Characterizing the metabotype and its persistency in lactating Holstein cows: An approach toward metabolic efficiency measures.

The variation in feed efficiency among dairy cows is due to differences in fermentation and digestion characteristics, but recent studies have suggested that various aspects of postabsorptive metabolic processes including heat production or the metabolizable energy for maintenance are more crucial. Thus, metabolic efficiency largely determines feed efficiency, but whether divergent feed efficient cows differ in O2 consumption and metabolic CO2 production, directly determining the metabolic rate has not been investigated. Therefore, the objective of the present study was to determine whether variation in ME intake (MEI), O2 consumption, and metabolic CO2 production account for the variation in metabolic efficiency of dairy cows and whether this effect persists across the lactation cycle. Eighteen cows with different German breeding value functional herd life were kept in freestalls with ad libitum access to a total mixed ration that was kept constant in composition throughout the first lactation. Cows were blood sampled and weighed at wk 5, 13, and 42 postpartum (pp) and transferred into respiration chambers. Animals were retrospectively clustered according to MEI, O2 consumption, and metabolic CO2 production, each normalized to metabolic body weight (mBW). Cluster analysis revealed 9 high metabolically efficient (high-Meff) and 9 low metabolically efficient cows. The high-Meff cows had greater MEI and feed conversion efficiency, produced less metabolic CO2 and methane, had a stronger negative energy balance, and tended to have a lower metabolic respiratory quotient. Further, high-Meff cows had lower residual MEI, less heat energy loss, and lower plasma glucose concentrations, but used a greater portion of body reserves instead of feed energy for milk synthesis, particularly at wk 5 and 13 pp. However, these group differences did not persist by wk 42 pp. Cow groups were not different in O2 consumption, milk yield, metabolizable energy for maintenance, or the efficiency of tissue utilization for milk synthesis, but high-Meff cows tended to have the lower German relative breeding value functional herd life, indicating a link between metabolic performance and productive lifespan. In conclusion, the use of a clustering approach involving MEI/mBW, O2/mBW, and CO2/mBW seems to be a promising method to differentiate cows with divergent metabolic efficiency but does not allow identifying an individual metabotype that persists across the whole lactation cycle.

Relationship between the fatty acid profile of hair and energy availability of lactating primiparous cows.

We hypothesised that a relationship would exist between hair fatty acids, especially C12:0, C14:0 and C16:0, and parameters of energy metabolism such as energy intake, energy mobilisation, and energy requirement for maintenance and milk performance. For this study, 11 primiparous German Holstein cows were available from which hair samples at weeks 6 and 8 of lactation were analysed. The average body weight of these animals was 558 ± 27 kg at calving and milk yield at 100-days in milk was 3,537 ± 529 kg. Feed intake and milk yield were measured daily. Body weight and back fat thickness were measured at calving and in weeks 2, 4, and 8 of lactation. Energy balance and energy utilisation were calculated until week 6 of lactation. Spearman's correlation coefficients were found to be significantly positive for the relationship between the percentage of C12:0 and C14:0 fatty acids in the hair in lactation week 8 and energy intake in weeks 5 and 6 (0.62 < r < 0.65, P < 0.05). If the animals are grouped according to their energy utilisation between weeks 1 and 6 into two groups higher (n = 6) or lower (n = 5) than the median, animals of the high energy utilising group had a higher energy intake. These animals had also higher percentages of the C12:0 fatty acid in their hair fat (week 6: 4.9% vs. 3.1%, P < 0.05; week 8: 4.3% vs. 2.9%, P = 0.05). Our hypothesis is supported, and this study justifies further investigation of the content of medium-chain fatty acids in hair samples as biomarkers for the metabolic status of a cow during early lactation.

Effects of long-term microalgae supplementation on muscle microstructure, meat quality and fatty acid composition in growing pigs.

