burst, and expression of CD18 and L-selectin, hemogram, cortisol and glucose concentrations, and haptoglobin concentration. Another subgroup of cows (n = 40/treatment) selected on wk 1 of each replicate was treated with chicken egg ovalbumin on d −21, −7, and 7 and had blood sampled weekly from d −21 to 21 for determination of immunoglobulin G anti-ovalbumin. All cows (n = 149) had blood sampled weekly for nonesterified fatty acid (NEFA) and β-hydroxybutyrate (BHBA) concentrations from d −21 to 21. Treatment did not affect percentage of PMNL positive for phagocytosis and oxidative burst (AIAO = 64.3 ± 2.9 vs. TRD = 64.3 ± 2.9%) and intensity of phagocytosis [AIAO = 2,910.82 ± 405.99 vs. TRD = 2,981.52 ± 406.87 geometric mean fluorescence intensity (GMFI)] and oxidative burst (AIAO = 7,667.99 ± 678.29 vs. TRD = 7,742.70 ± 682.91 GMFI). Similarly, treatment did not affect the percentage of PMNL expressing CD18 (AIAO = 96.3 ± 0.7 vs. TRD = 97.8 ± 0.7%) and L-selectin (AIAO = 44.1 ± 2.8 vs. TRD = 45.1 ± 2.8%) or the intensity of expression of CD18 (AIAO = 3,496.2 ± 396.5 vs. TRD = 3,598.5 ± 396.9 GMFI) and L-selectin (AIAO = 949.8 ± 22.0 vs. TRD = 940.4 ± 22.3 GMFI). Concentration of immunoglobulin G anti-ovalbumin was not affected by treatment (AIAO = 0.98 ± 0.05 vs. TRD = 0.98 ± 0.05 OD). The percentage of leukocytes classified as granulocyte (AIAO = 38.9 ± 1.5 vs. TRD 38.2 ± 1.5%) and the granulocyte:lymphocyte ratio (AIAO = 0.75 ± 0.04 vs. TRD = 0.75 ± 0.04) were not affected by treatment. Concentrations of cortisol (AIAO = 14.95 ± 1.73 vs. TRD = 18.07 ± 1.73 ng/mL), glucose (AIAO = 57.6 ± 1.5 vs. TRD = 60.0 ± 1.5 ng/mL), and haptoglobin (AIAO = 3.09 ± 0.48 vs. TRD = 3.51 ± 0.49 OD) were not affected by treatment. According to the current experiment, a stable prepartum grouping strategy does not improve innate immune parameters or antibody concentration compared with weekly prepartum regrouping.     

Intramammary challenge of lipopolysaccharide stimulates secretion of lingual antimicrobial peptide into milk of dairy cows

Lingual antimicrobial peptide (LAP) belongs to the β-defensin family in cattle and is found in bovine milk. However, it is unclear whether LAP is involved in the early immune response to mammary infection. The aim of the study was to investigate the changes of LAP concentration in milk after intramammary challenge with lipopolysaccharide (LPS), the gram-negative bacteria cell membrane component, in dairy cows. Milk was collected before and after LPS or phosphate-buffered saline (control) challenge every hour for 12 h on d 0 and twice daily from d 1 to 7. Somatic cell count (SCC), LAP concentration, and lactoperoxidase (LPO) activity in the milk were measured. Somatic cell count started to increase at 2 h postchallenge and remained high until d 5 (694 ± 187 × 10³ to >1,000 ± 0 × 10³ cells/mL at d 0; >1,000 ± 0 × 10³ cells/mL at d 1 to 3; 684 ± 194 × 10³ to 829 ± 108 × 10³ cells/mL at d 4; 527 ± 197 × 10³ to 656 ± 145 × 10³ cells/mL at d 5). Somatic cell count increased in the control cows, although the levels were lower compared with those in the LPS challenge group. The LAP concentration in milk increased significantly at 2 h post-LPS-challenge and was maintained at high levels until d 2 (8.6 ± 0.6 to 17.5 ± 2.3 nM). In the control cow infused with phosphate-buffered saline, there was no increase of LAP concentration in milk (5.1 ± 0.6 to 7.2 ± 0.8 nM). Increase of LPO activity in the milk was observed at 6 h after LPS challenge and continued until d 3 (4.7 ± 0.3 to 9.4 ± 1.1 U). No increase of LPO activity was observed in the milk of control cows. The increase and subsequent decrease in LAP concentration after LPS challenge occurred earlier than those of LPO activity. In multiparous cows with LPS infusion, there was a significantly negative relationship between the days leading to the basal levels in LAP concentration and LPO activity (r = −0.75). These results suggest that LPS induces secretion of LAP into milk within hours and that LPO may have a synergistic antimicrobial function with LAP in mammary glands of dairy cows.     

