Lactating Holstein Cows Research Articles

Effect of reproductive management programs that prioritized artificial insemination at detected estrus or timed artificial insemination on the reproductive performance of primiparous Holstein cows of different genetic merit for fertility

Our objective was to compare reproductive outcomes of primiparous lactating Holstein cows of different genetic merit for fertility submitted for insemination with management programs that prioritized artificial insemination (AI) at detected estrus (AIE) or timed AI (TAI). Moreover, we aimed to determine whether subgroups of cows with different fertility potential would present a distinct response to the reproductive management strategies compared. Lactating primiparous Holstein cows (n = 6 commercial farms) were stratified into high (Hi-Fert), medium (Med-Fert), and low (Lo-Fert) genetic fertility groups (FG) based on a Reproduction Index value calculated from multiple genomic-enhanced predicted transmitting abilities. Within herd and FG, cows were randomly assigned either to a program that prioritized TAI and had an extended voluntary waiting period (P-TAI; n = 1,338) or another that prioritized AIE (P-AIE; n = 1,416) and used TAI for cows, not AIE. Cows in P-TAI received first service by TAI at 84 ± 3 d in milk (DIM) after a Double-Ovsynch protocol, were AIE if detected in estrus after a previous AI, and received TAI after an Ovsynch-56 protocol at 35 ± 3 d after a previous AI if a corpus luteum (CL) was visualized at nonpregnancy diagnosis (NPD) 32 ± 3 d after AI. Cows with no CL visualized at NPD received TAI at 42 ± 3 d after AI after an Ovsynch-56 protocol with progesterone supplementation (P4-Ovsynch). Cows in P-AIE were eligible for AIE after a PGF2α treatment at 53 ± 3 DIM and after a previous AI. Cows not AIE by 74 ± 3 DIM or by NPD 32 ± 3 d after AI received P4-Ovsynch for TAI at 74 ± 3 DIM or 42 ± 3 d after AI. Binary data were analyzed with logistic regression, count data with Poisson regression, continuous data by ANOVA, and time to event data by Cox's proportional hazard regression. Pregnancy per AI (P/AI) to first service was greater for cows in the Hi-Fert (59.8%) than the Med-Fert (53.6%) and Lo-Fert (47.7%) groups, and for the P-TAI (58.7%) than the P-AIE (48.7%) treatment. Overall, P/AI for all second and subsequent AI combined did not differ by treatment (P-TAI = 45.2%; P-AIE = 44.5%) or FG (Hi-Fert = 46.1%; Med-Fert = 46.0%; Lo-Fert = 42.4%). The hazard of pregnancy after calving was greater for the P-AIE than the P-TAI treatment [hazard ratio (HR) = 1.27, 95% CI: 1.17 to 1.37)], and for the Hi-Fert than the Med-Fert (HR = 1.16, 95% CI: 1.05 to 1.28) and Lo-Fert (HR = 1.34, 95% CI: 1.20 to 1.49) groups. More cows in the Hi-Fert (91.2%) than the Med-Fert (88.4%) and Lo-Fert (85.8%) groups were pregnant at 200 DIM. Within FG, the hazard of pregnancy was greater for the P-AIE than the P-TAI treatment for the Hi-Fert (HR = 1.41, 95% CI: 1.22 to 1.64) and Med-Fert (HR = 1.28, 95% CI: 1.12 to 1.46) groups but not for the Lo-Fert group (HR = 1.13, 95% CI: 0.98 to 1.31). We conclude that primiparous Holstein cows of superior genetic merit for fertility had better reproductive performance than cows of inferior genetic merit for fertility, regardless of the type of reproductive management used. In addition, the effect of programs that prioritized AIE or TAI on reproductive performance for cows of superior or inferior genetic merit for fertility depended on the outcomes evaluated. Thus, programs that prioritize AIE or TAI could be used to affect certain outcomes of reproductive performance or management.

Impact of oral calcium bolus timing on milk production and health events in early lactation Holstein cows

Treatment of subclinical hypocalcemia by administration of an oral Ca bolus at 0 and 24 h post calving has shown limited suc­cess in increasing production and minimizing adverse health events. Our objective was to explore the effect of delayed oral Ca bolus supplementation on milk production and disease inci­dence and the impact of delayed oral Ca bolus supplementation on postpartum blood Ca dynamics. We conducted a random­ized controlled trial on multiparous Holstein cows (n = 998) from 4 herds in New York. At calving, cows were randomly assigned to 1 of 3 treatment groups: 1) no supplemental Ca at or around parturition (CON; n = 343), 2) an oral Ca bolus con­taining 43 g Ca at calving and 24 h later (BOL-C; n = 330), or 3) an oral Ca bolus containing 43 g Ca at 48 and 72 h post-calving (BOL-D; n = 325). We created generalized linear mixed models to analyze differences in serum total Ca (tCa) at 1 and 4 DIM and milk yield for the first 10 wk of lactation between treatment groups and multivariable Poisson regression models to analyze adverse event outcomes (metritis, displaced abomasum, died or sold, or a combination of the above) in the first 30 DIM. Serum tCa was lower at 1 than 4 DIM (P < 0.001; 1.91 [95% CI = 1.90, 1.93] mmol/L and 2.26 [95% CI = 2.24, 2.28] mmol/L, respectively), and there was no difference in tCa between treatment groups (P = 0.16). Milk yield increased by week (P < 0.001) and was not af­fected by treatment (P = 0.18; CON: 48.9 [95% CI = 48.1, 49.8] kg, BOL-C: 48.3 [95% CI = 47.5, 49.1] kg, BOL-D: 49.3 [95% CI = 48.5, 50.1] kg). A treatment by parity group interaction for milk yield (P = 0.005) showed that BOL-D parity 3 cows produced more milk than BOL-D cows in parity group 2 or 4+ as well as CON and BOL-C cows in parity groups 2, 3 or 4+. Incidence of adverse events did not differ between treatments (P = 0.39; CON: 9.5%, BOL-C: 8.1%, BOL-D: 11.5%). Our findings suggest that delaying oral Ca bolus supplementation has limited impact on blood Ca concentrations but may be beneficial to cohorts of cows as a tar­geted prophylactic supplement to support milk production.

