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Summary: Supplementation of dairy cows with trans‐10, cis‐12 conjugated linoleic acid (CLA) allows nutrient repartitioning despite an energy deficiency in early lactation, which might be a benefit for the immune system, too. In this study, we investigated potential nutrient sparing effects of CLA in early lactating cows with low plasma glucose concentrations exposed to an intramammary lipopolysaccharide (LPS) challenge. Fifteen multiparous Holstein cows were exposed to an intramammary LPS challenge in week 4 p.p. Eight cows (CLA) were supplemented daily with 70 g of lipid‐encapsulated CLA (6.8 g trans‐10, cis‐12 and 6.6 g of the cis‐9, trans‐11 CLA isomer; CLA) and seven cows with 56 g of control fat (CON). Blood samples were obtained every 30 min along with rectal temperature, heart and respiratory rate, and milk samples were taken hourly until 10 hr after the LPS application. Plasma was analysed for concentrations of glucose, free fatty acids, beta‐hydroxybutyrate (BHB), cortisol, insulin and glucagon. In milk, somatic cell count and activity of lactate dehydrogenase were determined. Initial plasma glucose concentration was lower in CLA than in CON. During the immunostimulation, CLA had higher glucose concentrations than CON, and BHB decreased distinctly in CLA, whereas CON cows maintained BHB concentration at a lower level. Body temperature in CLA increased earlier, the difference between peak and basal temperature was higher, and the decline thereafter occurred earlier. In conclusion, CLA supplementation of early lactating cows exposed to an intramammary LPS challenge affected local and systemic immune responses. We assume that CLA supplementation triggered glycogen storage. Cows supplemented with CLA provided more glucose and preferentially used BHB as an energy source during the immune response. The more intense metabolic and more concentrated endocrine responses support an immunomodulatory effect of CLA supplementation. [ABSTRACT FROM AUTHOR]

Summary: Feeding dairy cows indoors or on pasture affects not only labour, machinery and housing costs, but also animals’ performance and metabolism. This study investigates the effects of indoor feeding (IF) with a partial‐mixed ration (PMR) versus pasture‐based feeding (PF) on milk production, fertility, backfat thickness (BFT), body weight (BW) loss and energy metabolism of Brown Swiss (BS) dairy cows with similar genetic production potential. The IF herd consisted of 13 cows fed a PMR composed of maize and grass silage plus protein concentrate according to each cow's requirements. The PF herd consisted of 14 cows offered barn‐ventilated hay ad libitum after calving from January until March and grazed on semi‐continuous pastures during the vegetation period. The IF cows produced more energy‐corrected milk (ECM) per standard lactation (9,407 vs. 5,960 kg; p < .01), more milk fat (378 vs. 227 kg; p < .01) and milk protein (326 vs. 215 kg; p < .01). The calving interval (377 vs. 405 days; p < .01) and time empty (86 vs. 118 days; p < .01) were shorter in the PF compared to IF, possibly also due to different selection criteria for maintaining the respective seasonal calving rhythm. The empty body fat loss calculated according to BCS until its nadir was higher in IF cows (IF: 10.4 vs. PF: 4.8 MJ/day; p < .01), but no differences were noted in total body fat loss estimated via BFT (p = .24). However, PF had lower blood glucose concentration at all investigated time points, but no differences occurred in serum non‐esterified fatty acid and β‐hydroxybutyrate concentrations post‐partum. In conclusion, BS cows were equally well suited for the IF with PMR and the PF system investigated here without developing a prominent metabolic load despite differences in nutrient supply. As such, investigated BS dairy cows in our trial seem to have a high capacity for metabolic adaptation to different production systems. [ABSTRACT FROM AUTHOR]

