Induced hyperketonemia affects the mammary immune response during lipopolysaccharide challenge in dairy cows

Metabolic adaptations during negative energy and nutrient balance in dairy cows are thought to cause impaired immune function and hence increased risk of infectious diseases, including mastitis. Characteristic adaptations mostly occurring in early lactation are an elevation of plasma ketone bodies and free fatty acids (nonesterified fatty acids, NEFA) and diminished glucose concentration. The aim of this study was to investigate effects of elevated plasma β-hydroxybutyrate (BHBA) at simultaneously even or positive energy balance and thus normal plasma NEFA and glucose on factors related to the immune system in liver and mammary gland of dairy cows. In addition, we investigated the effect of elevated plasma BHBA and intramammary lipopolysaccharide (LPS) challenge on the mammary immune response. Thirteen dairy cows were infused either with BHBA (HyperB, n = 5) to induce hyperketonemia (1.7 mmol/L) or with a 0.9% saline solution (NaCl, n = 8) for 56 h. Two udder quarters were injected with 200 μg of LPS after 48 h of infusion. Rectal temperature (RT) and somatic cell counts (SCC) were measured before, at 48 h after the start of infusions, and hourly during the LPS challenge. The mRNA abundance of factors related to the immune system was measured in hepatic and mammary tissue biopsies 1 wk before and 48 h after the start of the infusion, and additionally in mammary tissue at 56 h of infusion (8 h after LPS administration). At 48 h of infusion in HyperB, the mRNA abundance of serum amyloid A (SAA) in the mammary gland was increased and that of haptoglobin (Hp) tended to be increased. Rectal temperature, SCC, and mRNA abundance of candidate genes in the liver were not affected by the BHBA infusion until 48 h. During the following LPS challenge, RT and SCC increased in both groups. However, SCC increased less in HyperB than in NaCl. Quarters infused with LPS showed a more pronounced increase of mRNA abundance of IL-8 and IL-10 in HyperB than in NaCl. The results demonstrate that an increase of plasma BHBA upregulates acute phase proteins in the mammary gland. In response to intramammary LPS challenge, elevated BHBA diminishes the influx of leukocytes from blood into milk, perhaps by via modified cytokine synthesis. Results indicate that increased ketone body plasma concentrations may play a crucial role in the higher mastitis susceptibility in early lactation.     

Hyperketonemia during lipopolysaccharide-induced mastitis affects systemic and local intramammary metabolism in dairy cows

Hyperketonemia interferes with the metabolic regulation in dairy cows. It is assumed that metabolic and endocrine changes during hyperketonemia also affect metabolic adaptations during inflammatory processes. We therefore studied systemic and local intramammary effects of elevated plasma β-hydroxybutyrate (BHBA) before and during the response to an intramammary lipopolysaccharide (LPS) challenge. Thirteen dairy cows received intravenously either a Na-dl-β-OH-butyrate infusion (n = 5) to achieve a constant plasma BHBA concentration (1.7 ± 0.1 mmol/L), with adjustments of the infusion rates made based on immediate measurements of plasma BHBA every 15 min, or an infusion with a 0.9% NaCl solution (control; n = 8) for 56 h. Infusions started at 0900 h on d 1 and continued until 1700 h 2 d later. Two udder quarters were challenged with 200 μg of Escherichia coli LPS and 2 udder quarters were treated with 0.9% saline solution as control quarters at 48 h after the start of infusion. Blood samples were taken at 1 wk and 2 h before the start of infusions as reference samples and hourly during the infusion. Mammary gland biopsies were taken 1 wk before, and 48 and 56 h (8 h after LPS challenge) after the start of infusions. The mRNA abundance of key factors related to BHBA and fatty acid metabolism, and glucose transporters was determined in mammary tissue biopsies. Blood samples were analyzed for plasma glucose, BHBA, nonesterified fatty acid, urea, insulin, glucagon, and cortisol concentrations. Differences were not different for effects of BHBA infusion on the mRNA abundance of any of the measured target genes in the mammary gland before LPS challenge. Intramammary LPS challenge increased plasma glucose, cortisol, glucagon, and insulin concentrations in both groups but increases in plasma glucose and glucagon concentration were less pronounced in the Na-dl-β-OH-butyrate infusion group than in controls. In response to LPS challenge, plasma BHBA concentration decreased in controls and decreased also slightly in the BHBA-infused animals because the BHBA concentration could not be fully maintained despite a rapid increase in BHBA infusion rate. The change in mRNA abundance of citrate synthase in LPS quarters was significant between the 2 treatment groups. The results indicate that elevated circulating BHBA concentration inhibits gluconeogenesis before and during immune response to LPS challenge, likely because BHBA can replace glucose as an energy source.     

