Liver phosphorus content in Holstein-Friesian cows during the transition period

Hepatic lipidosis and hypophosphatemia are frequently observed in high-yielding periparturient dairy cows. Objectives of this study were to investigate the association of the liver P content with the degree of liver fat accumulation and serum P concentration and to characterize the change in liver P content throughout the transition period. In a cross-sectional study, liver biopsies obtained from 33 Holstein-Friesian cows 14 d postpartum (p.p.) were assayed for total lipid (TLip), triacylglycerol, DNA, P, Mg, K, Na, and Ca content. Serum samples obtained at the time of biopsy were analyzed for indices of liver function and injury and the serum P concentration was determined. From this cross-sectional study, 6 cows were selected for a longitudinal study and liver tissue obtained from the 6 cows on d −65, −30, −14, 1, 14, 28, and 49 relative to calving was assayed. The amounts of P, K, Mg, Na, and Ca were expressed as amount in dry weight (DW), wet weight (WW), nonfat wet weight (NFWW), and indexed to DNA. In the cross-sectional study, P_DW and P_WW decreased with increasing TLip, whereas P_NFWW and P_DNA were independent of TLip. Values for P_DNA varied widely, whereas P_NFWW varied within a narrow range. Stepwise regression analysis revealed the strongest associations between P_DW and the amount of tissue water (partial R² = 0.74) and the log to the base 10 of triacylglycerol (partial R² = 0.05). The P_WW was associated with the log to the base 10 of triacylglycerol (partial R² = 0.20), but no associations were found for P_NFWW. These findings indicate that decreased electrolyte content in dry and wet liver tissue with increased liver lipid content is predominantly due to the decrease in tissue water and therefore the distribution volume of electrolytes. In the longitudinal study, P_DW, P_WW, and P_NFWW were decreased on d 14 p.p. Similar directional decreases were found for K, Mg, and Na, but P was the only electrolyte that was significantly decreased in liver tissue at d 14 p.p. This finding indicates that the P content of liver tissue decreases in early lactation due to a reduction in hepatocellular cytosol volume as well as a decrease in cytosolic P concentration, with the latter having biological relevance. The clinical significance of decreased cytosolic P concentration in the hepatocytes of dairy cows in early lactation remains to be determined.     

Influence of feeding a low-phosphorus diet on leucocyte function in dairy cows

Phosphorus depletion and hypophosphatemia have been described to interfere with immune function in rats and humans. In dairy cows, hypophosphatemia has been associated with muscle weakness and recumbency as well as with intravascular hemolysis resulting from increased osmotic fragility of erythrocytes, but so far, the influence of P depletion and hypophosphatemia on immune function has not been studied. Therefore, the aim of this study was to investigate whether P depletion and ensuing hypophosphatemia are associated with impaired granulocyte and lymphocyte function. Eight mid-lactation dairy cows were fed a P-deficient ration (0.2% P/kg of DM) for a period of 4 wk. The depletion phase was preceded by a 2-wk acclimatization period and followed by a 2-wk repletion phase, during which the same ration was supplemented with P to meet or exceed daily requirements. Blood samples were collected at the end of the acclimatization period, after 2 and 4 wk of P depletion, and at the end of the repletion phase. Plasma phosphate concentrations ([Pi]) were determined and white blood cells were counted and isolated. General immune function was investigated by performing a phagocytosis assay with Staphylococcus aureus and a lymphocyte stimulation test (LST) with concanavalin A and pokeweed mitogen. The plasma [Pi] decreased significantly, with the lowest values (mean 0.7 ± 0.2 mmol/L) occurring after 2 wk of depletion, although depletion was continued for another 2 wk. During repletion, plasma [Pi] increased above baseline concentrations. Granulocyte counts changed in parallel with plasma [Pi] over time, decreasing significantly at 2 wk after P depletion and increasing again thereafter. Granulocyte survival after phagocytosis was lowest after 4 wk of P depletion. Phagocytosis activity of surviving granulocytes determined by mean fluorescence intensity was higher, indicating that phagocytosis was not negatively influenced by P depletion. Lymphocyte stimulation showed a similar trend, with a decreasing stimulation index at the end of P depletion, but differences were not statistically significant. Data presented in this study indicate that hypophosphatemia leads to a decrease in granulocyte counts. Chronic P depletion impairs granulocyte survival during phagocytosis but not phagocytosis activity. Lymphocyte function is not influenced by P depletion.     

Phosphorus partitioning during early lactation in dairy cows fed diets varying in phosphorus content

The effect of dietary P content on P partitioning and excretion during early lactation was evaluated in 13 cows fed diets containing 0.34 (no supplementary P), 0.51, or 0.67% P. All cows were fed a common pre-partum total mixed ration (TMR) (0.28% P), followed by common TMR (0.51% P) for 7 d post-partum. On day 7, cows were randomly assigned to one of the three dietary P treatments. All treatment diets contained 16.6% CP, 15.2% ADF, 26.3% NDF, and 0.74% Ca. Total collections of milk, urine, and feces were conducted during weeks 3, 5, 7, 9, and 11 of lactation. Average body weight (591 kg), milk yield (47.9 kg/d), and DMI (25.2 kg/d) throughout the 10-wk trial were not affected by dietary P content. With increasing dietary P, however, linear increases in P intake (84.7, 135.2, and 161.5 g/d), fecal P (42.3, 87.5, and 108.6 g/d), urinary P (0.32, 1.28, and 3.90 g/d), and total P excretion (42.6, 88.8, and 112.5 g/d) were observed. Apparent P digestibility (49.0, 34.4, and 32.8% of P intake) decreased quadratically with increasing dietary P. Phosphorus balance was highly variable, but cows fed the 0.34% P diet were in negative P balance longer than were cows fed diets containing 0.51 or 0.67% P. With increasing dietary P, serum concentrations of inorganic P increased linearly, but serum Ca and Mg concentrations decreased. Increasing dietary P increased fecal and urinary P excretion in early lactation cows. Increased duration of negative P balance and changes in blood mineral concentrations suggest that cows fed low P diets mobilized more P from body reserves than cows on medium- and high-P diets.     