Summary: Supplementing conjugated linoleic acid (CLA) is supposed to spare glucose due to the milk fat‐depressing effect of the trans‐10, cis‐12 CLA isomer, and allows repartitioning nutrients despite an energy deficiency in early lactation. However, there is still a lack of knowledge in terms of the dynamic pattern of the glucose turnover in transition dairy cows. We hypothesized that dairy cows supplemented with CLA have an altered rate of glucose turnover and insulin sensitivity during early lactation. We conducted three consecutive hyperglycaemic clamps (HGC) in weeks −2, +2 and +4 relative to parturition in Holstein cows supplemented daily either with 70 g of lipid‐encapsulated CLA (6.8 g trans‐10, cis‐12 and 6.6 g of the cis‐9, trans‐11 CLA isomer; CLA; n = 11) or with 56 g of control fat (CON; n = 11). From week −3 up to week +4 relative to parturition, milk yield and dry matter intake (DMI) were recorded daily, while body weight (BW) and milk composition were obtained once weekly. Blood samples were taken once weekly and every 30 min during the HGC. Plasma was analysed for concentrations of glucose, fatty acids (FFA), beta‐hydroxybutyrate (BHB), insulin, triglycerides and cholesterol. The CLA supplementation did not affect performance and metabolic parameters except for BHB and cholesterol. Furthermore, insulin concentrations and insulin sensitivity were affected by treatment. During the HGC in early lactation, insulin response was lower and decrease in FFA and BHB greater compared with the HGC in week −2 although glucose target concentration achieved during the steady‐state period was similar for all three HGC. Our findings in terms of insulin and cholesterol suggest that body reserves are preserved through CLA feeding without restraining animal's performance. Furthermore, CLA effects on cholesterol and triglyceride concentrations indicated beneficial effects on hepatic lipid export contributing to an improved efficiency of prevailing metabolites in circulation. [ABSTRACT FROM AUTHOR]

Changes in ACTH challenge test characteristics in dairy cows changing their physiological status at different lactational stages and different feeding levels were not investigated in terms of repeatability yet. In 23 multiparous Holstein cows (10 cows fed a sole fresh herbage diet without concentrate, 13 cows fed with concentrate), three ACTH challenge tests were performed: once during pregnancy shortly prior to drying off ( T1), and in week 3 ( T2) and 8 ( T3) after parturition. Test characteristics were correlated to performance and metabolic parameters: DMI, BW, energy balance ( EB), plasma concentrations of free fatty acids ( NEFA) and beta-hydroxybutyrate ( BHB). Basal plasma cortisol concentrations were higher at T1 compared with T2 and T3 ( p < .05). The adrenal cortex sensitivity (expressed as total AUC ( AUCt) of cortisol response after ACTH application) was lowest at T2 compared with T1 and T3 ( p < .05). Ranking of the individual animals' responses was not repeatable between time points of the ACTH tests. Enhancing the energy deficiency during early lactation by omission of concentrate did not affect baseline cortisol concentrations in plasma, but decreased peak height at T2 ( p < .05). Baseline plasma cortisol concentrations were positively correlated with cortisol peak values after ACTH application, previous lactation performance, milk yield and BW ( p < .05). The AUCt was positively correlated with baseline cortisol concentrations, EB and DMI. Cortisol release after ACTH injection was lower in animals with high plasma concentrations of NEFA, BHB and with higher contents of fat and free fatty acids in milk ( p < .05). Cortisol peak height after ACTH administration was higher in cows with a more positive EB, higher DMI and lower plasma concentrations of NEFA and BHB. In summary, cortisol responses to ACTH challenges in this study were not repeatable in dairy cows changing their physiological status. [ABSTRACT FROM AUTHOR]

Herbage feeding with only little input of concentrates plays an important role in milk production in grassland dominated countries like Switzerland. The objective was to investigate the effects of a solely herbage-based diet and level of milk production on performance, and variables related to the metabolic, endocrine and inflammatory status to estimate the stress imposed on dairy cows. Twenty-five multiparous Holstein cows were divided into a control (C+, n = 13) and a treatment group (C−, n = 12), according to their previous lactation yield (4679-10 808 kg) from week 3 ante partum until week 8 post-partum (p.p.). While C+ received fresh herbage plus additional concentrate, no concentrate was fed to C− throughout the experiment. Within C+ and C−, the median of the preceding lactation yields (7752 kg) was used to split cows into a high ( HYC+, HYC−)- and low-yielding ( LYC+, LYC−) groups. Throughout the study, HYC+ had a higher milk yield (35.9 kg/d) compared to the other subgroups (27.2-31.7 kg/d, p < 0.05). Plasma glucose (3.51 vs. 3.72 mmol/l) and IGF-1 (66.0 vs. 78.9ng/ mL) concentrations were lower in HYC−/ LYC− compared to HYC+/ LYC+ cows (p < 0.05). Plasma FFA and BHBA concentrations were dramatically elevated in HYC− (1.1 and 1.6 mmol/l) compared to all other subgroups (mean values: 0.5 and 0.6 mmol/l, p < 0.05). Saliva cortisol, plasma concentrations of serum amyloid A ( SAA), haptoglobin (Hp), beta-endorphin ( BE) and activity of alkaline phosphatase ( AP) were not different between C+ and C−. In conclusion, herbage-fed high-yielding cows without supplementary concentrate experienced a high metabolic load resulting in a reduced performance compared to cows of similar potential fed accordingly. Low-yielding cows performed well without concentrate supplementation. Interestingly, the selected markers for inflammation and stress such as cortisol, Hp, SAA, BE and AP gave no indication for the metabolic load being translated into compromised well-being. [ABSTRACT FROM AUTHOR]