We investigated the effects of long-term microalgae supplementation (7% in a piglet diet and 5% in a fattening diet) on muscle microstructure and meat quality, including fatty acid composition in female Landrace pigs (n = 31). The major effects were muscle-specific increases in n-3 and n-6 polyunsaturated fatty acids (PUFA) concentrations, resulting in increased accumulation of docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA). Carcass traits and meat quality of longissimus thoracis muscle were not affected by the microalgae diet with the exception of reduced drip loss (p = 0.01) and increased protein proportion (p = 0.04). In addition, the microalgae diet resulted in a shift to a more oxidative myofibre type composition in semitendinosus but not longissimus thoracis muscle. In conclusion, microalgae supplementation offers a unique opportunity to enhance essential n-3 PUFA contents in pig meat. The results support small but coordinated changes in skeletal muscle phenotypic appearance and functionality.

Intravenous lipid infusion affects dry matter intake, methane yield, and rumen bacteria structure in late-lactating Holstein cows.

Increasing the dietary fat content of ruminant diets decreases methane (CH4) production. This effect is caused by the toxic properties of fatty acids on rumen microbial populations, coating of feed particles diminishing the accessibility for microbes, and a reduction in dry matter intake (DMI). The latter effect is caused by postabsorptive long-chain fatty acids eliciting anorexic signaling; however, whether circulating long-chain fatty acids affect rumen CH4 production alike is unknown. To approach this question, 5 rumen-cannulated Holstein cows in late lactation received 2 jugular catheters and were kept in respiration chambers to measure CH4 production and DMI for 48 h. In a crossover design, cows were intravenously infused with a 20% lipid emulsion (LIPO) or 0.9% NaCl (CON). The LIPO cows received 2.1 kg of triglycerides/d [0.152 ± 0.007 g of triglycerides/(kg of BW × h)-1] consisting of 12.1% palmitic acid, 4.2% stearic acid, 31.1% oleic acid, and 52.7% linoleic acid. Blood and rumen fluid samples were taken hourly during the day. Results showed that LIPO compared with CON infusion increased plasma triglyceride as well as free fatty acid and serotonin concentrations but reduced the proportion of de novo synthesized milk fatty acids (sum of C6 to C16). Daily CH4 production and DMI were lower, whereas daily CH4 yield (CH4/DMI) was greater in LIPO than CON cows, although CH4 yield decreased from d 1 to d 2 by 2 to 14% in LIPO-infused cows only. This effect was associated with a higher (acetate + butyrate)/propionate ratio, tending lower propionate concentrations between 24 and 34 h of infusion, reduced relative abundances of genera belonging to Succinivibrio, Ruminococcaceae, and Ruminiclostridium, and greater relative Bacteroidetes genus abundances in the rumen.

Sea Buckthorn Pomace Supplementation in the Diet of Growing Pigs-Effects on Fatty Acid Metabolism, HPA Activity and Immune Status.

There is evidence that sea buckthorn, as a source of n-3 polyunsaturated fatty acids (n-3 PUFA), possesses health-enhancing properties and may modulate neuroendocrine and immune functions. In the present study, we investigated the effect of sea buckthorn pomace (SBP) supplementation in the diet of growing German Landrace pigs on fatty acids in the blood and hypothalamus, peripheral immune parameters and mRNA expression of corticotropin-releasing hormone (CRH), mineralocorticoid receptor (MR) and glucocorticoid receptor (GR) in the hypothalamus and spleen. Pigs were fed diets supplemented with 12% of dried SBP or 0% SBP (control group) over an intervention period of eight weeks. The fatty acid profiles in blood plasma were significantly affected by SBP supplementation only for C18:2n-6 and n-6/n-3 PUFA ratio compared with the control group. SBP supplementation did not significantly affect the fatty acid concentrations in the hypothalamus. Furthermore, there were no significant differences in mRNA expression of CRH, MR and GR in the hypothalamus or of GR mRNA expression in the spleen. Concerning the immune status, the plasma IgG levels tended to be higher in SBP pigs, whereas the leukocyte distribution, mitogen-stimulated lymphocyte proliferation, and serum IgM levels remained unchanged. In conclusion, the SBP supplementation of the diet only caused moderate effects on fatty acid metabolism, but no significant effects on hypothalamic-pituitary-adrenal (HPA) activity and immunity in growing pigs. It seems that a beneficial effect of dietary n-3 PUFA on health and welfare is more likely to be expected during stressful situations.