Effects of dietary vitamin C on neutrophil function and responses to intramammary infusion of lipopolysaccharide in periparturient dairy cows

Neutrophil function and the severity and incidence of mastitis in dairy cows is related to the intake of many antioxidant nutrients. Because vitamin C is the major water-soluble antioxidant in mammals, we examined the effect of dietary vitamin C on neutrophil function and responses to intramammary infusion of lipopolysaccahride (LPS) in periparturient dairy cows. At 2 wk before anticipated calving, Holstein cows were fed diets that provided 0 (16 cows) or 30 (15 cows) g/d of supplemental vitamin C (phosphorylated ascorbic acid). Treatments continued until 7 d after cows received an infusion of 10 μg of LPS into one quarter of the mammary gland (on average, this occurred 32 d postcalving). Supplementation of vitamin C increased plasma concentrations of vitamin C at calving, but no differences were observed in samples taken 24 h postinfusion. Concentrations of vitamin C in milk (24 h postinfusion) and in neutrophils (calving and 24 h postinfusion) were not affected by treatment, but vitamin C concentrations in neutrophils isolated from milk were about 3 times greater than concentrations in blood neutrophils. The LPS infusion did not alter concentrations of vitamin C in plasma or milk, suggesting that the LPS model did not produce the same effects as a bacterial infection of the mammary gland with respect to antioxidant effects. Supplemental vitamin C had no effect on neutrophil phagocytosis or bacterial kill. Dietary vitamin C reduced the milk somatic cell count but did not affect the febrile response or milk production following LPS infusion.     

Induction of hyperlipidemia by intravenous infusion of tallow emulsion causes insulin resistance in Holstein cows

The objective was to test whether the induction of elevated blood nonesterified fatty acids (NEFA) by i.v. infusion of a tallow emulsion altered glucose tolerance and responsiveness to insulin in Holstein cows. Six non-lactating, nongestating Holstein cows were assigned to a crossover design. One cow was excluded before initiation of the experiment because of complications from mastitis. Treatments consisted of 11-h i.v. infusions of saline (control) or a 20% (wt/vol) triacylglycerol (TG) emulsion derived from tallow (tallow) to elevate plasma NEFA. Each period consisted of two 11-h infusions (INF1 and INF2), separated by 1 d in which cows were not infused. Intravenous glucose tolerance tests (IVGTT) and insulin challenges (IC) were performed 8 h after initiation of INF1 and INF2, respectively. The infusion of treatments continued during the 3 h of sampling for IVGTT and IC. Cows were fed every 4 h at a rate to meet energy requirements for 5 d prior to each period, and every 2 h during the first 8 h of infusions. Infusion of tallow induced hyperlipidemia by increasing plasma NEFA (295 ± 9 vs. 79 ± 7 μEq/L), serum TG (41.0 ± 6 vs. 11.4 ± 4.4 mg/dL), and glycerol (0.81 ± 0.09 vs. 0.23 ± 0.1 mg/dL) concentrations during INF1. During INF2, tallow treatment increased plasma NEFA (347 vs. 139 ± 18 μEq/L), serum TG (20.8 ± 4.6 vs. 13.1 ± 2.3 mg/dL), and glycerol (0.88 ± 0.04 vs. 0.31 ± 0.02 mg/dL) concentrations. Induction of hyperlipidemia impaired glucose clearance during IVGTT, despite the greater endogenous insulin response to the glucose infusion, leading to a lower insulin sensitivity index [0.29 vs. 1.88 ± 0.31 × 10⁻⁴ min⁻¹/(μIU/mL)]. Accordingly, hyperlipidemia impaired glucose clearance during IC (1.58 vs. 2.72%/min), reflecting lower responsiveness to insulin. These data show that induction of hyperlipidemia causes insulin resistance in Holstein cows by impairing both sensitivity and maximum responsiveness to insulin. The induction of insulin resistance by TG, NEFA, or both may increase the availability of glucogenic nutrients to the periparturient dairy cow. Yet excessive elevation of NEFA may potentially lead adipocytes to become more insulin resistant, further increasing plasma NEFA concentration and the risk of metabolic disorders.     

Antilipolytic and lipolytic effects of administering free or ruminally protected nicotinic acid to feed-restricted Holstein cows