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.     

Influence of parturition and diets enriched in n-3 or n-6 polyunsaturated fatty acids on immune response of dairy cows during the transition period

The objectives of this study were to evaluate the functional properties of immunocompetent cells in dairy cows fed diets enriched in n-3 or n-6 polyunsaturated fatty acids during the transition period. Six weeks before calving, 21 primiparous and 27 multiparous pregnant Holstein dairy cows were randomly allotted to 1 of 3 dietary fat treatments: calcium salts of palm oil (Megalac), micronized soybeans, or whole flaxseed, which are, respectively, rich in saturated, n-6, or n-3 fatty acids. On wk 6 and 3 before parturition, cows received a subcutaneous injection of ovalbumin to measure the antibody response in colostrum and serum. Colostrum samples were collected at the first milking after calving, and blood samples were taken 6, 3, and 1 wk before the expected calving date and 1, 3, and 6 wk after calving. Blood mononuclear cells were cultured to evaluate the proliferative response to concanavalin A and the in vitro productions of interferon-gamma, tumor necrosis factor-alpha, nitric oxide, and prostaglandin E2. The serum antibody response to ovalbumin was unaffected by dietary fatty acids, but the response was lower in primiparous cows than in multiparous cows. A significant diet x parity interaction indicated that colostral antibody level against ovalbumin was significantly higher in multiparous cows fed soybeans than in those fed flaxseed or Megalac; there was no difference among treatments for primiparous cows. The lymphocyte response to concanavalin A was lower in cows fed soybeans than in those receiving flaxseed or Megalac when the cells were incubated with autologous serum. The proliferative response of mononuclear cells incubated with autologous serum was suppressed in the 1st wk after calving in both primiparous and multiparous cows, and multiparous cows showed a higher response than primiparous cows throughout the experiment. There was a significant interaction between parity and diet as a result of a greater production of interferon-gamma by mononuclear cells incubated with autologous serum in multiparous cows than in primiparous cows fed flaxseed; there was no difference among cows fed the other diets. Interferon-gamma production was reduced around calving while the inverse was observed for productions of nitric oxide and tumor necrosis factor-alpha. Productions of nitric oxide, prostaglandin E2, and tumor necrosis factor-gamma were greater in primiparous cows than in multiparous cows. In conclusion, functional properties of lymphocytes and monocyte/macrophage lineage of dairy cows during the transition period are modulated by parturition and the composition of polyunsaturated fatty acids in the diet.     