Reproductive performance primarily depends on the female genotype in a two-factorial breeding experiment using high-fertility mouse lines.

Mouse models showing an improved fertility phenotype are barely described in the literature. In the present study, we further characterized two outbred mouse models that have been selected for the phenotype 'high fertility' for more than 177 generations (fertility lines (FL) 1 and 2). In order to delineate the impact of males and females on fertility parameters, we performed a two-factorial breeding experiment by mating males and females of the three different genotypes (FL1, FL2, unselected control (Ctrl)) in all 9 possible combinations. Reproductive performance, such as number of offspring per litter or total birth weight of the entire pup, mainly depends on the female genotype. Although the reproductive performance of FL1 and FL2 is very similar, their phenotypes differ. FL2 animals of both genders are larger compared to FL1 and control animals. Females of the control line delivered offspring earlier compared to FL1 and FL2 dams. Males of FL1 are the lightest and the only ones who gained weight during the two weeks mating period. To address whether this effect is correlated with differing serum androgen levels, we measured the concentrations of testosterone, dehydroepiandrosterone, 4-androstenedione, androstanediol and dihydrotestosterone in males of all three lines by GC-MS. We measured serum testosterone between 5.0 and 6.4 ng/mL, whereas the concentrations of the other androgens were at least one order of magnitude lower, with no significant differences between the lines. Our data indicate that reproductive outcome largely depends on the genotype of the female in a two-factorial breeding experiment and supports previous findings that the phenotype 'high fertility' is warranted by using different physiological strategies.

Field study analysis of the influences of deworming regimens and housing conditions on parasites and sperm output in 21 European boar studs.

The current study reports the parasitological results of a quality control audit in 21 European boar studs. Field investigations were performed over a 2-year period (2012-2013) during the winter and spring. From each stud, an average of 30 (range, 25-33) individual faecal samples and ejaculates from 615 randomly selected Pietrain boars were analysed. Statistical analysis revealed a significant effect (P < 0.0001) of deworming regimen (DR) × age class of boar (A) and housing condition (H) × A on the presence of parasites. A second model indicated a significant effect (P = 0.0262) of DR × H × A on the presence of parasites. Sperm output was significantly affected (P < 0.0001) by the DR. Based on this study, recommendations for deworming AI boars are proposed.

Effects of a 6-wk intraduodenal supplementation with quercetin on energy metabolism and indicators of liver damage in periparturient dairy cows.

Periparturient dairy cows experience metabolic challenges that result in a negative energy balance (EB) and a range of postpartum health problems. To compensate for the negative EB, cows mobilize fatty acids from adipose tissues, which can lead to fatty liver disease, a periparturient metabolic disorder. Flavonoids, such as quercetin (Q), are polyphenolic substances found in all higher plants and have hepatoprotective potential and the ability to prevent or reduce lipid accumulation in the liver. In ruminants, few studies on the metabolic effects of Q are available, and thus this study was conducted to determine whether Q has beneficial effects on EB, lipid metabolism, and hepatoprotective effects in periparturient dairy cows. Quercetin was supplemented intraduodenally to circumvent Q degradation in the rumen. Cows (n=10) with duodenal fistulas were monitored for 7wk. Beginning 3wk before expected calving, 5 cows were treated with 100mg of quercetin dihydrate per kilogram of body weight daily in a 0.9% sodium chloride solution for a total period of 6wk, whereas the control cows received only the sodium chloride solution. The plasma flavonoid levels were higher in the Q-treated cows than in the control cows. A tendency for higher postpartum (pp) than antepartum (ap) plasma flavonoid levels was observed in the Q-treated cows than in the controls, which was potentially caused by a reduced capacity to metabolize Q. However, the metabolic status of the Q-treated cows did not differ from that of the control cows. The pp increases in plasma aspartate aminotransferase and glutamate dehydrogenase activities were less in the Q-treated cows than in the control cows. The Q had no effect on energy expenditures, but from ap to pp the cows had a slight decline in respiratory quotients. Irrespective of the treatment group, the oxidation of fat peaked after calving, suggesting that the increase occurred because of an increased supply of fatty acids from lipomobilization. In conclusion, supplementation with Q resulted in lower pp plasma aminotransferase and glutamate dehydrogenase, which indicated reduced liver damage. However, the direct effects of Q on the liver and the implications for animal performance remain to be investigated.

