Endocrine and metabolic changes in transition dairy cows are affected by prepartum infusions of a serotonin precursor

Serotonin (5-HT) has been shown to be involved in calcium homeostasis, modulating calcium concentration in blood. In addition, 5-HT participates in a variety of metabolic pathways, mainly through the modulation of glucose and lipid metabolism. The hypothesis of the present study was that the prepartum administration of 5-hydroxy-l-tryptophan (5-HTP), a 5-HT precursor, would affect endocrine systems related to calcium homeostasis, and interact with other endocrine and metabolic pathways during the transition period. In this study, 20 Holstein dairy cows were randomly assigned to 2 experimental groups. Both groups received a daily i.v. infusion of 1 L of either 0.9% NaCl (control group; n = 10) or 0.9% NaCl containing 1 mg of 5-HTP/kg of BW (5-HTP group, n = 10). Infusions started d 10 before estimated parturition date and ended the day of parturition, resulting in a minimum of 4 d of infusion (8.4 ± 0.7 d of infusion). Until parturition, blood samples were collected before the daily infusions, and postpartum daily until d 7, and on d 30. Plasma concentrations of parathyroid hormone (PTH) were transiently increased at parturition and on d 1 in control cows. In the 5-HTP group PTH remained unchanged. The concentration of pyridinoline (PYD), an established marker for calcium release from the bone to the bloodstream, increased on d 1 postpartum only in the 5-HTP group. In control cows, PYD concentrations did not change on d 1 postpartum. Melatonin concentrations were slightly but significantly increased in the 5-HTP group compared with the control group. Insulin concentrations decreased in both groups postpartum. Before parturition, leptin concentrations decreased in both groups and remained at this level until d 30 postpartum. Plasma IgG concentrations decreased in both groups on d ?1 postpartum. Haptoglobin increased in both groups on d ?1 and remained at this level until d 7 postpartum. No differences between groups were observed for insulin, glucagon, IgG, leptin, adiponectin, and haptoglobin concentrations. The results obtained in the present study evidenced that 5-HT is regulating calcium homeostasis independent of PTH. The lack of treatment effects on IgG and on other hormones and metabolites indicates that 5-HTP did not affect these other metabolic pathways and the IgG concentration during the transition period.     

Short communication: Circulating serotonin is related to the metabolic status and lactational performance at the onset of lactation in dairy cows

Serotonin (5-hydroxytryptamine, 5-HT) affects many physiological functions because it is involved in glucose and lipid metabolism, calcium homeostasis, and regulation of lactation in dairy cows. This study aimed to examine physiological differences in serum 5-HT concentrations (high vs. low) and their association with metabolic status and milk production at the onset of lactation. Twelve multiparous Holstein dairy cows were milked within 4 h of calving, and blood and milk samples were collected at the first 6 subsequent milkings after parturition and at the evening milkings on d 5, 8, 10, and 14. Cows were retrospectively divided into 2 groups (6 cows/group): low serum 5-HT (LSS) and high serum 5-HT (HSS) according to their calculated areas under the curve (AUC) for serum 5-HT for the entire experimental period (cut-off: 46,000 ng/mL × 324 h). Concentrations of 5-HT, free fatty acids (FFA), β-hydroxybutyrate (BHB), glucose, calcium, and IGF-1 were measured in blood. Milk was analyzed for fat, protein, lactose, and 5-HT concentrations. Milk yield was recorded at each milking and energy-corrected milk yield was calculated. Serum 5-HT concentrations were higher in HSS than in LSS [AUC (ng/mL × 324 h): 57,830 ± 4,810 vs. 25,005 ± 5,930]. The amount of energy-corrected milk was lower in HSS than in LSS. The HSS group produced less colostrum and had decreased milk yield, specifically during the first 6 milkings. Concentrations of FFA, BHB, and glucose in plasma did not differ between groups. Concentrations of IGF-1 in serum were elevated in HSS compared with LSS throughout the experiment. Total circulating calcium concentrations in serum tended to be higher in HSS than in LSS. Milk fat and protein yields were decreased in HSS compared with LSS. Milk 5-HT decreased overall during the experimental period, with LSS maintaining higher 5-HT concentrations than HSS until d 14 of lactation. In conclusion, cows with high serum 5-HT concentrations showed a reduced metabolic load at the onset of lactation, concomitantly lower milk yield, and a reduced energy output via milk.     

Short communication: Mammary gland tight junction permeability after parturition is greater in dairy cows with elevated circulating serotonin concentrations