Immune response of postpartum dairy cows fed flaxseed

Thirty Holstein cows were allotted at calving to 10 groups of three cows blocked for similar calving dates to determine the effects of dietary fatty acids on functional properties of immunocompetent cells in early lactation and at breeding. Cows were assigned at calving to one of three isonitrogenous, isoenergetic, and isolipidic supplements based on either calcium salts of palm oil, Megalac, micronized soybeans, or whole flaxseed. On the day of AI and 20 d later, cows were injected with ovalbumin to measure the antibody response. Blood samples were taken at different times after calving (d 5, 21, 42, and 105) and after AI (d 0, 10, 20, and 40) for quantification of serum progesterone, fatty acids, and prostaglandin E2 concentrations. Isolated peripheral blood mononuclear cells were cultured to evaluate the proliferative response to concanavalin A and in vitro productions of interferon-gamma and prostaglandin E2. In general, feeding flaxseed increased serum omega-3 fatty acids concentration compared with feeding Megalac or soybeans, which decreased the omega-6 to omega-3 fatty acids ratio. There was a significant diet x day interaction for the proliferative response of mononuclear cells after calving and AI, indicating that cell responses from cows fed flaxseed were transiently reduced compared with those fed Megalac and soybeans. Moreover, during the breeding period, serum progesterone concentration was significantly greater in cows fed flaxseed compared with those fed Megalac, whereas serum concentration of prostaglandin E2 was significantly lower in cows fed flaxseed than in those fed Megalac or soybeans. Dietary treatments had no effect on the antibody response to ovalbumin and on in vitro productions of interferon-gamma and prostaglandin E2. However, interferon-gamma and prostaglandin E2 were impaired in the first 3 wk after parturition regardless of dietary treatment. These results suggest that changes in fatty acids, progesterone, and prostaglandins E2 concentrations in serum due to dietary treatment and physiological status influenced systemic immunity as shown by reduced proliferative response. However, other mechanisms must be considered and are discussed to explain dietary effect on lymphocyte proliferative response to mitogenic stimulation and other immune functions.     

Relationships between rumen lipopolysaccharide and mediators of inflammatory response with milk fat production and efficiency in dairy cows

The main objective of this study was to evaluate correlative relationships between rumen lipopolysaccharide (LPS) and mediators of acute phase response with milk fat yield and efficiency in dairy cows challenged with graded amounts of barley grain in the diet. An additional aim of the study was to quantify the intercow variation in relation to milk fat production and acute phase response in cows fed graded amounts of grain. Eight primiparous, lactating Holstein cows (60 d in milk) were assigned to 1 of the 4 total mixed rations containing barley grain at 0, 15, 30, and 45% (dry matter basis) in a replicated 4 × 4 Latin square design. Free rumen LPS, plasma acute phase proteins, and milk fat content were quantified in multiple samples collected on d 5 and 7 of the measurement periods shortly before the morning feeding. Results showed markedly greater concentrations of rumen LPS with increasing dietary grain level. The correlative analysis revealed strong negative relationships between rumen LPS and milk fat content and yield. The predictor variable of rumen LPS explained 69% of the variation during the milk fat reduction of the cows. The stronger depression in milk fat percentage was obtained when rumen LPS exceeded a threshold of 5,564 ng/mL, corresponding to a milk fat content of 3.39%. The increase in concentration of rumen LPS was also associated with declines in milk fat yield and 3.5% fat-corrected milk (R² = 0.50), as well as milk energy efficiency (R² = 0.43). The correlative analysis also indicated that the increase of plasma C-reactive protein (CRP) in response to higher grain feeding was associated with a linear decrease of milk fat content and yield (R² = 0.28 to 0.46). Furthermore, the statistical analysis revealed high percentages of intercow variation related to milk fat variables, as well as the responses of rumen LPS and plasma CRP. Taken together, the current results implicate rumen LPS and the host CRP response in the lowering of milk fat content and milk energy efficiency in dairy cows fed high-grain diets. Further research is warranted to understand the mechanism(s) by which rumen LPS and inflammatory responses to LPS lower milk fat synthesis and milk energy efficiency and to develop novel strategies for their prevention.     

