Wk Postpartum Research Articles

First postpartum ovulation, metabolites and hormones in follicular fluid and blood in transition dairy cows supplemented with a Saccharomyces cerevisiae fermentation product

We hypothesized that feeding a Saccharomyces cerevisiae fermentation product (SCFP) from −4 through +7 wk (calving = Day 0) facilitates early first postpartum ovulation and alters blood and follicular fluid concentrations of glucose, beta-hydroxybutyrate (BHB), free fatty acids (FFA), and steroid hormones favorable to subsequent fertility. Holstein cows were fed individually a SCFP product (n = 24) or served as controls (n = 23). Blood samples were collected at wk −4 and −2 from expected calving and at 1, 2, 5, and 7 wk postpartum to determine plasma concentrations of FFA and BHB. Early spontaneous ovulation (progesterone > 1 ng/mL or corpus luteum presence by postpartum median Day 33) or late ovulation was determined. Plasma FFA in weekly samples was not affected by SCFP supplementation, but FFA was greater (P < 0.01; week by ovulation status) in late compared with early ovulating cows during and after postpartum wk 2. Plasma BHB in weekly samples was greater (P = 0.03) in SCFP than control cows and tended (P = 0.06) to be greater in late than early ovulating cows. Cows were exposed to ovulation synchronization (GnRH, PGF2α, and GnRH on Days 33, 40, and 43 ± 3, respectively). Transvaginal dominant follicle aspiration was conducted at Day 50, 7 d after GnRH on Day 43. Metabolites (FFA, BHB, and glucose) and steroid hormones (progesterone, androstenedione, and estradiol) measured in follicular fluid and blood samples collected at aspiration revealed that androstenedione in serum was numerically less (P = 0.11) in SCFP-treated compared with control cows, whereas androstenedione in serum was less (P < 0.05) in late than early ovulating cows. Concentrations of BHB (r = 0.75) and glucose (r = 0.52) in follicular fluid were positively correlated (P < 0.01) with those in blood. Body weight at calving and Day 42 was less (P ≤ 0.05), and energy balance through Days 28 and 42 was more positive (P < 0.05) in early than late ovulating cows and in SCFP-supplemented compared with control cows (P < 0.05). Dry matter intake, daily milk yield, and yields of fat, protein, lactose, and total solids were less (P < 0.01) in early compared with late ovulating cows, whereas milk fat percentage was increased (P < 0.01) by SCFP supplementation. We conclude that elevated postpartum BHB and FFA in plasma, greater negative energy balance, and greater milk yield and components were associated with later postpartum ovulation, but metabolites and steroid hormones in blood and follicular fluid were unaffected by SCFP treatment or ovulation status except for androstenedione.

Effects of adding an automated monitoring device to the health screening of postpartum Holstein cows on survival and productive and reproductive performances

Automated monitoring devices (AMD) have become more affordable, and consequently more popular among dairy producers. We hypothesized that the addition of AMD-generated health alerts to a health-screening program improves survival, milk production, and reproductive success. In addition, we hypothesized that cows diagnosed with clinical disease that have AMD alerts are at greater risk of culling, lower milk production, and decreased risk of pregnancy than cows without AMD alerts. Holstein cows (nulliparous = 282, parous = 328) were enrolled at -60 ± 3 d (d 0 = calving), when they were fitted with an AMD and assigned randomly to 1 of 2 health-screening strategies: (1) control: AMD alerts not provided to farm personnel; and (2) automated device: AMD alerts provided to farm personnel. Twice daily, study personnel determined which cows had AMD alerts (health index ≤79, rumination <200 min/d, or difference between current rumination and the average of the 3 preceding days <0) and provided the information to farm personnel. Farm personnel examined cows at 3, 5, and 9 d in milk (DIM) and when daily milk yield decreased ≥25% on consecutive days. We detected no differences between health-screening strategies regarding morbidity (control = 49.7 ± 3.3%, automated device = 52.8 ± 3.2%), but the interaction between health-screening strategy and parity tended to be associated with the number of clinical diseases per cow (primiparous: control = 0.46 ± 0.06, automated device = 0.65 ± 0.07 cases/cow; multiparous: 0.88 ± 0.08, automated device = 0.86 ± 0.08 cases/cow). Cows enrolled in the automated device strategy were more likely to be treated with supportive therapy (64.4 ± 3.1 vs. 55.0 ± 3.2%), whereas primiparous cows in the automated device strategy were more likely to be treated with anti-inflammatory drugs than those in the control strategy (41.6 ± 4.7 vs. 23.8 ± 4.0%). Health-screening strategy did not affect survival or total milk yield up to 22 wk postpartum, but cows in the automated device strategy had reduced risk of pregnancy after the first 2 services (54.5 ± 3.0 vs. 46.2 ± 3.2%). Cows diagnosed with a clinical disease without AMD alerts had reduced risk of removal from the herd by 150 DIM (5.7 ± 2.0 vs. 19.0 ± 3.3%), greater risk of pregnancy after the first 2 services (49.6 ± 4.5 vs. 33.6 ± 3.9%), and greater milk by 22 wk postpartum (6.7 ± 0.2 vs. 5.3 ± 0.2 × 103 kg) than cows diagnosed with a clinical disease that had an AMD alert. Adding AMD-generated health alerts to the health screening of postpartum cows in a herd with an existing screening program did not improve survival, milk yield, or reproductive success. In addition, AMD alerts in cows diagnosed with a clinical disease may be indicative of the future success of such cows.