After parturition, the start of copious milk production in dairy cows requires the closure of tight junctions (TJ) to form the blood–milk barrier and prevent paracellular transfer of blood constituents into milk [e.g., lactate dehydrogenase (LDH) and serum albumin (SA)] and vice versa [e.g., appearance of α-lactalbumin (α-LA) in blood]. Serotonin (5-HT) has been demonstrated to alter tight junction permeability in the mammary gland. The present study investigated individual differences of TJ permeability of mammary epithelium at the beginning of lactation in relation to circulating 5-HT in dairy cows. Blood and milk samples were obtained from 11 multiparous Holstein dairy cows for the first time at 4 h after parturition, at the following 5 milkings, and at the evening milkings on d 5, 8, 10, and 14 of lactation. Retrospectively, cows were split into 2 groups according to their calculated areas under the curve of serum 5-HT during the entire experimental period: a high-serum 5-HT (HSS) group (5 cows) and a low-serum 5-HT (LSS) group (6 cows). The areas under the curve of serum 5-HT concentrations over the 324-h experimental period were 62 ± 2 × 10³ ng/mL in HSS and 25 ± 5 × 10³ ng/mL in LSS. Plasma α-LA concentration was greater in LSS than in HSS cows at the first milking, but no difference between groups was found from the second to sixth milking. Yield of α-LA in milk was lower in HSS than in LSS during the first 6 milkings postpartum, especially in colostrum. Concentrations of α-LA, IgG₁, and IgG₂ in milk did not differ between groups during the entire experiment except for higher IgG observed in LSS than in HSS at the second milking and for higher IgG₂ found in HSS compared with LSS on d 5. In contrast, SA concentrations and LDH activity in milk were lower in LSS compared with HSS cows during the first 6 milkings postpartum, particularly in colostrum. Milk somatic cell count was higher in HSS than in LSS throughout the study. Higher circulating 5-HT concentrations were associated with an increased transfer of the paracellularly transported SA, LDH, and somatic cell count, especially at the first milking, suggesting that 5-HT affects TJ permeability during closure of the blood–milk barrier at the onset of lactation. Furthermore, higher serum 5-HT concentrations were associated with a lower α-LA yield in milk. A consistent relationship with serum 5-HT concentrations was neither observed for the transfer of IgG₂ nor the primarily transcellular transport of IgG₁ during the first milkings after parturition.     

Interaction of 5-hydroxy-l-tryptophan and negative dietary cation-anion difference on calcium homeostasis in multiparous peripartum dairy cows

Hypocalcemia affects almost 50% of all dairy cows. Our laboratory has previously demonstrated that infusions of the serotonin precursor 5-hydroxy-l-tryptophan (5-HTP) increase circulating calcium concentrations in the Holstein transition cow. It is unknown whether feeding a negative dietary cation-anion difference (DCAD) diet alters the relationship between 5-HTP and hypocalcemia. The main objective of this study was to determine whether feeding a negative DCAD (?DCAD) diet before calving in conjunction with 5-HTP treatment could further diminish the magnitude of hypocalcemia at the time of calving. We used a randomized complete block design with a 2 × 2 factorial arrangement. Thirty-one multiparous Holstein cows were fed either a positive (+13 mEq/100 g) or negative (?13 mEq/100 g) DCAD diet 21 d before parturition and were intravenously infused daily with saline or 5-HTP (1 mg/kg) starting 7 d before the estimated date of parturition. Cows were blocked by parity and were randomly assigned to 1 of 4 treatment groups: positive DCAD plus saline, positive DCAD plus 5-HTP, negative DCAD plus saline, and negative DCAD plus 5-HTP, resulting in n = 8 per group. Total calcium (tCa), ionized calcium (iCa), and feed intake were recorded. The iCa was elevated prepartum in the ?DCAD/5-HTP group compared with the other treatment groups as well as on d 0 and 1 postpartum. Although differences in tCa were not significant across the pre- or postpartum periods, tCa was numerically higher on d 0 and significantly higher on d 1 in ?DCAD/5-HTP cows compared with all other groups. Prepartum the ?DCAD/5-HTP treatment group ate less than the other treatment groups; however, postpartum dry matter intake differences were not significant. These findings demonstrate that feeding a ?DCAD diet in conjunction with 5-HTP prepartum can increase postpartum circulating iCa concentrations and therefore diminish the magnitude of hypocalcemia at the time of parturition.     

Short communication: Timing of first milking affects serotonin (5-HT) concentrations

Hormonal signals differentially regulate the timing of parturition, as well lactogenesis and, potentially, colostrum formation in the mammary gland. Non-neuronal serotonin (5-HT) is a homeostatic regulator of the mammary gland. In the current study, we manipulated the timing of first milking to investigate its effects on serum 5-HT and calcium concentrations in the maternal and calf circulation, as well as in colostrum. Twenty-three cows were randomly assigned to a control (CON; n = 10) group, milked for the first time at 4 h postcalving, or a treatment (TRT; n = 13) group, milked for the first time approximately 1 d before calving in addition to 4 h postcalving. Maternal blood samples were collected for 4 d precalving, 3 times daily, and 1 blood sample was taken 4 h postcalving. Calf blood samples were collected 4 (before first colostrum feeding) and 12 h after birth, and at 3 wk of age. Calves from both treatments were fed colostrum from their respective mothers. Serum 5-HT concentrations were greater in CON cows and decreased significantly in TRT cows after milking was initiated precalving (951 vs. 524 ± 111 ng/mL, respectively). Cow serum calcium concentrations were affected by time, beginning to decrease 1 d precalving until 4 h postcalving, but this drop in serum calcium was more pronounced in TRT cows. Serum 5-HT and calcium concentrations were negatively correlated (r = −0.57) for the CON cows and positively correlated (r = 0.6) for the TRT cows. Maternal calcium and 5-HT decreased similarly due to precalving milking. Calcium and 5-HT concentrations were greater in colostrum collected from TRT cows milked precalving. Overall, calves had higher circulating 5-HT concentrations than cows, and calves born to TRT cows had increased 5-HT concentrations compared with the CON. Precalving milking could affect 5-HT synthesis within the mammary gland and therefore affect maternal 5-HT and calcium concentrations. Further research is needed in ruminants to assess the extent of 5-HT placental transfer, its role on pre- and postnatal development of the calf, the importance of its presence in colostrum, and potential long-term effects on calf health.     