Supplementation with rumen-protected methionine or choline during the transition period influences whole-blood immune response in periparturient dairy cows

Methionine, together with Lys, is the most limiting AA for milk production in dairy cows. Besides its crucial role in milk production, Met and its derivate metabolites (e.g., glutathione, taurine, polyamines) are well-known immunonutrients in nonruminants, helping support and boost immune function and activity. In the present study, the effects of Met or choline, as its precursor, were investigated using an ex vivo whole blood challenge. The study involved 33 multiparous Holstein cows (from a larger cohort with a factorial arrangement of treatments) assigned from d ?21 to +30 relative to parturition to a basal control (CON) diet, CON plus rumen-protected Met (MET, Smartamine M, Adisseo NA, Alpharetta, GA) at a rate of 0.08% of dry matter, or CON plus rumen-protected choline (CHOL, ReaShure, Balchem Inc., New Hampton, NY) at 60 g/d. Blood was sampled on d ?15, ?7, 2, 7, and 20 for ex vivo lipopolysaccharide (LPS) challenge, and on d 1, 4, 14, and 28 relative to parturition for phagocytosis and oxidative burst assays. The MET cows had greater energy-corrected milk production and milk protein content. Overall, IL-6 response to LPS increased around parturition, whereas IL-1β remained constant, casting doubt on the existence of systemic immunosuppression in the peripartal period. Supplementation with MET dampened the postpartal blood response to LPS (lower IL-1β), while improving postpartum neutrophil and monocyte phagocytosis capacity and oxidative burst activity. In contrast, CHOL supplementation increased monocyte phagocytosis capacity. Overall, the data revealed a peripartal immune hyper-response, which appeared to have been mitigated by MET supplementation. Both MET and CHOL effectively improved immune function; however, MET affected the immune and antioxidant status before parturition, which might have been beneficial to prepare the cow to respond to metabolic challenges after parturition. These results provide insights on potential differences in the immunomodulatory action of methionine and choline in dairy cows. As such, the effects observed could have implications for ration formulation and dietary strategies.     

Effect of postruminal glutamine supplementation on immune response and milk production in dairy cows

Seventeen multiparous Holstein cows were used to examine the effect of an increased duodenal supply of Gln on immune function and production. Cows received continuous abomasal infusions of water (control: n = 8) or 300 g/d of Gln (n = 9) for 21 d starting within 48 h of calving. There were nonsignificant increases in milk and milk protein yields in response to Gln supplementation. Glutamine treatment had no effect on plasma glucose, nonesterified fatty acids (NEFA), or β-hydroxybutyrate (BHBA) concentrations but did tend to increase plasma urea N concentration. The Gln treatment resulted in an increase of 108 μM in the plasma Gln concentration. Total essential AA concentrations decreased with the Gln treatment, whereas total nonessential AA concentrations were unaffected. T Lymphocyte proliferation did not differ between the control and Gln-treated cows. Treatment had no effect on the relative abundance of CD8 T cells but did increase the abundance of CD4 T cells. Cytokine production, as measured by IFN-γ concentration determined in vitro in concanavalin-A-stimulated peripheral blood mononuclear cells, was similar between the treatments. Over the first 3 wk following calving, Gln supplementation had limited effects on milk production, metabolic parameters, and immune function.     

Metabolites and immune variables associated with somatic cell counts of primiparous dairy cows

The main objective of this study was to investigate associations between serum concentrations of several blood variables related to metabolic and immunological status around calving, and udder health measured as milk somatic cell counts (SCC), Box-Cox transformed to bcSCC, at first test-milking in 287 primiparous cows in 20 Swedish dairy herds. Possible systematic effects of breed and age at calving on blood profiles were also investigated. Ordinary linear regression models, with robust standard errors and adjusting for clustering within herds, were used to investigate associations between blood variables and bcSCC. Hierarchical linear regression models, with herd as random factor, were used to investigate systematic effects on blood variables. The results showed that greater concentrations of β-hydroxybutyrate (BHBA) and glucose before calving were associated with lesser bcSCC at first test-milking, whereas greater concentrations of nonesterified fatty acids (NEFA) before calving and greater delta NEFA (describing the difference in concentrations before and after calving) were associated with greater bcSCC at first test-milking. In addition, greater α-tocopherol concentrations in the period −5 to +5 d relative to calving were associated with lesser bcSCC at first test-milking, whereas greater concentrations of collectin of 43 kDa (CL-43) postpartum (1 to 21 d after calving) were associated with greater bcSCC. Postpartum concentrations of conglutinin and haptoglobin were also associated with bcSCC, but not independently of each other. Moreover, significant breed differences were observed for insulin, urea nitrogen, conglutinin, cholesterol, NEFA, and CL-43, the latter 3 as an interaction with period. Overall, cows of the Swedish Red breed had greater concentrations of insulin, cholesterol, urea nitrogen, and conglutinin, and lesser concentrations of NEFA and CL-43 than cows of the Swedish Holstein breed. Age at calving as main effect was significantly associated with BHBA, glucose, insulin, NEFA, urea nitrogen, and conglutinin. Heifers calving at >27 mo of age had greater BHBA and NEFA values, and lesser glucose, insulin, and urea nitrogen values compared with heifers calving at <27 mo. Heifers calving at an age <25 mo had greater conglutinin and urea nitrogen values, and lesser NEFA values compared with heifers calving at >25 mo. The results show that there are several associations among metabolites, immunological variables, and udder health of primiparous cows, but also that these variables vary between breeds and between cows of different age at first calving.     