Enrichment of chocolate byproducts and protected fats with zinc to partially replace corn in diets of early lactation Holstein cows

Abstract The present experiment aimed to evaluate the effect of partial replacement of corn with chocolate byproducts, protected fats, or their mixture supplemented with zinc in the diet of lactating cows on feed utilization and lactation performance for 90 days. Fifty multiparous Holstein cows (565±25 kg BW, 3±1 parity, 7±1 days in milk, and a previous milk production of 35±2.9 kg/d), were randomly assigned to 5 treatments in a completely randomized design. The control diet contained (per kg DM): 412 g concentrate feed mixture, 412 g corn silage, and 176 g berseem hay. The control diet was supplemented with 1 g of zinc daily (Zinc diet). In the other diets, each kg of crushed corn grain was replaced with (DM basis) 600 g chocolate byproducts (CH diet), 400 g protected fats (PF diet), or 500 g of chocolate byproducts and protected fats mixture (1:1 DM basis) (CHPF diet). Both amounts of protected fats and chocolate byproducts had the same energy concentration as 1 kg of corn. The total mixed ration was prepared and distributed using a horizontal mixer system after mixing for 20 min. Chemical analysis showed that the replacement minimally affected the composition of the total mixed rations. Additionally, treatments did not affect feed intake. Compared to the control treatment, the zinc treatment did not affect milk production, milk composition, or feed efficiency. Increased (P<0.05) daily production of milk and component yields, and feed efficiency as well as fat concentration were observed by the CH, PF and CHPF treatments. Without affecting other measured blood parameters or hematocrit, treatments increased (P<0.05) the concentrations of total proteins, albumin, glucose, cholesterol, triglyceride, and zinc in the blood of cows. Additionally, the CH, PF, and CHPF treatments increased (P<0.01) the digestibility of nutrients compared to the control treatment. It is concluded that the addition of zinc to the control did not affect cow performance; however, its addition to chocolate byproducts, protected fats, or their mixture to partially replace corn improved the performance of cows.

Effects of Sexed and Conventional Semen Use and Calf Sex on Milk Yield Parameters, Body Weight and Milk Electrical Conductivity in Holstein Cows

This study was designed to determine the effects of semen type and calf gender on milk yield parameters, milk electrical conductivity and milk flow values and postpartum maternal body weight. A total of 265 Holstein cows in the first lactation were used in the study. The effect of sperm type on maternal body weight levels after calving and milk electrical conductivity values was significant (P0.05). Heifers inseminated with sexed semen had higher 100-days milk yield, milk peak yield and postpartum maternal body weight in the first lactation. Milk peak duration was higher in heifers inseminated with conventional semen (P0.05). In conclusion, heifers inseminated with sexed semen had higher maternal body weights, 100-days milk yield, milk peak yields and milk electrical conductivity levels in the first lactation than inseminated with conventional semen

Targeted reproductive management for lactating Holstein cows: Reducing the reliance on exogenous reproductive hormones

Adoption of automated monitoring devices (AMD) affords the opportunity to tailor reproductive management according to the cow's needs. We hypothesized that a targeted reproductive management (TRM) would reduce the use of reproductive hormones while increasing the percentage of cows pregnant 305 d in milk (DIM). Holstein cows from 2 herds (n = 1,930) were fitted with an AMD at 251.0 ± 0.4 d of gestation. Early-postpartum estrus characteristics (EPEC; intense estrus = heat index ≥70; 0 = minimum, 100 = maximum) of multiparous cows were evaluated at 40 (herd 1) or 41 (herd 2) DIM and EPEC of primiparous cows were evaluated at 54 (herd 1) or 55 (herd 2) DIM. Control cows received the first artificial insemination at fixed time (TAI; primiparous, herd 1 = 82 and herd 2 = 83 DIM; multiparous, herd 1 = 68 and herd 2 = 69 DIM) following the Double-Ovsynch (DOV) protocol. Cows enrolled in the TRM treatment were managed as follows: (1) cows with at least one intense estrus were inseminated upon AMD detected estrus for 42 d and, if not inseminated, were enrolled in the DOV protocol; and (2) cows without an intense estrus were enrolled in the DOV protocol at the same time as cows in the control treatment. Control cows were re-inseminated based on visual or patch aided detection of estrus, whereas TRM cows were re-inseminated as described for control cows with the aid of the AMD. Cows received a GnRH injection 27 ± 3 d after insemination and, if diagnosed as nonpregnant, completed the 5-d Cosynch protocol and received TAI 35 ± 3 d after insemination. Among cows in the TRM treatment, 55.8 and 42.9% of primiparous and multiparous cows, respectively, received the first insemination in spontaneous estrus. The interaction between treatment and parity affected pregnancy 67 d after the first AI (primiparous: control = 37.6%, TRM = 27.4%; multiparous: control = 41.0%, TRM = 44.7%). The TRM treatment increased re-insemination in estrus (control = 48.3%, TRM = 70.5%). Pregnancy 67 d after re-inseminations tended to be affected by the interaction between treatment and EPEC (no intense estrus: control = 25.3%, TRM = 32.0%; intense estrus: control = 32.9%, TRM = 32.2%). The interaction between treatment and EPEC affected pregnancy by 305 DIM (no intense estrus: control = 80.8%, TRM = 88.2%; intense estrus: control = 87.1%, TRM = 86.1%). Treatment did not affect the number of reproductive hormone treatments among cows that had not had an intense estrus (control = 10.5 ± 0.3, TRM = 9.1 ± 0.2 treatments/cow), but cows in the TRM treatment that had an intense estrus received fewer reproductive hormone treatments than cows in the control treatment (2.0 ± 0.1 vs. 9.6 ± 0.2 treatments/cow). Selecting multiparous cows for first AI in estrus based on EPEC reduced the use of reproductive hormones without impairing the likelihood of pregnancy to first AI. The use of AMD for re-insemination expedited the establishment of pregnancy among cows that did not display an intense estrus early postpartum.