Effects of restricted dietary phosphorus supply during the dry period on productivity and metabolism in dairy cows

Phosphorus in bovine nutrition is under ongoing scrutiny because of concerns with excessive amounts of P excreted in manure contributing to environmental pollution. Feeding rations with excessive P content, however, still remains common practice, particularly during the transition period, as limited P supply in late gestation and early lactation is thought to present a risk for health and productivity of high-yielding dairy cows. The objectives of this study were to investigate the effect of restricted P supply during the last 4 wk of pregnancy on Ca and P homeostasis during the transition period in high-yielding dairy cows, and to identify possible effects on metabolism and productivity throughout the following lactation. Thirty late-pregnant multiparous dairy cows were randomly assigned to either a dry cow diet with low (LP) or adequate P (AP) content [0.16 and 0.30% P in dry matter (DM), respectively] to be fed in the 4 wk before calving. After calving all cows received the same ration with adequate P content (0.46% P in DM). Blood, milk, and liver tissue samples were obtained during the dry period and the following lactation, DM intake (DMI), body weight, milk production, and disease occurrence were monitored. Plasma was assayed for the concentrations of P, Ca, Na, and K, metabolic parameters, and liver enzyme activities. Liver tissue was analyzed for mineral, triglyceride, cholesterol, and water contents. Repeated-measures ANOVA was used to identify treatment, time, and treatment × time interaction effects. Cows fed LP had lower plasma P concentrations ([Pi]) than AP cows during restricted P feeding, reaching a nadir of 1.1 mmol/L immediately before calving. After calving, plasma [Pi] of LP cows was at or above the level of AP cows and within the reference range for cattle. Symptoms assumed to be associated with hypophosphatemia were not observed, but plasma Ca was higher from 1 wk before to 1 wk after calving in LP cows, which was associated with a numerically lower incidence of clinical and subclinical hypocalcemia in LP cows. Both treatments had a similar 305-d milk yield (12,112 ± 1,298 kg for LP and 12,229 ± 1,758 kg for AP cows) and similar DMI. Plasma and liver tissue biochemical analysis did not reveal treatment effects on energy, protein, or lipid metabolism. The results reported here indicate that restricted dietary P supply during the dry period positively affected the Ca homeostasis of periparturient dairy cows but did not reveal negative effects on DMI, milk production, or metabolic activity in the following lactation. Restriction of P during the dry period was associated with hypophosphatemia antepartum but neither exacerbated postparturient hypophosphatemia, which is commonly observed in fresh cows, nor was associated with any clinical or subclinical indication of P deficiency in early lactation.     

Effects of supplementing a Saccharomyces cerevisiae fermentation product during the transition period on rumen fermentation of dairy cows fed fresh diets differing in starch content

The objective of this study was to evaluate the effects of supplementing a Saccharomyces cerevisiae fermentation product (SCFP; NutriTek, Diamond V, Cedar Rapids, IA) during the transition period (d −28 ± 3 to 23 ± 3 relative to calving) on rumen fermentation and mRNA abundance of genes in the rumen epithelium of fresh cows (d 1 to 23 ± 3 after calving) fed diets differing in starch content. Eighteen ruminally cannulated multiparous Holstein cows were fed diets with SCFP (n = 9) or without (CON; n = 9) throughout the experiment. All cows were fed a common basal controlled-energy close-up diet (1.43 Mcal/kg, net energy for lactation; 13.8% starch) before calving. Cows within each treatment (CON or SCFP) were fed either a low-starch (LS; 22.1% starch) or high-starch (HS; 28.3% starch) diet during the fresh period. Cows were assigned to treatment after balancing for parity, body condition score, and expected calving date. Rumen pH was measured continuously for 72 h starting on d −10, −3, 1, 7, and 21 relative to calving date. Rumen papillae were collected on d −10 and 21 relative to calving. Supplementation of SCFP had no effect on rumen pH during d −10 to −8, but mean rumen pH tended to be higher (6.64 vs. 6.49) for SCFP cows than for CON cows during d −3 to −1. Feeding SCFP decreased the range of rumen pH variation compared with CON within the HS group during both d 7 to 9 (1.08 vs. 1.38) and d 21 to 23 (1.03 vs. 1.30) after calving. In addition, nadir rumen pH tended to be higher (5.64 vs. 5.44) and duration of pH below 5.8 tended to be shorter (116 vs. 323 min/d) for the SCFP group than for the CON group during d 21 to 23 after calving. Supplementation of SCFP increased the mRNA abundance of insulin-like growth factor-6 (1.10 vs. 0.69) before calving and decreased the mRNA abundance of putative anion transporter isoform 1 (1.12 vs. 2.27) after calving. Nadir rumen pH tended to be higher during d 1 to 3 (5.63 vs. 5.41) for LS cows than for HS cows, but rumen pH was not affected by dietary starch content during other time periods. Dietary starch content had no effect on mRNA abundance of genes in the rumen epithelium after calving. These results suggest that supplementation of SCFP may reduce the range of variation in rumen pH in fresh cows fed HS diets and the duration of subacute ruminal acidosis by the end of the fresh period regardless of dietary starch content and that decreasing dietary starch content during the fresh period may reduce the decrease in rumen pH immediately after parturition.     