Short communication: Supplementation of colostrum and milk with 5-hydroxy-l-tryptophan affects immune factors but not growth performance in newborn calves

In ruminants, colostrum is the main source of immunoglobulins for the newborn animal, conferring immune protection until the immune system becomes active and able to synthesize its own immunoglobulins. Serotonin (5-HT), a biogenic amine derived from tryptophan, has stimulatory effects on many physiological processes, including components of the innate (mastocytes, eosinophils, and natural killer cells) and adaptive (T and B lymphocytes) immune systems. Based on the known effects of 5-HT on the immune system, we hypothesized that increased concentrations of 5-HT, through administration of its precursor 5-hydroxy-l-tryptophan (5-HTP), may positively affect development of the calf's immune system and therefore support health and growth performance during the first weeks of life. Eighteen calves were randomly assigned to 1 of 2 experimental groups (control and 5-HTP), resulting in n = 9 per treatment group. Both groups received 2 colostrum meals from a common pool of colostrum. Thereafter, calves were fed milk replacer twice daily for 30 d. In the 5-HTP group, colostrum and milk replacer were supplemented with 1.5 mg of 5-HTP/kg of birth weight during the first 15 d after birth. Body weight was recorded at birth and on d 5, 10, 15, and 30 after birth. Blood samples were collected every morning (0800 h) before feeding from birth until d 5 and then on d 7, 9, 11, 13, 15, and 30 after birth. Serum 5-HT concentrations were increased as a consequence of the 5-HTP supplementation. Plasma immunoglobulin G concentrations did not differ between groups throughout the experimental period. The blood mRNA abundance of several factors related to the innate and adaptive immune system [nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB), serum amyloid A-1 (SAA1), chemokine C-C motif ligand 5 (CCL5), cyclooxygenase 2 (PTGS2), haptoglobin (HP), and IL-1β] were increased in calves supplemented with 5-HTP. Supplementation of 5-HTP did not affect any of the measured metabolites (fatty acids and glucose) or minerals (calcium and magnesium) or milk feed intake, feed conversion ratio, and growth. In conclusion, 5-HTP supplementation induced an increase of 5-HT concentrations in blood and caused an increase in mRNA abundance of several factors related to the innate and adaptive immune systems, which might increase the protection of the calf against external agents.     

Association of peripartum plasma insulin concentration with milk production,colostrum insulin levels,and plasma metabolites of Holstein cows

The main objective of this study was to assess associations between plasma insulin concentration around parturition and production in Holstein cows. Primiparous and multiparous cows (n = 267) were enrolled. Blood samples were collected within 12 h after parturition (d 0), and on d 3 and 10 after calving. In addition, blood samples were collected 7 d before (?7 d) the expected date of parturition and colostrum samples were collected within 8 h after parturition from a subset of cows to measure insulin concentration (n = 47). All samples were harvested from 0630 to 1100 h and were used to quantify insulin, nonesterified fatty acids (NEFA), and β-hydroxybutyrate. The plasma concentrations of insulin on d ?7 and 0 were not correlated with insulin levels in colostrum. Cows were grouped according to plasma insulin concentration based on the median as low insulin (L-INS) or high insulin (H-INS) on d 0 (median = 0.35 ng/mL; range 0.2 to 1.2), 3 (median = 0.32 ng/mL; range 0.2 to 1.6), and 10 (median = 0.30 ng/mL; range 0.2 to 0.8). We detected that cows in the L-INS group on d 0 (L-INS = 0.57 ± 0.02; H-INS = 0.49 ± 0.02 mmol/L), d 3 (L-INS = 0.56 ± 0.02; H-INS = 0.49 ± 0.02 mmol/L), and d 10 (L-INS = 0.61 ± 0.03; H-INS = 0.55 ± 0.03 mmol/L) had higher NEFA concentrations compared with cows in the H-INS group. Compared with H-INS cows, milk yield was higher for cows classified as L-INS on d 0 (L-INS = 40.75 ± 0.69; H-INS = 38.41 ± 0.64 kg) and d 10 (L-INS = 40.95 ± 0.74; H-INS = 38.66 ± 0.64 kg). Moreover, fat-corrected milk was higher for cows classified as L-INS on d 0 (L-INS = 40.59 ± 2.36; H-INS = 37.73 ± 2.31 kg) and d 10 (L-INS = 41.00 ± 2.42; H-INS = 38.65 ± 2.28 kg) compared with H-INS cows, and energy-corrected milk was higher for L-INS cows compared with H-INS cows regardless of the day (d 0, L-INS = 44.50 ± 0.70 vs. H-INS = 41.67 ± 0.64 kg; d 3, L-INS = 43.65 ± 0.74 vs. H-INS = 40.88 ± 0.72 kg; d 10, L-INS = 44.09 ± 0.73 vs. H-INS = 40.55 ± 0.68 kg). We conclude that low plasma insulin concentration during early lactation is associated with higher milk yield in the long term.     