Metabolic Heat Stress Adaption in Transition Cows: Differences in Macronutrient Oxidation between Late-Gestating and Early-Lactating German Holstein Dairy Cows.

High ambient temperatures have severe adverse effects on biological functions of high-yielding dairy cows. The metabolic adaption to heat stress was examined in 14 German Holsteins transition cows assigned to two groups, one heat-stressed (HS) and one pair-fed (PF) at the level of HS. After 6 days of thermoneutrality and ad libitum feeding (P1), cows were challenged for 6 days (P2) by heat stress (temperature humidity index (THI) = 76) or thermoneutral pair-feeding in climatic chambers 3 weeks ante partum and again 3 weeks post-partum. On the sixth day of each period P1 or P2, oxidative metabolism was analyzed for 24 hours in open circuit respiration chambers. Water and feed intake, vital parameters and milk yield were recorded. Daily blood samples were analyzed for glucose, ß-hydroxybutyric acid, non-esterified fatty acids, urea, creatinine, methyl histidine, adrenaline and noradrenaline. In general, heat stress caused marked effects on water homeorhesis with impairments of renal function and a strong adrenergic response accompanied with a prevalence of carbohydrate oxidation over fat catabolism. Heat-stressed cows extensively degraded tissue protein as reflected by the increase of plasma urea, creatinine and methyl histidine concentrations. However, the acute metabolic heat stress response in dry cows differed from early-lactating cows as the prepartal adipose tissue was not refractory to lipolytic, adrenergic stimuli, and the rate of amino acid oxidation was lower than in the postpartal stage. Together with the lower endogenous metabolic heat load, metabolic adaption in dry cows is indicative for a higher heat tolerance and the prioritization of the nutritional requirements of the fast-growing near-term fetus. These findings indicate that the development of future nutritional strategies for attenuating impairments of health and performance due to ambient heat requires the consideration of the physiological stage of dairy cows.

A phenotypical approach to the effects of production traits, parturition, puerperium and body condition on commencement of luteal activity in high yielding dairy cows.

The interval from calving to commencement of luteal activity (CLA) was determined by progesterone measurements from milk samples obtained once a week until the 14th week post-partum in 513 German Holstein cows in first to third parity. Milk samples were analyzed by an "on-farm" device (eProCheck(®), Minitüb, Germany) and simultaneously by RIA. The objective of this study was to examine the effect of milk yield, protein content and body condition of a cow on the CLA post-partum. Milk progesterone concentrations of "on-farm" measurements correlated with measurements done by the RIA-method significantly (r=0.72; P<0.001). Within the analyzed herd the interval from calving until the first rise of progesterone averaged 5.6±2.4 weeks. The 100-days milk yield was not associated with CLA. Cows with a milk protein content at 1st milk recording of ≤3.5% revealed first luteal activity 1.3±0.3 weeks later than cows that had a content of >3.75% protein (P<0.01). Furthermore cows with assisted calving or dystocia presented significantly later CLA than cows which required no help during the calving process (P<0.05). The change in back fat thickness from 1st to 2nd milk recording had a significant influence on CLA (P<0.05). In conclusion the phenotypic impact of milk yield on fertility cannot be confirmed regarding to CLA. The negative energy balance after calving, caused by the high milk yields, is more detrimental for the cyclical activity as was shown by the parameters milk protein content and change in BFT.