The effects of birth and calving seson and birth year on milk production and composition in first lactating cows

The aim of this study was to investigate the effects of birth and calving season (spring, summer, autumn, and winter (astronomical)) and birth year on milk production and composition (content of lactose, fat, protein, and total fat and protein (kg, %)) of Holstein first lactating cows. The analysed data based on milk production records of the first lactation Holstein cows (n=158 and n=246 on farm 1 and farm 2, resp.) recorded within the years of 2013 to 2017 on two farms in west part of Slovakia. The birth season of heifers had no significant effect on the milk composition and production. The significant effect of birth year was observed on all parameters evaluated, except on total fat and protein content (%). Milk yield, depending on the birth year, increased from 7793.31±1629.31 kg (in cows born in year 2013) to 9628.76±1386.45 kg (in cows born in year 2017). Similarly, the content (kg) of all components evaluated was higher in cows born in year 2017 than in cows born in year 2013. The influence of the farm was evident in all parameters evaluated, except for total fat and protein and lactose content (%). Milk production and milk composition of first-lactation cows appear to be influenced by factors other than calving and birth season. The management practices implemented on the evaluated farms could be one of the influencing factors

Assessment of milk and blood enzymes in Holstein cows with subclinical mastitis

The objective of this study was to determine the activities of lactate dehydrogenase (LDH), alkaline phosphatase (ALP), gamma-glutamyl transferase (GGT), aspartate aminotransferase (AST) and alanine aminotransferase (ALT) in the milk and blood of lactating Holstein cows and investigate their association with subclinical mastitis (SCM). This study also aims to evaluate the impact of SCM and its severity based on California mastitis test (CMT) scores on the above mentioned enzymes, as well as to test the efficiency of their use as an early predictive diagnostic tool. Thirty multiparous, lactating dairy Holstein cows were examined using CMT and according to the obtained results classified into four groups; control group 1 (SCM-free = 10, score = 0), group 2 (weak positive = 8, score +1), group 3 (positive = 7, score +2) and group 4 (strong positive = 5, score +3). The milk LDH, ALP, and GGT activities were significantly higher in groups 3 and 4 compared to group 2 and the control group. LDH activity was significantly higher in group 2 compared to the control group. The mean blood LDH and ALP activities in SCM groups were significantly higher than in the control group. ALP blood activity was significantly higher in group 4 compared to group 2. The blood GGT activity was significantly higher in group 4 compared to other groups. There was a significant positive association between milk LDH, ALP, and GGT enzymes as well as the blood enzymes (ALP and GGT) in SCM cows. The results showed that subclinical mastitis causes considerable changes in the activities of enzymes in milk and blood of infected cows.

Abundance of proteins and genes associated with nutrient signaling, protein turnover, and transport of amino acids and glucose in fetuses from lactating Holstein cows

Availability of nutrients in maternal circulation and abundance of nutrient transporters, metabolic enzymes, and nutrient-responsive proteins in fetal tissues coordinate growth. To begin characterizing these mechanisms, we evaluated the abundance of nutrient signaling genes and proteins in bovine fetal tissues. Liver, entire intestine, and semitendinosus muscle were harvested from fetuses (4 female, 2 male) collected at slaughter from 6 clinically-healthy multiparous Holstein dairy cows (167 ± 7 days in milk, 37 ± 6 kg milk/d, 100 ± 3 d gestation). Data were analyzed using PROC MIXED in SAS 9.4. Among proteins measured, abundance of the amino acid (AA) utilization and insulin signaling proteins p-AKT and p-mTOR was greater (P < 0.01) in liver and intestine. The abundance of p-EEF2 (translation elongation) and SLC2A4 (glucose uptake) was greater (P < 0.05) in liver relative to intestine and muscle suggesting this organ has a greater capacity for anabolic processes. In contrast, among mTOR signaling genes, the abundance of IRS1 was greatest (P < 0.01) in muscle and lowest in the intestine, whereas, abundance of AKT1 and mTOR was greater (P < 0.01) in intestine and muscle than liver. Abundance of the protein degradation-related genes UBA1, UBE2G1, and TRIM63 was greater (P < 0.01) in muscle than intestine and liver. Among nutrient transporters, abundance of glucose transporters SLC5A1 and SLC2A2 was greatest (P < 0.01) in the intestine than liver and muscle. Several AA transporters had greater (P < 0.01) abundance in the intestine or liver compared with muscle. Overall, these molecular analyses highlighted important biological differences on various aspects of metabolism in fetal tissues.