The objectives were to determine effects of 12 hourly infusions of different quantities of nicotinic acid (NA) on plasma nonesterified fatty acid (NEFA; experiment 1) and whether longer (108 h) continuous infusions of NA could induce sustained reductions of plasma NEFA (experiment 2) in nonlactating, nongestating Holstein cows that were feed restricted. Experiment 1 was a 5 × 5 Latin square with 6-d periods and 9 recovery days between each period. Each period consisted of 5 d of partial feed restriction to increase plasma NEFA concentration. Treatments were abomasal infusions of 0, 0.25, 0.5, 1, or 3 mg of NA/h per kilogram of body weight (BW), infused as hourly boluses for 12 h, starting 4 d after initiation of partial feed restriction. Plasma NEFA was decreased for the highest dose: from 448 μEq/L to 138 ± 75 μEq/L at 1 h after the first bolus of 3 mg of NA/h per kilogram of BW. This initial reduction in plasma NEFA concentration was followed by an increase in concentration at 2, 3, and 4 h relative to initiation of infusions. Plasma NEFA then decreased to 243 μEq/L 6 h after initiation of treatments and remained low until termination of infusions. A rebound in plasma NEFA concentration occurred at 3 and 4 h after termination of infusion for cows that received 3 mg of NA/h per kilogram of BW. Experiment 2 was a 5 × 5 Latin square with 7-d periods and 9 recovery days between each period. Each period consisted of 5 d of partial feed restriction to increase plasma NEFA concentration. Treatments were continuous abomasal infusion of 0, 0.5, 1, or 3 mg of free NA/h per kilogram of BW for 4.5 d starting at feed restriction or 0.5 mg of NA/h per kilogram of BW infused directly into the rumen in a form protected from microbial degradation. The ruminal administration of protected NA was initiated 2 d before abomasal infusions and initiation of feed restriction to establish steady postruminal delivery of NA by start of abomasal infusions. Plasma NEFA was approximately 70 μEq/L before initiation of feed restriction and increased to 509, 587, 442, 850, and 108 μEq/L at 4.5 d for cows that received 0, 0.5 (protected NA), 0.5 (free NA), 1, and 3 mg of NA/h per kilogram of BW, respectively. An antilipolytic response was achieved with the highest abomasal dose, which maintained plasma NEFA concentration lower than the control group. An increase in plasma NEFA concentration was observed after termination of infusions for cows that received 1 and 3 mg of NA/h per kilogram of BW. Plasma NEFA was 1,900 μEq/L at 4 h after termination of infusion for cows receiving 1 mg of NA/h per kilogram of BW and 1,360 μEq/L at 5 h after termination of infusion for cows receiving 3 mg of NA/h per kilogram of BW. In nongestating, nonlactating cows it is unlikely that a dose of NA exists that will reduce plasma NEFA concentration and prevent the rebound that occurs following termination of NA administration.     

Effects of intravenous glucose infusion and nutritional balance on serum concentrations of nonesterified fatty acids, glucose, insulin, and progesterone in nonlactating dairy cows

The objective of this study was to evaluate serum concentrations of nonesterified fatty acids, glucose, insulin, and progesterone in nonlactating dairy cows according to nutritional balance and glucose infusion. Ten nonlactating, ovariectomized Gir × Holstein cows were stratified by body weight (BW) and body condition score (BCS) on d −28 of the study, and randomly assigned to 1) negative nutrient balance (NB) or 2) positive nutrient balance (PB). From d −28 to d 0, cows were allocated according to nutritional treatment (5 cows/treatment) into 2 low-quality pastures with reduced forage availability. However, PB cows individually received, on average, 3 kg/cow per day (as-fed) of a concentrate during the study. All cows had an intravaginal progesterone releasing device inserted on d −14, which remained in cows until the end of the study. Cow BW and BCS were assessed again on d 0. On d 0, cows within nutritional treatment were randomly assigned to receive, in a crossover design containing 2 periods of 24 h each, 1) intravenous glucose infusion (GLU; 0.5 g of glucose/kg of BW, as a 5% glucose solution administered, on average, at 32 mL/min over a 3-h period), or 2) intravenous saline infusion (SAL; 0.9% solution infused on average at 32 mL/min over a 3-h period). Prior to the beginning of each period, all cows were fasted for 12 h. Blood samples were collected, relative to the beginning of the infusion, at −12 and −11.5 h (beginning of fasting), and at −0.5, 0, 0.5, 1, 2, 3, 4, 5, and 6 h. Following the last blood collection of period 1, cows received (PB) or not (NB) concentrate and were returned to their respective pastures. Changes in BCS and BW were greater in NB cows compared with PB cows (−0.60 and −0.25 ± 0.090 for BCS, respectively; −22.4 and 1.2 ± 6.58 kg for BW, respectively). Cows receiving GLUC had greater glucose concentrations from 0.5 to 3 h relative to infusion compared with SAL cows. Insulin concentrations were greater in PB cows assigned to GLUC compared with SAL cohorts at 0.5 and 3 h following infusion, whereas NB cows assigned to GLUC had greater insulin concentrations compared with SAL cohorts at 0.5, 1, 2, and 3 h. Progesterone concentrations were greater in PB cows assigned to GLUC at 2, 3, and 4 h following infusion compared with SAL cohorts. In conclusion, the effects of glucose infusion on serum concentrations of insulin and progesterone in nonlactating dairy cows were dependent on cow nutritional status.     