Low and high dietary protein:carbohydrate ratios during pregnancy affect materno-fetal glucose metabolism in pigs.

Inadequate dietary protein during pregnancy causes intrauterine growth retardation. Whether this is related to altered maternal and fetal glucose metabolism was examined in pregnant sows comparing a high-protein:low-carbohydrate diet (HP-LC; 30% protein, 39% carbohydrates) with a moderately low-protein:high-carbohydrate diet (LP-HC; 6.5% protein, 68% carbohydrates) and the isoenergetic standard diet (ST; 12.1% protein, 60% carbohydrates). During late pregnancy, maternal and umbilical glucose metabolism and fetal hepatic mRNA expression of gluconeogenic enzymes were examined. During an i.v. glucose tolerance test (IVGTT), the LP-HC-fed sows had lower insulin concentrations and area under the curve (AUC), and higher glucose:insulin ratios than the ST- and the HP-LC-fed sows (P < 0.05). Insulin sensitivity and glucose clearance were higher in the LP-HC sows compared with ST sows (P < 0.05). Glucagon concentrations during postabsorptive conditions and IVGTT, and glucose AUC during IVGTT, were higher in the HP-LC group compared with the other groups (P < 0.001). (13)C glucose oxidation was lower in the HP-LC sows than in the ST and LP-HC sows (P < 0.05). The HP-LC fetuses were lighter and had a higher brain:liver ratio than the ST group (P < 0.05). The umbilical arterial inositol concentration was greater in the HP-LC group (P < 0.05) and overall small fetuses (230-572 g) had higher values than medium and heavy fetuses (≥573 g) (P < 0.05). Placental lactate release was lower in the LP-HC group than in the ST group (P < 0.05). Fetal glucose extraction tended to be lower in the LP-HC group than in the ST group (P = 0.07). In the HP-LC and LP-HC fetuses, hepatic mRNA expression of cytosolic phosphoenolpyruvate carboxykinase (PCK1) and glucose-6-phosphatase (G6PC) was higher than in the ST fetuses (P < 0.05). In conclusion, the HP-LC and LP-HC sows adapted by reducing glucose turnover and oxidation and having higher glucose utilization, respectively. The HP-LC and LP-HC fetuses adapted via prematurely expressed hepatic gluconeogenic enzymes.

Describing temperament in an ungulate: a multidimensional approach.

Studies on animal temperament have often described temperament using a one-dimensional scale, whereas theoretical framework has recently suggested two or more dimensions using terms like "valence" or "arousal" to describe these dimensions. Yet, the valence or assessment of a situation is highly individual. The aim of this study was to provide support for the multidimensional framework with experimental data originating from an economically important species (Bos taurus). We tested 361 calves at 90 days post natum (dpn) in a novel-object test. Using a principal component analysis (PCA), we condensed numerous behaviours into fewer variables to describe temperament and correlated these variables with simultaneously measured heart rate variability (HRV) data. The PCA resulted in two behavioural dimensions (principal components, PC): novel-object-related (PC 1) and exploration-activity-related (PC 2). These PCs explained 58% of the variability in our data. The animals were distributed evenly within the two behavioural dimensions independent of their sex. Calves with different scores in these PCs differed significantly in HRV, and thus in the autonomous nervous system's activity. Based on these combined behavioural and physiological data we described four distinct temperament types resulting from two behavioural dimensions: "neophobic/fearful--alert", "interested--stressed", "subdued/uninterested--calm", and "neoophilic/outgoing--alert". Additionally, 38 calves were tested at 90 and 197 dpn. Using the same PCA-model, they correlated significantly in PC 1 and tended to correlate in PC 2 between the two test ages. Of these calves, 42% expressed a similar behaviour pattern in both dimensions and 47% in one. No differences in temperament scores were found between sexes or breeds. In conclusion, we described distinct temperament types in calves based on behavioural and physiological measures emphasising the benefits of a multidimensional approach.

An extension of PPLS-DA for classification and comparison to ordinary PLS-DA.