Effect of liquid flavor supplementation of the diet on dairy cows in the transition period

A 9-wk trial was conducted to study the performance of 24 Holstein cows during the transition period (3 wk prepartum to 6 wk postpartum). Cows were assigned to either a control or liquid-flavored (0.52 mL/kg of feed) total mixed ration in a randomized complete block design. The diets contained corn silage, alfalfa haylage, cottonseed, and a grain mix based on ground corn and soybean meal. Cows were fed to ensure 10% orts, and the diet provided (on a dry matter basis) 13% crude protein, 32% acid detergent fiber, 44% neutral detergent fiber, and 1.54 Mcal/kg of NEL prepartum and 17.5% crude protein, 30% acid detergent fiber, 40% neutral detergent fiber, and 1.57 Mcal/kg of NEL postpartum. An additional 2.3 kg of alfalfa hay was fed during the first 5 d postpartum. Weekly means of dry matter intake (DMI), milk yield, milk protein, milk fat, SNF, somatic cell counts, and body weight (BW) were analyzed using a repeated measures procedure. There was no effect of treatment on these variables, and least squares means were 16.9 and 15.7 kg/d for DMI, 38 and 35.3 kg/d for milk yield, 3.10 and 3.11% for milk protein, 3.69 and 3.74% for milk fat, 8.37 and 8.16% for SNF, 1.99 x 10(5) and 4.33 x 10(5) for somatic cell count, and 631 and 651 kg for BW for cows fed control and flavored diets, respectively. Individual cow daily DMI data were fitted to an exponential model describing pre- and postpartum feed consumption [DMI = a - b x e(-c x t), where DMI was measured in kg, a = asymptotic DMI, b = potential fractional increase in DMI, c = fractional rate of increase in DMI, and t = days prior to calving or days in milk]. Fractional rates of increase in DMI were similar: 0.139 and 0.123/d for control and flavored diets, respectively. Data for both groups were separately analyzed using multiple regression with 3.5% fat-corrected milk as the dependent variable and BW and DMI as independent variables. More BW was mobilized per unit increase in 3.5% fat-corrected milk in cows fed the control than in cows fed the flavored diet. Cows fed the control diet tended to be in more negative energy balance during early lactation than cows fed the flavored diet. It was concluded that feeding flavor improved energy balance of cows in early lactation and may reduce the risk of health or reproductive problems.     

Effect of dietary phosphorus deprivation on leukocyte function in transition cows

Phosphorus depletion and hypophosphatemia have been described to hamper immune function in different species, an effect barely studied in dairy cows commonly developing hypophosphatemia in early lactation. Dietary P deprivation in mid lactating dairy cows was associated with a decline of the number of granulocytes and impaired granulocyte survival, whereas the phagocytic activity remained unaffected. The objective of the study reported here was to determine the effect of P deprivation on the leukocyte function of periparturient dairy cows. Eighteen multiparous and late pregnant dairy cows were randomly assigned to either a treatment group that was offered a markedly P-deficient diet or a control group receiving the same ration with adequate P content. The study consisted of a 2-wk acclimation period that was followed by a P deprivation period extending from 4 wk before to 4 wk after parturition and a P repletion period of 2 wk thereafter. Blood samples for leukocyte counts and leukocyte function analysis were obtained at the end of the acclimation period, after 2 wk of P deprivation, within the first week of lactation, at the end of the P depletion period and after 2 wk of dietary P supplementation. Blood samples for biochemical analysis were obtained weekly. Immune function was assessed by means of a phagocytosis assay and a lymphocyte stimulation test. Dietary P deprivation resulted in pronounced and sustained hypophosphatemia. Time effects were observed on the counts of different leukocyte fractions, the relative number of phagocytic granulocytes, the degree of phagocytosis, and the lymphocyte proliferation. Differences between P-deprived and control cows were only identified for the degree of phagocytosis that was lower in P-deprived cows compared with control cows. The correlation and regression analyses, however, revealed positive associations of the plasma phosphate concentration and the granulocyte count, the relative number of phagocytic granulocytes, and the degree of phagocytosis at the end of the dietary P deprivation when P depletion was most severe. The results of the study reported here indicate a mild negative effect of pronounced and sustained hypophosphatemia on the granulocyte count and the phagocytic activity of granulocytes in transition dairy cows. The clinical relevance of this effect for health and productivity of dairy cows remains to be determined.     

Relationships between endometritis and metabolic state during the transition period in pasture-grazed dairy cows

