The effect of increasing dose levels of a novel consensus bacterial 6-phytase variant (PhyG) on growth performance, digestibility and retention of minerals and bone ash was evaluated in weaned piglets fed diets containing varying levels of phytate-bound phosphorus (PP) and no added inorganic phosphate (iP). Treatments comprised: a wheat, corn, barley and soybean meal-based negative control diet (NC), reduced in digestible phosphorus (P) (iP free), calcium (Ca), energy and digestible amino acids vs NRC 2012 requirements, formulated at three PP levels (Low, 2.40 g/kg; Medium, 2.90 g/kg; High, 3.40 g/kg); the NC supplemented with PhyG at 250, 500, 1,000, 2,000 or 4,000 phytase units (FTU)/kg or with a comparator phytase (PhyB) at 1,000 FTU/kg. A completely randomized design employed 144 DanBred [[EQUATION]] Pi males (8 piglets per treatment, initial body weight [BW] 9.2 ± 1.0 kg). Piglets were adapted for 10 d in floor-pens (2 pigs per pen) and 5 d in metabolism crates (individually), followed by 5 d of fecal collection. Pigs were slaughtered on d 20 to collect ileal digesta and femurs. Dietary PP level had no effect on BW, BW gain, feed intake or apparent ileal digestibility (AID) of P or Ca, bone ash or strength. Total tract digestibility (ATTD) and retention of P at d 20 were reduced in Medium compared with Low PP diets (-6.4% and -6.6%, respectively; P < 0.05) whereas ATTD and retention of Ca were reduced in Medium or High vs Low PP diets (P < 0.05). Body weight (d 10 and 20), d 0 to 20 BW gain, AID P, ATTD and retention of P and Ca and ATTD gross energy increased exponentially with increasing PhyG dose level (P < 0.05) regardless of PP level; ATTD and retention of P were improved (above NC) by 44.7 and 45.3%, and by 55.9 and 56.1%, with PhyG at 1,000 and 4,000 FTU/kg, respectively, and at 1,000 FTU/kg were higher with PhyG than with PhyB across all PP levels (ATTD P +4.3% on average, P < 0.05). In conclusion, PhyG increased growth performance, total tract digestibility and retention of minerals in weaned piglets regardless of dietary PP level, and at 1,000 FTU/kg PhyG was more efficacious at improving ATTD of P than PhyB. Negative effects of Medium or High PP levels on ATTD of P and Ca were ameliorated by increasing the dose level of the novel phytase.

The natural polymer chitin is an abundant source for valuable N-acetylchitooligosaccharides and N-acetylglucosamine applicable in several industries. The endochitinase Chit36-TA from Trichoderma asperellum was recombinantly expressed in Komagataella phaffii for the enzymatic degradation of chitin from unused insect exuviae into N-acetylchitooligosaccharides. Chit36-TA was purified by Ni–NTA affinity chromatography and subsequently biochemically characterized. After deglycosylation, the endochitinase had a molecular weight of 36 kDa. The optimum pH for Chit36-TA was 4.5. The temperature maximum of Chit36-TA was determined to be 50 °C, while it maintained > 93