Classification studies are widely applied, e.g. in biomedical research to classify objects/patients into predefined groups. The goal is to find a classification function/rule which assigns each object/patient to a unique group with the greatest possible accuracy (classification error). Especially in gene expression experiments often a lot of variables (genes) are measured for only few objects/patients. A suitable approach is the well-known method PLS-DA, which searches for a transformation to a lower dimensional space. Resulting new components are linear combinations of the original variables. An advancement of PLS-DA leads to PPLS-DA, introducing a so called 'power parameter', which is maximized towards the correlation between the components and the group-membership. We introduce an extension of PPLS-DA for optimizing this power parameter towards the final aim, namely towards a minimal classification error. We compare this new extension with the original PPLS-DA and also with the ordinary PLS-DA using simulated and experimental datasets. For the investigated data sets with weak linear dependency between features/variables, no improvement is shown for PPLS-DA and for the extensions compared to PLS-DA. A very weak linear dependency, a low proportion of differentially expressed genes for simulated data, does not lead to an improvement of PPLS-DA over PLS-DA, but our extension shows a lower prediction error. On the contrary, for the data set with strong between-feature collinearity and a low proportion of differentially expressed genes and a large total number of genes, the prediction error of PPLS-DA and the extensions is clearly lower than for PLS-DA. Moreover we compare these prediction results with results of support vector machines with linear kernel and linear discriminant analysis.

Enhanced sensitivity of skeletal muscle growth in offspring of mice long-term selected for high body mass in response to a maternal high-protein/low-carbohydrate diet during lactation.

AIM: To investigate the effects of a high-protein/low-carbohydrate diet fed to mice of different genotypes during pregnancy and/or lactation on offspring skeletal muscle growth and metabolism. METHODS: Pregnant mice from strains selected for high body mass (DU6) or endurance running performance (DUhLB) and from an unselected control strain (DUK) were fed iso-energetic diets containing 20 % (C) or 40 % protein and low carbohydrate (HP) from mating to weaning at day 21 of age. At birth, offspring were cross-fostered resulting in different exposure to maternal prenatal-preweaning diets (C-C, HP-C, C-HP, HP-HP). Rectus femoris muscle of male mice (n = 291) was examined at day 23, 44, 181 and 396 of age for cellular growth and metabolism. RESULTS: At day 23 of age, body and muscle growth was retarded by 30-40 % (P < 0.0001) in response to the C-HP and HP-HP, but not to the HP-C diet, due to reduced fibre size (P < 0.0001) but not fibre number. DNA was highly reduced in DU6, less in DUhLB, but not in DUK muscle (strain × diet; P < 0.0001). Despite some compensation, muscle growth was still impaired (P < 0.001) in adulthood (day 44; day 181), but at senescence only in DU6 mice (strain × diet; P < 0.05). Only at weaning, isocitrate and lactate dehydrogenase activities were increased or decreased (P < 0.0001), respectively, without influence on fibre type composition. CONCLUSION: A high-protein/low-carbohydrate diet fed to dams during lactation, but not during pregnancy, retards skeletal muscle growth in offspring with greater response of a heavy, obese compared with a physically fit and a control genotype and causes a transient shift towards oxidative versus glycolytic muscle metabolism.

The cognitive capabilities of farm animals: categorisation learning in dwarf goats (Capra hircus).

The ability to establish categories enables organisms to classify stimuli, objects and events by assessing perceptual, associative or rational similarities and provides the basis for higher cognitive processing. The cognitive capabilities of farm animals are receiving increasing attention in applied ethology, a development driven primarily by scientifically based efforts to improve animal welfare. The present study investigated the learning of perceptual categories in Nigerian dwarf goats (Capra hircus) by using an automated learning device installed in the animals' pen. Thirteen group-housed goats were trained in a closed-economy approach to discriminate artificial two-dimensional symbols presented in a four-choice design. The symbols belonged to two categories: category I, black symbols with an open centre (rewarded) and category II, the same symbols but filled black (unrewarded). One symbol from category I and three different symbols from category II were used to define a discrimination problem. After the training of eight problems, the animals were presented with a transfer series containing the training problems interspersed with completely new problems made from new symbols belonging to the same categories. The results clearly demonstrate that dwarf goats are able to form categories based on similarities in the visual appearance of artificial symbols and to generalise across new symbols. However, the goats had difficulties in discriminating specific symbols. It is probable that perceptual problems caused these difficulties. Nevertheless, the present study suggests that goats housed under farming conditions have well-developed cognitive abilities, including learning of open-ended categories. This result could prove beneficial by facilitating animals' adaptation to housing environments that favour their cognitive capabilities.