The primary objective of this study was to identify relationships between endometritis and metabolic state during the calving transition and early lactation periods. A subset of mixed age and breed dairy cows (n = 78) from a seasonal, pasture-grazed herd of 389 cows was examined. The selected cows were grouped as having endometritis at d 42 postpartum or being unaffected by endometritis. Endometritis was defined as >6% (upper quartile) of uterine nucleated cells being polymorphonuclear cells (H-PMN; n = 38); unaffected by endometritis was defined as ≤1% of nucleated cells being polymorphonuclear (L-PMN; n = 40). Milk yield was determined at each milking, and milk composition (fat and protein) was determined at 2-wk intervals. Blood samples collected on d −14, 0 (d of calving), 4, 7, 14, 28, and 42 were analyzed for indicators of energy status (nonesterified fatty acids, glucose, and urea), liver function (albumin, globulin, glutamate dehydrogenase, and aspartate aminotransferase), inflammation (haptoglobin), and mineral status (Ca and Mg). Samples collected weekly from d 21 to 63 or 70 were analyzed for progesterone content. The postpartum anovulatory interval was defined to end on the first day postpartum that plasma progesterone concentration was ≥1 ng/mL. A greater percentage of H-PMN cows failed to ovulate before d 63 or 70 (34%) compared with L-PMN cows (10%), although the proportions of cows ovulating within either polymorphonuclear group was similar through d 56 postpartum. Plasma concentrations of albumin and the albumin:globulin ratio were consistently lower in H-PMN cows. Plasma Mg was lower, whereas glutamate dehydrogenase and aspartate aminotransferase were higher, in H-PMN cows during early lactation compared with L-PMN cows. Circulating metabolites indicative of energy status (nonesterified fatty acids, glucose, and urea) were not different between polymorphonuclear groups. Among 3- to 5-yr-old cows, daily milk yield for the first 42 d after calving was lower for H-PMN cows than for L-PMN cows. Among cows >5 yr old, protein percentage was lower in H-PMN cows compared with L-PMN cows. In summary, endometritis at 42 d postpartum in the herd studied was associated with an increased likelihood of remaining anovulatory. These cows had lower albumin concentrations throughout the calving transition period, perhaps indicating impaired liver function, with lower plasma Mg and evidence of hepatocellular damage in early lactation. Similar profiles of nonesterified fatty acids and glucose indicated that energy status was not a risk factor for endometritis.     

Coordination of lipid droplet-associated proteins during the transition period of Holstein dairy cows

Dairy cows often experience negative energy balance with the onset of lactation, and severe or prolonged negative energy balance can contribute to declines in overall fitness. Energy stores, in the form of adipose tissue triacylglycerides, are mobilized during times of energy deficit, and recent research has implicated several proteins associated with the lipid droplet as lipolytic regulators. The objective of this study was to determine if these novel proteins associated with lipolytic regulation are altered with the changing metabolic demands of lactation. Weekly blood samples were collected from 26 Holstein cows from 21 d before expected parturition through 28 d postpartum, and again at 150 d postpartum. Serum nonesterified fatty acids, glycerol, and β-hydroxybutyrate were measured. Energy balance was calculated from daily feed intake and milk yield, weekly body weight, and monthly milk component measurements. Adipose tissue biopsies were taken 21 d before expected parturition (−21 d) and at 5, 21, and 150 d postpartum. Semiquantitative Western blotting was used to measure abundance of hormone-sensitive lipase (HSL), phosphorylated HSL, perilipin, phosphorylated perilipin (PPLIN), adipose triglyceride lipase (ATGL), and comparative gene identity-58 (CGI-58). Abundance of ATGL was less at 5 and 21 d in milk (DIM) compared with −21 and 150 DIM, even though cows were in negative energy balance and experiencing increased rates of lipolysis in early lactation. In contrast, phosphorylated HSL and PPLIN increased with increasing lipolysis immediately after parturition. Additionally, PPLIN was negatively correlated with milk yield at 5, 21, and 150 d postpartum, and negatively correlated with feed intake and energy balance at 21 d postpartum. This result is consistent with the hypothesis that phosphorylation of perilipin is responsive to signals for increased triaclyglyceride mobilization. Finally, a consistent negative correlation between abundance of perilipin and CGI-58 proteins was observed throughout the transition period. These results confirm that novel lipolytic proteins in adipose tissue are regulated at the level of protein abundance and phosphorylation during the periparturient period and into mid lactation.     

Indicators of resilience during the transition period in dairy cows: A case study

The transition period is a demanding phase in the life of dairy cows. Metabolic and infectious disorders frequently occur in the first weeks after calving. To identify cows that are less able to cope with the transition period, physiologic or behavioral signals acquired with sensors might be useful. However, it is not yet clear which signals or combination of signals and which signal properties are most informative with respect to disease severity after calving. Sensor data on activity and behavior measurements as well as rumen and ear temperature data from 22 dairy cows were collected during a period starting 2 wk before expected parturition until 6 wk after parturition. During this period, the health status of each cow was clinically scored daily. A total deficit score (TDS) was calculated based on the clinical assessment, summarizing disease length and intensity for each cow. Different sensor data properties recorded during the period before calving as well as the period after calving were tested as a predictor for TDS using univariate analysis of covariance. To select the model with the best combination of signals and signal properties, we quantified the prediction accuracy for TDS in a multivariate model. Prediction accuracy for TDS increased when sensors were combined, using static and dynamic signal properties. Statistically, the most optimal linear combination of predictors consisted of average eating time, variance of daily ear temperature, and regularity of daily behavior patterns in the dry period. Our research indicates that a combination of static and dynamic sensor data properties could be used as indicators of cow resilience.     