Effect of inhibiting the lactogenic signal at calving on milk production and metabolic and immune perturbations in dairy cows

During the periparturient period, the abrupt increase in energy demand for milk production often induces metabolic and immunological disturbances in dairy cows. Our previous work has shown that reducing milk output by milking once a day or incompletely in the first few days of lactation reduces these disturbances. The aim of this study was to reduce metabolic and immunological disturbances by limiting milk production during the first week of lactation by inhibiting the lactogenic signal driven by prolactin. Twenty-two fresh cows received 8 i.m. injections of the prolactin-release inhibitor quinagolide (QUIN; 2 mg) or water as a control (CTL). The first injection was given just after calving, and the subsequent 7 injections were given every 12 h. Milk production was measured until d 28 after calving. Blood samples were taken from d 1 (calving) to d 5 and then on d 7, 10, 14, 21, and 28 to measure concentrations of urea, phosphorus, calcium, glucose, nonesterified fatty acids (NEFA), β-hydroxybutyrate, and prolactin. Other blood samples were taken on d 2, 5, 10, and 28 to analyze oxidative burst, phagocytosis, and the effect of the serum on the lymphoproliferation of peripheral blood mononuclear cells from donor cows. Blood prolactin concentration was lower from d 2 to 5 but higher from d 10 to 28 in the QUIN cows than in the CTL cows. Milk production was lower from d 2 to 6 in the QUIN cows than in the CTL cows (24.3 ± 6.4 and 34.8 ± 4.1 kg/d on average, respectively). We observed no residual effect of quinagolide on milk production after d 6. During the first week of lactation, blood glucose and calcium concentrations were higher and β-hydroxybutyrate concentration was lower in the QUIN cows than in the CTL cows. Blood NEFA, urea, and phosphorus concentrations were not affected by the treatment. At d 2 and 5, the phagocytosis ability of polymorphonuclear leukocytes was not affected by treatment; however, quinagolide injection enhanced the proportion of cells that entered oxidative burst, The mitogen-induced proliferation of peripheral blood mononuclear cells was greater when they were incubated with serum harvested from the CTL cows and was negatively correlated with the NEFA concentration in the serum. Reducing the prolactin peak at calving was effective in reducing milk production during the first week of lactation without compromising the dairy cow's overall productivity. Slowing the increase in milk production allowed a more gradual transition from pregnancy to lactation and led to a reduction in metabolic stress and an improvement in some immune system aspects during this period.     

Effects of supplementing Saccharomyces cerevisiae fermentation products to dairy cows from the day of dry-off through early lactation

The scope of this experiment was to study the effects of Saccharomyces cerevisiae fermentation product (SCFP; NutriTek, Diamond V) on milk yield, milk composition, somatic cell count, rumination activity, and immunometabolic profile (inflammation) of dairy cows during the peripartum period. Postpartum inflammation severity was evaluated as the liver functionality index (LFI). The LFI is based on profiles of specific blood inflammatory markers in the first month of lactation. We hypothesized that SCFP could increase the rumination time in dairy cows. Treatments were control (CTR; no supplement, n = 17) or SCFP (19 g/d of NutriTek, n = 17) included into a pellet delivered at robotic milking unit. Treatments were fed from d −60 to 42 relative to parturition. Cows were fed the same basal rations formulated to pre- or postpartum requirements. Cows were voluntarily milked with robotic milking unit. Blood samples were collected at d −60, −28, −7, 7, and 28 relative to parturition. To study the effect of the treatment and severity of inflammation during periparturient period on subsequent cow performance, cows were retrospectively divided into 2 groups based on their LFI score: low (LLFI) and high (HLFI). Thus, LFI grouping and supplementation treatment groups were as follows: LLFI-CTR, LLFI-SCFP, HLFI-CTR, HLFI-SCFP. Data were analyzed with ANOVA using a mixed model for repeated measures; the model included the effect of the diet, LFI group, time relative to parturition, and their interaction. The nonesterified fatty acids concentrations were greatest at d 7 of lactation for LLFI-CTR compared with other groups. No other differences in plasma metabolites were observed. The LLFI-CTR cows had a greater reduction of body condition score from d −7 until 28 relative to parturition compared with other groups. Somatic cell counts were not different among groups, with averages of 175, 169, 384, and 126 × 1,000 cells/mL for the HLFI-CTR, HLFI-SCFP, LLFI-CTR, and LLFI-SCFP group, respectively, regardless of day. However, the LLFI-CTR had greater somatic cell count on d 42 compared with other groups. During the week before parturition, the LLFI-CTR group had reduced rumination time of 46 min compared with the other 3 groups. However, the minutes of rumination per day was only different between LLFI-CTR and the LLFI-SCFP groups. Milk production of cows was different for LFI scores as follows: 50.2 versus 46.7 kg/d for HLFI and LLFI, respectively. Interestingly, there were no differences of milk production due to supplementation treatment of the HLFI cows. However, the LLFI-SCFP group produced 49.1 kg/d compared with 44.3 kg/d of the LLFI-CTR group during the first month of lactation. Milk composition did not differ throughout the experimental period for the 4 groups of cows. In conclusion, SCFP supplementation assisted cows experiencing low LFI to maintain milk production, somatic cell count, and plasma nonesterified fatty acid concentrations similar to cows with high LFI.     