High-protein-low-carbohydrate diet during pregnancy alters maternal plasma amino acid concentration and placental amino acid extraction but not fetal plasma amino acids in pigs.

A high protein-low-carbohydrate diet during pregnancy can cause intra-uterine growth restriction. However, its impact during pregnancy on maternal, umbilical and fetal plasma amino acid (AA) profiles is unknown. A maternal high-protein (30 %)-low-carbohydrate (HP-LC) diet was compared with isoenergetic standard (12·1 % crude protein; ST) and low-protein (6·5 %)-high-carbohydrate (LP-HC) diets fed to nulliparous pregnant sows to examine changes in AA concentrations in maternal, venous and arterial umbilical and fetal plasma in mid and late pregnancy. At 64 and 94 days of pregnancy (dp), sows underwent Caesarean section, and maternal, umbilical and fetal plasma samples were collected. The HP-LC diet mainly affected maternal plasma AA concentrations. Plasma concentrations of Ile and Val were increased and those of Ala, Glu and Gly were decreased (P ≤ 0·05) in HP-LC compared with ST sows at 64 and 94 dp. The LP-HC diet decreased fetal plasma Glu concentration compared with the ST diet at 94 dp. Substantial AA catabolism was reflected by increased (P ≤ 0·05) maternal and fetal plasma urea concentrations with the HP-LC compared with the ST and LP-HC diets at 94 dp. Fractional placental extraction of Val was higher whereas those of Ala, Gln and Glu were lower in the HP-LC compared with the ST sows at 64 and 94 dp (P ≤ 0·05). Reduced fetal mass at 94 dp was accompanied by reduced fetal extraction of Lys and Pro in the HP-LC group (P ≤ 0·05). In conclusion, a maternal HP-LC diet during pregnancy altered maternal plasma composition of many AA and modified placental AA extraction to compensate for imbalanced maternal nutrient intake.

Intrauterine growth retarded progeny of pregnant sows fed high protein:low carbohydrate diet is related to metabolic energy deficit.

High and low protein diets fed to pregnant adolescent sows led to intrauterine growth retardation (IUGR). To explore underlying mechanisms, sow plasma metabolite and hormone concentrations were analyzed during different pregnancy stages and correlated with litter weight (LW) at birth, sow body weight and back fat thickness. Sows were fed diets with low (6.5%, LP), adequate (12.1%, AP), and high (30%, HP) protein levels, made isoenergetic by adjusted carbohydrate content. At -5, 24, 66, and 108 days post coitum (dpc) fasted blood was collected. At 92 dpc, diurnal metabolic profiles were determined. Fasted serum urea and plasma glucagon were higher due to the HP diet. High density lipoprotein cholesterol (HDLC), %HDLC and cortisol were reduced in HP compared with AP sows. Lowest concentrations were observed for serum urea and protein, plasma insulin-like growth factor-I, low density lipoprotein cholesterol, and progesterone in LP compared with AP and HP sows. Fasted plasma glucose, insulin and leptin concentrations were unchanged. Diurnal metabolic profiles showed lower glucose in HP sows whereas non-esterified fatty acids (NEFA) concentrations were higher in HP compared with AP and LP sows. In HP and LP sows, urea concentrations were 300% and 60% of AP sows, respectively. Plasma total cholesterol was higher in LP than in AP and HP sows. In AP sows, LW correlated positively with insulin and insulin/glucose and negatively with glucagon/insulin at 66 dpc, whereas in HP sows LW associated positively with NEFA. In conclusion, IUGR in sows fed high protein:low carbohydrate diet was probably due to glucose and energy deficit whereas in sows with low protein:high carbohydrate diet it was possibly a response to a deficit of indispensable amino acids which impaired lipoprotein metabolism and favored maternal lipid disposal.