The link between energy availability, turnover of energy substrates and the onset of inflammation in dairy cows is complex and poorly investigated. To clarify this, plasma inflammatory variables were measured in mid-lactating dairy cows allocated to three groups: hyperinsulinemic hypoglycaemic clamp, induced by insulin infusion (HypoG, n = 5); hyperinsulinemic euglycaemic clamp, induced by insulin and glucose infusion (EuG; n = 6); control, receiving a saline solution infusion (NaCl; n = 6). At 48 h after the start of i.v. infusions, two udder quarters per cow were challenged with 200 μg of E. coli lipopolysaccharide ( LPS). Individual blood samples were taken before clamps, before LPS challenge (i.e. 48 h after clamps) and 6.5 h after. At 48 h, positive acute phase proteins (pos APP) did not differ among groups, whereas albumin and cholesterol (index of lipoproteins), negative APP (neg APP), were lower (p < 0.05) in EuG compared to NaCl and HypoG. The concentration of IL-6 was greater in EuG (p < 0.05) but only vs. HypoG. At 6.5 h following LPS challenge, IL-6 increased in the NaCl and EuG clamps (p < 0.05), while TNF- α increased (p < 0.05) in the EuG only. Among the pos APP, haptoglobin markedly increased in EuG (p < 0.05), but not in NaCl (p = 0.76) and in HypoG; ceruloplasmin tended to decline during LPS challenge, the reduction was significant when all animals were considered (p < 0.05). Conversely, all the neg APP showed a marked reduction 6.5 h after LPS challenge in the three groups. In conclusion, EuG caused an inflammatory status after 48-h infusion (i.e. decrease of neg APP) and induced a quicker acute phase response (e.g. marked rise of TNF- α, IL-6) after the intramammary LPS challenge. These data suggest that the simultaneous high availability of glucose and insulin at the tissue-level makes dairy cows more susceptible to inflammatory events. In contrast, HypoG seems to attenuate the inflammatory response. [ABSTRACT FROM AUTHOR]

The aim was to study the variation in metabolic responses in early-lactating dairy cows ( n = 232) on-farm that were pre-selected for a high milk fat content (>45 g/l) and a high fat/protein ratio in milk (>1.5) in their previous lactation. Blood was assayed for concentrations of metabolites and hormones. Liver was measured for mRNA abundance of 25 candidate genes encoding enzymes and receptors involved in gluconeogenesis (6), fatty acid β-oxidation (6), fatty acid and triglyceride synthesis (5), cholesterol synthesis (4), ketogenesis (2) and the urea cycle (2). Two groups of cows were formed based on the plasma concentrations of glucose, non-esterified fatty acids (NEFA) and β-hydroxybutyric acid (BHBA) (GRP+, high metabolic load; glucose <3.0 m m, NEFA >300 μ m and BHBA >1.0 m m, n = 30; GRP−, low metabolic load; glucose >3.0 m m, NEFA <300 μ m and BHBA <1.0 m m, n = 30). No differences were found between GRP+ and GRP− for the milk yield at 3 weeks post-partum, but milk fat content was higher (p < 0.01) for GRP+ than for GRP-. In week 8 post-partum, milk yield was higher in GRP+ in relation to GRP− (37.5 vs. 32.5 kg/d; p < 0.01). GRP+ in relation to GRP− had higher (p < 0.001) NEFA and BHBA and lower glucose, insulin, IGF-I, T3, T4 concentrations (p < 0.01). The mRNA abundance of genes related to gluconeogenesis, fatty acid β-oxidation, fatty acid and triglyceride synthesis, cholesterol synthesis and the urea cycle was different in GRP+ compared to GRP− (p < 0.05), although gene transcripts related to ketogenesis were similar between GRP+ and GRP−. In conclusion, high metabolic load post-partum in dairy cows on-farm corresponds to differences in the liver in relation to dairy cows with low metabolic load, even though all cows were pre-selected for a high milk fat content and fat/protein ratio in milk in their previous lactation. [ABSTRACT FROM AUTHOR]