Lying behavior and activity during the transition period of clinically healthy grazing dairy cows

Lying behavior and activity may provide useful information for the prediction of an imminent calving and the health of transition dairy cows; however, it is important first to understand what constitutes typical lying behavior and activity because this has not been defined for grazing dairy cows during the transition period. Our objective was to describe changes in lying behavior and activity in grazing dairy cows during the transition period using varying phenotypes typical of commercial dairy herds under grazing systems. Behavior data from IceTag or IceQube (IceRobotics, Edinburgh, Scotland) triaxial accelerometers were collected for 310 cow parities from multiparous, mixed-age (mean ± standard deviation; 4.5 ± 1.65 yr), and mixed-breed [Holstein-Friesian (HF), n = 216; and HF × Jersey, n = 94] grazing dairy cows from 4 parent experiments. The IceTags or IceQubes captured lying and activity data during the transition period (−21 to 34 d relative to calving) to allow the calculation of daily lying time (h/d), daily lying bouts (LB; no./d), mean LB duration (min/bout), and the number of steps taken (steps/d). Lying behavior and activity were analyzed using a repeated measures ANOVA during 3 periods: PRE (−21 to −3 d), POST (3 to 21 d), and the day of calving (d 0). Lying time was lower on d 0 (7.25 h/d) compared with PRE and POST lying times (10.3 and 8.58 h/d, respectively), with more frequent LB on d 0 (12.9 no./d) compared with the PRE and POST daily LB (8.15 vs. 7.74 no./d). Cows took more steps POST (4,424 steps/d) compared with d 0 and PRE (4,105 and 2,289 steps/d, respectively). Regression analysis determined that daily lying time decreased substantially from −3 to 0 d (slope = −1.03 ± 0.07 h/d) and from −2 and −1 d for daily LB (slope = 5.09 ± 0.54 no./d), which may be due to the calving event itself but also reflect restlessness. Daily lying time, daily LB, LB duration, and number of steps taken were substantially altered at the time of the calving event in grazing dairy cows. Cows were more active, spent less time lying, and took more steps postcalving compared with precalving, and it appears that this behavior may largely be due to activity associated with twice daily milking. Mean lying behavior and activity measures were more highly variable across individuals than across groups. Information available via activity monitors may contribute to the improvement of individual management of transition dairy cows, and this research provides a benchmark for typical changes in behavior during the transition period in grazing systems.     

Effect of protein level and methionine supplementation on dairy cows during the transition period

Cows experience a significant negative protein balance during the first 30 d of lactation. Given the functional effects of AA on health, especially in challenging periods such as calving, higher levels of protein and specific AA in the diet may act to improve health and feed intake. The response of dairy cows to 3 protein supplementation strategies during the transition period and through the first 45 d in milk was evaluated. The final data set had 39 Holstein cows blocked based on parity (primiparous vs. multiparous) and expected calving and randomly assigned within each block to one of 3 dietary treatments: low protein (LP), high protein (HP), or high protein plus rumen-protected methionine (HPM). Treatments were offered from d ?18 ± 5 to 45 d relative to parturition. Pre- and postpartum diets were formulated for high metabolizable protein (MP) supply from soybean meal, and HP and HPM provided higher MP balance than LP. Preplanned contrasts were LP versus HP+HPM and HP versus HPM. Significance was declared at P ≤ 0.05 and trends at 0.05 < P ≤ 0.10. Cows fed HP and HPM had greater fry matter intake (DMI) prepartum than LP (+2 kg/d), and there was a trend for greater DMI with HPM than with HP (+1.6 kg/d). Body weight and condition score before and after calving did not differ among treatments. High protein (HP and HPM) tended to increase milk yield during the first 45 d of lactation (+1.75 kg/d), increased milk lactose content and urea-N in milk and plasma, tended to increase blood BHB 14 d postpartum, and tended to reduce milk/DMI compared with LP. Blood concentrations of calcium at calving and of glucose, and nonesterified fatty acids pre- and postpartum did not differ. High protein induced lower concentration of plasma IL-1 at calving and lowered blood lymphocytes 21 d postpartum, suggestive of a reduced inflammatory status compared with LP. The concentrations of IL-10, tumor necrosis factor alpha, and other hemogram variables did not differ among treatments. Addition of rumen-protected methionine to the HP diet did not alter milk yield but increased fat and total solids concentrations. The rumen-protected methionine had no effect on blood metabolites and immunity markers, with the exception of increased pre-partum insulin concentrations. The data indicate that dairy cows around calving respond positively to an increase in the supply of MP and to rumen-protected methionine supplementation of the HP diet by increasing intake and improving immune status.     

Dietary structural to nonfiber carbohydrate concentration during the transition period in grazing dairy cows

A study was conducted to evaluate the potential effects of altering the proportion of dietary structural carbohydrate (SC) relative to nonfiber carbohydrate (NFC) pre- and postpartum on milk production and the circulating concentrations of hormones and metabolites. Dietary treatments were arranged as a 2 × 2 factorial, with 68 multiparous cows assigned to isoenergetic diets [114 MJ of metabolizable energy (ME)/cow per d] precalving; diets were either fresh pasture and pasture silage (PreP) or pasture and pasture silage supplemented with 3 kg dry matter (DM)/cow per d of a corn- and barley-based concentrate for 36 d prepartum (PreC). Final treatments were 13 or 32% DM NFC, respectively. After calving, cows within each prepartum diet were assigned to isoenergetic diets (179 MJ of ME/cow per d) differing in their SC and NFC content. Postcalving diets were either fresh pasture and pasture silage (PostP) or pasture and pasture silage supplemented with 5 kg DM/cow per d of a corn- and barley-based concentrate (PostC) until 35 d in milk. Final treatments were 18 and 38% DM NFC, respectively. Relative to day of calving (d 0), blood samples were collected at least weekly from d −28 to d 35. During the prepartum period, PreC cows had lower plasma urea, albumin, insulin-like growth factor-I, and Ca concentrations, but greater nonesterified fatty acid and Mg concentrations. There were no evident effects of prepartum diet on body weight or body condition score, milk yield, or milk composition. During the postpartum period, PostC cows had lower concentrations of plasma urea, β-hydroxybutyrate, and Ca, but greater concentrations of nonesterified fatty acids, glucose, insulin-like growth factor-I, and Mg. Postpartum metabolic differences in PostC cows were associated with increased milk protein production and reduced milk fat (yield and %). Results do not support a periparturient metabolic benefit to altering the SC to NFC ratio precalving, but imply an altered rumen fermentation, gluconeogenesis, and milk composition when dietary SC to NFC ratio is altered postcalving.     