Effects of metritis treatment strategies on health, behavior, reproductive, and productive responses of Holstein cows

Our objectives were to compare the effects of ceftiofur crystalline free acid (CCFA) and ampicillin trihydrate (AMP) treatments of cows diagnosed with metritis on uterine health, behavior, reproductive, and productive responses. A controlled randomized clinical trial was designed. Metritis was defined as vaginal discharge (VD) = 5 (fetid, watery, red/brown) within 21 d in milk (DIM) and rectal temperature (RT) <39.5°C, whereas VD = 5 and RT ≥39.5°C was defined as puerperal metritis. On the day of diagnosis (d 0), cows were paired by parity and severity of metritis (metritis vs. puerperal metritis) and assigned randomly to the AMP and CCFA treatments. Cows enrolled in the AMP (n = 308) treatment were moved to a nonsalable-milk pen, where they were treated once daily for 5 d, and were moved back to their original pen 72 h after the last treatment (d 7). Cows enrolled in the CCFA (n = 310) treatment remained in their original pen and received 2 treatments of CCFA, 72 h apart. Rectal temperature was measured daily from d 0 to 6 and on d 11. Vaginal discharge was evaluated on d 4, 6, and 11 to assess cure. Cure was defined as the absence of treatment with additional antimicrobial before experiment d 11, VD <5, and RT <39.5°C. Cows were examined at 28 ± 3 DIM for purulent VD (PVD) and at 35 ± 3 DIM for cytological endometritis. Pregnancy was diagnosed at 40 ± 3 and 60 ± 7 d after first and second artificial inseminations. Cure of metritis did not differ between treatments on d 11 (AMP = 64.6 ± 3.1, CCFA = 63.5 ± 3.1%). Cows treated with AMP had greater RT from experiment d 1 to 6 compared with cows treated with CCFA (AMP = 39.1 ± 0.02, CCFA = 39.0 ± 0.02°C). Cows in the AMP treatment had greater prevalence of PVD at 28 ± 3 DIM (AMP = 82.6 ± 2.3, CCFA = 74.4 ± 2.7%) and tended to have greater prevalence of cytological endometritis at 35 ± 3 DIM (AMP = 77.8 ± 6.2 vs. CCFA = 61.7 ± 7.5%) than CCFA-treated cows. Treatment did not affect the hazard of pregnancy among multiparous cows; however, among primiparous cows, CCFA treatment reduced the hazard of pregnancy and increased the median days to pregnancy (AMP = 145 vs. CCFA = 169 d). Finally, average daily milk yield up to 14 wk postpartum was not affected by treatment (AMP = 38.0 ± 0.4, CCFA = 37.5 ± 0.4 kg). We conclude from the current experiment that CCFA was more effective in reducing RT and improving uterine health of metritic cows; however, the improved hazard of pregnancy of primiparous cows treated with AMP is important and warrants further investigation.

Seasonal heat load is more potent than the degree of body weight loss in dysregulating immune function by reducing white blood cell populations and increasing inflammation in Holstein dairy cows

We investigated the effects of seasonal heat stress and the degree of body weight (BW) loss postpartum (PP) on immune and inflammatory markers in transition dairy cows. Blood sampled twice weekly during the transition period (3 wk prepartum to 3 wk PP) from 12 peripartum Holstein cows in summer and 12 in winter was analyzed for plasma nonesterified fatty acids, the oxidative stress marker malondialdehyde (MDA), haptoglobin, and tumor necrosis factor-α (TNF-α). Complete blood counts and white blood cell (WBC) subpopulations (CD4, CD8, CD25, WC1, and CD335) were examined weekly during the transition period. Adipose tissue biopsies were taken at 7 d PP from summer-calving cows. In cows calving during seasonal heat stress, plasma TNF-α concentrations were 4.9-fold higher prepartum and 5.1-fold higher PP, and MDA concentrations were higher prepartum and PP than in winter. Hematocrit was lower prepartum and tended to be lower PP in summer compared with winter, and after adjusting to hematocrit values fewer WBC prepartum (-17%) and PP (-22%), lower monocytes prepartum (-37%) and PP (-49%), and fewer neutrophils (-25%) PP were found in summer compared with winter-calving cows. The percentages of cytotoxic T cells (CD8) and natural killer cells (CD335) were lower, as well as prepartum gamma delta-T cells (WC1) in summer compared with winter-calving cows. Moreover, lower red blood cell counts prepartum and mean platelet volume and red blood cell distribution width prepartum and PP, as well as higher hemoglobin prepartum and PP, were found in summer than winter-calving cows. In summer, cows that lost more BW PP had higher plasma MDA and TNF-α concentrations, fewer basophil numbers prepartum, a decrease in the percentage of cells expressing theIL-2 receptor (CD25), increased ratio of T helper cells (CD4):CD8, and a tendency for increased WC1 percentages in blood prepartum, as well as a higher percentage of WC1 PP and higher abundance of TNF-α and lower lipopolysaccharide-binding protein in adipose tissue compared with cows that lost less BW PP. In conclusion, seasonal heat stress during the peripartum period was associated with lower WBC counts coupled with increased levels of subacute inflammation, indicating dysregulation of immune function, whereas the degree of BW loss had minor effects. This suggests that heat load is a more potent stressor associated with immune function than the variation in the degree of BW loss PP in high-yielding peripartum dairy cows.