Summary Thirty-two multiparous Holstein cows were used to investigate the effects of chromium- l-methionine (Cr-Met) supplementation and dietary grain source on metabolic indices throughout the periparturient period. Cows were fed a total mixed ration with the concentrate portion based on ground barley (barley based diet, BBD) - or ground corn (corn-based diet, CBD) from 21 days before anticipated calving through 28 days after calving. The Cr-Met was supplemented at dosages of 0 or 0.08 mg of Cr/kg of metabolic body weight throughout the experiment. Thus, treatments were in a 2 (Cr-Met) × 2 (grain sources) factorial arrangement. Plasma glucose, non-esterified fatty acids, triglyceride, very low-density lipoprotein, blood urea nitrogen (BUN), insulin, glucagon, cortisol and insulin-like growth factor-I (IGF-I) concentrations were not affected by chromium supplementation on calving day (d 0). However, there was a trend for decreased β-hydroxybutyrate (BHBA) and increased cholesterol on d 0 in cows fed the Cr supplemented diet. On d 21 postpartum (wk 3 p.p.), plasma glucagon concentration tended to be greater in cows fed the Cr supplemented diet. However, other plasma metabolite and hormone measures were not affected by Cr supplementation in wk 3 p.p. There was no effect of grain source on d 0 plasma metabolic and endocrine measures. In wk 3 p.p., a significant difference was found only for plasma concentration of glucagon that was higher for cows fed the BBD compared with the CBD. There was an interaction of Cr-Met supplementation and grain source in wk 3 p.p. plasma concentration of BUN to decrease in CBD and increase in BBD, supplemented with Cr-Met. No Cr by grain interactions were observed for other plasma metabolic variables on d 0 and in wk 3 p.p. These results indicate that Cr-Met supplementation and substituting barley grain with corn throughout the transition period have only moderate effects on metabolic and endocrine parameters. [ABSTRACT FROM AUTHOR]

Summary Grass may have a differential impact on the metabolism of the dairy cow, depending on the grazing system applied. In this study, the hypothesis was tested that metabolism of grazed vs. zero-grazed dairy cows is differently regulated throughout the vegetation period. The study included three experimental periods (p1, p2 and p3) of 14 days each, and two treatments [grazing from pasture, PASTURE, n = 9; or zero-grazing in a free-stall barn (BARN, n = 9)]. Blood and liver samples were collected at the end of each period when the cows were on average 64, 120 and 197 DIM. Concentrations of metabolites and hormones, and activities of various enzymes were determined in plasma. Liver samples were measured for mRNA abundance of genes encoding enzymes and nuclear receptors involved in metabolic pathways. PASTURE cows had higher plasma concentrations of T3, BHB, and total protein than BARN cows across periods (p < 0.05). BARN cows had higher concentrations of NEFA (in p1) and urea (in p1 and p2) in relation PASTURE cows (p < 0.05), and had higher mRNA abundance of liver-X-receptor-α and glycerol-3-phosphate-acyltransferase across periods (p < 0.05). The results confirm the hypothesis that metabolism is different between BARN and PASTURE cows throughout the vegetation period, and show that the observed differences are mainly reflected in parameters of lipid metabolism. [ABSTRACT FROM AUTHOR]