Supplementation with N-carbamoylglutamate during the transition period improves the function of neutrophils and reduces inflammation and oxidative stress in dairy cows

The aim of this study was to investigate the effects of N-carbamoylglutamate (NCG) supplementation during the transition period on the functions of blood polymorphonuclear neutrophils (PMN), inflammation, and oxidative stress in dairy cows. Thirty multiparous Chinese Holstein dairy cows at wk 4 before parturition were blocked into 2 groups by parity, body weight, and milk yield of previous lactation, and randomly allocated to 2 dietary treatments of basal diet supplemented without (control, n = 15) or with 20 g/d per cow of NCG (NCG, n = 15). The supplementation was carried out from d ?21 to 21 relative to calving. Health incidents (mastitis, retained placenta, and lameness) were recorded, and blood samples were collected at d ?21, ?7, 0 (the calving date), 7, and 21 relative to parturition and analyzed for variables related to inflammation and oxidative stress. In addition, whole blood was collected at d 7 to isolate PMN and used for analysis of the expression of functional genes and from d ?21 to 21 for determination of weekly hematological parameters. The number of lymphocytes was greater at d 7 in the blood of NCG cows. The plasma level of malondialdehyde was lower in the NCG group, and blood reactive oxygen species were lower at d 7, whereas total antioxidant capacity tended to be greater in the NCG group and glutathione peroxidase tended to be higher at d 21 in cows fed NCG, suggesting that NCG supplementation improved antioxidation in cows. In addition, the concentration of serum amyloid A was lower in NCG-fed animals during the postpartum stage. Blood concentrations of IL6 and tumor necrosis factor-α were lower and tended to be lower in NCG-fed animals at d 7, respectively. Meanwhile, the concentrations of IL6 tended to be lower in NCG-fed animals at d 21. Furthermore, the expression of S100A9 and MMP9 in the PMN was lower and tended to be lower, respectively, whereas the expression of ITGB2, XBP1 tended to be higher and expression of CLEC6A was higher in NCG-fed cows. Overall, our results indicated that supplementation with NCG during the transition period showed the beneficial effects on animal health, by improving PMN functions and alleviating inflammation status and oxidative stress in dairy cows.     

Feeding rumen-protected lysine altered immune and metabolic biomarkers in dairy cows during the transition period

This experiment was conducted to determine the effects of feeding rumen-protected lysine (RPL; AjiPro-L Generation 3, Ajinomoto Health and Nutrition North America Inc.) from −26 ± 4.6 d prepartum (0.54% RPL of dietary dry matter intake) to 28 d postpartum (0.39% RPL of dietary dry matter intake) on immunometabolic status and liver composition in dairy cows. Seventy-five multiparous Holstein cows, blocked by parity, previous 305-d mature-equivalent milk production, expected calving date, and body condition score during the far-off dry period were assigned to 1 of 4 dietary treatments in a randomized, complete block design with a 2 × 2 factorial arrangement of treatments. Treatments prepartum consisted of total mixed ration top dressed with RPL (PRE-L) or without RPL (PRE-C), and postpartum treatments consisted of total mixed ration top dressed PRE-L prepartum and postpartum, PRE-L prepartum and PRE-C postpartum, PRE-C prepartum and PRE-L postpartum, and PRE-C prepartum and postpartum in 300 g of molasses. Blood samples were taken on −7 ± 0.5, 0 ± 0.5, 7 ± 0.9, 14 ± 0.9, and 28 ± 0.5 d relative to calving. Whole blood samples were taken on −14 ± 0.5, −7 ± 0.5, 7 ± 0.9, and 14 ± 0.9 d relative to calving for oxidative burst and phagocytic capacity of monocytes and neutrophils. Liver samples were collected via a biopsy on −12 ± 4.95 and 13 ± 2.62 d relative to calving and analyzed for liver composition (triacylglyceride and carnitine concentrations), mRNA expression of hepatic genes, and protein abundance. Protein abundance was calculated by normalizing intensity bands for a specific protein with glyceraldehyde-3-phosphate dehydrogenase. Concentrations of haptoglobin and glutathione peroxidase activity in plasma were lower at d 0 for cows in PRE-L (102 µg/mL and 339 nmol/min per mL, respectively) compared with cows in PRE-C (165 µg/mL and 405 nmol/min per mL, respectively). Oxidative burst capacity in monocytes tended to be greater on d 7 postpartum for cows in PRE-L (65.6%) than cows in PRE-C (57.5%). Additionally, feeding RPL altered the mRNA expression in liver tissue prepartum [decreased INSR (insulin receptor), CPT1A (carnitine palmitoyltransferase 1A), and IL1B (interleukin 1 β)] and postpartum [increased IL8 (interleukin 8), EHMT2 (euchromatic histone lysine methyltransferase 2), TSPO (translocator protein), and SLC3A2 (solute carrier family 3 member 2); and decreased SLC7A1 (solute carrier family 7 member 1), SOD1 (superoxide dismutase 1), and SAA3 (serum amyloid A 3)] compared with cows not consuming RPL]. Additionally, cows in the PRE-C prepartum and PRE-L postpartum treatment tended to have greater protein abundance of mTOR postpartum compared with the PRE-C prepartum and postpartum treatment. Protein abundance of SLC7A7 (solute carrier family 7 member 7) pre- and postpartum tended to be greater and BBOX1 (gamma-butyrobetaine dioxygenase 1) tended to be less when RPL was consumed prepartum. In conclusion, cows that consumed RPL during the transition period had molecular changes related to liver composition, enhanced liver function indicated by greater total protein and albumin concentrations in plasma, and improved immune status indicated by decreased haptoglobin, glutathione peroxidase activity, and immune related mRNA expression.     