Endocrine and metabolic responses to glucose, insulin, and adrenocorticotropin infusions in early-lactation dairy goats of high and low milk yield

This experiment aimed to examine endocrine and metabolic responses to glucose, insulin, and adrenocorticotropin (ACTH) infusions in early-lactation dairy goats of different levels of milk production (LMP). Goats were grouped as either high (HY; 4.0 L/d, n = 13) or low milk yield (LY; 2.4 L/d, n = 13). Individual milk yield (L/d) and dry matter intake (DMI; kg/d) were measured daily. Concentration (mM) of glucose, fatty acids, and β-hydroxybutyrate, percent of milk fat and protein, body weight (BW; kg), and body condition score (BCS) were assessed weekly (from 2-6 wk postpartum). An intravenous glucose tolerance test (IVGTT), an insulin tolerance test (ITT), and an ACTH stimulationtest were carried out at 43, 44, and 45 ± 0.7 d in milk, respectively. The HY goats had greater milk yield (+67%), energy-corrected milk (ECM; +70%), DMI (+28%), ratio of ECM output to metabolic BW (+67%), and feed efficiency (+25%), but lesser BCS than LY goats (2.4 vs. 2.6). The DMI (% of BW) was moderately correlated with ECM (r = 0.70) and negatively correlated with BCS (r = -0.57). At the time of the IVGTT, HY goats had lesser basal insulin and glucose than LY goats. However, results from IVGTT and ITT indicate that the sensitivity of peripheral tissues to insulin was unaffected by LMP. Compared with LY, HY goats had lesser insulin secretion (-52%) and greater insulin clearance rate (+47%) after glucose infusion. The ITT and ACTH stimulation test results show that both the growth hormone response to insulin and the cortisol response to ACTH were unaffected by LMP. Also, basal plasma concentrations of GH and cortisol were not correlated with glucose and fatty acids concentrations or any performance traits. Collectively, our results suggest that differences between HY and LY goats, concerning milk yield and feed efficiency, were probably more closely related to differences in insulin secretion and clearance than to differences in peripheral tissue responsiveness to the effects of catabolic and anabolic hormones.

Acute phase proteins and markers of oxidative status in water buffalos during the transition from late pregnancy to early lactation

The transition period, from pregnancy to lactation, implies comprehensive metabolic and endocrine changes including a systemic inflammatory reaction and oxidative stress around calving in dairy cows. The aim of the present study was a longitudinal characterization of the serum concentration of acute phase proteins (APP), i.e., haptoglobin (Hp), serum amyloid A (SAA) and acidic glycoprotein (AGP), as well as of markers for oxidative stress in another large dairy animal, i.e. water buffalo, during the transition from late pregnancy to early lactation. As indicators of oxidative status, derivatives of reactive oxygen metabolites (dROM), ferric reducing ability (FRAP), thiobarbituric acid reactive substances (TBARS), and advanced oxidation protein products (AOPP) were determined in serum. Indicators for metabolic stress included nonesterified fatty acids (NEFA), ß-hydroxybutyrate (BHB) and adiponectin. Bovine specific ELISA methods for Hp and adiponectin were adapted and validated for their application to water buffalo samples. Blood samples were collected weekly from 11 pluriparous water buffalo cows (lactation number 4.6 ± 1.6; daily milk yield 9.0 ± 1.9 kg; means ± SD) from 6 weeks (wk) ante partum (ap) until 8 wk post partum (pp). The maximum concentrations of Hp were observed in wk 1 pp, followed by a decrease towards values lower than before calving starting from wk 3 pp. The concentrations of SAA also peaked in wk 1 pp and then returned to basal values. The AGP serum concentrations increased suddenly from the first to the second wk pp and remained elevated for all the observation period.Indicators of oxidative status which changed in concentration during the transition period were dROM, AOPP and the oxidative stress index (OSi) (dROM/FRAP ratio). Briefly, dROM and AOPP values were lower pp as compared to ap, and OSi was largely following the pattern of dROM due to the constant FRAP values. The TBARS values did not change during the observation period.From the metabolic indicators, adiponectin was not changing with time, whereas greater NEFA and BHB values were observed ap than pp. The time course of NEFA and of some indicators for oxidative status (dROM, OSi and AOPP) point to greater metabolic load in late pregnancy as compared with the first wk of lactation - contrary to the common situation in dairy cows. Both BHB and NEFA values remained below the thresholds applied for dairy cows to define subclinical or clinical ketosis, thus indicating that the buffaloes included in this study were not under metabolic stress. The increase in concentration of the APP around calving supports the concept that an inflammatory reaction is a physiological epiphenomenon of the onset of lactation in water buffalos that is independent of metabolic stress.

Prepartum Fat Mobilization in Dairy Cows with Equal Body Condition and Its Impact on Health, Behavior, Milk Production and Fertility during Lactation.

Simple SummaryAn excess of lipolysis and subsequent increase on non-esterified fatty acids concentrations may impair animal health, welfare, and productivity after calving. In this study, we evaluated the effect of fat mobilization in dairy cows with a recommended body condition score at the beginning of the close-up period on blood indicators of health, incidence of diseases, behavior, milk production, and fertility during postpartum. An increased fat mobilization in dairy cows with an equal body condition score modified the inflammatory and oxidative stress responses during the early postpartum without impairing their health status and fertility. Moreover, behavior and milk production were sensitive indicators that reflected the negative effects of the excess of prepartum fat mobilization through lactation.The objective of this study was to evaluate the effect of two levels of fat mobilization at the close-up period in dairy cows with an equal body condition score (BCS = 3.0) on the circulating concentrations of metabolic, inflammatory, and oxidative stress biomarkers, incidence of diseases, behavior, milk production, and fertility during the postpartum. Late-gestation multiparous Holstein cows (n = 59) with a body condition score of 3.0 (5-point scale) were enrolled at the beginning of the close-up period and then were followed during the entire lactation. Cows were retrospectively allocated into two groups: animals with prepartum non-esterified fatty acids concentration over 0.3 mmol/L were categorized as high fat mobilization (HFM) (n = 26), and below this threshold as low fat mobilization (LFM) (n = 33). Blood samples were collected 21 d before expected calving and once weekly for 3 wk postpartum in order to analyze β-hydroxybutirate, haptoglobin, fibrinogen, total proteins, and malondialdehyde. Health was observed daily for 21 d postpartum. Behavioral data was collected with an accelerometer and milk production and fertility were obtained from the farm records. An increased fat mobilization in dairy cows with equal BCS modified the inflammatory and oxidative stress responses during the early postpartum without impairing their health status and fertility. Moreover, milk production and behavior were markedly affected by excessive prepartum fat mobilization through lactation.