Summary Blood plasma and hepatic parameters were identified that describe the differences between metabolically robust or vulnerable dairy cows grouped according to their past health status. Data from a field study on dairy cows were used from which metabolically challenged dairy cows were selected that had a milk fat percentage of >4.5 mg/g and a fat to protein ratio of >1.5 in their previous early lactation. The selected cows were either classified as metabolically robust or vulnerable based on the occurrence of various metabolic and (re)production disorders in their previous lactations. Blood and liver tissue samples were collected in week 3 ante partum (a.p.) (−3wk), in week 4 (+4wk) and in week 13 (+13wk) post-partum (p.p.). Plasma concentrations of metabolites and hormones and mRNA expression of genes involved in metabolic pathways in the liver were used as variables for a two-group discriminant analysis (DA). Average discriminant scores (centroids) were different (p < 0.05) in −3wk, +4wk and in +13wk. In −3wk, significant variables that best explained the differences between metabolically robust and vulnerable cows were parity, plasma triglycerides, glucose and mRNA abundance of carnitine palmitoyltransferase 2 (CPT2). In addition, based on the classification matrix, 69% of the dairy cows were correctly classified. In +4wk, identified significant parameters were parity, plasma glucose and urea, and 67% of the cows were correctly classified. In +13wk, significant variables that explained the differences between the groups were parity, mRNA abundance of acyl-CoA synthetase long-chain 1 and CPT1, and 66% of the cows were correctly classified. In conclusion, the identified variables may distinguish from metabolically challenged cows, those cows that had a poorer health performance in their previous lactations. [ABSTRACT FROM AUTHOR]

Summary Dairy cows with high and low plasma non-esterified fatty acid (NEFA) concentrations in early lactation were compared for plasma parameters and mRNA expression of genes in liver and subcutaneous adipose tissue. The study involved 16 multiparous dairy cows with a plasma NEFA concentration of >500 μmol/l [ n = 8, high NEFA (HNEFA)] and <140 μmol/l [ n = 8, low NEFA (LNEFA)] in the first week post-partum (pp). Blood samples, adipose and liver tissues were collected on day 1 (+1d) and at week 3 pp (+3wk). Blood plasma was assayed for concentrations of metabolites and hormones. Subcutaneous adipose and liver tissues were analysed for mRNA abundance by real-time qRT-PCR encoding parameters related to lipid metabolism. Results showed that mean daily milk yield and milk fat quantity were higher in HNEFA than in LNEFA cows (p < 0.01), and the NEB was more negative in HNEFA than in LNEFA in +3wk too (p < 0.05). HNEFA cows had slightly lower (p < 0.1) insulin concentrations than LNEFA cows across the study period, and the body condition score decreased more from +1d to +3wk in HNEFA than in LNEFA (p = 0.09). The mRNA abundance of genes in the liver related to fatty acid oxidation (carnitine palmitoyltransferase 2 and very long chain acyl-coenzyme A dehydrogenase) and ketogenesis (3-hydroxy-3-methylglutaryl-coenzyme A synthase 2) were lower in HNEFA than in LNEFA cows. No differences between the two groups were observed for mRNA expression of genes in adipose tissue. The number of calculated significant correlation coefficients (moderately strong) between parameters in the liver and in adipose tissue was nearly similar on +1d, and higher for HNEFA compared with LNEFA cows in +3wk. In conclusion, dairy cows with high compared with low plasma NEFA concentrations in early lactation show differentially synchronized mRNA expression of genes in adipose tissue and liver in +3wk that suggests a different orchestrated homeorhetic regulation of lipid metabolism. [ABSTRACT FROM AUTHOR]

Summary The objective was to study changes in plasma leptin concentration parallel to changes in the gene expression of lipogenic- and lipolytic-related genes in adipose tissue of dairy cows around parturition. Subcutaneous fat biopsies were taken from 27 dairy cows in week 8 antepartum (a.p.), on day 1 postpartum (p.p.) and in week 5 p.p. Blood samples were assayed for concentrations of leptin and non-esterified fatty acids (NEFA). Subcutaneous adipose tissue was analysed for mRNA abundance by real-time qRT-PCR encoding for leptin, adiponectin receptor 1 (AdipoR1), adiponectin receptor 2 (AdipoR2), hormones-sensitive lipase (HSL), perilipin (PLIN), lipoprotein lipase (LPL), acyl-CoA synthase long-chain family member 1 (ACSL1), acetyl-CoA carboxlyase (ACC), fatty acid synthase (FASN) and glycerol-3-phosphate dehydrogenase 2 (GPD2). Body weight and body condition score of the cows were lower after parturition than before parturition. The calculated energy balance was negative in week 1 and 5 p.p., with higher negative energy balance in week 1 p.p. compared with that in week 5 p.p. On day 1 p.p., highest concentrations of NEFA (353.3 μmol/l) were detected compared with the other biopsy time-points (210.6 and 107.7 μmol/l, in week 8 a.p., and week 5 p.p. respectively). Reduced plasma concentrations of leptin during p.p. when compared with a.p. would favour increasing metabolic efficiency and energy conservation for mammary function and reconstitution of body reserves. Lower mRNA abundance of ACC and FASN expression on day 1 p.p. compared with other biopsy time-points suggests an attenuation of fatty acid synthesis in subcutaneous adipose tissue shortly after parturition. Gene expression of AdipoR1, AdipoR2, HSL, PLIN, LPL, ACSL1 and GPD2 was unchanged over time. [ABSTRACT FROM AUTHOR]