Accuracy of milk ketone bodies from flow-injection analysis for the diagnosis of hyperketonemia in dairy cows

The objectives of this study were (1) to determine the correlations between blood β-hydroxybutyrate (BHBA) and milk components [BHBA, acetone, fat, protein, and fat:protein (F:P) ratio], and (2) to establish optimal thresholds for milk components to predict hyperketonemia in dairy cows. Data on 163 cows from 37 herds were used in this cross-sectional study. Herds were visited once during the study period, and cows between 2 and 90 d in milk were blood sampled within 4 h of milk sampling for the Dairy Herd Improvement test. Blood BHBA concentrations were measured using a cow-side electronic meter, Precision Xtra, which was considered the gold standard test in this study. Milk BHBA and acetone concentrations were measured in Dairy Herd Improvement milk samples by flow-injection analysis; whereas, milk fat and protein were tested using Fourier transform infrared spectroscopy. Hyperketonemia was defined by a blood BHBA concentration ≥1.4 mmol/L. The prevalence of hyperketonemia (based on blood BHBA values) in this study population was 21.0%. Pearson correlation coefficients between blood BHBA and milk BHBA, acetone, fat, protein, and F:P ratio were 0.89, 0.73, 0.21, 0.04, and 0.17, respectively. Receiver operating characteristic curves were generated and thresholds for each individual milk component were determined based on the maximal sum of sensitivity and specificity. Optimal threshold values for hyperketonemia were milk BHBA ≥0.20 mmol/L, acetone ≥0.08 mmol/L, fat ≥4.2%, and F:P ratio ≥1.3. Based on these thresholds, milk BHBA and acetone had greater sensitivity (84 and 87%, respectively) and greater specificity (96 and 95%, respectively) than the other milk components (fat, protein, and F:P). Series and parallel testing slightly improved the accuracy of milk BHBA and acetone values to predict hyperketonemia. A multivariable model that accounted for milk BHBA and milk acetone values simultaneously had the highest accuracy of all tested models for predicting hyperketonemia. These results support that milk BHBA and milk acetone values from flow-injection analysis are accurate diagnostic tools for hyperketonemia in dairy cows and could potentially be used for herd-level hyperketonemia surveillance programs.     

Effects of intrauterine infusion of Escherichia coli lipopolysaccharide on uterine health,resolution of purulent vaginal discharge,and reproductive performance of lactating dairy cows