Hepatic Global Transcriptomic Profiles of Holstein Cows According to Parity Reveal Age-Related Changes in Early Lactation.

Cows can live for over 20 years, but their productive lifespan averages only around 3 years after first calving. Liver dysfunction can reduce lifespan by increasing the risk of metabolic and infectious disease. This study investigated the changes in hepatic global transcriptomic profiles in early lactation Holstein cows in different lactations. Cows from five herds were grouped as primiparous (lactation number 1, PP, 534.7 ± 6.9 kg, n = 41), or multiparous with lactation numbers 2-3 (MP2-3, 634.5 ± 7.5 kg, n = 87) or 4-7 (MP4-7, 686.6 ± 11.4 kg, n = 40). Liver biopsies were collected at around 14 days after calving for RNA sequencing. Blood metabolites and milk yields were measured, and energy balance was calculated. There were extensive differences in hepatic gene expression between MP and PP cows, with 568 differentially expressed genes (DEGs) between MP2-3 and PP cows, and 719 DEGs between MP4-7 and PP cows, with downregulated DEGs predominating in MP cows. The differences between the two age groups of MP cows were moderate (82 DEGs). The gene expression differences suggested that MP cows had reduced immune functions compared with the PP cows. MP cows had increased gluconeogenesis but also evidence of impaired liver functionality. The MP cows had dysregulated protein synthesis and glycerophospholipid metabolism, and impaired genome and RNA stability and nutrient transport (22 differentially expressed solute carrier transporters). The genes associated with cell cycle arrest, apoptosis, and the production of antimicrobial peptides were upregulated. More surprisingly, evidence of hepatic inflammation leading to fibrosis was present in the primiparous cows as they started their first lactation. This study has therefore shown that the ageing process in the livers of dairy cows is accelerated by successive lactations and increasing milk yields. This was associated with evidence of metabolic and immune disorders together with hepatic dysfunction. These problems are likely to increase involuntary culling, thus reducing the average longevity in dairy herds.

Effect of substituting grass-clover silage with maize silage for dairy cows on nutrient digestibility, rumen metabolism, enteric methane emission and total carbon footprint

The aim of this experiment was to determine the optimal ratio between maize silage and grass-clover silage in lactating dairy cow diets. The evaluation was based on nutrient digestibility, rumen metabolism and total carbon footprint (CF). Carbon footprint included enteric methane emission and CF from feed production, transportation, processing, and C sequestration in the soil, based on a Life Cycle Assessment (LCA) approach. Four diets varying in proportions of grass-clover silage and maize silage (100:0, 67:33, 33:67, and 0:100 on dry matter (DM) basis, respectively) were fed ad libitum to 4 lactating Holstein cows (a fifth cow was involved later in the study) fitted with ruminal, duodenal, and ileal canulae in a 4 × 4 Latin square design. Nutrient digestibility was measured using chromium oxide and titanium oxide as flow markers, and methane and CO2 emissions were measured in open-circuit respiration chambers on 5 consecutive days for each cow in each period. Data were analyzed with treatment and period as fixed effects and cow as random effect. DM intake (DMI) increased linearly (P = 0.01) with increased proportion of maize silage in the feed ration. Under the conditions in the present study, diets had only minor effects on digestibility of nutrients and there was no influence on rumen pH. Total VFA concentration decreased linearly with increased level of maize silage, but there were only minor effects on molar proportions of VFAs in the rumen. However, there was a linear increase in duodenal flow of microbial DM with increasing level of maize silage. The cows produced 30.7, 29.4. 28.4, and 26.2 L of methane/kg of DMI on diets 100:0, 66:33, 33:66, and 0:100, respectively (P = 0.02 for linear effect), which is equivalent to a 15% decrease, when all grass-clover silage was replaced by maize silage. Despite the significant reduction in enteric methane emission with increased proportion of maize silage, only a 4% decrease in total CF per kg DMI was found, when including CF from feed production, transportation, processing, manure, energy use and C sequestration in the soil, when using a LCA approach. If direct Land Use Change (LUC) or indirect LUC was included in estimation of CF, the reduction in CF when replacing all grass-clover silage was also 4–6%. However, the total CF was highly dependent on assumptions related to yield of grass-clover and maize.

Expression of genes for transport of water and angiogenesis, as well as biochemical biomarkers in Holstein dairy cows during the ovsynch program