Milk 13C and 15N discriminations as biomarkers of feed efficiency and energy status in early lactation cows

Measuring feed utilization efficiency (FUE, milk production / feed intake) and energy status of dairy cows is essential to optimize dairy management and reduce feed waste, but FUE measurement is laborious and costly for dairy farms, particularly when cows are fed in groups. Our study was to investigate whether milk 13C and 15N stable isotope discriminations (naturally existing isotope; milk δ13C or δ15N – diet δ13C or δ15N) were used as biomarkers to estimate FUE, energy balance, and milk production of early lactation cows. Ten multiparous lactating cows with similar body weight (mean ± SD; 691 ± 18.0 kg) and parity (1.6 ± 0.5) were fed a homogeneous diet from 4 to 11 wk postpartum. Body fat had 1.5‰ less δ13C (P < 0.001) and muscle had similar δ13C compared to animal diet. Muscle had more than 2‰ greater δ15N than the diet (P < 0.0001). Milk 13C discrimination was reduced (P = 0.04) and milk 15N discrimination was increased (P < 0.0001) as lactation progressed postpartum. In early lactation, milk 13C discrimination was positively correlated with net energy balance (NEl) (linear; R2 = 0.49), and milk 15N discrimination was negatively correlated with NEl balance (linear; R2 = 0.56). Meanwhile, milk 13C discrimination was negatively and linearly (R2 = 0.30) correlated with feed carbon utilization efficiency, and milk 15N discrimination was positively and linearly (R2 = 0.55) correlated with feed nitrogen utilization efficiency. For milk production, milk 13C discrimination was negatively and linearly correlated with the milk yield (R2 = 0.44), milk fat yield (R2 = 0.40) and lactose yield (R2 = 0.46). Milk 15N discrimination was positively and linearly correlated with the milk protein yield (R2 = 0.49). Collectively, our data revealed that milk 13C and 15N discriminations could be as biomarkers to estimate the FUE, energy status, and milk production of early lactating cows. Potentially, measuring milk isotopic discrimination may be also applied to understand energy and nutrient partitioning of internal C and N reservoirs (body fat and muscle) for supporting milk synthesis in early lactation.

Discovery of different metabotypes in overconditioned dairy cows by means of machine learning

Using data from targeted metabolomics in serum in combination with machine learning (ML) approaches, we aimed at (1) identifying divergent metabotypes in overconditioned cows and at (2) exploring how metabotypes are associated with lactation performance, blood metabolites, and hormones. In a previously established animal model, 38 pregnant multiparous Holstein cows were assigned to 2 groups that were fed differently to reach either high (HBCS) or normal (NBCS) body condition score (BCS) and backfat thickness (BFT) until dryoff at -49 d before calving [NBCS: BCS < 3.5 (3.02 ± 0.24) and BFT < 1.2 cm (0.92 ± 0.21), mean ± SD; HBCS: BCS > 3.75 (3.82 ± 0.33) and BFT > 1.4 cm (2.36 ± 0.35)]. Cows were then fed the same diets during the dry period and the subsequent lactation, and maintained the differences in BFT and BCS throughout the study. Blood samples were collected weekly from 7 wk antepartum (ap) to 12 wk postpartum (pp) to assess serum concentrations of metabolites (by targeted metabolomics and by classical analyses) and metabolic hormones. Metabolic clustering by applying 4 supervised ML-based classifiers [sequential minimal optimization (SMO), random forest (RF), alternating decision tree (ADTree), and naïve Bayes-updatable (NB)] on the changes (d 21 pp minus d 49 ap) in concentrations of 170 serum metabolites resulted in 4 distinct metabolic clusters: HBCS predicted HBCS (HBCS-PH, n = 13), HBCS predicted NBCS (HBCS-PN, n = 6), NBCS predicted NBCS (NBCS-PN, n = 15), and NBCS predicted HBCS (NBCS-PH, n = 4). The accuracies of SMO, RF, ADTree, and NB classifiers were >70%. Because the number of NBCS-PH cows was low, we did not consider this group for further comparisons. Dry matter intake (kg/d and percentage of body weight) and energy intake were greater in HBCS-PN than in HBCS-PH in early lactation, and HBCS-PN also reached a positive energy balance earlier than did HBCS-PH. Milk yield was not different between groups, but milk protein percentage was greater in HBCS-PN than in HBCS-PH cows. The circulating concentrations of fatty acids (FA) increased during early lactation in both groups, but HBCS-PN cows had lower concentrations of β-hydroxybutyrate, indicating lower ketogenesis compared with HBCS-PH cows. The concentrations of insulin, insulin-like growth factor 1, leptin, adiponectin, haptoglobin, glucose, and revised quantitative insulin sensitivity check index did not differ between the groups, whereas serum concentrations of glycerophospholipids were lower before calving in HBCS-PH than in HBCS-PN cows. Glycine was the only amino acid that had higher concentration after calving in HBCS-PH than in HBCS-PN cows. The circulating concentrations of some short- (C2, C3, and C4) and long-chain (C12, C16:0, C18:0, and C18:1) acylcarnitines on d 21 pp were greater in HBCS-PH than in HBCS-PN cows, indicating incomplete FA oxidation. In conclusion, the use of ML approaches involving data from targeted metabolomics in serum is a promising method for differentiating divergent metabotypes from apparently similar BCS phenotypes. Further investigations, using larger numbers of cows and farms, are warranted for confirmation of this finding.