This study evaluated the effect of a multi-enzyme complex on growth performance in weaned pigs fed a wheat-barley-rye-based diet containing fiber-rich by-products and with a low level of highly digestible protein and energy ingredients. Two experimental runs were performed, each with 96 (DE x DL) × Pi pigs [24–26 days of age; 8.0 ± 0.06 kg initial body weight (BW)] assigned to two treatments in 24 pens per run (4 pigs/pen, 2 males and 2 females; 12 pens/treatment/run) in a randomized block design. Pigs were allocated to treatment on the day of weaning. Treatments comprised: 1) a nutritionally adequate wheat-barley-rye-soybean meal-based diet with added wheat bran and rapeseed meal, containing 750 FTU/kg of a commercial Buttiauxella sp. phytase (ALT), and 2) the same diet supplemented with a multi-enzyme containing 4,000 U/kg xylanase, 200 U/kg β-glucanase, 2,000 U/kg protease and 100 U/kg amylase (ALT+multi-enzymes). Diets were formulated in two phases (Pre-starter, d 0–14; starter, d 15–42) and fed ad libitum as pelleted diets for 42 days. The analyzed content of crude fiber and neutral detergent fiber in ALT diets were, respectively, 4.0% and 11.5% in pre-starter, and 4.2% and 14.8% in starter phase. Body weight was measured at the start and end of each phase, feed intake was determined weekly. Data were analyzed by one-way ANOVA using IBM SPSS 21 including run as a random effect. During pre-starter phase, average daily gain (ADG) and average daily feed intake (ADFI) were greater in pigs fed ALT+multi-enzymes vs. ALT, by 8.4% (232 vs 214 g/day) and 6.7% (300 vs. 281 g/day), respectively, although this was not statistically significant. During starter phase, pigs fed ALT+multi-enzymes exhibited increased ADG vs. ALT (+35 g/day or 5.5%; P < 0.05). For the overall period (d 0 to 42), pigs fed ALT+multi-enzyme were, on average, 1.3 kg heavier than pigs fed ALT (P < 0.05), ADG was increased by +32 g/day (4.4%; P < 0.05) and FCR tended (P = 0.07) to be reduced (1.48 vs 1.44). Total feed costs per kg BW gain, exclusive of the cost of the enzymes, tended to be reduced in ALT+multi-enzymes vs. ALT, (-0.014 USD/kg BW gain, overall; P = 0.09). These results demonstrate that the multi-enzyme supplemented on top of phytase improved growth performance. Given the established mode of action of the individual enzymes, this may have been mediated by enhanced digestibility and utilization of nutrients in the ALT diet. In conclusion, supplementing a high-fiber mixed diet containing by-products and low inclusion of highly digestible energy and protein ingredients with a multi-enzyme complex improved growth performance in weaned pigs and led to economic benefit.

Duckweed is gaining attention in animal nutrition and is considered as a potential alternative protein source for broiler chickens. In order to evaluate the nutritional value of duckweed, three individual batches were investigated. They consisted of a mixture of Lemna minuta and Lemna minor (A, 17.5% crude protein), Spirodela polyrhiza (B, 24.6% crude protein) and Lemna obscura (C, 37.0% crude protein). Treatment diets contained 50% batch A, 50% batch B, and 25, 50 and 75% of batch C. All diets were fed to broiler chickens (Ross 308) from an age of 21 to 27 days. Diets with a share of 50 and 75% of batch C led to decreased feed intake (109.3 and 74.9 g/day, respectively) compared to the control. Standardized ileal digestibility of crude protein and amino acids differed significantly between duckweed batches, at values for methionine between 49.9 and 90.4%. For all amino acids, batch A consistently had the lowest and batch C the highest digestibility. Batches had different tannin contents of 2943, 2890 and 303 mg/kg for batches A, B and C, respectively. The apparent ileal digestibility of phosphorus differed significantly between all batches (50.8-78.9%). Duckweed can be used as a protein feed for broiler chickens. However, a defined and stable biomass composition optimized for the requirements of broiler chickens is needed.

Reports of the underlying mechanisms of dietary grape extract (GE) in overcoming weaning challenges in piglets have been partly inconsistent. Furthermore, evaluations of the effects of GE at weaning in comparison to those of widely used therapeutic antibiotics have been scarce. To explore the mode of action of GE in selected tissues and plasma, we evaluated gut morphology, antioxidant and inflammation indices. Accordingly, 180 weaning piglets were allocated to three treatment groups: negative control (NC), NC and antibiotic treatment for the first 5 days of the trial (positive control, PC), and NC and GE (entire trial). The villus surface was positively affected by GE and PC on day 27/28 of the trial in the jejunum and on day 55/56 of the trial in the ileum. In the colon, NC tended (p < 0.10) to increase crypt parameters compared to PC on day 55/56. The PC group tended (p < 0.10) to increase catalase activity in the ileum and decrease Cu/Zn-SOD activity in the jejunum, both compared to NC. There were no additional effects on antioxidant measurements of tissue and plasma, tissue gene expression, or plasma acute-phase proteins. In conclusion, GE supplementation beneficially affected the villus surface of the small intestine. However, these changes were not linked to the antioxidant and anti-inflammatory properties of GE.