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

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

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

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

Short communication: Circulating serotonin (5-HT) concentrations on day 1 of lactation as a potential predictor of transition-related disorders

The monoamine serotonin (5-hydroxytryptamine; 5-HT) has been described as a homeostatic regulator of lactation. Recently, our laboratory determined that 5-HT is involved in the regulation of calcium and glucose homeostasis during the transition period in rodents. More specifically, we demonstrate that 5-HT is responsible for calcium mobilization from bone and upregulation of hepatic gluconeogenic enzymes and mammary gland glucose transporters. Our objective was to investigate the correlation between circulating 5-HT concentrations and circulating ionized calcium, parathyroid hormone-related protein (PTHrP), and glucose concentrations on d 1 postpartum. We also investigated the correlation between circulating 5-HT and milk fever and ketosis incidence and severity in multiparous Holstein cows at the onset of lactation. Blood samples were collected from 42 multiparous cows on d 1 of lactation and analyzed for 5-HT, calcium, glucose, and PTHrP. Milk fever (determined subjectively for each cow on d 1 postpartum) and ketosis incidence and severity (scale 1 to 4, determined objectively for each cow during the first 10 d postpartum) were recorded for all animals. Serum 5-HT was positively correlated with serum calcium and with plasma PTHrP (r > 0.37). Serum 5-HT was negatively correlated with milk fever incidence and with ketosis severity (most severe ketosis incidence recorded during the first 10 d postpartum; r < -0.33). Serum calcium and plasma glucose concentrations were negatively correlated with milk fever and ketosis severity, respectively (r < -0.39). These data indicate that 5-HT potentially plays a role in the regulation of calcium and glucose homeostasis during the transition period in cattle, which we previously demonstrated in rodents. Increased circulating concentrations of 5-HT might decrease milk fever at the onset of lactation and ketosis severity during the first 10 d postpartum in dairy cows. Understanding this physiological axis could help describe the underlying mechanisms associated with these periparturient metabolic disorders in dairy cows.     

Evaluation of 2 different treatment procedures after calving to improve harvesting of high-quantity and high-quality colostrum

The objective of this study was to evaluate 2 different treatment procedures at the first milking after calving to increase colostrum quantity and to improve colostrum quality in dairy cows. We hypothesized that either exogenous treatment with oxytocin or the presence of the calf at first milking would lead to higher colostrum quantity and higher IgG concentration. The study was conducted from October to December 2017 on a commercial dairy farm in Germany. A total of 567 cows at the time of calving were enrolled, but for the final analyses only 521 animals were considered. The cows were randomly assigned on a daily basis into 1 of 3 groups: (1) control group (n = 177), (2) application of 20 IU of oxytocin i.m. (OXY; n = 163), and (3) presence of the calf (CA; n = 181) before and during milking. Cows in the control and oxytocin group had no contact with their calves after calving and were milked in a separate milking parlor. Cows in the oxytocin group were injected with 20 IU of oxytocin i.m. 3 min before manual stimulation. For cows in the third group, the calf was placed into a calf cart and located in front of the cow 3 min before manipulation of the cow. Colostrum quantity was determined by a digital hanging scale. The colostrum quality was assessed with digital Brix refractometry and ELISA. To evaluate the effect of 2 different treatment procedures, a generalized linear mixed model was constructed using SPSS (SPSS Inc., IBM, Ehningen, Germany). The mean (±SE) colostrum quantity was 4.17 ± 0.30 kg. The treatment procedures and the harvesting time after calving had no effect on colostrum quantity. Parity, calf birth weight, and calving time affected colostrum quantity. Cows in second parity had the lowest quantity of colostrum (3.74 ± 0.37 kg) compared with cows in parity 1 (4.75 ± 0.34 kg) and cows in parity 3 or greater (4.75 ± 0.38 kg). Cows calving during the night (2200 until 0600 h; 4.93 ± 0.37 kg) had the highest quantity of colostrum compared with cows calving in the morning (0600 until 1400 h; 4.17 ± 0.38 kg) or afternoon (1400 until 2200 h; 4.14 ± 0.34 kg). Regarding colostrum quality, 48% of the colostrum samples contained ≥50 mg of IgG/mL. The mean IgG concentration was 54.6 ± 2.80 mg of IgG/mL. Colostrum quality was affected by the treatment procedures, colostrum quantity, parity, calving time, harvesting time after calving, and the calving day during the week. Both treatment procedures (i.e., OXY with mean IgG concentration results of 57.0 mg of IgG/mL and CA with 56.0 mg of IgG/mL) resulted in higher IgG concentrations in colostrum compared with the control group (50.7 mg of IgG/mL). With increasing colostrum quantity, the colostrum quality decreased in primiparous and multiparous cows. A longer time lag between calving and milking negatively affected the colostrum quality. Concentration of IgG was higher for cows in parity 3 or greater (64.6 ± 2.59 mg of IgG/mL) compared with cows in parity 1 (48.5 ± 2.86 mg of IgG/mL) and cows in parity 2 (50.7 ± 2.89 mg of IgG/mL). Cows calving during the night had greater IgG concentrations (60.4 ± 2.92 mg of IgG/mL) compared with cows calving in the morning (51.9 ± 2.98 mg of IgG/mL) or afternoon (51.3 ± 2.71 mg of IgG/mL). Harvesting colostrum on quieter days, such as Sundays, resulted in higher IgG concentrations (61.4 ± 3.70 mg of IgG/mL). The assessment by Brix refractometry resulted in a mean result of 26.0 ± 0.20% Brix. Treatment procedures and the harvesting time after calving had no effect on colostrum quality. A negative association was observed between colostrum quantity and quality in primiparous and multiparous cows determined by Brix refractometry. Brix readings were greater for cows in parity 3 or higher (27.7 ± 0.26% Brix) compared with cows in parity 1 (25.3 ± 0.30% Brix) and cows in parity 2 (25.0 ± 0.32% Brix). In conclusion, the treatment procedure for the first milking is irrelevant to improve the quantity of colostrum. Both treatment procedures, however, increased IgG concentrations as determined by ELISA.     