The use of a radiotelemetric ruminal bolus to detect body temperature changes in lactating dairy cattle

The objective of this study was to validate the efficacy of a radiotelemetric bolus (RTB) to detect changes in ruminal temperature resulting from (1) systemic illnesses that are associated with febrile responses and (2) subacute ruminal acidosis (SARA). Eight rumen-fistulated, lactating Holstein cows (586 ± 37 kg of body weight, 106 ± 18 d in milk) were used in a replicated 4 × 4 Latin square design with a 2 × 2 factorial arrangement. Each period consisted of 21 d. The factors were 2 diets, a moderate forage:concentrate [MFC; 52:48; % of dry matter (DM)] or a high forage:concentrate (HFC; 65:35, % of DM) total mixed ration, and a challenge with a single intramammary injection of lipopolysaccharide (LPS; 100 μg derived from Escherichia coli 0111:B4) or no LPS (sterile saline). Thus, the 4 resulting treatments were (1) MFC with LPS challenge, (2) MFC with saline, (3) HFC with LPS challenge, and (4) HFC with saline. Cows were fed at 0800 and 1400 h daily. Cows received the intramammary injections at 0900 h of d 21. Ruminal pH and ruminal temperature were also measured on d 21 every minute via an indwelling logging system that resided in the ventral sac of the rumen and via a radiotelemetric bolus that resided in the reticulum. Vaginal temperature was also recorded every minute via temperature loggers. Prior to LPS injection, the duration of rumen pH below 5.6 (indicative of SARA) was higher in cows receiving MFC than cows receiving HFC (148 ± 24 and 62 ± 24 min/d, respectively). The temperature measured at the same time via RTB was higher for MFC than HFC cows (167 ± 21 vs. 104 vs. 21 min/d above 38.8 °C, respectively). The following day, cows challenged with LPS showed signs of mastitis within the injected quarters, depressed DM intake, decreased milk yield, and a peak vaginal temperature of 41.3 ± 0.1 °C 5.5 h after the LPS injection. The RTB system successfully detected a fever response parallel to that measured by the vaginal loggers but temperature peak detected by RTB was, on average, 0.5 °C lower than that detected by the vaginal logger. Although the RTB system was able to detect a temperature response to the diet effect before LPS challenge, it was unable to detect this effect during the LPS challenge, likely because cows receiving the LPS challenge had decreased feed consumption. In conclusion, radiotelemetry has the potential to improve the detection of SARA and fever on farm.     

The use of nicotinic acid to induce sustained low plasma nonesterified fatty acids in feed-restricted Holstein cows

The objectives were to determine the effects of nicotinic acid (NA) on blood metabolites (experiment 1) and whether successive doses of NA could induce sustained reductions of plasma nonesterified fatty acids (NEFA; experiment 2) in feed-restricted, nonlactating Holstein cows. Experiment 1 was a single 4 × 4 Latin square with 1-wk periods. Each period consisted of 2.5 d of feed restriction to increase plasma NEFA and 4.5 d of ad libitum feeding. Treatments were abomasal administration of 0, 6, 30, or 60 mg of NA/kg of body weight (BW), given as a single bolus 48 h after initiation of feed restriction. Plasma NEFA concentration decreased from 546 μEq/L to 208 ± 141 μEq/L at 1 h after the infusion of 6 mg of NA/kg of BW, and to less than 100 ± 148 μEq/L at 3 h after the abomasal infusion of the 2 highest doses of NA. A rebound occurred after the initial decrease of plasma NEFA concentration. The rebound lasted up to 9 h for the 30-mg dose of NA, and up to 6 h for the 6-mg dose. Experiment 2 was a randomized complete block design with 3 treatments and 6 cows. Starting at 48 h of feed restriction, cows received 9 hourly abomasal infusions of 0, 6, or 10 mg of NA/kg of BW. Plasma NEFA concentrations decreased from 553 μEq/L ± 24 immediately before the initiation of treatments to <100 μEq/L during hourly infusions of 6 or 10 mg of NA/kg. Data suggest that the maximal antilipolytic response was achieved with the lowest dose of NA. A rebound of NEFA started 2 to 3 h after NA infusions were terminated. In both experiments, the NEFA rebound period coincided with increases in insulin and no change or increased glucose concentrations, suggesting a state of insulin resistance induced by elevated NEFA. This model for reducing plasma NEFA concentration by abomasal infusions of NA can be used to study the metabolic ramifications of elevated vs. reduced NEFA concentrations. The data demonstrate potential benefits and pitfalls of using NA to regulate plasma NEFA and prevent lipid-related metabolic disorders.     