Influence of corn supplementation to beef cows during mid- to late-gestation: maternal feed intake, body condition, plasma metabolites, and calf growth

To determine the effect of supplementing dry-rolled corn during mid- to late-gestation in cows fed low-quality forage on feed intake, body weight (BW), and body condition score (BCS), plasma metabolites, and neonatal growth, 47 multiparous cows (661 ± 7.8 kg BW, 5.2 ± 0.1 BCS, 7.5 ± 0.2 yr of age) predominately of Angus breeding and carrying male calves were assigned randomly to supplement treatments: 0% (unsupplemented; n = 24) or 0.2% of BW of dry-rolled corn (supplemented; DM basis; n = 23) from d 110 to 265 of gestation. All cows were fed the same basal forage consisting of grass hay, wheat straw, and sugar beet concentrated separator byproduct. Feed intake was monitored individually from d 110 of gestation until d 21 post-calving. Cows on the supplemented treatment were individually allowed access to 1 Insentec feeder containing corn per pen. Body weight and BCS were determined every 28 d until d 240 of pregnancy, at parturition, and at 3 wk postpartum. Jugular blood samples also were collected every 28 d during the gestation period. Calves were weighed at birth, 3 wk postpartum, and at weaning (d 168). Data were analyzed using the MIXED procedure of SAS. Corn supplementation increased (P ≤ 0.008) BW and BCS change during gestation and tended (P = 0.07) to decrease gestation length. Corn supplementation decreased (P = 0.002) dry matter intake (DMI) of the basal forage but did not influence (P = 0.84) total DMI (forage + corn) during gestation. Plasma glucose was not influenced (P = 0.12) by supplemental treatment and plasma urea N and non-esterified fatty acids were greater (P ≤ 0.02) in unsupplemented than supplemented cows. Body weight and BCS change, and DMI in cows from birth until 21 d of lactation were not influenced (P ≥ 0.27) by supplementation treatment. Corn supplementation during gestation did not affect (P = 0.76) calf birth weight. At 3 wk postpartum calves from supplemented cows were heavier (P = 0.04) but corn supplementation did not influence BW (P = 0.76) at weaning. Overall, supplementation of dry-rolled corn at 0.2% of BW during mid- to late-gestation likely did not elicit large enough differences in energy status to have lasting effects on calf BW. However, corn supplementation at low levels during gestation may be useful as a substitute for forage to improve energy status of the cow while decreasing the need for forage.

Effect of stearic or oleic acid on milk performance and energy partitioning when fed in diets with low and high rumen-active unsaturated fatty acids in early lactation.

This experiment was conducted to determine the effects of stearic acid (SA; C18:0) or rumen-protected oleic acid (OA; C18:1 cis-9) on milk performance and energy partitioning of early lactation cows when supplemented in diets with low and high level of rumen unsaturated fatty acids (RUFA). In low RUFA experiment (LRUFA), FA supplement rich in either SA or calcium salts OA was added to a basal diet with a low concentration of RUFA (0.75% vs. 1.4%, LRUFA-SA vs. LRUFA-OA). In high RUFA experiment (HRUFA), 2% soybean oil was added to the diet fed in the LRUFA experiment. In each experiment, 30 multiparous cows were blocked by parity and predicted transmitting ability for milk yield and were randomly fed 1 of 2 treatment diets from 2 to 13 wk postpartum. In the LRUFA experiment, LRUFA-SA had 2.4 kg/d more dry matter intake (DMI) (P < 0.01), 3.8 kg/d more energy-corrected milk (P < 0.01), and 0.3% units more milk fat percentage (P < 0.01) and 0.2 kg/d more milk fat yield (P < 0.01). Dietary treatments did not affect body weight, energy balance, and energy intake partitioning into milk, maintenance, and body tissues (P > 0.1). In the HRUFA experiment, HRUFA-SA had 1.4 kg/d more DMI (P = 0.03) but similar milk and milk components yields (P > 0.1). HRUFA-SA had a tendency to gain more body weight (P = 0.07) and had more positive energy balance (P = 0.01) and decreased gross feed efficiency (milk yield/DMI) (P = 0.01). Consistently, HRUFA-SA increased intake energy partitioning into body tissues (P = 0.02) and decreased energy partitioning into milk (P = 0.01). In summary, SA supplementation had more DMI relative to OA, but the effects on milk and milk fat production were different and affected by the level of RUFA in the basal diet. In application, SA supplementation was more effective to improve milk production when included in the basal diet with the low RUFA.

