Effect of rumen-protected branched-chain amino acid supplementation on production- and energy-related metabolites during the first 35 days in milk in Holstein dairy cows

Essential AA are critical for multiple physiological processes. Branched-chain AA (BCAA) supplementation has beneficial effects on body weight, lipogenesis, and insulin resistance in several species. The BCAA are used for milk and body protein synthesis as well as being oxidized by the tricarboxylic acid cycle to produce ATP during catabolic states. The objective was to evaluate the effect of rumen-protected BCAA (375 g of 27% l-Leu, 85 g of 48% l-Ile, and 91 g of 67% l-Val) with or without propylene glycol (PG) oral administration on milk production, dry matter intake, nonesterified fatty acids, β-hydroxybutyrate, and plasma urea nitrogen during the first 35 d in milk (DIM) in dairy cattle. Multiparous Holstein cows were enrolled in blocks of three 28 d before expected calving and assigned randomly to either the control or 1 of 2 treatments. The control (n = 26) received 200 g/d of dry molasses, the BCAA treatment (n = 23) received BCAA mixed with 200 g/d of dry molasses from calving until 35 DIM, and the BCAA plus PG (BCAAPG) treatment (n = 25) received BCAA mixed with 200 g/d of dry molasses from calving until 35 DIM plus 300 mL of PG once daily from calving until 7 DIM. Postpartum, dry matter intake least squares means (LSM; 95% confidence interval) were 20.7 (19.9, 21.7), 21.3 (20.4, 22.3), and 21.9 (20.9, 22.8) kg for control, BCAA, and BCAAPG, respectively. Milk yield (1–35 DIM) LSM were 41.7 (39.4, 44.0), 42.7 (40.3, 45.0), and 43.7 (41.4, 46.0) kg for control, BCAA, and BCAAPG, respectively. Energy-corrected milk LSM were 50.3 (46.8, 53.7), 52.4 (48.9, 55.8), and 52.9 (49.5, 56.4) kg for control, BCAA, and BCAAPG, respectively. Milk urea nitrogen LSM in milk for control, BCAA, and BCAAPG were 8.60 (8.02, 9.22), 9.70 (9.01, 10.45), and 9.75 (9.08, 10.47) mg/dL. Plasma urea nitrogen concentrations LSM for control, BCAA, and BCAAPG were 8.3 (7.7, 8.9), 10.1 (9.4, 10.9), and 9.6 (9.4, 10.3) mg/dL, respectively. The numbers of plasma samples classified as hyperketonemia were 77, 44, and 57 in control, BCAA, and BCAAPG, respectively. The BCAA supplementation increased plasma urea nitrogen and milk urea nitrogen, free valine concentration in plasma, and decreased hyperketonemia events during the postpartum period.     

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

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

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

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

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.     

Prepartal standing behavior as a parameter for early detection of postpartal subclinical ketosis associated with inflammation and liver function biomarkers in peripartal dairy cows

A degree of negative energy balance is commonly experienced by cows during early lactation. This physiological state, if pronounced or prolonged, leads to partial oxidation of nonesterified fatty acids as an energy source and, consequently, increasing blood β-hydroxybutyrate (BHB) concentrations and potentially development of ketosis in postpartal dairy cows. Twenty-four multiparous Holstein cows received a common prepartal and postpartal diet. Cows were fitted with an accelerometer mounted laterally on the distal left hind leg using vet wrap from ?30 to 15 d relative to parturition. A retrospective analysis was performed using the postpartal BHB data at 8 time points from 0 to 15 d in milk measured with the Precision Xtra (Abbott Diabetes Care, Alameda, CA). Cows with an average blood BHB <1.4 mmol/L were designated nonketotic (NONKET; n = 12), and those with ≥1.4 mmol/L were designated ketotic (KET; n = 12). A total of 8 samples per cow were used for this analysis. Subsequent analyses of behavioral patterns and blood biomarkers were performed using this group effect. On average, blood BHB reached subclinical levels (1.4 ± 0.3 mmol/L; mean ± standard error of the mean) at 3 d postpartum for all cows in this study. Behavioral patterns were obtained from accelerometer data, and correlation analysis was performed between these behaviors such as standing and lying time from ?30 to 3 d relative to parturition and blood BHB concentration at 3 d postpartum. The strongest correlation was obtained between standing time at 3 d before calving and blood BHB at 3 d postpartum. Dry matter intake was greater (ca. 3 kg/d) in NONKET cows than in KET cows. An interaction of group × time for milk yield resulted in an overall increase of 5.7 kg/d in NONKET cows in comparison with KET. The blood concentrations of biomarkers for liver function (γ-glutamyltransferase and glutamic-oxaloacetic transaminase), inflammation (IL-6), and metabolism (nonesterified fatty acids) were increased at various time points in KET cows in comparison with NONKET during the transition period. Overall, lower bilirubin in NONKET cows than in KET further confirmed an impaired liver function in the latter group of cows. Our findings revealed the potential for establishing correlations between prepartal behavioral patterns derived from accelerometer data and postpartal subclinical ketosis, and further confirming the latter by physiological alterations in biomarkers related to inflammation and liver function. Our data also indicate that cows with a predisposition to postpartal subclinical or clinical ketosis will remain standing for fewer hours during the days leading to parturition, which decreased DMI, and this condition was further reflected in lower milk yield.     