Changes in expression of genes associated with angiogenesis and transport of water by cells, as well as biomarkers of oxidative stress were determined at specific times during the ovsynch protocol to synchronize estrus and breed Holstein dairy cows. Blood samples were taken from 82 lactating Holstein cows at the time of the 1st GnRH injection (G1), 7 days later at the time of the PGF2a (PG) injection, and 48 h after the PGF2a treatment when the second injection of GnRH was administered (G2). The serum was analyzed for malondialdehyde (MDA), reduced glutathione (GSH), glutathione peroxidase (GPX), nitric oxide (NO), catalase (CAT), and total antioxidant capacity (TAC). The expression of vascular endothelial growth factor (VEGF), vascular endothelial growth factor receptor 2 (VEGFR2), endothelial nitric oxide synthase (eNOS3), aquaporin 3 (AQP3), and AQP4 mRNAs in peripheral blood mononuclear cells (PBMCs) was analyzed. The number of copies of each of the mRNAs was quantified using qPCR. Pregnancy status was determining at 32 ± 3 days after insemination using an ultrasound “Sonoscape-5V″ model. Receiver operating curves (ROC) were used to assess the sensitivity and specificity of the biochemical parameters in serum to predict establishment of p The expression of MDA, GPX, and Catalase changed (P < 0·05) between G1, PG and G2 phases of the ovsynch protocol with higher levels at PG than at G1 and G2. The highest levels of NO were detected at G2. The ROC analyses identified NO, TAC and CAT as the most sensitive and specific biomarker for pregnancy with areas under the curve being 0.875 (P < 0.0001), 0.843 (P < 0.03), 0.833 (P < 0.017), sensitivity being 75.3, 42.86, and 26.27%, and specificity being 90, 90 and 85% respectively. The expression for VEGF, VEGFR2, eNOS3, AQP3, and AQP4 mRNAs was upregulated at PG compared to G1 and G2 phases of the ovsynch protocol. The results suggest that following the first injection of GnRH, there is an increase in expression of VEGF, VEGFR2, eNOS3, AQP3, and AQP4 mRNAs by the time of the PGF2a injection and then expression decreased. Further, ROC analyses identified increases in NO, TAC and CAT as the most sensitive and specific biomarkers with the greatest potential to predict establishment of pregnancy in Holstein cows.

Effects of the timing of protein infusion on the daily rhythms of milk synthesis and plasma hormones and metabolites in dairy cows

Milk synthesis exhibits a daily rhythm that is modified by the timing of feed intake. However, it is unknown how specific nutrients entrain this daily rhythm. Amino acids have an important role in milk synthesis, and may have a role in entrainment of mammary circadian rhythms. The objective of this study was to determine the effects of intestinally absorbed protein on daily rhythms of milk and milk component synthesis and key plasma hormones and metabolites. Nine lactating Holstein cows were assigned to 1 of 3 treatment sequences in a 3 × 3 Latin square. Treatments included abomasal infusions of 500 g/d of sodium caseinate either continuously throughout the day (CON), for 8 h/d from 0900 to 1700 h (DAY), or for 8 h/d from 2100 to 0500 h (NGT). Cows were milked every 6 h during the final 8 d of each period. A 24-h rhythm was fit to data using cosine analysis and the amplitude and acrophase were determined. Night infusion of protein decreased the daily milk yield and milk protein yield by 8.2% and 9.2%, respectively. Milk fat yield was increased 5.5% by DAY and milk fat concentration was increased 8.8% by NGT. Milk yield exhibited a daily rhythm in all treatments, with NGT increasing the amplitude of the daily rhythm 33% compared with CON. Milk fat concentration fit a daily rhythm in CON and NGT, but not DAY, whereas milk protein concentration fit a daily rhythm in CON and DAY, but not NGT. Moreover, DAY abolished the daily rhythm of plasma glucose concentration, but induced rhythms of plasma insulin and nonesterified fatty acid concentrations. Results suggest that feeding increased protein levels during the early part of the day may increase milk fat yield and modify energy metabolism through increased daily variation in insulin-stimulated lipid release, but additional research focused on feeding multiple diets across the day is required.

Reproductive performance of lactating dairy cows with elevated milk β-hydroxybutyrate levels during first 6 weeks of lactation

Although there is evidence that ketosis negatively affects fertility, the effect of late and early ketosis on the reproductive performance of lactating cows has not been systematically investigated. The aim of this study was to evaluate the association between time and amplitude of elevated milk BHB (EMB) occurring within 42 d in milk (DIM) and subsequent reproductive performance of lactating Holstein cows. The dairy herd information data of 30,413 cows with 2 test-day milk BHB recordings during early lactation periods 1 and 2 (5-14 and 15-42 DIM, respectively) assessed as negative (<0.15 mmol/L), suspect (0.15-0.19 mmol/L), or positive (≥0.2 mmol/L) for EMB were used in this study. Based on the time and amplitude of milk BHB, cows were grouped into 7 groups: (1) healthy cows negative in both periods 1 and 2 were classified as NEG; (2) suspect in period 1 and negative in period 2: EARLY_SUSP; (3) suspect in period 1 and suspect/positive in period 2: EARLY_SUSP_Pro; (4) positive in period 1 and negative in period 2: EARLY_POS; (5) positive in period 1 and suspect/positive in period 2: EARLY_POS_Pro; (6) negative in period 1 and suspect in period 2: LATE_SUSP; and (7) negative in period 1 and positive in period 2: LATE_POS. The overall prevalence of EMB within 42 DIM was 27.4%, with the highest prevalence being EARLY_SUSP (10.49%). Cows in EARLY_POS and EARLY_POS_Pro, but not other EMB categories, had a longer interval from calving to first service compared with NEG cows. For the reproductive parameters, first service to conception interval, days open and calving interval, cows in all EMB groups except EARLY_SUSP had longer intervals compared with NEG cows. These data indicate that there is a negative association between EMB within 42 d and reproductive performance after the voluntary waiting period. The intriguing findings of this study are the unaltered reproductive performance of EARLY_SUSP cows, and the negative association between late EMB and reproductive performance. Hence, monitoring and prevention of ketosis during the first 6 wk of lactation is necessary to optimize reproductive performance of lactating dairy cows.