Effects of prepartum 2,4-thiazolidinedione on metabolism and performance in transition dairy cows

Thiazolidinediones (TZD) are potent synthetic ligands for peroxisome proliferator-activated receptor-γ that have been shown previously to reduce plasma nonesterified fatty acids and increase peripartal dry matter intake (DMI) in dairy cows. Data from Holstein cows (n = 36) entering their second or greater lactation were used to determine whether late prepartum administration of TZD would affect periparturient metabolism, milk production, and ovarian activity. Cows were administered 0, 2.0, or 4.0 mg of TZD/kg of BW by intrajugular infusion once daily from 21 d before expected parturition until parturition. Plasma samples were collected daily from 22 d before expected parturition through 21 d postpartum and twice weekly from wk 4 through 9 postpartum. In response to increasing TZD dosage, plasma nonesterified fatty acid concentrations decreased linearly during the postpartum period (d 0 to +21: 348, 331, 268 ± 31 μEq/L, respectively). Plasma concentrations of glucose were highest in cows administered 4.0 mg of TZD/kg of BW during the peripartum and postpartum periods (d −7 to +7: 57.9, 57.8, 61.1 ± 0.8 mg/dL and d 0 to +21: 51.6, 49.3, 54.7 ± 1.1 mg/dL, respectively). Plasma concentrations of β-hydroxybutyrate were increased during the peripartum period by TZD administration (9.6, 9.9, 10.2 ± 0.3 mg/dL) but were not affected during the postpartum period. Plasma insulin was not affected by treatment during any time period. Postpartum liver triglyceride content was decreased linearly (11.0, 10.4, 4.2 ± 1.6%) and glycogen content was increased linearly (2.16, 2.38, 2.79 ± 0.19%) by prepartum TZD administration. Prepartum TZD administration linearly increased DMI during the peripartum period (d −7 to +7: 16.1, 17.2, 17.3 ± 0.5 kg/d). Cows administered TZD prepartum maintained higher postpartum body condition scores than control cows (wk 1 through 9: 2.77, 2.89, 3.02 ± 0.05). There was no effect of prepartum TZD on milk yield; however, yields of 3.5% fat-corrected milk (52.2, 54.6, 48.0 ± 1.6 kg/d) and most other milk components were decreased in cows that received 4.0 mg of TZD/kg of BW prepartum. Prepartum TZD administration linearly decreased the number of days to first ovulation (29.3, 28.3, 19.0 ± 3.6 d). These results suggest that prepartum administration of TZD improves metabolic health and DMI of periparturient dairy cows and may decrease reliance on body fat reserves during early lactation.     

Effects of short dry periods on performance and metabolic status in Holstein dairy cows

To evaluate effects of different dry period lengths on milk yield, milk composition, and energy balance of dairy cows, 122 multiparous and primiparous Holstein dairy cows were used in a completely randomized experimental design with 56-, 42-, and 35-d dry period lengths. Actual dry period lengths for respective treatments (TRT) were 56 ± 5.1 d, 42 ± 2.1 d, and 35 ± 2.7 d. Overall, cows in the 42- and 56-d TRT gained more body condition than those in 35-d TRT during the dry period; however, postpartum body condition score did not change substantially among the TRT. Although from 3 to 210 DIM, differences were not detected in the milk yield of multiparous cows between the 35- and 56-d TRT, primiparous cows in the 35-d TRT produced less milk than those in 56-d TRT. In primiparous cows, the milk production at wk 9, 10, and 11 of lactation was lower in the 35-d compared with the 56-d TRT. Primiparous cows in the 35-d compared with the 56-d TRT produced less milk protein. In the 35-d TRT, serum triglyceride concentration was greater in primiparous cows than in multiparous cows during the peripartum period. Among primiparous cows, those in the 56-d TRT had greater concentrations of nonesterified fatty acids than those in the 35-d TRT during the peripartum period. No significant differences were observed in concentrations of serum glucose, insulin, and insulin-like growth factor-I during early lactation among TRT. There was also no difference among TRT for incidence of metabolic disorders. Thus, this study indicates that shortening the dry period to 35 d may be beneficial in multiparous and overconditioned cows, but not in primiparous cows.     

Acute phase proteins in bovine milk in an experimental model of Staphylococcus aureus subclinical mastitis

The objectives were to establish the origin of 2 acute phase proteins in milk during subclinical bovine mastitis and to characterize the relationship between those proteins in milk and blood. Haptoglobin (Hp) and mammary-associated serum amyloid A (M-SAA3) appear in milk during mastitis, whereas Hp and serum amyloid A increase in serum during mastitis. The concentrations of these proteins were determined in an experimental model using a field strain of Staphylococcus aureus to induce subclinical mastitis in dairy cows. The expression of mRNA coding for these proteins was assessed and the presence of M-SAA3 in mammary tissues was determined using immunocytochemistry. Increases of M-SAA3 and Hp in milk occurred within 12 h of Staphylococcus aureus infusion, with peak concentrations occurring 3 d after infusion of the bacteria. The increase of acute phase proteins in milk (15 h) preceded the increase in serum concentrations of both proteins (24 h). Expression of mRNA for M-SAA3 and Hp increased in both mammary and hepatic tissues 48 h after infusion of the mammary glands. In mammary tissue, the increase of M-SAA3 mRNA was greater than the increase in Hp mRNA expression, whereas in hepatic tissue, the increase in M-SAA3 mRNA was less than that for Hp mRNA. Immunocytochemistry demonstrated that M-SAA3 protein was present within secretory epithelial cells at significantly higher levels in infected mammary glands than in control tissues. These proteins, which have host defense and antibacterial activities, may play a significant role in the early response to invasion of mammary tissues by pathogenic bacteria.     