Randomized controlled trial investigating the effects of a breastfeeding relaxation intervention on maternal psychological state, breast milk outcomes, and infant behavior and growth

Biological signaling and communication between mothers and infants during breastfeeding may shape infant behavior and feeding. This signaling is complex and little explored in humans, although it is potentially relevant for initiatives to improve breastfeeding rates. The aim of this study was to investigate physiological and psychological aspects of mother-infant signaling during breastfeeding experimentally, testing the effects of a relaxation intervention on maternal psychological state, breast milk intake, milk cortisol levels, and infant behavior and growth. Primiparous breastfeeding mothers and full-term infants were randomly assigned to receive relaxation therapy [intervention relaxation group; n=33 (RG)] or to the control group [n=31 (CG); no relaxation therapy] at 2 wk postpartum. Both groups received standard breastfeeding support. Home visits were conducted at 2 (HV1), 6 (HV2), 12 (HV3) and 14 (HV4) wk to measure maternal stress and anxiety, breast milk intake and milk cortisol, and infant behavior and growth. RG mothers had lower stress scores postintervention than the CG (HV3 ∆=-3.13; 95% CI: -5.9, -0.3) and lower hindmilk cortisol at HV1 (∆=-44.5%; 95% CI: -76.1%, -12.9%) but not at HV2. RG infants had longer sleep duration (∆=82 min/d; 95% CI: 16, 149 min/d) at HV2 and higher gains in weight and body mass index standardized deviation score than the CG infants (∆=0.76; 95% CI: 0.3, 1.22; and ∆=0.59; 95% CI: 0.09, 1.1, respectively). RG infants had a mean milk intake at HV3 that was 227 g/d higher than that of the CG infants (P=0.031) after controlling for gender and milk intake at HV1. The trial shows the effectiveness of a simple relaxation intervention for improving maternal and infant outcomes and identifies some potential signaling mechanisms for investigation in future and larger studies, especially in settings where mothers are more stressed, such as those with preterm or low birth weight infants. This trial was registered at clinicaltrials.gov as NCT01971216.

Comparative analyses of estimated and calorimetrically determined energy balance in high-yielding dairy cows

Our aim was to compare the energy balance estimated (EBest) according to equations published by various energy feeding systems (German Society for Nutrition Physiology, French National Institute for Agricultural Research, and US National Research Council) and the EB calculated by use of calorimetrically measured heat production (EBhp) of 20 high-yielding (≥10,000 kg/305 d) German Holstein cows at -4 (pregnant, nonlactating) and 2 wk (early lactation) relative to parturition. In addition to heat production, feed and water intake, physical activity (including standing-lying behavior), body weight, body condition score, body temperature, plasma concentrations of fatty acids and β-hydroxybutyrate, milk yield, and milk composition were measured to characterize the metabolic status. The EBhp was balanced [2.74 ± 4.09 MJ of metabolizable energy (ME)/d; ±standard error] before calving, but strongly negative (-84.7 ± 7.48 MJ of ME/d) at wk 2 of lactation. At both time points, EBhp and EBest differed significantly. On average, the equations overestimated the antepartum EB by 33 MJ of ME/d and underestimated the postpartum negative EB by 67 MJ of ME/d, respectively. Because the same ME intake and energy-corrected milk values were used for calculation of EBest and EBhp in our study, we considered that the factors (0.488 to 0.534 MJ of ME/kg0.75) currently used to calculate the ME requirements for maintenance probably underestimate the needs of high-yielding dairy cows, particularly during early lactation. In accord, heat production values determined under standard conditions of thermoneutrality and locomotion restriction amounted to 0.76 ± 0.02 MJ of ME/kg0.75 (4 wk antepartum) and 1.02 ± 0.02 MJ of ME/kg0.75 (2 wk postpartum), respectively. The expected positive correlation between EBhp and DMI was observed in pregnant cows only; however, a bias of 26 MJ of ME/d between mean actual energy intake and ME intake predicted according to German Society for Nutrition Physiology was found in cows at wk 4 antepartum. At both investigated time points, mobilization of tissue energy reserves (reflected by plasma fatty acid concentration) was related to EBhp. In early lactating cows, metabolic body weight (kg0.75) and the percentage of milk fat showed the strongest correlation (correlation coefficient = -0.70 and -0.73) to EBhp. Our findings must be taken into account when experimental data are interpreted because the true energy status might be significantly overestimated when EBest is used.

Effects of pegbovigrastim administration on periparturient diseases, milk production, and reproductive performance of Holstein cows