Gene expression of adipose factors, which may be part of the mechanisms that underlie insulin sensitivity, were studied in dairy cows around parturition. Subcutaneous fat biopsies and blood samples were taken from 27 dairy cows in week 8 antepartum (a.p.), on day 1 postpartum (p.p.) and in week 5 p.p. In the adipose tissue samples, mRNA was quantified by real-time reverse transcription polymerase chain reaction for tumour necrosis factor alpha (TNFα), insulin-independent glucose transporter (GLUT1), insulin-responsive glucose transporter (GLUT4), insulin receptor, insulin receptor substrate 1 (IRS1), insulin receptor substrate 2 (IRS2), regulatory subunit of phosphatidylinositol-3 kinase (p85) and catalytic subunit of phosphatidylinositol-3 kinase. Blood plasma was assayed for concentrations of glucose, β-hydroxybutyric acid, non-esterified fatty acids (NEFA) and insulin. Plasma parameters followed a pattern typically observed in dairy cows. Gene expression changes were observed, but there were no changes in TNFα concentrations, which may indicate its local involvement in catabolic adaptation of adipose tissue. Changes in GLUT4 and GLUT1 mRNA abundance may reflect their involvement in reduced insulin sensitivity and in sparing glucose for milk synthesis in early lactation. Unchanged gene expression of IRS1, IRS2 and p85 over time may imply a lack of their involvement in terms of insulin sensitivity dynamics. Alternatively, it may indicate that post-transcriptional modifications of these factors came into play and may have concealed an involvement. [ABSTRACT FROM AUTHOR]

Liver tissue was collected from eight random dairy cows at a slaughterhouse to test if gene expression of pyruvate carboxylase (PC), mitochondrial phosphoenolpyruvate carboxykinase (PEPCKm) and cytosolic phosphoenolpyruvate carboxykinase (PEPCKc) is different at different locations in the liver. Obtained liver samples were analysed for mRNA expression levels of PC, PEPCKc and PEPCKm and subjected to the MIXED procedure of SAS to test for the sampled locations with cow liver as repeated subject. Additionally, the general linear model procedure (GLM) for analysis of variance was applied to test for significant differences for mRNA abundance of PEPCKm, PEPCKc and bPC between the livers. In conclusion, this study demonstrated that mRNA abundance of PC, PEPCKc and PEPCKm is not different between locations in the liver but may differ between individual cows. [ABSTRACT FROM AUTHOR]

This study tested the hypothesis that adrenaline has different effects on blood plasma glucose, lactate and nonesterified fatty acid (NEFA) concentrations in early and mid lactation and that the feeding of crystalline fat (consisting of triglycerides) and free fatty acids (FFA), compared with a starch-rich ration, modifies the effects of adrenaline in dairy cows. Glucose, lactate and NEFA responses to intravenously infused adrenaline [15 μg/kg × (min) for 10 min] were studied in weeks 9 and 19 of lactation in cows whose diets were supplemented with crystalline fat or FFA or a starch-rich ration. Concentrations of glucose, lactate and NEFA rapidly increased up to 15 min after the start of adrenaline infusions. Glucose responses to adrenaline were comparable in weeks 9 and 19 of lactation, whereas the lactate responses were smaller, and those of NEFA were greater in week 9 than in week 19 of lactation. As the feeding of crystalline fat, FFA or a starch-rich ration did not modify the effects of adrenaline on glucose, lactate or NEFA, the target tissue sensitivity and/or responsiveness to adrenaline had obviously not been changed. [ABSTRACT FROM AUTHOR]