Relationship between intramammary infection and antibody concentrations in Jersey and Holstein colostrum

Feeding calves a high-quality and antibody-rich colostrum is an important management practice for supporting calf health and productivity. Colostrum quality and antibody concentrations are highly variable between cows and among quarters within a cow. Intramammary infections often occur during the time of colostrum formation; however, it is unknown if these infections ultimately affect colostrum quality and antibody concentrations. The objective of this study was to determine if antibody concentrations and Brix percentage in colostrum from infected mammary glands (quarters) differed from uninfected. In 2 cross-sectional studies, colostrum samples were aseptically collected at first milking from 110 Holstein and 89 Jersey cows at 3 Holstein and 4 Jersey commercial dairy farms in Ohio. A total of 771 quarter samples were collected, underwent bacteriological culture, and were measured for Brix percentage with a digital refractometer. When 1 infected and 1 uninfected quarter existed among the fore or rear quarters within a cow, IgG₁, IgG₂, IgA, and IgM antibody concentrations were determined via ELISA for the paired quarters (n = 82). Overall, for Holstein cows, Brix percentages were greater in multiparous than primiparous cows (30.5 vs. 23.7 ± 2.1 SEM), but an opposite pattern was observed for Jersey cows (24.3 vs. 27.2 ± 1.2 SEM). Uninfected quarters in both Holstein and Jersey multiparous cows had greater Brix percentage than colostrum from infected quarters; this pattern was absent for Holstein and Jersey primiparous cows. For Holstein cows, concentrations of IgG₁, IgG₂, and IgA were greater in multiparous cows than primiparous cows; quarter-infection status did not significantly influence antibody concentrations. For Jersey samples, antibody concentrations did not differ between primiparous and multiparous cows and were not significantly affected by quarter-infection status. The results of these works indicate that infection status at parturition does not markedly affect colostrum antibody concentrations and quality, and that other factors at the local level of the mammary gland more greatly influence colostrogenesis and antibody transport into the mammary gland during colostrogenesis.     

Metabolic responses of lactating dairy cows to 14-day intravenous infusions of glucagon.

Twenty cows were assigned at parturition to two groups to study metabolic effects of continuous intravenous infusions of glucagon. Groups were control cows and cows treated with glucagon at 10 mg/d for 14 d starting at d 21 postpartum. Daily blood samples and nine liver biopsies were taken from d 7 to 49 postpartum. Plasma glucagon increased six- to seven-fold during infusions of treated cows. Plasma insulin was increased heterogeneously by glucagon infusions. Plasma glucose increased 11.5 and 9.0 mg/dl during wk 1 and 2 of glucagon infusions. No other plasma metabolites tested (nonesterified fatty acids, beta-hydroxybutyrate, and urea N) were affected by glucagon infusions. Liver glycogen decreased by d 2 of glucagon infusion but was repleted to preinfusion values by d 7 and increased to 169% of the preinfusion baseline values at 3 d after cessation of glucagon. Milk production decreased transiently during glucagon infusions. Both milk production and milk protein percentage decreased during glucagon infusion, which could imply a decreased availability of amino acids for milk protein synthesis. Feed intakes did not increase during glucagon infusions, which was in contrast to the control group. Results indicated that glucagon infusions caused liver glycogenolysis initially and probably enhanced gluconeogenesis but glucagon did not appear to increase lipolysis from adipose tissue in these early lactating dairy cows.     

Hepatic effects of rumen-protected branched-chain amino acids with or without propylene glycol supplementation in dairy cows during early lactation