The aim of this study was to evaluate the effects of treating Holstein cows with pegbovigrastim on periparturient diseases, milk production, and reproductive performance while exploring the mode of action of an immunomodulatory protein. Cows were randomly allocated to 1 of 2 treatments, untreated control (CTR, n = 423) and pegbovigrastim (PEG, n = 417). At 7 d from the anticipated calving date (d -7), cows allocated to PEG received a subcutaneous injection of 15 mg of pegylated recombinant bovine granulocyte colony stimulating factor (pegbovigrastim injection, Imrestor, Elanco Animal Health, Greenfield, IN). A second injection was administered within 24 h after calving (d 0). Blood samples were obtained from a subset of cows (CTR, n = 103; PEG, n = 102) at -7 and 0, 3, 7, and 14 d relative to parturition. Samples were used for hemogram and quantification of haptoglobin, nonesterified fatty acids, β-hydroxybutyrate, and trace and macro minerals. Vaginal cytobrush was performed on the same subset cows at d 0, 7, and 14 to assess the relative neutrophil count. Additionally, colostrum samples were collected to measure IgG, IgM, IgA, and lactoferrin concentrations. Postpartum disease occurrence was recorded from calving until 30 d in milk (DIM). Weekly milk yield was recorded for the first 12 wk after calving. Cows treated with PEG had a 3- to 4-fold increase in circulating polymorphonuclear leukocyte, lymphocyte, and monocyte numbers, with a peak at 3 d after treatment followed by a gradual decline, but the counts remained significantly greater compared with CTR at 14 DIM. The administration of PEG did not affect the incidence of clinical and subclinical mastitis, retained fetal membranes, metritis, puerperal metritis, and endometritis. Primiparous cows treated with PEG tended to have lower odds of developing hyperketonemia than CTR [odds ratio (OR) = 0.57, 95% confidence interval (CI) = 0.23 to 1.42]. Cows treated with PEG had higher odds of being diagnosed with lameness within 30 DIM compared with CTR (OR = 1.79, 95% CI = 1.16 to 2.76); however, we found no significant differences by 60 DIM. Treatment with PEG increased the odds of displaced abomasum (OR = 8.27, 95% CI = 1.02 to 66.6). Cows treated with PEG had higher odds of being diagnosed with 1 or more clinical diseases compared with CTR cows (OR = 1.39, 95% CI = 1.02 to 1.90). We observed no differences in linear scores or milk composition between treatments. Furthermore, primiparous cows treated with PEG produced more milk than CTR primiparous cows during the first 12 wk postpartum (PEG = 37.51 ± 0.66; CTR = 35.91 ± 0.65 kg), but no differences were observed on energy-corrected milk. Treatment did not alter reproductive performance; additionally, cows diagnosed with metritis or puerperal metritis and treated with PEG tended to have higher proportion of neutrophils in the vaginal mucosa when compared with CTR metritic cows. Although PEG treatment increased circulating polymorphonuclear leukocyte, monocyte, and lymphocyte numbers, as expected, it was detrimental to cow health because it increased morbidity.

MILK COMPOSITION OF INDIAN RHINOCEROS ( RHINOCEROS UNICORNIS) AND CHANGES OVER LACTATION.

The objective of this study was to determine the major nutrient composition of Indian rhinoceros milk ( Rhinoceros unicornis) over the first 13 mo of an 18-mo lactation period and to compare the results to those of previous studies on rhinoceros, African elephant ( Loxodonta africana), and horse milk ( Equus ferus caballus). The following parameters were measured: dry matter (DM), crude ash (ASH), crude protein (CP), ether extract (EE), nitrogen-free extract (NFE; calculated), lactose, calcium (Ca), phosphorus (P), magnesium (Mg), fatty acids (FAs), and gross energy (GE). DM, ASH, CP, and EE were determined with a proximate analysis, lactose with infrared spectroscopy and an enzymatic method, minerals with an autoanalyzer, FA with gas chromatography, and GE with bomb calorimetry. Milk samples were collected from two Indian rhinoceros cows from Zoo Basel. Rhino A gave birth to her third calf on 10 September 2012; three samples were collected and analyzed (colostrum, milk 1 wk and 2 wk postpartum). Rhino B gave birth to her eighth calf on 05 October 2013; samples were collected and 15 were chosen for the analyses (from colostrum to 13 mo postpartum). The composition of rhino B's colostrum was 13.8% DM (wet-weight basis), 4.8% ASH, 61.8% CP, 0.7% EE, 32.6% NFE, 26.7% lactose, 0.59% Ca, 0.54% P, 0.2% Mg (DM basis), and 20.3 MJ GE/kg DM. Rhino B's sample collected 13 mo postpartum averaged 8.0% DM (wet-weight basis), 3.6% ASH, 16.3% CP, 1.8% EE, 78.3% NFE, 84.7% lactose, 0.54% Ca, 0.48% P, 0.09% Mg (on DM basis), and 17.43 MJ GE/kg DM. The main FAs in rhino B's and rhino A's samples were C10 : 0, C12 : 0, C16 : 0, C18 : 1n9c, and C18 : 2n6c. Milk of the Indian rhinoceros is low in fat and protein but high in lactose, which is comparable to the milk composition of other rhinoceros species and horses, but not African elephants.

Serum hepatokines in dairy cows: periparturient variation and changes in energy-related metabolic disorders

BackgroundDuring peripartum period, dairy cows are highly susceptible to energy metabolism disorders such as fatty liver and ketosis. Angiopoietin-like protein 4 (ANGPTL4) and fibroblast growth factor 21 (FGF21), known as hepatokines, play important roles in lipid metabolism. The purposes of our study were to evaluate variations of serum ANGPTL4 and FGF21 concentrations in periparturient dairy cows and changes in these serum analyte concentrations of energy-related metabolic disorders in early lactation dairy cows.This study was divided into two experiments. Experiment I: Blood parameters were measured in healthy periparturient Holstein cows from 4 wk antepartum to 4 wk postpartum (n = 219). In this experiment, weekly blood samples were obtained from 4 wk before the expected calving date through 4 wk after calving. Experiment II: Blood parameters were measured in healthy cows (n = 30) and cows with clinical ketosis (n = 29) and fatty liver (n = 25) within the first 4 wk of lactation. In the present study, all blood samples were collected from the coccygeal vein in the early morning before feeding.ResultsSerum ANGPTL4 and FGF21 concentrations peaked at parturition, and declined rapidly over the following 2 wk Serum ANGPTL4 and FGF21 concentrations were positively correlated with serum non-esterified fatty acids (NEFA) concentration (r = 0.856, P = 003; r = 0.848, P = 0.004, respectively). Cows with clinical ketosis and fatty liver had significantly higher serum ANGPTL4 and FGF21 concentrations than healthy cows (P < 0.01).ConclusionSerum ANGPTL4 and FGF21 concentrations were elevated during peripartum period, suggesting that energy balance changes that were associated with parturition contributed significantly to these effects. Although FGF21 and ANGPTL4 could play important roles in the adaptation of energy metabolism, they may be involved in the pathological processes of energy metabolism disorders of dairy cows in the peripartum period.