Effect of transition cow health and estrous expression detected by an automated activity monitoring system within 60 days in milk on reproductive performance of lactating Holstein cows

The objective of this observational study was to evaluate the association of transition cow health and estrous expression, detected by an automated activity monitoring system (Smarttag Neck, Nedap Livestock Management), with reproductive performance in lactating Holstein cows. A total of 3,750 lactating Holstein cows (1,563 primiparous cows and 2,187 multiparous cows) from a commercial dairy farm in Slovakia calving from January 2020 until July 2021 were enrolled on an ongoing basis. Activity data were recorded from d 7 until d 60 postpartum. Within this observational period, cows were classified into 3 categories: (1) no estrus event (Estrus0), (2) 1 estrus event (Estrus1), or (3) 2 or more estrus events (Estrus2+). Transition cow health was assessed by farm personnel within the first 30 d in milk (DIM) using standard operating procedures. Generalized linear mixed models were used to analyze continuous and categorical data. Cox proportional hazard models were used for time to event data. The overall prevalence of anestrus was 20.8%. Multiparous cows had a greater risk for anestrus compared with primiparous cows [odds ratio (OR) = 1.4]. Cows with stillbirth (OR = 1.76), retained placenta (OR = 2.19), puerperal metritis (OR = 1.48), or subclinical ketosis (OR = 1.51) had a greater risk for anestrus. In addition, cows calving in summer (OR = 0.82), autumn (OR = 0.38), or winter (OR = 0.56) had a higher incidence of anestrus than cows calving in spring. Estrous expression from d 7 until d 60 postpartum was associated with estrous duration (DU) and estrous intensity at first artificial insemination (AI). Cows in Estrus0 had the shortest DU at first postpartum AI (9.4 ± 0.18 h) compared with cows in Estrus1 (10.5 ± 0.13 h) and Estrus2+ (11.4 ± 0.12 h). Cows in Estrus2+ had a longer DU at first postpartum AI compared with cows in Estrus1. For Estrus0, Estrus1, and Estrus2+ cows, pregnancy per AI at first service was 42.5%, 50.9%, and 55.4%, respectively. Estrous expression from d 7 until d 60 postpartum was associated with time to first AI and time to pregnancy. Compared with Estrus0 cows, Estrus1 [hazard ratio (HR) = 1.43] and Estrus2+ cows (HR = 1.62) had an increased hazard of being inseminated within 100 DIM. Compared with Estrus2+, Estrus1 cows had a reduced hazard of being inseminated within 100 DIM (HR = 0.89). Compared with Estrus0 cows, Estrus1 (HR = 1.24) and Estrus2+ cows (HR = 1.46) had an increased hazard of becoming pregnant within 200 DIM. Median DIM to pregnancy were 121, 96, and 92 for Estrus0, Estrus1, and Estrus2+ cows, respectively. In conclusion, cows with transition cow disorders (i.e., stillbirth, retained placenta, puerperal metritis, or subclinical ketosis) had a greater chance for anestrus compared with healthy cows. Cows in Estrus0 had reduced estrous expression at first AI and inferior reproductive performance compared with cows that displayed estrous activity from d 7 until d 60.     

Transition cow nutrition and management strategies of dairy herds in the northeastern United States: Part I—Herd description and performance characteristics