Essential amino acids (EAA) are critical for multiple physiological processes. Branched-chain amino acid (BCAA) supplementation provides energy substrates, promotes protein synthesis, and stimulates insulin secretion in rodents and humans. Most dairy cows face a protein and energy deficit during the first weeks postpartum and utilize body reserves to counteract this shortage. The objective was to evaluate the effect of rumen-protected BCAA (RP-BCAA; 375 g of 27% l-leucine, 85 g of 48% l-isoleucine, and 91 g of 67% l-valine) with or without oral propylene glycol (PG) administration on markers of liver health status, concentrations of nonesterified fatty acids (NEFA) and β-hydroxybutyrate (BHB) in plasma, and liver triglycerides (TG) during the early postpartum period in dairy cows. Multiparous Holstein cows were enrolled in blocks of 3 and randomly assigned to either the control group or 1 of the 2 treatments from calving until 35 d postpartum. The control group (n = 16) received 200 g of dry molasses per cow/d; the RP-BCAA group (n = 14) received RP-BCAA mixed with 200 g of dry molasses per cow/d; the RP-BCAA plus PG (RP-BCAAPG) group (n = 16) received RP-BCAA mixed with 200 g of dry molasses per cow/d, plus 300 mL of PG, once daily from calving until 7 d in milk (DIM). The RP-BCAA and RP-BCAAGP groups, on average (± standard deviation), were predicted to receive a greater supply of metabolizable protein in the form of l-Leu 27.4 ± 3.5 g/d, l-Ile 15.2 ± 1.8 g/d, and l-Val 24.2 ± 2.4 g/d compared with the control cows. Liver biopsies were collected at d 9 ± 4 prepartum and at 5 ± 1 and 21 ± 1 DIM. Blood was sampled 3 times per week from calving until 21 DIM. Milk yield, dry matter intake, NEFA, BHB, EAA blood concentration, serum chemistry, insulin, glucagon, and liver TG and protein abundance of total and phosphorylated branched-chain ketoacid dehydrogenase E1α (p-BCKDH-E1α) were analyzed using repeated measures ANOVA. Cows in the RP-BCAA and RP-BCAAPG groups had lower liver TG and lower activities of aspartate aminotransferase and glutamate dehydrogenase during the first 21 DIM, compared with control. All cows, regardless of treatment, showed an upregulation of p-BCKDH-E1α at d 5 postpartum, compared with levels at 21 d postpartum. Insulin, Met, and Glu blood concentration were greater in RP-BCAA and RP-BCAAPG compared with control during the first 35 DIM. Therefore, the use of RP-BCAA in combination with PG might be a feasible option to reduce hepatic lipidosis in dairy cows during early lactation.     

Amplifying local serotonin signaling prior to dry-off hastens mammary gland involution and redevelopment in dairy cows

The monoamine serotonin (5-hydroxytryptamine, 5-HT) has been reported to inhibit milk protein gene expression and increase mammary epithelial cell (MEC) tight junction permeability after milk stasis. We hypothesized that increasing serotonin synthesis and signaling within the mammary epithelium before milk stasis would increase systemic and local involution markers, and downregulate the expression of milk protein and tight junction during involution, leading to more efficient tissue growth during the redevelopment phase. Herein, we examined the outcomes of increasing local mammary 5-HT synthesis before milk stasis on involution biomarkers, mammary gland microstructure, and gene and protein expression during the dry period. Multiparous Holstein cows were administered intramammary infusions (via the teat canal) of sterile water (CON, 4 mL/teat, n = 7) or 5-hydroxy-l-tryptophan (5-HTP, serotonin precursor, 20 mg/teat, n = 7) once daily for 5 d before dry-off (d 0). Blood, milk, and mammary secretions were collected and analyzed for components and metabolites. Mammary secretions were collected 12 h after the last milking and on d 1 to 4 during the dry period at 1200 h. Mammary gland biopsies were performed on d 4 (i.e., involution phase) and d 36 (i.e., redevelopment phase) of the dry period for histological and molecular evaluation. Milk protein and tight junction gene expression was quantified via real-time PCR. Hematoxylin and eosin staining, immunohistochemistry (Ki67), and immunofluorescence (serotonin, cleaved caspase 3) were performed to visualize tissue microstructure and to quantify serotonin intensity and cell turnover. Data were analyzed in SAS (SAS Institute Inc.) using 2-way ANOVA. After d 0, mammary secretions of 5-HTP cows had increased concentrations of 5-HT, lactoferrin, and bovine serum albumin. On d 1, 5-HTP cows had greater α-lactalbumin concentrations in plasma relative to CON. Serotonin intensity was increased in the mammary tissue of 5-HTP cows on d 4, relative to CON. On d 4, milk protein and tight junction gene expression was downregulated, MEC number was reduced, and cleaved caspase 3 protein was greater in mammary tissue of 5-HTP cows, relative to CON. On d 36, milk protein genes were upregulated, and the lumen:outer alveolar area and Ki67-positive cells were increased in the mammary tissue of 5-HTP cows, relative to CON. Amplifying serotonin signaling in the mammary epithelium before milk stasis at dry-off achieves greater apoptosis, leading to a reduction in MEC, allowing for greater cell proliferation, which results in more MEC during the redevelopment phase preceding the onset of lactation.     

Jugular arginine infusion relieves lipopolysaccharide-triggered inflammatory stress and improves immunity status of lactating dairy cows

The objective of this study was to evaluate the effects of jugular l-Arg infusion on performance and immune function during lipopolysaccharide (LPS)-induced inflammation of lactating dairy cows. Eight Holstein cows (multiparous, 608.8 ± 31.5 kg) at mid-lactation were randomly assigned to 5-d jugular infusions of control (saline), Arg (3 g/h), LPS (0.033 μg/kg per h), and LPS + Arg (0.033 μg/kg per h of LPS and 3 g/h of Arg) in a replicated 4 × 4 Latin square design with 4 infusion periods separated by 10-d noninfusion periods. Jugular solutions of saline, Arg, LPS, and LPS + Arg were continuously infused using peristaltic pumps for approximately 6 h/d during infusion periods. Milk yield was measured on each day of the infusion period. Milk samples were obtained on the last 2 d of each infusion period, and blood samples were obtained on the last day of each infusion period before infusion (0 h) and at 3 and 6 h. We found that the jugular LPS infusion significantly increased serum concentrations of IL-1β, IL-6, tumor necrosis factor, inducible nitric oxide synthase, and lipopolysaccharide binding protein, whereas Arg attenuated the increase in IL-6 and inducible nitric oxide synthase levels and tended to decrease the lipopolysaccharide binding protein level. Arginine alleviated the decrease in dry matter intake and milk fat yield and the increase of somatic cell count induced by LPS. Total casein in milk was decreased during the LPS-induced inflammation period, and jugular Arg infusion significantly increased the content of total casein. In contrast, lactalbumin in milk increased during the LPS-induced inflammation period, whereas jugular Arg infusion significantly decreased the content of lactalbumin. The concentrations of plasma Gly, Thr, Ile, Leu, Arg, Phe, and total free AA were significantly decreased by LPS treatment, but Arg attenuated this tendency. These results indicated that jugular Arg infusion (18 g/d) has protective effects on relieving inflammatory stress and improving immunity status triggered by LPS. In conclusion, Arg could attenuate inflammatory stress and improve milk performance of lactating dairy cows. This protective effect may be due to the ability of Arg to suppress LPS effects and improve immunity status.     