A meta-analysis was conducted to evaluate the effect of a multi-enzyme complex (MEC) on growth performance in weaned piglets [initial body weight (BW) of 7.6±1.2 kg]. Database was constructed using 4 unpublished studies, contributing 306 data points. The dietary treatments consisted of a conventional diet (CC) with highly digestible nutrients, a high-fiber, low-digestible protein diet (NC), and the NC supplemented with MEC. All diets were corn/wheat/soybean meal based, formulated to meet or exceed NRC specification for weaned piglets. The NC contained ingredients such as barley, distiller's dried grains with solubles, wheat middling, rye and rapeseed meal depending on the geographic location of the study. In all studies, MEC provided 4,000 U/kg Xylanase, 200 U/kg β-glucanase, 100 U/kg Amylase, and 2,000 U/kg Protease. All diets contained 750 FTU/kg of a commercial Buttiauxella phytase. Piglets were fed experimental diets over 2 phases for 42 d, from d 1 of weaning. Growth performance was recorded bi-weekly. Data were analyzed using Mixed Model by JMP 14.0. The average daily gain of piglets fed MEC supplemented diets was increased by 7.4% vs NC (P = 012) during phase 1. The final BW (d 42) was improved by 1.1 kg (4.1%; P = 0. 037) in piglets fed MEC supplemented diets compared with NC. Respective improvements (P < 0.05) in feed conversion ratio during phase 1 and overall were 11% (16 points) and 4.4% (7 points) in MEC supplemented diets when compared with NC, which indicates better nutrient digestibility. In conclusion, supplementing a high fiber weaner diets with a multi-enzyme complex improved final BW and feed efficiency in piglets.

This study intended to classify ad libitum–fed calves according to their milk replacer (MR) meal size using the K-means clustering approach. This study aimed to investigate the effects of MR meal size on feed intake, growth performance, and blood metabolic and hormones of ad libitum MR-fed calves. German Holstein calves (16 male and 16 female) were studied from birth until d 77 of age. All calves received first colostrum (2.5 kg) milked from their dams within 2 h after birth. Subsequent colostrum meals (subsequent 4 meals until 2.5 d of age; 2 meals/d) and MR (125 g of powder/L; 21.7% crude protein, 18.6% crude fat) were fed ad libitum by teat bucket until d 10 ± 2 of age. Afterward, calves were housed in group pens with automatic feeders for MR (maximum of 25 L/d) and concentrate from 10 ± 3 d of age. Half of the calves received MR supplemented with butyrate to improve growth performance. Milk intake was stepped down to 2 L/d from wk 9 to 10, and 2 L/d of MR were offered until the end of the study. On d 1, 2, 4, and 7, and then weekly until wk 11 of age, blood samples were collected for measurement of metabolites and hormones related to energy metabolism and growth. The K-means cluster analysis on the MR meal size data collected from the automatic feeder resulted in 3 clusters (n = 14, n = 12, and n = 6). Two clusters with a sufficient cluster size (n = 14 and n = 12) were included for further statistical analysis using repeated measures mixed-model ANOVA. In both clusters, butyrate supplementation was equally distributed and failed to affect a difference in MR meal size. Cluster 1 showed calves with higher MR meal size (HI; 2.2 ± 0.11 L/visit of MR) and cluster 2 with lower meal size (LO; 1.8 ± 0.07 L/visit of MR) supplemented MR without (HIB−; n = 6; LOB−, n = 7) or with 0.33% calcium-sodium butyrate (HIB+; n = 6; LOB+, n = 7). Dry matter intake of MR did not differ between HI and LO, but intakes of concentrate and total dry matter tended to be greater in HI than in LO and increased more distinctly in HI than in LO at the end of the study. The average daily gain (g/d) was greater in HI than in LO. Plasma concentrations of total protein (g/L), albumin (g/L), glucose (mmol/L), urea (mmol/L), insulin (µg/L), and glucagon (ng/L) were higher, and the concentrations of insulin-like growth factor I tended to be higher, in HI than in LO calves. Plasma β-hydroxybutyrate was higher in LO than in HI at d 63 and lower in calves fed MR with butyrate at d 77. In conclusion, clustering analysis discriminates 2 main groups of calves with different MR meal size and indicates an effect of MR meal size on solid feed intake, growth performance, and metabolic changes.