Involvement of skeletal muscle protein, glycogen, and fat metabolism in the adaptation on early lactation of dairy cows.

During early lactation, high-yielding dairy cows cannot consume enough feed to meet nutrient requirements. As a consequence, animals drop into negative energy balance and mobilize body reserves including muscle protein and glycogen for milk production, direct oxidation, and hepatic gluconeogenesis. To examine which muscle metabolic processes contribute to the adaptation during early lactation, six German Holstein cows were blood sampled and muscle biopsied throughout the periparturient period. From pregnancy to lactation, the free plasma amino acid pattern imbalanced and plasma glucose decreased. Several muscle amino acids, as well as total muscle protein, fat, and glycogen, and the expression of glucose transporter-4 were reduced within the first 4 weeks of lactation. The 2-DE and MALDI-TOF-MS analysis identified 43 differentially expressed muscle protein spots throughout the periparturient period. In early lactation, expression of cytoskeletal proteins and enzymes involved in glycogen synthesis and in the TCA cycle was decreased, whereas proteins related to glycolysis, fatty acid degradation, lactate, and ATP production were increased. On the basis of these results, we propose a model in which the muscle breakdown in early lactation provides substrates for milk production by a decoupled Cori cycle favoring hepatic gluconeogenesis and by interfering with feed intake signaling.

Proteome and radioimmunoassay analyses of pituitary hormones and proteins in response to feed restriction of dairy cows.

The hypothalamic-pituitary system controls homeostasis during feed energy reduction. In order to examine which pituitary proteins and hormone variants are potentially associated with metabolic adaptation, pituitary glands from ad libitum and energy restrictively fed dairy cows were characterized using RIA and 2-DE followed by MALDI-TOF-MS. We found 64 different spots of regulatory hormones: growth hormone (44), preprolactin (16), luteinizing hormone (LH) (1), thyrotropin (1), proopiomelanocortin (1) and its cleavage product lipotropin (1), but none of these did significantly differ between feeding groups. Quantification of total pituitary LH and prolactin concentrations by RIA confirmed the results obtained by proteome analysis. Also, feed energy restriction provoked increasing non-esterified fatty acid, decreasing prolactin, but unaltered glucose, LH and growth hormone plasma concentrations. Energy restriction decreased the expression of glial fibrillary acidic protein, triosephosphate isomerase, purine-rich element-binding protein A and elongation factor Tu, whereas it increased expression of proline synthetase co-transcribed homolog, peroxiredoxin III, ß-tubulin and annexin A5 which is involved in the hormone secretion process. Our results indicate that in response to feed energy restriction the pituitary reservoir of all posttranslationally modified hormone forms remains constant. Changing plasma hormone concentrations are likely attributed to a regulated releasing process from the gland into the blood.

Finding biomarker signatures in pooled sample designs: a simulation framework for methodological comparisons.

Detection of discriminating patterns in gene expression data can be accomplished by using various methods of statistical learning. It has been proposed that sample pooling in this context would have negative effects; however, pooling cannot always be avoided. We propose a simulation framework to explicitly investigate the parameters of patterns, experimental design, noise, and choice of method in order to find out which effects on classification performance are to be expected. We use a two-group classification task and simulated gene expression data with independent differentially expressed genes as well as bivariate linear patterns and the combination of both. Our results show a clear increase of prediction error with pool size. For pooled training sets powered partial least squares discriminant analysis outperforms discriminance analysis, random forests, and support vector machines with linear or radial kernel for two of three simulated scenarios. The proposed simulation approach can be implemented to systematically investigate a number of additional scenarios of practical interest.