The objectives of the current experiment were to evaluate the effects of intrauterine infusion of Escherichia coli lipopolysaccharide (LPS) in cows diagnosed with purulent vaginal discharge (PVD) on intrauterine cell population, resolution of PVD, uterine health, and reproductive performance. Jersey cows (n = 3,084) were examined using the Metricheck device to diagnose PVD at 35 ± 6 d postpartum. Purulent vaginal discharge was defined as the presence of purulent (≥50% pus) discharge detectable in the vagina. Of the 310 cows positive for PVD, 267 cows were enrolled in the current experiment. To ensure proper timing of treatment and collection of samples, only 9 PVD-positive cows were treated per day. Selected cows were balanced at 35 ± 6 d postpartum for lactation number, body condition score, and milk yield and were randomly assigned to receive an intrauterine infusion of 20 mL of phosphate-buffered saline (PBS; control, n = 87), 20 mL of PBS with 150 µg LPS (LPS150, n = 91), or 20 mL of PBS with 300 µg of LPS (LPS300, n = 89). Uterine cytology was performed immediately before treatment and 1, 2, and 7 d after treatment to evaluate the effect of LPS treatment on intrauterine cell population. Cows were examined with the Metricheck device at 7 and 28 d after treatment to evaluate the effects of treatment on resolution of PVD. Reproductive status was recorded up to 200 d postpartum. Cows diagnosed with PVD had greater incidence of twinning, dystocia, retained placenta, and metritis after calving than cows without PVD. Count of polymorphonuclear leukocytes (PMNL) in uterine cytology 1, 2, and 7 d after intrauterine infusion was not statistically different among treatments. From d 0 to 1, however, PMNL count in uterine cytology of PBS cows increased by 5%, whereas the PMNL count in uterine cytology of LPS150 and LPS300 cows increased by 54 and 48%, respectively. Treatment did not affect the likelihood of cows being diagnosed with PVD 7 and 28 d after intrauterine infusion. Cows without PVD and LPS150 cows were more likely to be pregnant after the first postpartum AI than PBS cows. After the second postpartum AI, cows without PVD were more likely to be pregnant than PBS and LPS300 cows. Hazard of pregnancy up to 200 d postpartum was decreased for PBS and LPS300 cows compared with cows without PVD, and it tended to be decreased for LPS150 cows compared with cows without PVD. Intrauterine treatment with 150 µg of E. coli LPS of cows diagnosed with PVD improved likelihood of pregnancy after the first postpartum AI, but further research is needed to elucidate the mechanism by which LPS treatment improved fertility.     

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.     

Milk supplemented with dried seaweed affects the systemic innate immune response in preweaning dairy calves

Intact seaweed or seaweed extracts are used as feed supplements to improve the gut microbiome in young animals. Seaweeds provide functional polysaccharides, and they are a good source of vitamins, minerals, and phenolic compounds, all of which are relevant for immune system development. However, literature on the effects of dried seaweed supplementation on immune system development is limited, especially in calves. This experiment aimed to study the effect of feeding milk supplemented with Ulva lactuca, Ascophyllum nodosum, or Saccharina latissima on the systemic immune status of preweaning dairy calves. Forty male Holstein calves with birth body weight 41 ± 4 kg and plasma Brix percentage ≥8.7% at d 2 after birth were used in this study. Calves were fed 4 L of cow milk twice a day (total 8 L/d). From d 2 to d 28, calves in the control group (n = 10) received milk without seaweed supplementation. Over the same period, experimental calves received milk supplemented with Ulva lactuca (SW1; n = 10), Ascophyllum nodosum (SW2; n = 10), or Saccharina latissima (SW3, n = 10). Dried and ground seaweeds were offered at a daily allowance of 50 g/8 L of milk (i.e., approximately 5% inclusion rate on a dry matter basis). Blood samples were collected from a jugular vein on d 2, 4, 7, 14, 21, and 28 after birth. Plasma concentrations of total protein, albumin, immunoglobulins, and acute-phase proteins (i.e., serum amyloid A, fibrinogen, and haptoglobin) were measured. We detected no differences in average daily gain, plasma immunoglobulins, albumin, or total protein. However, the contrast analysis revealed that plasma concentrations of fibrinogen (SW1 and SW2) and serum amyloid A (SW2 and SW3) were significantly higher in the seaweed groups compared with the control group. We also found a tendency for high plasma haptoglobin in the seaweed groups (SW1 and SW2) compared with the control group. Differences in acute-phase protein concentrations could be partially explained by the large differences in micromineral intake between control and seaweed-supplemented calves. Feeding milk supplemented with dried seaweed increased plasma concentrations of variables related to the innate immune response in preweaning dairy calves.     