Naturally occurring mastitis effects on timing of ovulation, steroid and gonadotrophic hormone concentrations, and follicular and luteal growth in cows

The effects of naturally occurring subclinical chronic or clinical short-term mastitis on time of ovulation, plasma steroid and gonadotropin concentrations, and follicular and luteal dynamics were examined in 73 lactating Holstein cows. Cows were sorted by milk somatic cell count and bacteriological examination into an uninfected group (n = 22), a clinical mastitis group (n = 9; events occurring 20 ± 7 d before the study), and a subclinical chronic mastitis group (n = 42). In addition, uninfected and mastitic cows were further sorted by their estrus to ovulation (E-O) interval. About 30% of mastitic cows (mainly subclinical) manifested an extended E-O interval of 56 ± 9.2 h compared with 28 ± 0.8 h in uninfected cows and 29 ± 0.5 h in the other 70% of mastitic cows. In mastitic cows with extended E-O interval, the concentration of plasma estradiol at onset of estrus was lower than that of uninfected cows or mastitic cows that exhibited normal E-O intervals (3.1 ± 0.4, 5.8 ± 0.5, and 5.5 ± 0.5 pg/mL, respectively). The disruptive effect of mastitis on follicular estradiol probably does not involve alterations in gonadotropin secretion because any depressive effects of mastitis on pulsatile LH concentrations were not detected. Cortisol concentrations did not differ among groups. The preovulatory LH surge in mastitic cows with delayed ovulation varied among individuals, being lower, delayed, or with no surge noted compared with the normal LH surge exhibited by uninfected cows or mastitic cows with normal E-O interval (6.8 ± 0.7 ng/mL). The diameter of the second-wave dominant follicle was larger and the number of medium follicles was smaller in uninfected and subclinical cows with normal intervals compared with subclinical cows with extended intervals (13.4 ± 0.5 vs. 10.9 ± 0.9 mm, and 3.8 ± 0.2 vs. 6.7 ± 0.14 follicles, respectively). Mid-luteal progesterone concentrations were similar in uninfected and mastitic cows. These results indicate for the first time that around 30% of cows with subclinical chronic mastitis exhibit delayed ovulation that is associated with low plasma concentrations of estradiol and a low or delayed preovulatory LH surge.     

Effect of milking frequency and dosing interval on the pharmacokinetics of cephapirin after intramammary infusion in lactating dairy cows

The objective was to determine the effect of milking frequency and dosing interval on pharmacokinetics of cephapirin after intramammary infusion. Six healthy Holstein cows were administered cephapirin (200 mg) into 1 rear mammary gland after each of 2 milkings. Cows were milked twice daily (2×) and dosed at a 12-h interval or 3 times daily (3×) and dosed at an 8- or 16-h interval. A duplicated Latin square design allowed each cow to receive all 3 frequency-dose treatments, with intervening washout periods. Concentrations of cephapirin (CEPH) and desacetylcephapirin (DAC) in milk from the treated glands were determined at each milking after infusion using liquid chromatography-mass spectrometry. Data were fitted using 1- and 2-compartment pharmacokinetic models, as well as a noncompartmental model. Cephapirin was rapidly metabolized to DAC in the mammary gland, with DAC being the predominant agent in milk until 48 h after infusion. Pharmacokinetics of CEPH and DAC were similar for all treatment groups, with a 1-compartment model providing a better fit than a 2-compartment model in most instances. Milking frequency did not affect the length of time that milk CEPH concentration exceeded MIC₅₀ or MIC₉₀ values (the minimum inhibitory antimicrobial concentration needed to inhibit 50 or 90% of microbial activity, respectively) for common mastitis pathogens, except that cows milked 3× and dosed at a 16-h interval maintained inhibitory concentrations approximately 8 h longer than those dosed at an 8-h interval. Time for milk CEPH concentration to reach the FDA tolerance did not differ among treatment groups [mean ± SD; 68 ± 20, 66 ± 22, and 57 ± 18 h after last treatment for cows treated at 12, 16, and 8 h, respectively]. Mean residence time for CEPH in the mammary gland was linearly and negatively associated with the volume of milk produced. Calculated CEPH concentration in composite milk from all 4 mammary glands was below the FDA tolerance in all cows by 96 h after the last infusion, which is the labeled withholding time for the preparation used. Our findings suggest that this CEPH preparation, which is labeled for 2 doses 12 h apart, could be administered at a 16-h interval in herds milking 3×, with no significant effect on inhibitory concentrations in milk or withdrawal time; extended withdrawal times would be prudent for cows with very low milk production. Further investigation is needed to determine if milking frequency affects CEPH pharmacokinetics in cows with clinical mastitis.     