Enteric diseases in piglets, such as post-weaning diarrhea (PWD), often require antibiotic treatment of the entire litter. Grape polyphenols may help overcome PWD and thereby reduce the need for antibiotics. The potential of a grape extract (GE; continuous in-feed supplementation) on performance of weaning piglets, compared with both negative (NC; corn-based diet) and positive control (PC; NC + in-feed antibiotic (amoxicillin) in a therapeutic dosage for day 1–day 5 post weaning) was assessed. Apparent total tract digestibility (ATTD) and microbial metabolites were also evaluated on two sampling points (day 27/28 and day 55/56). We assigned 180 weaning piglets (6.9 ± 0.1 kg body weight (BW)) to 6 male and 6 female pens per treatment with 5 piglets each. Animals from PC showed higher BW on day 13 compared with NC and GE, and a tendency for higher BW on day 56 (p = 0.080) compared to NC. Furthermore, PC increased the average daily feed intake in the starter phase (day 1–day 13), and the average daily gain in the early grower phase (day 14–day 24). Overall, GE improved the ATTD at the same level as PC (ash, acid-hydrolyzed ether extract), or at a higher level than PC (dry matter, organic matter, gross energy, crude protein, P). There were no effects on microbial metabolites apart from minor trends for lactic acid and ammonia. Dietary inclusion of GE may have beneficial effects compared to therapeutic antibiotics, as frequently used at weaning.

Dairy calves are able to digest and metabolize large amounts of milk or milk replacer when milk is freely available (1). However, ad libitum milk-or milk replacer-fed calves largely variate in their feeding behaviour and individual feed intake during the preweaning period (2).The aim of the present study was to detect possible similarities between milk feeding behaviour in specific subgroups of calves with free access to milk replacer (MR) and to explore how it can affect feed intake, growth performance, and blood metabolites and hormones. The study intends to classifying ad libitum-fed calves according to their individual voluntary MR intake using a clustering approach.

The objective of the current study was to characterize muscle and blood serum acylcarnitine (AcylCN) profiles and to determine the mRNA abundance of muscle carnitine acyltransferases in periparturient dairy cows with high (HBCS) and normal body condition (NBCS). Fifteen weeks antepartum, 38 pregnant multiparous Holstein cows were assigned to 2 groups that were fed differently to reach the targeted BCS and backfat thickness (BFT) until dry-off at -49 d before calving (HBCS: BCS >3.75 and BFT >1.4 cm; NBCS: <3.5 and <1.2 cm). Thereafter, both groups were fed identical diets. Blood samples and biopsies from the semitendinosus muscle were collected on d -49, 3, 21, and 84 relative to calving. Actual BCS at d -49 were 3.02 ± 0.24 and 3.82 ± 0.33 (mean ± SD) for NBCS and HBCS, respectively. In both groups, serum profiles showed marked changes during the periparturient period, with decreasing concentrations of free carnitine and increasing concentrations of long-chain AcylCN. Compared with NBCS, HBCS had greater serum long-chain AcylCN in early lactation, which may point to an insufficient adaptation of their metabolism in response to the metabolic load of fatty acids around parturition. The muscle concentrations of C5-, C9-, C18:1-, and C18:2-AcylCN were lower and those of C14:2-AcylCN were greater in HBCS than in NBCS cows. The mRNA abundance of carnitine palmitoyltransferase (CPT)1, muscle isoform (CPT1b) and CPT2 increased from d -49 to early lactation (d 3, d 21), followed by a decline to nearly antepartum values by d 84; this change was not affected by group. In conclusion, over-conditioning around calving seems to be associated with mitochondrial overload, which can result in incomplete fatty acid oxidation in dairy cows.

Abstract Background Fibre sources as feed components with specific physical characteristics like a high swelling capacity (SC), viscosity and water-binding capacity (WBC) have been discussed to affect sow behaviour and to have long-term effects on lactational performance. The present studies aim to analyse the effects of different fibre sources in diets for sows on behaviour in gestation, reproductive performance as well as piglet development. Methods Twenty-eight feedingstuffs (four grain varieties, 16 by-products, three oilseeds and five leguminous plants) were compared concerning swelling capacity, viscosity and water binding capacity to select fibre sources with optimal physical characteristics. Following this a digestibility study was carried out with eight castrates for determining digestibilities of gross energy, crude protein, crude fibre, crude fat and crude ash. Additionally, a practical feeding experiment during gestation was performed with 96 sows of Danish genetics. Two supplements for sows with different fibre sources were composed, namely a control diet (based on wheat bran and lignocellulose) and a test diet containing sugar beet pulp, alfalfa, rapeseed meal, soybean hulls, grape pomace and lignocellulose. Six pens with eight sows each were video-monitored for 2 weeks (evaluation of interactions and fights). Furthermore, the animals were subjected to weekly scoring to count skin injuries. To check the fibre effect on reproductive performance and piglet development, the body condition development of the sows as well as the number and weight of live and stillborn piglets, litter weight- and weaning weight of the litters were recorded. Results Digestibility of crude fibre increased significantly in the experimental group (58.8% ± 3.3 vs. 49.0% ± 4.3, p = 0.01). At the sow trial, there was a tendency to observe less aggressive interactions and fewer fights in sows in the fibre group without significance. No significant differences could be measured between the two groups concerning performance parameters of sows and piglets. Conclusion Only changing the fibre source in a gestational diet does not have significant effects on the sows´ behaviour and performance of sows and piglets in lactation. It should be investigated how the amount of fibre can be increased without having any negatives effects on the performance so that the positive effects on the behaviour of the sows become more obvious.