Our objective was to describe management and herd characteristics of the transition period on freestall dairy herds in the northeastern United States using an on-farm survey and prospective cohort design. Enrolled herds (n = 72) had a median of 900 milking cows (range: 345–2,900) and a rolling herd average of 12,674 kg (standard deviation ± 1,220 kg), and 87.2% (n = 82/94) of fresh pens were milked at least 3×/d. The prevalence of herds with ≥15% of sampled cows with elevated concentrations of nonesterified fatty acids prepartum (≥0.27 mmol/L, 2–14 d before parturition) and postpartum [primiparous: ≥0.60 mmol/L, multiparous: ≥0.70 mmol/L, 3–14 d in milk (DIM)], β-hydroxybutyrate postpartum (≥1.2 mmol/L, 3–14 DIM), and haptoglobin postpartum (≥1 g/L, 0–12 DIM) was 51%, 51%, 51%, and 57%, respectively. In most herds, cows were moved to a calving pen when showing signs of labor (73.6%; n = 53/72) instead of 0 to 3 d before expected calving (26.4%, n = 19/72). Cows remained in the calving or maternity pen for a median (range) time of 2 (0–24) h after parturition before moving to the next pen. Primiparous cows remained in the first pen moved to after parturition for a longer period than multiparous cows [median (range) days: 12 (1.5–25) vs. 6 (1.5–22)]. Approximately 20% of herds had routine vaccinations administered in the maternity or calving pen, first pen after parturition, or both. Almost all herds (n = 69/72) performed fresh cow health checks; however, only 53% (n = 38/72) locked up all fresh cows daily. More herds housed primiparous and multiparous cows in separate pens during the far-off dry (65.3%; n = 47/72) and high-lactation (81.9%; n = 59/72) periods compared with the close-up dry (31.9%; n = 23/72) and fresh periods (27.8%; n = 20/72). At least half of the pens observed during the far-off dry, close-up dry, and fresh periods had a stocking density <100%. Approximately one-third of pens observed during the far-off dry period had feed pushed up ≤4×/d compared with approximately 15 to 20% of pens observed during the close-up dry, fresh, and high-lactation periods. More than half of the total mixed ration samples acquired from the far-off and close-up dry period visits had greater than the recommended proportion of particles in the 19-mm screen of the Penn State Particle Separator. The results of this observational study illustrated the range of management practices used in freestall herds in this region and lay the groundwork for future hypothesis-driven studies using this sampled population.     

Genetic analyses of blood β-hydroxybutyrate predicted from milk infrared spectra and its association with longevity and female reproductive traits in Holstein cattle

Ketosis is one of the most prevalent and complex metabolic disorders in high-producing dairy cows and usually detected through analyses of β-hydroxybutyrate (BHB) concentration in blood. Our main objectives were to evaluate genetic parameters for blood BHB predicted based on Fourier-transform mid-infrared spectra from 5 to 305 d in milk, and estimate the genetic relationships of blood BHB with 7 reproduction traits and 6 longevity traits in Holstein cattle. Predicted blood BHB records of 11,609 Holstein cows (after quality control) were collected from 2016 to 2019 and used to derive 4 traits based on parity number, including predicted blood BHB in all parities (BHBp), parity 1 (BHB1), parity 2 (BHB2), and parity 3+ (BHB3). Single- and multitrait repeatability models were used for estimating genetic parameters for the 4 BHB traits. Random regression test-day models implemented via Bayesian inference were used to evaluate the daily genetic feature of BHB variability. In addition, genetic correlations were calculated for the 4 BHB traits with reproduction and longevity traits. The heritability estimates of BHBp, BHB1, BHB2, and BHB3 ranged from 0.100 ± 0.026 (± standard error) to 0.131 ± 0.023. The BHB in parities 1 to 3+ were highly genetically correlated and ranged from 0.788 (BHB1 and BHB2) to 0.911 (BHB1 and BHB3). The daily heritability of BHBp ranged from 0.069 to 0.195, higher for the early and lower for the later lactation periods. A similar trend was observed for BHB1, BHB2, and BHB3. There are low direct genetic correlations between BHBp and selected reproductive performance and longevity traits, which ranged from ?0.168 ± 0.019 (BHBp and production life) to 0.157 ± 0.019 (BHBp and age at first calving) for the early lactation stage (5 to 65 d). These direct genetic correlations indicate that cows with higher BHBp (greater likelihood of having ketosis) in blood usually have shorter production life (?0.168 ± 0.019). Cows with higher fertility and postpartum recovery, such as younger age at first calving (0.157 ± 0.019) and shorter interval from calving to first insemination in heifer (0.111 ± 0.006), usually have lower BHB concentration in the blood. Furthermore, the direct genetic correlations change across parity and lactation stage. In general, our results suggest that selection for lower predicted BHB in early lactation could be an efficient strategy for reducing the incidence of ketosis as well as indirectly improving reproductive and longevity performance in Holstein cattle.     