Effect of rumen-protected B vitamins and choline supplementation on health,production, and reproduction in transition dairy cows

The objectives were to determine the effects of a rumen-protected blend of B vitamins and choline (RPBC) on the incidence of health disorders, milk yield, and reproduction in early lactation and the effects on gene expression and liver fat infiltration. A randomized controlled trial in 3 commercial dairy herds (n = 1,346 cows with group as the experimental unit; experiment 1) and a university research herd (n = 50 cows with cow as the experimental unit; experiment 2) evaluated the use of 100 g/cow per d of commercially available proprietary RPBC supplement (Transition VB, Jefo, St. Hyacinthe, Quebec, Canada), or a placebo, fed 3 wk before to 3 wk after calving. In experiment 2 liver biopsies were taken at 4 and 14 ± 1 d in milk to measure triacylglycerol concentrations and expression of 28 genes selected to represent relevant aspects of liver metabolism. Treatment effects were assessed using multivariable mixed logistic regression models for binary health and reproductive outcomes; linear regression models for milk yield, dry matter intake, and liver outcomes; and survival analysis for time insemination and pregnancy. In experiment 1, treatment did not have an effect on the incidence of hyperketonemia (blood β-hydroxybutyrate ≥ 1.2 mmol/L; cumulative incidence to 3 wk postpartum of 28 to 30%), clinical health disorders, or udder edema. The prevalence of anovulation at 8 wk postpartum was 11% in the treatment group and 23% in the control but did not differ statistically given group-level randomization. Pregnancy at first insemination (33 and 35%) and median time to pregnancy to 200 d in milk (96 and 97 d) were not different between treatment and control, respectively. No difference was observed between treatment groups in milk yield or components through the first 3 Dairy Herd Improvement Association test days (44 kg/d in both groups, accounting for parity and components). In experiment 2, there were no differences between treatment groups in feed intake. Mean blood β-hydroxybutyrate was lower at wk 3 in RPBC (0.6 vs. 0.9 ± 0.12 mmol/L) with no difference between treatments for mean blood concentrations of fatty acids (wk ?1 or 1) and β-hydroxybutyrate at wk 1 or 2. The gene for acyl-CoA oxidase 1 (ACOX1) had lower mRNA abundance in RPBC with no difference between treatments for the other genes, but the expression of half of the genes assessed differed with days in milk. Liver triacylglycerol was lower in primiparous cows at 4 d in milk in RPBC (2.0 vs. 4.4 ± 1.2%) but not at 14 d in milk (2.2 vs. 3.2 ± 0.97%) with no treatment effect in multiparous cows (4.6 ± 0.8%). Accounting for parity, days in milk, fat and protein percentages, repeated test days, and a random effect of cow, no significant difference was observed between treatments in milk yield across the first 3 Dairy Herd Improvement Association tests (41.2 ± 1.3 in RPBC vs. 38.0 ± 1.4 kg/d in control). Under the diet and management conditions of the field study including low prevalence of clinical health disorders, in experiment 1 we did not detect a benefit of RPBC, but in experiment 2 liver fat content decreased in primiparous cows.     