Effect of dietary seaweed (Ascophyllum nodosum) supplementation on milk mineral concentrations, transfer efficiency, and hematological parameters in lactating Holstein cows

This study investigated the effect of feeding seaweed (Ascophyllum nodosum) to dairy cows on milk mineral concentrations, feed-to-milk mineral transfer efficiencies, and hematological parameters. Lactating Holstein cows (n = 46) were allocated to 1 of 2 diets (n = 23 each): (1) control (CON; without seaweed) and (2) seaweed (SWD; replacing 330 g/d of dried corn meal in CON with 330 g/d dried A. nodosum). All cows were fed the CON diet for 4 wk before the experiment (adaptation period), and animals were then fed the experimental diets for 9 wk. Samples included sequential 3-wk composite feed samples, a composite milk sample on the last day of each week, and a blood sample at the end of the study. Data were statistically analyzed using a linear mixed effects model with diet, week, and their interaction as fixed factors; cow (nested within diet) as a random factor; and data collected on the last day of the adaptation period as covariates. Feeding SWD increased milk concentrations of Mg (+6.6 mg/kg), P (+56 mg/kg), and I (+1,720 μg/kg). It also reduced transfer efficiency of Ca, Mg, P, K, Mn, and Zn, and increased transfer efficiency of Mo. Feeding SWD marginally reduced milk protein concentrations, whereas there was no effect of SWD feeding on cows' hematological parameters. Feeding A. nodosum increased milk I concentrations, which can be beneficial when feed I concentration is limited or in demographics or populations with increased risk of I deficiency (e.g., female adolescents, pregnant women, nursing mothers). However, care should also be taken when feeding SWD to dairy cows because, in the present study, milk I concentrations were particularly high and could result in I intakes that pose a health risk for children consuming milk.

Use of milk proteins as biomarkers of changes in the rumen metaproteome of Holstein cows fed low-fiber, high-starch diets

Dietary levels of undegraded neutral detergent fiber (uNDF240) and rumen-fermentable starch (RFS) can affect the rumen microbiome and milk composition. The objective of the study is to investigate the use of milk proteins as biomarkers of rumen microbial activity through a comparative evaluation of the rumen microbial and milk protein profiles produced by Holstein cows fed diets with varying contents of physically effective uNDF240 (peuNDF240) and RFS. Eight ruminally cannulated lactating Holstein cows were included in a larger study as part of a 4 × 4 Latin square design with 4 28-d periods to assess 4 diets varying in peuNDF240 and RFS content. For this experiment, cows received one of 2 dietary treatments: (1) low-peuNDF240, high-RFS (LNHR) diet or (2) high-peuNDF240, low-RFS (HNLR) diet. Within each period, rumen fluid samples were collected from each cow on d 26 (1400 h) and d 27 (0600 h and 1000 h), and milk samples were collected from each cow on d 25 (2030 h), d 26 (0430 h, 1230 h, and 2030 h), and d 27 (0430 h and 1230 h). Microbial proteins were isolated from each rumen fluid sample. For milk samples, milk proteins were fractionated, and the whey fraction was subsequently isolated. Isolated proteins within each rumen fluid or milk sample were isobarically labeled and analyzed by liquid chromatography-tandem mass spectrometry. Product ion spectra acquired from rumen fluid samples were searched using SEQUEST against 71 composite databases. In contrast, product ion spectra acquired from milk samples were searched against the Bos taurus database. Data were analyzed using the PROC MIXED procedure in SAS 9.4 to assess the effect of diet and time of sampling. To increase stringency, the false discovery rate-adjusted P-value (PFDR) was also calculated to account for multiple comparisons. Using the mixed procedure, a total of 129 rumen microbial proteins were quantified across 24 searched microbial species. Of these, the abundance of 14 proteins across 9 microbial species was affected due to diet and diet × time interaction, including 7 proteins associated with energetics pathways. Among the 159 quantified milk proteins, the abundance of 21 proteins was affected due to the diet and diet × time interaction. The abundance of 19 of these milk proteins was affected due to diet × time interactions. Of these, 16 proteins had the disparity across diets at the 0430 h sampling time, including proteins involved in host defense, nutrient synthesis, and transportation, suggesting that biological shifts resulting from diet-induced rumen changes are not diurnally uniform across milkings. The concentration of lipoprotein lipase (LPL) was statistically higher in the milk from the cows fed with the LNHR diet, which was numerically confirmed with an ELISA. Further, as determined by ELISA, the LPL concentration was significantly higher in the milk from the cows fed with the LNHR diet at 0430 h sampling point, suggesting that LPL concentration may indicate dietary carbohydrate-induced ruminal changes. The results of this study suggest that diet-induced rumen changes can be reflected in milk in a diurnal pattern, further highlighting the need to consider sampling time points for using milk proteins as a representative biomarker of rumen microbial activity.

Partial replacement of corn with chocolate byproducts or protected fat in diets of early lactating Holstein cows: feed utilization and lactational performance

This study determined the effects of the partial replacement of corn with chocolate byproducts or protected fat on the performance of cows. Forty multiparous Holstein cows were assigned to four treatments in a completely randomized design for 90 d. The control diet was a complete diet containing 41.2% concentrate feed mixture, 41.2% corn silage, and 17.6% berseem hay. In the other experimental diets, each kg of crushed corn grain in the control diet (DM basis) was replaced with 600 g chocolate byproducts (CH), 400 g protected fat (PF), or 500 g of a mixture of chocolate byproducts and protected fat (1:1 DM basis) (CHPF). Both amounts of PF and CH had the same energy concentration as 1 kg of corn. The experimental treatments did not affect total intake; however, they increased (p < 0.001) daily production of milk, milk fat content, and feed efficiency. Moreover, treatments increased (p < 0.05) blood total protein, albumin, glucose, cholesterol, and triglyceride. The CH, PF, and CHPF treatments increased (p < 0.01) nutrient digestibility. It is concluded that replacing 1 kg of corn in the diet of lactating Holstein cows with PF at 400 g, followed by CHPF at 500 g, and CH at 600 g enhanced milk production and feed efficiency.