Effect of rumen-protected niacin on lipid metabolism, oxidative stress, and performance of transition dairy cows

The objective of this study was to evaluate the effect of a rumen-protected niacin product (RPN; 65% nicotinic acid; NiaShure, Balchem Corp., New Hampton, NY) on lipid metabolism, oxidative stress, and performance of transition dairy cows. Thirty nonlactating multiparous Holstein cows in late gestation were paired according to expected calving date and randomly assigned to 12 g/cow per day of RPN product or to an unsupplemented control (CON) diet. Treatment diets were fed from 21 d before expected calving through 21 d after parturition. Blood samples were taken on d -21, -14, -7, 1, 7, 14, and 21 relative to calving for plasma nonesterified fatty acid (NEFA), β-hydroxybutyrate (BHBA), glucose, and superoxide dismutase (SOD) analyses. Liver samples were taken by biopsy on d 1 and 21 relative to calving for triglyceride (TG) analysis. Data were analyzed for a randomized complete block design with repeated measures. Pre- and postpartum dry matter intake, milk yield, and protein were unaffected by treatment. Milk fat percentage (5.08 vs. 4.44%) and somatic cell score (3.93 vs. 2.48) were reduced for RPN. Treatment × time interactions were observed for energy-corrected milk (ECM) and fat-corrected milk (FCM) yields; RPN reduced ECM and FCM yields by 8.5 and 8.9 kg/cow per day, respectively, in the first week of lactation. Although body weight and condition score decreased during the experimental period, no differences due to treatment were observed. However, calculated postpartum energy balance tended to be improved for RPN because of the reduction in ECM yield. Time and treatment × time effects were observed for plasma NEFA. On d 1 postpartum, NEFA reached 1,138±80 μEq/L for CON compared with 698±80 μEq/L for RPN. Cows supplemented with RPN tended to have lower plasma NEFA concentrations than CON cows on d 7 and 14 postpartum. Plasma BHBA, glucose, and SOD and liver TG concentrations were unaffected by treatment. In conclusion, supplementation with 12 g/cow per day of the RPN product provided a bioavailable source of niacin that modified lipid metabolism but did not affect milk yield over the first 3 wk of lactation or oxidative stress of transition dairy cows.     

Elevation of blood β-hydroxybutyrate concentration affects glucose metabolism in dairy cows before and after parturition

Recent studies in mid- and late-lactation dairy cows showed that β-hydroxybutyrate (BHB) infusion had a considerable effect on glucose metabolism and immune response during intramammary lipopolysaccharide challenge. The objective of the present study was to infuse BHB during the dry period and after parturition to investigate the effects of elevated plasma BHB concentrations on metabolism and endocrine changes in transition dairy cows. The hypothesis tested was that regulation of glucose metabolism would change at different physiological stages and an additional elevation of BHB concentration would alter glucose concentration. Multiparous Holstein cows in wk ?2 (antepartum, a.p.; n = 6) and wk +2 (postpartum, p.p.; n = 8) relative to calving were infused (4 h from 0800 to 1200 h) with a BHB solution to increase plasma BHB concentration to 1.5 to 2.0 mmol/L (HyperB). The same period the next day without any infusion was considered the control period (CON). Blood samples were taken 1 h before the start of infusion as reference samples and every 30 min during the following 6 h (4 h of infusion and 2 h after infusion) in the HyperB and CON periods, and analyzed for glucose, BHB, insulin, and glucagon concentrations. During the steady state period (the latter 2 h of the 4-h infusion), plasma BHB concentration reached 1.87 ± 0.05 mmol/L (a.p.) and 1.93 ± 0.05 mmol/L (p.p.) in HyperB compared with 0.55 ± 0.06 mmol/L (a.p.) and 0.64 ± 0.04 mmol/L (p.p.) in CON, respectively. The 4-h average BHB infusion rate was 12.4 ± 1.0 and 13.3 ± 0.9 μmol/kg of BW per minute in wk ?2 and +2, respectively. Infusion of BHB caused a decrease of plasma glucose concentrations relative to preinfusion levels both before and after parturition, although basal glucose concentrations were different before and after calving. Infusion of BHB increased plasma insulin concentrations a.p. but not p.p., despite a higher basal insulin concentration before than after parturition. These findings show that effects of hyperketonemia on plasma glucose concentrations are similar before and after calving but that endocrine adaptation to hyperketonemia differs before and after parturition. We assume that BHB is a metabolic key regulator in early lactating dairy cows and may affect glucose concentration by further pathways such as gluconeogenesis and altered lipolysis.     

Short communication: Associations between blood glucose concentration,onset of hyperketonemia,and milk production in early lactation dairy cows

The objectives of this study were to describe the associations between hypoglycemia and the onset of hyperketonemia (HYK) within the first 6 wk of lactation, to evaluate the effects of body condition score at calving on glucose concentration, and to study the effects of hypoglycemia on milk production. A total of 621 dairy cows from 6 commercial dairy farms in Germany were enrolled between 1 and 4 d in milk (DIM). Cows were tested twice weekly using an electronic handheld meter for glucose and β-hydroxybutyrate (BHB), respectively, for a period of 42 d. Hypoglycemia was defined as glucose concentration ≤2.2 mmol/L. Hyperketonemia was defined as a BHB concentration ≥1.2 mmol/L. The onset of HYK was described as early onset (first HYK event within the first 2 wk postpartum) and late onset (first HYK event in wk 3 to 6 postpartum). The effect of ketosis status on blood glucose within 42 DIM was evaluated using a generalized linear mixed model. No effect was observed of HYK on glucose concentration in primiparous cows. Multiparous cows with early-onset HYK had a lower glucose concentration (?0.21 mmol/L) compared with nonketotic cows. Overall, primiparous cows had a lower prevalence and incidence of hypoglycemia than multiparous cows. Hypoglycemia in multiparous cows was associated with higher first test-day milk production and 100 DIM milk production. In conclusion, hypoglycemia mainly occurred in multiparous cows with early-onset HYK, whereas primiparous cows were at a lower risk for hypoglycemia.     