Effects of supplementing a Saccharomyces cerevisiae fermentation product during the periparturient period on performance of dairy cows fed fresh diets differing in starch content

The objective of this study was to evaluate the effects of supplementing a Saccharomyces cerevisiae fermentation product (SCFP; NutriTek, Diamond V, Cedar Rapids, IA) during the periparturient period (d ?28 ± 3 to 44 ± 3 relative to calving) on dry matter intake (DMI), milk production, apparent total-tract nutrient digestibility, and postpartum ovarian activity of dairy cows fed fresh diets varying in starch content. From d 28 ± 3 before the expected calving date until d 44 ± 3 after calving, 117 Holstein cows were fed diets with SCFP (SCFP; n = 59) or without (control, CON; n = 58). A common, basal, controlled-energy close-up diet (net energy for lactation: 1.43 Mcal/kg; 13.8% starch) was fed before calving. Cows within each treatment (CON or SCFP) were fed either a low- (LS; 22.1% starch) or high-starch (HS; 28.3% starch) diet from d 1 to 23 ± 3 after calving (fresh period), resulting in 4 treatment groups: LS-CON (n = 30), LS-SCFP (n = 29), HS-CON (n = 28), and HS-SCFP (n = 30). All cows were fed the HS diets from d 24 ± 3 to 44 ± 3 after calving (post-fresh period). Cows were assigned to treatment balanced for parity, body condition score, body weight, and expected calving date. Milk yield was higher for cows fed the LS diets compared with those fed the HS diets during the fresh period (34.1 vs. 32.1 kg/d), whereas DMI and 3.5% fat-corrected milk yield (FCM) were not affected by dietary starch content, and LS cows tended to lose more body condition than HS cows (?0.42 vs. ?0.35 per 21 d) during the fresh period. Overall DMI during the close-up and fresh periods did not differ between SCFP and CON cows. However, SCFP supplementation transiently increased DMI on d 1 (13.0 vs. 11.9 kg/d) and 5 (15.5 vs. 14.1 kg/d) after calving compared with CON. During the post-fresh period, SCFP cows tended to eat less than CON cows (19.8 vs. 20.6 kg/d) but had similar 3.5% FCM (44.9 vs. 43.6 kg/d), resulting in greater feed efficiency for SCFP cows (FCM/DMI; 2.27 vs. 2.13). Neither starch content of fresh diets nor SCFP supplementation affected the interval from calving to first ovulation or the incidence of double ovulation. These findings suggest that feeding low-starch diets during the fresh period can increase milk production of dairy cows during the fresh period, and that supplementation of SCFP may increase feed intake around calving and feed efficiency in the post-fresh period.     

Milk production of dairy cows fed wet corn gluten feed during the dry period and lactation

An experiment was conducted with 36 primiparous and 40 multiparous Holstein cows to examine the effects of feeding wet corn gluten feed (WCGF) on 305-d milk production, dry matter (DM) intake, body condition score (BCS), and health. The experimental treatments included: 1) control—WCGF not fed (n = 27); 2) WCGF-L—cows received diets containing WCGF (38% DM basis) during lactation (n = 23); and 3) WCGF-DL—cows received diets containing WCGF (38% DM basis) during the dry period and lactation (n = 26). During the dry period, cows consuming WCGF were observed to have a significant gain in BCS (0.07 ± 0.06) compared with a loss in BCS in cows fed the control diet (control = −0.11 ± 0.06 and WCGF-L = −0.04 ± 0.06). During lactation, there were no differences by treatment on BCS. Cows consuming WCGF during lactation consumed more feed compared with the control: 25.4, 23.8, and 21.2 ± 0.76 kg/d for WCGF-L, WCGF-DL, and the control, respectively. Milk production was higher for cows consuming WCGF: 35.0, 34.7, and 31.1 ± 2.1 kg/d for WCGF-L, WCGF-DL, and the control, respectively. No differences were found in either DM intake or actual milk yield between the WCGF-L and WCGF-DL treatments, indicating that prepartum diets did not influence lactational performance. The WCGF diets resulted in significant reductions in the concentration of milk fat (3.94, 3.74, and 4.15 ± 0.08% for WCGF-L, WCGF-DL, and the control, respectively), but because total milk yield was increased, there were no differences in total milk fat yield. In addition, 3.5% of fat-corrected milk tended to be affected by diet: 38.9, 36.3, and 34.7 ± 1.93 kg/d for WCGF-L, WCGF-DL, and the control, respectively. The increasing effect of DM intake and milk yield in cows consuming WCGF resulted in a similar efficiency of 3.5% fat-corrected milk production for all treatments, averaging 1.5 ± 0.09. Total protein yields were significantly higher for cows consuming WCGF diets during lactation: 1.15, 1.10, 1.00 ± 0.06 kg/d for WCGF-L, WCGF-DL, and the control, respectively. These results indicate that diets may be formulated to contain as much as 37.5% WCGF (DM basis).     