Presynchronization with prostaglandin F2α and gonadotropin-releasing hormone simultaneously improved first-service pregnancy per artificial insemination in lactating Holstein cows compared with Presynch-14 when combined with detection of estrus

This study aimed to determine the effect of 2 simple breeding strategies combining artificial insemination (AI) after detection of estrus (AIED) and timed AI (TAI) on first-service fertility in lactating Holstein cows. Weekly, lactating Holstein cows (n = l,049) between 40 and 46 d in milk (DIM) were randomly assigned to initiate 1 of 2 breeding strategies for first service: Presynch-14 and PG+G. Presynch-14 is a presynchronization strategy with 2 PGF2α treatments 14 d apart with the last PGF2α 14 d before the initiation of the Ovsynch protocol. Cows treated with PG+G receive a simpler presynchronization program that uses PGF2α and GnRH simultaneously 7 d before Ovsynch. In both treatments, cows detected in standing estrus by tail chalk at any time ≥55 DIM were inseminated, and treatment was discontinued (n = 525). Cows completing treatment received TAI from 78 to 84 DIM (n = 526). In a subgroup of cows that received TAI, blood was collected (n = 163) to assess circulating concentrations of progesterone, and ultrasonographic evaluations of ovaries were performed on the day of first GnRH of Ovsynch (n = 162) and PGF2α of Ovsynch (n = 122). The proportion of cows that received TAI was greater for PG+G compared with Presynch-14 (63.5 vs. 31.9%), which increased DIM at first service for cows treated with PG+G compared with Presynch-14 (75.5 ± 0.4 vs. 68.7 ± 0.4). For cows receiving TAI, the ovulatory response to first GnRH of Ovsynch (73.8 vs. 48.8%) and the proportion of cows with functional corpora lutea (92.6 vs. 73.1%) were greater for PG+G than Presynch-14. Cows treated with PG+G had greater overall pregnancy per AI (P/AI) 42 ± 7 d after AI (40.2 vs. 33.6%) and calving per AI (32.1 vs. 25.2%) than Presynch-14. For cows receiving AIED, treatment did not affect P/AI 42 ± 7 d after AI. However, for cows receiving TAI, PG+G increased P/AI compared with Presynch-14 (44.6 vs. 35.2%). Overall, cows receiving TAI had greater P/AI 42 ± 7 d after AI (42.5 vs. 31.5%) and calving per AI (34.1 vs. 23.7%) and decreased pregnancy loss (16.8 vs. 25.2%) than cows receiving AIED. In summary, PG+G increased the proportion of cows receiving TAI and the DIM at first service, P/AI, and calving per AI compared with Presynch-14 when both TAI programs were combined with AIED.

Genetic relationships between weight loss in early lactation and daily milk production throughout lactation in Holstein cows

After calving, high-yielding dairy cows mobilize body reserves for energy, sometimes to the detriment of health and fertility. This study aimed to estimate the genetic correlation between body weight loss until nadir and daily milk production (MY24) in first- (L1) and second-lactation (L2) Holstein cows. The data set included 859,020 MY24 records and 570,651 daily raw body weight (BWr) phenotypes from 3,989 L1 cows, and 665,361 MY24 records and 449,449 BWr phenotypes from 3,060 L2 cows, recorded on 36 French commercial farms equipped with milking robots that included an automatic weighing platform. To avoid any bias due to change in digestive content, BWr was adjusted for variations in feed intake, estimated from milk production and BWr. Adjusted body weight was denoted BW. The genetic parameters of BW and MY24 in L1 and L2 cows were estimated using a 4-trait random regression model. In this model, the random effects were fitted by second-order Legendre polynomials on a weekly basis from wk 1 to 44. Nadir of BW was found to be earlier than reported in the literature, at 29 d in milk, and BW loss from calving to nadir was also lower than generally assumed, close to 29 kg. To estimate genetic correlations between body weight loss and production, we defined BWL5 as the loss of weight between wk 1 and 5 after calving. Genetic correlations between BWL5 and MY24 ranged from -0.26 to 0.05 in L1 and from -0.11 to 0.10 in L2, according to days in milk. These moderate to low values suggest that it may be possible to select for milk production without increasing early body mobilization.

Association of activity and subsequent fertility of dairy cows after spontaneous estrus or timed artificial insemination