Hepatic lipid metabolism in transition dairy cows fed flaxseed

Thirty-three Holstein cows averaging 687 kg of body weight were allotted 6 wk before the expected date of parturition to 11 groups of 3 cows blocked within parity for similar calving dates to determine the effects of feeding different sources of fatty acids on blood parameters related to fatty liver and profile of fatty acids in plasma and liver. Cows were fed lipid supplements from 6 wk before the expected date of parturition until d 28 of lactation. Cows within each block were assigned to 1 of 3 isonitrogenous and isoenergetic dietary supplements: control with no added lipids (CO); unsaturated lipids supplied as whole flaxseed (FL; 3.3 and 11.0% of the dry matter in prepartum and postpartum diets, respectively); and saturated lipids supplied as Energy Booster (EB; 1.7 and 3.5% of the DM in prepartum and postpartum diets, respectively). Diets EB and FL had similar ether extract concentrations. Multiparous cows fed EB had lower dry matter intake and milk production, higher concentrations of nonesterified fatty acids and β-hydroxybutyrate in plasma and triglycerides (TG) and total lipids in liver, and lower concentrations of plasma glucose and liver glycogen than those fed FL and CO. Production of 4% fat-corrected milk was similar among treatments. Multiparous cows fed FL had the highest liver concentrations of glycogen on wk 2 and 4 after calving and lowest concentrations of TG on wk 4 after calving. Liver C16:0 relative percentages in multiparous cows increased after calving whereas those of C18:0 decreased. Relative percentages of liver C16:0 were higher in wk 2 and 4 postpartum for multiparous cows fed EB compared with those fed CO and FL; those of C18:0 were lower in wk 4 postpartum for cows fed EB compared with those fed CO and FL. Liver C18:1 relative percentages of multiparous cows increased after calving and were higher in wk 4 for cows fed EB compared with those fed CO and FL. The inverse was observed for liver C18:2 relative percentages. In general, diets had more significant effects on plasma concentrations of nonesterified fatty acids, β-hydroxybutyrate, and glucose and liver profiles of fatty acids, TG, total lipids, and glycogen of multiparous than primiparous cows. These data suggest that feeding a source of saturated fatty acids increased the risk of fatty liver in the transition cow compared with feeding no lipids or whole flaxseed. Feeding flaxseed compared with no lipids or a source of saturated fatty acids from 6 wk before calving could be a useful strategy to increase liver concentrations of glycogen and decrease liver concentrations of TG after calving, which may prevent the development of fatty liver in the transition dairy cow.     

Direct-fed microbial supplementation and health and performance of pre- and postpartum dairy cattle: a field trial

A double-blind field trial was conducted on a commercial dairy to study the effects of feeding a direct-fed microbial (DFM) product consisting of 2 strains of Enterococcus faecium plus Saccharomyces cerevisiae yeast on prepartum and postpartum performance of Holstein cows. Treatments consisted of the normal pre- and post-fresh TMR supplemented with the DFM (2 g/cow per d) or a placebo. Treatments started approximately 10 d prepartum and continued until about 23 d in milk (DIM). A total of 366 Holstein cows were enrolled in 1 of 2 placebo groups or 2 DFM-supplemented groups. Groups were enrolled consecutively, starting with the placebo treatment. Sample size was limited to 4 groups because the cooperating dairy prematurely terminated the study due to increased health problems in one of the groups. Blood samples were taken during the prefresh period between 2 and 10 d prior to calving and at weekly intervals from 3 to 23 DIM. Blood concentrations of nonesterified fatty acids before calving and β-hydroxy-butyrate after calving were not affected by treatment. Supplementation with the DFM product increased milk fat percentage for the first lactation cows and increased milk protein percentage for the second and greater lactation cows during the first 85 DIM. Second-lactation cows fed the DFM product received fewer antibiotic treatments before 85 DIM than cows receiving the placebo. This validated the dairy producer's concern that cows consuming one of the diets (revealed to be the placebo diet after the study was completed) were experiencing more health problems. Most measures of milk yield were numerically increased by supplementation with the DFM product. However, differences in milk yield were not significant. Key covariates for main study outcomes included milk yield in the previous (first) lactation, body condition score prior to calving, days spent in the maternity pen, and stocking density of the pre-fresh pen.     