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.     

Natural antibodies related to energy balance in early lactation dairy cows

The objectives of this study were to determine the presence of natural antibodies (NAb) in plasma and milk of individual dairy cows and to study the relation between NAb concentrations and energy balance (EB) and dietary energy source. Cows (n = 76) were fed a mainly glucogenic, lipogenic, or a mixture of both diets (50:50 dry matter basis) from wk 3 before the expected calving date until wk 9 postpartum. Diets were isocaloric (net energy basis) and equal in intestinal digestible protein. Blood and milk were sampled weekly. Liver biopsies were taken in wk −2, 2, 4, and 6 relative to calving. Data are expressed as LSM ± SEM. The NAb titers are expressed as the ²log values of the highest dilution giving a positive reaction. The NAb concentration in plasma binding either keyhole limpet hemocyanin (KLH) or Escherichia coli lipopolysaccharide (LPS) increased with parity. The NAb concentration binding KLH was greater for cows fed the glucogenic diet (9.63 ± 0.08) compared with the lipogenic diet (9.26 ± 0.08). In milk, cows fed the glucogenic diet had smaller NAb concentrations binding KLH (3.98 ± 0.18) and LPS (2.88 ± 0.17) compared with cows fed the mixed diet (KLH: 4.93 ± 0.18; LPS: 3.70 ± 0.17). The NAb concentration in plasma had a positive relation with energy balance variables: EB, dry matter intake, milk yield, and plasma cholesterol, whereas NAb concentration in milk had a negative relation with energy balance variables: EB, dry matter intake, and plasma cholesterol. Additionally, NAb concentrations in milk had a positive relation with plasma nonesterified fatty acid concentration and milk fat and protein percentage. There was a tendency for a positive relation of NAb concentration binding LPS in plasma and somatic cell count in milk. No significant relations were detected between NAb concentrations in milk or plasma and plasma β-hydroxybutyrate concentration and liver triacyl glyceride content. In conclusion, NAb are present in both milk and plasma of dairy cows peripartum and NAb concentrations increase with parity. Furthermore, our data indicate that a negative energy balance in dairy cows in early lactation can be associated with compromised innate immune function as indicated by decreased NAb concentration in plasma.     

Production and health of cows given monensin prepartum and a high-energy diet postpartum

The object of this study was to evaluate the impact of monensin administration on the early lactation performance of cows maintained on a high-energy diet, and on health traits during the transition period. Cows (n = 168; parity 3.3 ± 1.4, initial body condition score 3.1 ± 0.08, and milk yield of 34.3 kg/d ± 0.9 for multiparous cows in the preceding lactation) were divided into control and monensin treatment groups. A controlled-release capsule supplying 335 mg of monensin/d for 95 d was inserted into the rumen of monensin-treated cows 30 d before the expected calving. Blood samples were obtained 2 h after feeding on d 14 prepartum and on d 7, 14, and 50 postpartum. Plasma glucose concentration was 3% higher (58 ± 0.5 vs. 56.4 ± 0.5 mg/dL) and β-hydroxybutyric acid was 17% (6.7 ± 0.3 vs. 8.0 ± 0.3 mg/dL) lower in monensin-treated than control cows. Plasma glucose was 10% higher (60.0 ± 0.6 vs. 54.5 ± 0.3 mg/dL) and β-hydroxybutyric acid was 16% lower (6.8 ± 0.3 vs. 7.9 ± 0.2 mg/dL) in primiparous than multiparous cows. Plasma nonesterified fatty acid concentration (measured only in primiparous cows) was 17% lower (287 ± 15 vs. 336 ± 17 μEq/L) in treated than in control cows. Rate of ketosis incidence was 60% lower (8 vs. 21%) in monensin-treated than in control cows, and the proportion of control cows that required a supply of glucogenic precursors was 3-fold higher than in monensin-treated cows. The body condition score was 3.1 ± 0.05, 2.7 ± 0.05, and 2.4 ± 0.05 on d 60 prepartum and d 7 and 50 postpartum, respectively, and was not affected by treatment. During the first 5 mo of lactation, milk yield was 7% higher (37.6 ± 0.6 vs. 35.2 ± 0.6 kg/d) in monensin-treated cows than in control cows. Our results showed that monensin administration, as a controlled-release capsule in prepartum cows, can be beneficial, even if these cows are maintained on a high-energy diet during the subsequent lactation.     