The objective of this observational study was to evaluate the association between increased physical activity at first artificial insemination (AI) and subsequent pregnancy per AI (P/AI) in lactating Holstein cows following spontaneous estrus or a timed AI (TAI) protocol. We also wanted to identify factors associated with the intensity of activity increase (PA) captured by automated activity monitors (AAM) and fertility. Two experiments were conducted, in which cows either were inseminated based on the alert of the AAM system (AAM cows) or received TAI following a 7-d Ovsynch protocol (TAI cows) if not inseminated within a farm-specific period after calving. Experiment 1 included 2,698 AI services from AAM cows and 1,042 AI services from TAI cows equipped with the Smarttag Neck (Nedap Livestock Management) from a dairy farm in Slovakia (farm 1). In the second experiment, 6,517 AI services from AAM cows and 1,226 AI services from TAI cows fitted with Heatime (Heatime Pro; SCR Engineers Ltd.) from 8 dairy farms in Germany (farms 2-9) were included. Pregnancy diagnosis was performed on a weekly basis by transrectal ultrasound (farms 1, 3, 7, 8) or by transrectal palpation (farms 2, 4-6, 9). Estrous intensity was represented by the peak value of the change in activity. In experiment 1, PA was categorized into low (x-factor 0-20) and high (x-factor 21-100) PA, and in experiment 2 into low (activity change = 35-89) and high (activity change = 90-100) PA. In TAI cows from both experiments, PA was additionally categorized into cows with no AAM alert. Data were analyzed separately for AAM and TAI cows using multinomial logistic regression models for PA in TAI cows and logistic regression models for PA in AAM cows and P/AI in both groups. In experiment 1, P/AI of AAM cows was greater for AI services performed with conventional frozen semen (57.6%) compared with sexed semen (47.2%), whereas type of semen only tended to be associated with P/AI in TAI cows (54.4% conventional frozen semen vs. 48.9% sexed semen). In experiment 2, P/AI was greater for fresh semen (AAM cows: 44.4% vs. TAI cows: 44.2%) compared with conventional frozen semen (AAM cows: 37.6% vs. TAI cows: 34.6%). In both experiments, pregnancy outcomes were associated with PA. In experiment 1, AAM cows with high PA (55.1%) had greater P/AI than cows with low PA (49.8%). Within TAI cows, cows with no alert (38.8%) had reduced P/AI compared with cows with low (54.2%) or high PA (61.8%). In experiment 2, AAM cows with high PA (45.8%) had greater P/AI compared with cows with low PA (36.4%). Timed AI cows with no alert (27.4%) had decreased P/AI compared with cows with low (41.1%) or high (50.8%) PA. The greatest risk factors for high PA were parity (experiment 1) and season of AI (except for TAI cows from experiment 1). We conclude that high PA at the time of AI is associated with greater odds of pregnancy for both AAM and TAI cows. In both experiments, about 2 thirds of AAM cows (experiment 1: 69.9% and experiment 2: 70.7%) reached high PA, whereas only approximately one-third or less of TAI cows (experiment 1: 37.3% and experiment 2: 23.6%) showed high PA. Although we observed similar results using 2 different AAM systems for the most part, risk factors for high PA might differ between farms and insemination type (i.e., AAM vs. TAI).

Increased dietary vitamin D3 and calcium partially alleviate heat stress symptoms and inflammation in lactating Holstein cows independent of dietary concentrations of vitamin E and selenium

Twelve multiparous Holstein cows (42.2 ± 5.6 kg of milk/d; 83 ± 27 d in milk) were used in a split-plot design testing the effects of mineral and vitamin supplementation on the time course of animal performance, metabolism, and inflammation markers during heat stress. The main plot was the average concentrations of dietary vitamin E and Se (adequate: 11.1 IU/kg of vitamin E and 0.55 mg/kg of Se, and high: 223 IU/kg of vitamin E and 1.8 mg/kg of Se, respectively). Within each plot, cows were randomly assigned to (1) heat stress (HS) with adequate concentrations of vitamin D3 and Ca (1,012 IU/kg and 0.73%, respectively), (2) HS with high concentrations of vitamin D3 and Ca (HS+D3/Ca; 3,764 IU/kg and 0.97%, respectively), or (3) pair-feeding (PF) in thermoneutrality with adequate concentrations of vitamin D3 and Ca (1,012 IU/kg and 0.73% Ca) in a Latin square design with 14-d periods and 7-d washouts. The highest rectal temperature was recorded at 1700 h for HS (39.4°C; mean of d 1 to 14), being 1.2 and 0.8°C greater than for PF and HS+D3/Ca, respectively. Respiratory rate and water intake were higher in HS (73 breaths/min and 115 L/d, respectively) relative to PF (28 breaths/min and 76 L/d). Heat stress decreased dry matter intake progressively, reaching a nadir on d 5 to 7 (33% reduction) and was not different between treatments. Milk yield decreased progressively in all treatments, but remained greater in PF relative to HS from d 3 to 14 (10%), whereas HS and HS+D3/Ca were not different. Milk fat, protein, and lactose concentrations and yields were lower in HS relative to PF from d 3 to 14, but not different between HS and HS+D3/Ca. Relative to PF, preprandial insulin concentrations were increased in HS, whereas plasma nonesterified fatty acids were decreased on d 7 and 14. Plasma lipopolysaccharide-binding protein concentrations increased in HS cows on d 7 and 14, respectively, relative to PF, whereas they were reduced in HS + D3/Ca on d 14. Plasma C-reactive protein, tumor necrosis factor-α, and fecal calprotectin were increased in HS relative to both PF and HS+D3/Ca on d 7 and 14. Rectal temperature was positively associated with plasma lipopolysaccharide-binding protein (r = 0.72), tumor necrosis factor-α (r = 0.74), C-reactive protein (r = 0.87), and with milk somatic cells (r = 0.75). Plasma 8-hydroxy-2-deoxyguanosine concentrations presented a 3-way interaction, where 8-hydroxy-2-deoxyguanosine was lower in HS than in PF on d 7 and 14, and lower in HS+D3/Ca relative to HS on d 14 in the adequate vitamin E and Se treatment, but no effects were observed in the high vitamin E and Se group. Plasma superoxide dismutase concentrations increased over time, and were higher in HS relative to PF on d 14, whereas HS+D3/Ca was similar to HS. Heat stress markedly reduced milk production and milk components while increasing markers of leaky gut and inflammation. In contrast, vitamin D3 and Ca supplementation reduced hyperthermia (d 7-14), markers of leaky gut, and inflammation independent of dietary concentrations of vitamin E and Se.