Controlled trial of the effect of negative dietary cation-anion difference on postpartum health of dairy cows

The objective of this study was to assess the effects of feeding negative dietary cation-anion difference (DCAD) dry cow diets on postpartum health. Cows from 4 commercial dairy farms in Ontario, Canada, were enrolled in a pen-level controlled trial from November 2017 to April 2019. Close-up pens (1 per farm), with cows 3 wk before expected calving, were randomly assigned to a negative DCAD [TRT; ?108 mEq/kg of dry matter (DM); target urine pH 6.0–6.5] or a control diet (CON; +105 mEq/kg of DM with a placebo supplement). Each pen was fed TRT or CON for 3 mo (1 period) then switched to the other treatment for the next period, with 4 periods per farm. Urine pH was measured weekly until calving, and body condition score (BCS) was measured at enrollment and at 5 wk postpartum. Data from 15 experimental units [8 TRT and 7 CON, with 1,086 (TRT: n = 681; CON: n = 405) observational units (cows)] that received the assigned diet for >1 wk were included. The incidence of milk fever (MF), retained placenta (RP), metritis, hyperketonemia (blood β-hydroxybutyrate >1.2 mmol/L, measured weekly in wk 1 and 2), clinical mastitis within 30 DIM (MAST), displaced abomasum (DA) within 30 d in milk (DIM), purulent vaginal discharge (PVD, assessed once at wk 5), and number of disease events (≥1 or ≥2) were analyzed with logistic regression models with treatment, parity, BCS, and their interactions, accounting for pen-level randomization and clustering of animals within farm with random effects, giving 10 degrees of freedom to test treatment effects. Multiparous cows fed TRT had greater blood calcium between 1 and 4 DIM than multiparous cows fed CON, and the prevalence of subclinical hypocalcemia (total Ca ≤2.14 mmol/L) was lesser when fed TRT compared with CON (d 1: 73 ± 6% vs. 93 ± 4%; d 2: 65 ± 7% vs. 90 ± 5%), with no differences between treatments detected in primiparous cows. We detected interactions of treatment and BCS at enrollment for MF in multiparous cows and of treatment and parity for ≥2 disease events. Overconditioned (BCS ≥3.75) multiparous cows had reduced incidence of MF when fed TRT (TRT: 2 ± 1%, vs. CON: 13 ± 8%). We detected no treatment effects on RP, metritis, hyperketonemia, or PVD incidence. Cows fed TRT had lesser incidence of DA (1.7 ± 0.7% vs. 3.6 ± 1.6%) and tended to have lesser incidence of MAST compared with CON (1.8% ± 0.6% vs. 4.4 ± 1.4%). No treatment effect was detected on ≥1 disease events (TRT: 38 ± 7%, vs. CON: 42 ± 8%); however, multiparous cows on TRT were less likely to have ≥2 disease events than cows on CON (14 ± 4% vs. 23 ± 6%). Under commercial herd conditions, feeding prepartum diets with negative DCAD improved several measures of postpartum health.     

Effect of prepartum exercise on lying behavior,labor length,and cortisol concentrations

The objective of this study was to assess the effect of exercise and pasture turnout on lying behavior, labor length, and cortisol concentrations around the time of parturition in dairy cows. Twenty-nine primiparous and 31 multiparous, pregnant, nonlactating Holstein (n = 58) and Jersey × Holstein cross (n = 2) dairy cows were assigned to control (n = 20), exercise (n = 20), or pasture (n = 20) treatments at dry-off using rolling enrollment. Control cows remained in the dry cow group pen. Exercise cows were removed from the dry cow group pen 5 times per week and walked for 1.4 ± 0.1 h at 1.88 ± 0.58 km/h. Pasture cows were moved to an outdoor paddock 5 times per week for 1.8 ± 0.3 h/d. Cows were housed in deep-bedded sand freestalls in a naturally ventilated, 4-row freestall barn. Cows were moved into maternity pens on the day of projected calving or when cows displayed signs that calving was imminent (restlessness, raised or lifted tail, ruptured amniotic sac, or swollen vulva), and treatments were discontinued. Cameras continuously recorded cows from entry into the pen until farm staff noted a calf, and one observer continuously watched video for two visually observable periods throughout the calving process: time from initial observation of amniotic sac to initial observation of calf's feet, and time from initial observation of calf's feet to full expulsion of calf. Assisted calvings were excluded. Accelerometers were attached to the rear fetlocks of cows 3 d before dry-off and removed 14 d postpartum. Activity was summarized by day for the 7 d before and after delivery time recorded from video observation into lying time (hours per day), lying bout frequency (bouts per day), lying bout duration (minutes per bout), and steps (number per day). Plasma total cortisol concentration was measured on d 0 and 3 postpartum and determined by a radioimmunoassay procedure using a commercially available kit. Data were analyzed using mixed linear model. During calving, time from appearance of the amniotic sac to appearance of the calf's feet was longer for pasture cows compared with control. Control cows engaged in fewer lying bouts and less overall lying time compared with pasture and exercise cows. Cortisol concentrations were higher on the day of calving compared with 3 d later, regardless of treatment. Understanding the effects of lying alterations around calving and increases in labor period length may allow for physical activity recommendations for late-gestation dairy cows.     