Prepartum 2,4-thiazolidinedione alters metabolic dynamics and dry matter intake of dairy cows

Thiazolidinediones (TZD) are potent, synthetic ligands for peroxisome proliferator activated receptor-gamma (PPAR-γ) that reduce plasma nonesterified fatty acids (NEFA) and potentiate the action of insulin in peripheral tissues of several species. Holstein cows (n = 9) entering their second or greater lactation were used to determine whether late prepartum administration of TZD would affect periparturient metabolism and milk production. Cows were limit-fed a total mixed ration (TMR) during the prepartum period to provide no more than 130% of predicted energy requirements. During the postpartum period cows were fed a common TMR for ad libitum intake. Cows were administered either 2,4-TZD (4.0 mg/kg of body weight) or saline (control) by intrajugular infusion once daily from 25 d before expected parturition until parturition. Plasma samples were collected daily from 26 d before expected parturition through 7 d postpartum. Plasma NEFA concentrations decreased during the prepartum period (d −21 to −1; 70 vs. 83 ± 4 μEq/L) and tended to be decreased during the peripartum period (d −7 to d +7; 113 vs. 205 ± 32 μEq/L) due to prepartum TZD administration. Plasma concentrations of glucose were not affected by treatment; however, plasma β-hydroxybutyrate concentrations decreased in TZD-treated cows (8.6 vs. 10.7 ± 1.7 mg/dL) as parturition approached, and plasma insulin concentrations increased during the peripartum period (0.65 vs. 0.38 ± 0.07 ng/mL). Postpartum liver triglyceride and glycogen content was not affected by treatment. Prepartum TZD administration tended to increase dry matter intake during the peripartum and postpartum periods (16.6 vs. 14.6 ± 0.8 kg/d and 20.0 vs. 17.2 ± 1.2 kg/d, respectively). Milk yield for the first 30 d postpartum and milk composition measured on d 8 postpartum were not affected by treatment. There was no effect of prepartum TZD administration on insulin-dependent glucose utilization assessed using insulin challenge during either the prepartum or postpartum periods. These results suggest that administration of TZD during the late prepartum period has the potential to improve metabolic health and DMI of periparturient dairy cows and warrants further investigation.     

Supplemental choline for prevention and alleviation of fatty liver in dairy cattle

Two experiments were conducted to evaluate if supplementing rumen-protected choline (RPC; Reashure, Balchem Encapsulates, Slate Hill, NY) could prevent or alleviate fatty liver in dairy cattle. The first experiment evaluated the effect of supplementing RPC on hepatic triacylglycerol (TAG) accumulation during fatty liver induction. Twenty-four dry cows between 45 to 60 d prepartum were paired by body weight (BW) and body condition score (BCS) and randomly assigned to control or supplementation with 15 g of choline as RPC/d. From d 0 to 6, before treatment application, all cows were fed 1.4 kg/d of concentrate and forage ad libitum. Samples of blood and liver, obtained during the pretreatment period, were used for covariate adjustment of blood metabolites and liver composition data. During fatty liver induction (d 7 to 17), cows were fed 1.4 kg/d of concentrate with or without supplementation with RPC, and forage intake was restricted, so cows consumed 30% of the total energy requirements for pregnancy and maintenance. Supplementation with RPC during fatty liver induction did not affect plasma glucose and plasma β-hydroxybutyrate (BHBA) concentration but did decrease plasma nonesterified fatty acid (NEFA; 703 vs. 562 μEq/L, SE = 40) and liver TAG accumulation (16.7 vs. 9.3 μg/μg of DNA, SE = 2.0). In the second experiment, we evaluated the effect of supplementing RPC on the clearance of liver TAG when cows were fed ad libitum after the induction of fatty liver by feed restriction. Twenty-eight cows between 45 and 60 d prepartum were paired according to BCS and BW and assigned to treatments. Fatty liver was induced by feeding 1.4 kg/ d of concentrate (without RPC) and restricting forage intake, so cows consumed 30% of maintenance and pregnancy energy requirements for 10 d. From d 11 to 16, after feed restriction, cows were fed forage ad libitum and 1.4 kg/d of concentrate with or without RPC. Treatments were not applied during fatty liver induction; however, following feed restriction, liver for cows assigned to control and RPC treatments contained 6.8 and 12.7 μg of TAG/μg of DNA, respectively. Measurements obtained before treatment served as covariates for statistical analysis. During the depletion phase, plasma glucose, BHBA, and NEFA were not affected by treatment. Liver TAG, expressed as covariate adjusted means, was 6.0 and 4.9 μg/μg of DNA (SE = 0.4) on d 13, and 5.0 and 1.5 μg/μg of DNA (SE = 0.9) on d 16 for control and RPC, respectively. Rumen-protected choline can prevent and possibly alleviate fatty liver induced by feed restriction.