Secretion and mammary gland uptake of prolactin in dairy cows during lactogenesis

Mammary arteriovenous differences of prolactin concentration and net mammary uptake of prolactin from blood were quantified near parturition in 9 dairy cows. Six cows were milked once daily for at least 6 d before parturition, and prepartum lactogenesis occurred in 3 of 6 cows. Prepartum milking 2 or more d before parturition abruptly increased secretion of prolactin into blood, but milkings within 1 d before or after parturition did not increase prolactin secretion. Concentrations of prolactin in whole milk sampled over 8 d before parturition (64.5 ng/ml) were substantially greater than those occurring several days after parturition (19 ng/ml). Milk prolactin concentrations were unaffected by the successful induction of prepartum lactogenesis, which greatly increased prepartum yields of milk (2 to 8 kg/milking). Therefore, the alveolar lumenal content of prolactin was greatest in pregnant cows with prepartum lactogenesis. This enhanced content of intraalveolar prolactin before parturition was associated with an absence of mammary uptake of prolactin immediately prior to ejection of the prolactin-containing milk from the alveoli. However, prolactin uptake was quickly restored to about 2 micrograms/min per half udder shortly after milk ejection. During the prepartum period, an enhanced intraalveolar reservoir of 200 to 400 micrograms prolactin, due to induction of prepartum lactogenesis, appears to saturate temporarily all putative sites for uptake of prolactin from blood.     

Effects of feeding rumen-protected methionine pre- and postpartum in multiparous Holstein cows: Lactation performance and plasma amino acid concentrations

Objectives were to evaluate the effect of feeding rumen-protected methionine (RPM) in pre- and postpartum total mix ration (TMR) on lactation performance and plasma AA concentrations in dairy cows. A total of 470 multiparous Holstein cows [235 cows at University of Wisconsin (UW) and 235 cows at Cornell University (CU)] were enrolled approximately 4 wk before parturition, housed in close-up dry cow and replicated lactation pens. Pens were randomly assigned to treatment diets (pre- and postpartum, respectively): UW control (CON) diet = 2.30 and 2.09% of Met as percentage of metabolizable protein (MP) and RPM diet = 2.83 and 2.58% of Met as MP; CU CON = 2.22 and 2.19% of Met as percentage of MP, and CU RPM = 2.85 and 2.65% of Met as percentage of MP. Treatments were evaluated until 112 ± 3 d in milk (DIM). Milk yield was recorded daily. Milk samples were collected at wk 1 and 2 of lactation, and then every other week, and analyzed for milk composition. For lactation pens, dry matter intake (DMI) was recorded daily. Body weight and body condition score were determined from 4 ± 3 DIM and parturition until 39 ± 3 and 49 DIM, respectively. Plasma AA concentrations were evaluated within 3 h after feeding during the periparturient period [d ?7 (±4), 0, 7 (±1), 14 (±1), and 21 (±1); n = 225]. In addition, plasma AA concentrations were evaluated (every 3 h for 24 h) after feeding in cows at 76 ± 8 DIM (n = 16) and within 3 h after feeding in cows at 80 ± 3 DIM (n = 72). The RPM treatment had no effect on DMI (27.9 vs. 28.0 kg/d) or milk yield (48.7 vs. 49.2 kg/d) for RPM and CON, respectively. Cows fed the RPM treatment had increased milk protein concentration (3.07 vs. 2.95%) and yield (1.48 vs. 1.43 kg/d), and milk fat concentration (3.87 vs. 3.77%), although milk fat yield did not differ. Plasma Met concentrations tended to be greater for cows fed RPM at 7 d before parturition (25.9 vs. 22.9 µM), did not differ at parturition (22.0 vs. 20.4 µM), and were increased on d 7 (31.0 vs. 21.2 µM) and remained greater with consistent concentrations until d 21 postpartum (d 14: 30.5 vs. 19.0 µM; d 21: 31.0 vs. 17.8 µM). However, feeding RPM decreased Leu, Val, Asn, and Ser (d 7, 14, and 21) and Tyr (d 14). At a later stage in lactation, plasma Met was increased for RPM cows (34.4 vs. 16.7 µM) consistently throughout the day, with no changes in other AA. Substantial variation was detected for plasma Met concentration (range: RPM = 8.9–63.3 µM; CON = 7.8–28.8 µM) among cows [coefficient of variation (CV) > 28%] and within cow during the day (CV: 10.5–27.1%). In conclusion, feeding RPM increased plasma Met concentration and improved lactation performance via increased milk protein production.