Frequency of feed delivery affects the behavior of lactating dairy cows

The objectives of this study were to examine how frequency of feed delivery affects 1) the behavior of group-housed and group-fed dairy cows and 2) the extent of feed sorting. These objectives were tested in two experiments. In each experiment, 48 lactating Holstein cows, split into groups of 12, were subjected to each of 2 treatments (over 10-d periods) in a cross-over design. The treatments for the first experiment were 1) delivery of feed once per day (1×) and 2) delivery of feed twice per day (2×). Treatments for the second experiment were 1) delivery of feed 2× and 2) delivery of feed four times per day (4×). For the 1×, 2×, and 4× treatments, feed was pushed up 3, 2, and 0 times per day, respectively. For both experiments, cows had 0.6 m of feeding space; one cows was allowed per lying stall. Time-lapse video was used to quantify the feeding and lying behavior, as well as the aggressive behavior displayed at the feed bunk by the cows. Changes in NDF content of the TMR throughout the day were used to determine the extent of feed sorting by the cows. In both experiments, increased frequency of feed provision increased, as well as changed, the distribution of daily feeding time. The changes in distribution of feeding time resulted in cows having more equal access to feed throughout the day. Frequency of feed delivery had no effect on the daily lying time of the cows or the daily incidence of aggressive interactions at the feed bunk. However, subordinate cows were not displaced as frequently when fed more often. For all treatments, in both experiments, the NDF content of the TMR present in the feed bunk increased throughout the day, indicating that sorting of the feed had occurred. Further, the amount of sorting of the feed was reduced by increasing the frequency of feed delivery from 1× to 2×. These results indicate that frequent delivery of feed improves access to feed for all cows, particularly during peak feeding periods when fresh feed is provided, and reduces the amount of feed sorting.     

Decreased insulin response in dairy cows following a four-day fast to induce hepatic lipidosis

Negative energy balance has been implicated in the development of fatty liver, insulin resistance, and impaired health in dairy cows. A 4-d fasting model previously was reported to increase liver triglycerides more than 2.5-fold. The purpose of the present study was to evaluate insulin response in this fasting model. Nonlactating, nonpregnant Holstein cows were fasted for 4 d (6 cows) or fed continuously as control cows (4 cows). Samples were collected 5 d before fasting, during fasting, and immediately after the 4-d fast, 8 d after the fast, and 16 d after the fast. Fasted cows had greater liver triglyceride content (49.4 vs. 16.2 mg/g, wet-weight basis) at the end of the fasting period compared with control cows. Fasted cows also had increased plasma nonesterified fatty acid (NEFA) concentrations (1.24 vs. 0.21 mmol/L) and increased plasma β-hydroxybutyrate (BHBA) concentrations at the end of the fasting period. Liver triglyceride, plasma NEFA, and plasma BHBA in fasted cows returned to prefasting concentrations by the end of the experiment. Plasma glucose concentrations were not affected by fasting. Plasma insulin concentrations were decreased (6.3 vs. 14.1 μU/mL) and insulin-stimulated blood glucose reduction was decreased (24.9 vs. 48.6%) in the fasted cows compared with control cows at the end of the fast, indicating reduced insulin response. Insulin response was negatively correlated with plasma NEFA and liver triglycerides. Decreased insulin response may be an important complication of negative energy balance and hepatic lipidosis.     

Time of feed delivery affects the feeding and lying patterns of dairy cows

The objective of this experiment was to determine whether it is the return from milking or delivery of fresh feed that has the greater effect on the daily patterns of feeding and lying behavior of dairy cattle. Forty-eight lactating Holstein cows were subjected to each of 2 treatments in a 2 x 2 cross-over design replicated over time. The treatments were 1) milking and feed delivery times coinciding and 2) feed delivery 6 h after milking. Cows were milked twice daily at 0500 and 1700 h. An electronic monitoring system was used to measure the time spent at the feed alley. Time-lapse video was used to quantify the lying time and incidence of aggressive displacements of the cows at the feed alley. Cows increased their total daily feeding time by 12.5% when fed 6 h after milking. This change was driven by an 82% increase in feeding time during the first hour immediately following the delivery of fresh feed and a 26% decrease in feeding time during the first hour after milking. The delivery of feed 6 h after milking did not change the daily lying time of the cows, but did decrease the latency to lie down after milking by 20 min. The reduction in feeding time after milking and decreased latency to lie down resulted in a tendency for less aggressive interactions at the feed alley after the cows returned from milking. These results indicate that the delivery of fresh feed has a greater impact on stimulating feeding behavior than does the return from milking and that changes in feeding management can affect both the feeding and lying behavior of dairy cows.