Short communication: Temporal changes in the skin morphology of dairy cows during the periparturient period

Management of dairy cow productivity requires monitoring of their nutritional status by visual observation. It has been suggested that changes in hair coat appearance are among the indicators of nutritional state in dairy cows. Temporal changes in the skin morphology in cows, however, have not been reported. In this study, we examined the changes in the skin of dairy cows that occur during the peripartum period. Seven pluriparous cows were used. Skin samples were collected at 28 d before the due date and 28 d and 56 d after calving for morphological examination. Hair follicle width was 108.8 ± 5.9 µm (±SD) in the dry period, 95.5 ± 5.5 µm at 28 d after calving, and 104.2 ± 5.3 µm at 56 d postpartum. The percentages of anagen hair follicles during these 3 periods were 41.4 ± 3.4, 18.5 ± 3.4, and 32.3 ± 3.3%, respectively. The corresponding sebaceous gland sizes were 8,362.0 ± 707.6, 7,800.0 ± 831.4, and 9,186.8 ± 962.6 µm², respectively. Hair follicle width was positively correlated with percentage of anagen hair follicles. The thickness of epidermal and proliferation rate of epidermal cell were also correlated. However, the hair follicle width, sebaceous gland size and cell proliferation rate, and thickness and proliferation rate of epidermal cells did not show any marked changes.     

Different durations of whole raw soybean supplementation during the prepartum period: Measures of cellular immune function in transition cows

The objective of this study was to evaluate different durations of whole raw soybean (WS) supplementation (diet rich in n-6 fatty acid) during the prepartum period on cellular immune function of dairy cows in the transition period and early lactation. Thirty-one Holstein cows were used in a completely randomized design and assigned to 4 experimental groups (G) [G90, G60, G30, and G0 (control)] supplemented with a diet containing 12% of WS from 90, 60, 30 and 0 d relative to the calving date, respectively. Cows were dried off 60 d before the expected calving date. After parturition, all cows were fed a diet containing 12% of WS until 84 DIM. Blood samples were collected before the morning feeding (d ?56 ± 2, ?28 ± 2, ?14 ± 2, ?7 ± 2, at the day of partum, 7 ± 2, 14 ± 2, 28 ± 2, and 56 ± 2 relative to parturition). Cell phenotyping and phagocytosis assays were carried out using monoclonal antibodies and flow cytometry technique. Duration of WS supplementation linearly increased the percentage of blood CD3⁺ cells, as well as increased the percentage of blood CD8⁺ cells in the postpartum period, notably in G30, whereas the lowest values were observed in G0. Further, the duration of WS supplementation linearly increased the reactive oxygen species median fluorescence intensity of CH138⁺ cells after phagocytizing Staphylococcus aureus in the postpartum period. Longer periods of WS supplementation linearly increased phagocytosis median fluorescence intensity of CH138⁺ cells in the prepartum period of cows. Duration of WS supplementation linearly increased the percentage of blood CD14⁺ cells producing reactive oxygen species when stimulated either by Staph. aureus or Escherichia coli in the postpartum period. In conclusion, longer periods of WS supplementation during late lactation and the dry period (beginning on d 90 of the expected calving date) alter the leukocyte population and improve neutrophil immune response in the postpartum period with no detrimental effects on cow performance.     

Reducing prepartum urine pH by supplementing anionic feed ingredients: Effects on physiological and productive responses of Holstein × Gir cows