Short communication: Associations of udder edema with health, milk yield, and reproduction in dairy cows in early lactation

Udder edema (UE) is a common condition of cows around calving, but its effects are not well characterized. The objectives of this study were to determine the associations of UE with the incidence of health disorders and with milk yield and reproduction in dairy cows in early lactation. On 3 commercial farms, UE was scored weekly on 1,346 cows, on a scale of 0 to 3, from 1 wk before calving to 3 wk after calving. Among cows with complete UE scores, 30% never had edema, 12% had edema only prepartum, 11% had it only postpartum, and 48% had edema prepartum and in at least 1 wk postpartum. Udder edema was associated with a greater incidence of clinical mastitis before 30 d in milk (5 vs. 2%). Subclinical ketosis (blood β-hydroxybutyrate ≥1.2 mmol/L) was more prevalent at wk 2 (11 vs. 6%) postpartum among cows with UE. No association was observed of UE with other diseases or culling in early lactation. In a subset of 912 cows with complete UE and 3 test-days of milk yield data, differences were observed in yield at test d 1 among UE categories. Cows with UE only prepartum produced less milk (39.9 kg/d) than cows with UE postpartum only (42.4 kg/d) and cows with UE both prepartum and postpartum (41.6 kg/d), none of which differed from cows without UE (40.9 kg/d). Udder edema was not associated with the prevalence of anovulation, or the time to or probability of pregnancy at first insemination, yet to 300 d in milk, cows that had UE postpartum had a shorter time from calving to pregnancy than cows without UE. The associations of UE with health and productivity are mixed, and the mechanisms underlying UE and its effects merit further investigation.

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

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

Short communication: Relationship between body condition score and plasma adipokines in early-lactating Holstein dairy cows

We hypothesized that plasma adipokine concentrations of early-lactation dairy cows are related to body condition score (BCS) at calving and to markers of metabolic status of the cow. As part of a larger study with 117 multiparous Holstein dairy cows, which had high BCS (BCS >4.0) or normal BCS (3.25-3.5) at calving, 22 cows were randomly selected (n = 11 per group) to be enrolled in this study. Cows were divided into 2 groups based on their BCS at calving: (1) normal BCS with BCS of 3.35 ± 0.13 (mean ± SD) and (2) high BCS cows with BCS of 4.14 ± 0.17. The 22 selected animals did not have a clinically diagnosed health problem after calving. Blood samples were taken right after calving (d 1) and before morning feeding on d 8, 15, and 21 postpartum concurrently with body condition scoring for all cows. Blood samples were analyzed for plasma adiponectin, leptin, tumor necrosis factor-α, and IL-6. The mean BCS remained highest in high-BCS cows during the first 21 d in milk. Leptin concentrations decreased progressively for all cows after calving. However, differences in BCS at calving were not related to leptin concentrations. Adiponectin, IL-6, and tumor necrosis factor-α concentrations were neither influenced by days in milk nor BCS after calving. Leptin and the leptin-to-adiponectin ratio did not show any correlation at any time point during the first 21 d in milk with plasma concentrations of nonesterified fatty acids or β-hydroxybutyrate, which are considered as markers of metabolic status. Only for IL-6 at d 8 did we find a strong correlation with metabolic status indicators. In conclusion, plasma adipokine concentrations during the first 3 wk postpartum were not related to BCS in lactating Holstein cows that were clinically healthy at calving.

Lactating Canadian Women Consuming 1000 µg Folic Acid Daily Have High Circulating Serum Folic Acid Above a Threshold Concentration of Serum Total Folate

Consumption of high-dose folic acid supplements is common throughout pregnancy and lactation in several countries, including Canada, Brazil, and the United States, and may lead to high levels of circulating unmetabolized folic acid. The objective of the study was to characterize serum and whole-blood folate forms in Canadian lactating women regularly consuming a daily high-dose folic acid supplement. One-hundred and seventeen Canadian lactating women aged between 18 and 42 y, with a geometric mean ± SD prepregnancy body mass index (kg/m2) of 23.1 ± 1.2, were enrolled in a vitamin D supplementation trial between 13 and 22 wk of gestation. As part of the trial, the women received a daily multivitamin containing 1000 µg folic acid throughout pregnancy and lactation until 8 wk postpartum. At 8 wk postpartum, serum folate forms, including folic acid and RBC total folate, were determined from nonfasted blood samples. Differences in median folate vitamer concentrations among quintiles of serum total folate status were assessed by the Wald test and quantile regression methods. A breakpoint in the relation between serum folic acid and serum total folate was modeled with the use of the segmented package in R. Median serum total folate concentration among participants was 79.3 nmol/L (5th-95th percentile 30.7-186 nmol/L) and median RBC folate concentration was 2790nmol/L (5th-95th percentile 1330-4850nmol/L). There was a breakpoint in the relation between serum total folate and serum folic acid at 78.5 nmol/L (95% CI: 67.9, 89.1 nmol/L), below which serum folic acid was not associated with serum total folate, and above which serum folic acid increased 0.78 nmol/L (95% CI: 0.70, 0.86 nmol/L; P<0.001) for each 1 nmol/L increase in serum total folate. These data demonstrate the potential for high serum folic acid concentrations proportional to overall folate concentrations in lactating women with serum total folate >80 nmol/L taking high-dose supplemental folic acid. This study was registered at clinicaltrials.gov as NCT01112891.