Changes in feeding, drinking, and standing behavior of dairy cows during the transition period

The aims of this study were to determine how measures of feeding, drinking, and standing behavior change over the period around calving, to derive objective meal and drinking bout criteria for transition cows, and to describe the within-cow consistency of these behavioral measures. Fifteen transition dairy cows were monitored from 10 d before until 10 d after calving. Meal criteria were calculated for each cow in both the pre- and postcalving periods and varied among cows from 5.3 to 105.2 min. There was a tendency for the average number of meals per day to be higher after calving than before calving, but time spent eating declined from 87 to 62 min/d from the precalving to the postcalving period. Time spent drinking before calving averaged 5.5 min/d and increased gradually after calving to an average of 6.8 min/d. Total daily standing times remained reasonably similar over the transition period but were highest at around calving (14.4 h) and lowest during the precalving period (12.3 h). On the day of calving, there was a dramatic increase in the number of standing bouts (21.8 bouts) compared with the pre- and postcalving averages of 11.7 and 13.1 bouts, respectively. In summary, changes in feeding behavior may help account for the well-documented changes in feed intake during transition. Documented changes in standing behavior suggest that cow comfort may be particularly important during the time around calving.     

Metabolism and lactation performance in dairy cows fed a diet containing rumen-protected fat during the last twelve weeks of gestation

Effects of dietary fat supplementation prepartum on liver lipids and metabolism in dairy cows are contradictory. Thus, we examined in 18 German Holstein cows (half-sib; first lactation 305-d milk yield >9,000 kg) whether dietary fat:carbohydrate ratio during the last trimester of gestation affects lipid metabolism and milk yield. The diets were formulated to be isoenergetic and isonitrogenous but differed in rumen-protected fat (FD; 28 and 46.5 g/kg of dry matter during far-off and close-up dry period; mainly C16:0 and C18:1) and starch concentration [carbohydrate diet (CD); 2.3 times as much starch as FD]. Diets were given ad libitum starting 12 wk before expected parturition. After parturition all cows were fed a single lactation diet ad libitum for 14 wk. With the FD treatment, dry matter intake was depressed prepartum, milk yield during first 4 wk of lactation was lower (36.9 vs. 41.0 kg/d), and postpartum energy balance during this period was more negative. During the first 4 wk, cows in the FD group had lower lactose percentage and yield but higher milk fat, whereas milk protein and fat yield as well as energy-corrected milk did not differ. Between wk 5 and 14, milk fat and milk protein percentage was lower in CD than in FD. Milk fat C14:0 was lower and C16:1 was higher in the FD group. For FD cows, plasma triacylglycerol, nonesterified fatty acids, and cholesterol concentrations were higher prepartum, whereas plasma β-hydroxybutyrate and glucose concentrations were lower. During the first 10 d after parturition, plasma triacylglycerol concentration was higher in FD, and prepartum plasma glucose and cholesterol differences persisted during the first 14 wk of lactation. Irrespective of prepartum nutrient composition, concentrations of plasma leptin and subcutaneous fat leptin mRNA decreased between −10 d to +10 d relative to parturition, and liver lipids and glycogen reached maximum and minimal values, respectively, 10 d after parturition. Acetyl-coenzyme A carboxylase α mRNA abundance in subcutaneous fat decreased between −10 d to +1 d relative to parturition by 97%, whereas it was generally much lower in the liver and remained at a low level until wk 14 of lactation. In conclusion, feeding a diet containing rumen-protected fat during late lactation and dry period until calving negatively affected dry matter intake, energy balance, and milk yield during subsequent lactation, did not change acetyl-coenzyme A carboxylase α mRNA abundance in subcutaneous fat, and was not beneficial for liver lipid accumulation.     

Lower pregnancy losses in lactating dairy cows fed a diet enriched in alpha-linolenic acid

The objectives were to determine if a diet enriched in α-linolenic acid (ALA) would influence ovarian function, early embryo survival, conception rates, and pregnancy losses in lactating dairy cows. Beginning 28 d before breeding, Holstein cows (55 ± 22 d postpartum; mean ± SD) were assigned to diets supplemented with either rolled flaxseed (FLAX; 56.7% ALA, n = 62) or rolled sunflower seed (SUNF; 0.1% ALA, n = 59) to provide approximately 750 g of oil/d. Diets continued for 32 d after timed artificial insemination (TAI, d 0) following a Presynch/Ovsynch protocol. Barley silage- and barley grain-based TMR were formulated to meet or exceed National Research Council requirements. Metabolizable protein and net energy for lactation concentrations were similar in the 2 diets. Based upon a mean dry matter intake of 22 kg/d, cows fed FLAX or SUNF consumed > 410 g or < 1 g of ALA, respectively. Pregnancy was confirmed by ultrasound 32 d after TAI. Nonpregnant cows were placed on a second Ovsynch regimen and reinseminated 42 d after first TAI, and received oilseeds for 32 d after second TAI. Relative to prediet levels, FLAX increased the ALA content of milk by 187%. Ovarian ultrasonography was performed in 8 cows per diet; the mean diameter of ovulatory follicles was larger in cows fed FLAX compared with SUNF (16.9 ± 0.9 vs. 14.1 ± 0.9 mm), but follicle number, corpus luteum size, and plasma progesterone concentrations remained unaffected. Presumptive conception (progesterone < 1 ng/mL on d 0 and > 1 ng/mL on d 21) rates to first TAI were greater in FLAX than in SUNF (72.6 vs. 47.5%). Pregnancy losses were lower in cows fed FLAX (9.8%) compared with those fed SUNF (27.3%). Including flaxseed in the ration of dairy cows increased the size of the ovulatory follicle and reduced pregnancy losses.