This study compared physiological and productive parameters in 3/4 Holstein × 1/4 Gir dairy cows receiving a prepartum concentrate containing ammonium chloride to reduce urine pH near 7.0 (CON; n = 17), or a commercial anionic supplement to reduce urine pH near 6.0 (SUPP; n = 17). Nonlactating, multiparous, pregnant cows were assigned to receive SUPP or CON beginning 21 d before expected date of calving. Cows were maintained in a single drylot pen with ad libitum access to corn silage, and individually received their prepartum concentrate once daily (0800 h) before calving. Cows from both treatments completely consumed their concentrate allocation within 30 min after feeding. Cow body weight and body condition score were recorded once weekly, urine pH measured every 3 d, and blood samples collected on d ?21, ?14, ?9, ?6, and ?3 relative to expected calving date. After calving (d 0), cows were moved to an adjacent drylot pen with ad libitum access to water and a total mixed ration, and were milked twice daily (0600 and 1700 h). Cow body weight and body condition score were recorded once weekly and individual milk production was recorded daily until 30 d in milk (DIM). Blood samples were collected before each milking during the first 5 DIM, as well as at 6, 9, 16, 23, and 30 DIM before the morning milking. Based on actual calving dates, cows received SUPP or CON for (mean ± standard error) 19.2 ± 1.2 and 19.0 ± 0.9 d before calving, respectively. Urine pH was less in SUPP versus CON cows during the last 15 d of gestation (6.12 vs. 7.15, respectively). Milk yield during the first 5 DIM and throughout the experimental period was greater in SUPP versus CON cows (by 20 and 14%, respectively), whereas serum Ca concentrations did not differ between treatments during the first 5 DIM. Serum concentrations of fatty acids were greater in SUPP versus CON cows 3 d before and at calving (by 52 and 22%, respectively), whereas SUPP cows had lower serum glucose and cortisol concentration at calving (by 23 and 27%, respectively). Hence, the SUPP treatment decreased prepartum urine pH near 6.0 in Holstein × Gir dairy cows without depressing concentrate intake compared with CON, although total dry matter intake was not evaluated to fully investigate feed intake responses. Moreover, the SUPP treatment transiently affected serum glucose, fatty acids, and cortisol concentrations near the time of calving, and resulted in greater milk yield during the initial 30 DIM compared with CON.     

Effect of nutritional immunomodulation and heat stress during the dry period on subsequent performance of cows

Heat stress in dairy cows during the dry period impairs milk yield in the next lactation. Feeding OmniGen-AF (OG; Phibro Animal Health Corp., Teaneck, NJ) to lactating cows during heat stress may increase dry matter intake (DMI) and lowers respiration rate (RR) and rectal temperature (RT), but the effects in dry cows are not known. We hypothesized that OG supplementation before, during, and after the dry period (approximately 160 d total) would overcome the effects of heat stress and improve cow performance in the next lactation. Cows were randomly assigned to OG or control (placebo) treatments for the last 60 d in milk (DIM), based on mature-equivalent milk yield in the previous lactation. Cows were dried off 45 d before expected calving and randomly assigned to heat stress (HT) or cooling (CL) treatments. Thus, cows received dietary supplementation during late lactation before they were exposed to either CL or HT. After dry-off, treatment groups included heat stress with placebo (HT, only shade, 56 g/d of placebo, n = 17), HT with OG supplementation (HTOG, 56 g/d of OG, n = 19), cooling with placebo (CL, shade, fans, and soakers, 56 g/d of placebo, n = 16), and CL with OG supplementation (CLOG, 56 g/d of OG, n = 11). After parturition, all cows were kept under the same CL system and management, and all cows continued to receive OG or control treatment until 60 DIM. Cooling cows during the dry period reduced afternoon RT (CL vs. HT; 38.9 ± 0.05 vs. 39.3 ± 0.05°C) and RR (CL vs. HT; 45 ± 1.6 vs. 77 ± 1.6 breaths/min). Respiration rate was also decreased by OG supplementation under HT conditions (HTOG vs. HT; 69.7 ± 1.6 vs. 77.2 ± 1.6 breaths/min). An interaction was observed between OG supplementation and HT; HTOG cows tended to have lower morning RT compared with HT cows. During the dry period, OG reduced DMI relative to control cows. Birth weight was greater in calves from CL cows (CL vs. HT; 40.6 ± 1.09 vs. 38.7 ± 1.09 kg). No differences were detected among treatments in hematocrit, total protein, and body condition score. Cows offered CLOG, CL, and HTOG treatments had greater body weight during the dry period (794.9 ± 17.9, 746.8 ± 16.7, and 762.9 ± 14.9 kg, respectively) than HT cows (720 ± 16.2 kg). Gestation length was approximately 4 d longer for CL cows compared with HT cows. Cows offered CLOG, CL, and HTOG treatments produced more milk (41.3 ± 1.6, 40.7 ± 1.6, and 40.5 ± 1.6 kg/d, respectively) than HT treatment (35.9 ± 1.6 kg/d). Body weight after parturition and DMI were evaluated up to 60 DIM and averaged 661.5 ± 15.8 and 19.4 ± 0.7 kg/d, respectively, with no differences observed among treatments. These results confirm that exposure of dry cows to heat stress negatively affects milk yield in the subsequent lactation. Active cooling of dry cows and OG supplementation can reduce the negative effects of heat stress in the dry period on subsequent performance.