Optimal intestinal development in gilts is a prerequisite for a better farrowing and a higher feed intake in lactation, and these are important facets in improving animal welfare for sows in a high-performance situation. The present study aimed to evaluate the effects of a fiber-rich diet on these topics. Studies with 20 male Topigs castrates and a performance study with 38 female siblings were conducted. Pigs in both of the groups were offered either a control diet (CG) or a high fiber diet (FG) during the second half of the rearing phase (~97 kg BW) until 7 days a.p. At the time of the insemination of the gilts (body weight >170 kg), the castrates were slaughtered and the morphological parameters of the gastrointestinal tract were taken. In the performance study, the feed intake as well as the number and weight of the live and stillborn piglets and the weaning weight of the litters were recorded. The results indicated that the high fiber diet with a greater swelling capacity was beneficial to the development of the gastrointestinal tract (stomach weight CG = 714 g, FG = 812 g, p = 0.007), increased the litter weight (CG =15.9 kg, FG = 20.6 kg, p = 0.045) and resulted in a higher feed intake during lactation overall (CG = 196 kg, FG = 211 kg, p = 0.035).

This study applied a quantitative proteomics approach along with bioinformatics analyses to investigate changes in the plasma proteome of normal and overconditioned dairy cows during the transition period. Fifteen weeks before their anticipated calving date, 38 multiparous Holstein cows were selected based on their current and previous body condition scores (BCS) and allocated to either a high or a normal BCS group (19 cows each). They received different diets until dry-off to reach targeted differences in BCS and back fat thickness (BFT) until dry-off. At dry-off, normal BCS cows had a BCS <3.5 (minimum, 2.75) and BFT <1.2 cm (minimum, 0.58), and the high BCS cows had a BCS >3.75 (maximum, 4.50) and BFT >1.4 cm (maximum, 2.90). The proteomics study used a subset of 5 animals from each group. These cows were selected based on their circulating concentrations of fatty acids (FA) on d 14 postpartum and β-hydroxybutyrate (BHB) on d 21 postpartum, representing the greater or the lower extreme values within their BCS group, respectively. The high BCS subset (HE-HBCS) had 4.50 < BCS > 3.75, FA = 1.17 ± 0.46 mmol/L, and BHB = 2.15 ± 0.42 mmol/L (means ± SD), and the low BCS subset (LE-NBCS) had 3.50 < BCS > 2.75, FA = 0.51 ± 0.28 mmol/L, and BHB = 0.84 ± 0.17 mmol/L. Plasma samples from d -49, +7, and +21 relative to parturition were used for proteome profiling by applying the quantitative tandem mass tags (TMT) approach. Nondepleted plasma samples were subjected to reduction and digestion and then labeled with TMT 10plex reagents. High-resolution liquid chromatography-tandem mass spectrometry analysis of TMT-labeled peptides was carried out, and the acquired spectra were analyzed for protein identification and quantification. In total, 254 quantifiable proteins (criteria: 2 unique peptides and 5% false discovery rate) were identified in the plasma samples. From these, 24 differentially abundant proteins (14 more abundant, 10 less abundant) were observed in the LE-NBCS cows compared with the HE-HBCS cows during the transition period. Plasma α-2-macroglobulins were more abundant in HE-HBCS versus LE-NBCS cows at d +7 and +21. Gene Ontology enrichment analyses of differentially abundant proteins revealed that the acute inflammatory response, regulation of complement activation, protein activation cascade, and regulation of humoral immune response were the most enriched terms in the LE-NBCS group compared with the HE-HBCS group. In addition, we identified 24 differentially abundant proteins (16 in the LE-NBCS group, and 8 in the HE-HBCS group) during the transition period. The complement components C1q and C5 were less abundant, while C3 and C3d were more abundant in LE-NBCS compared with HE-HBCS cows. Overall, overconditioning around calving was associated with alterations in protein pathways related to acute inflammatory response and regulation of complement and coagulation cascades in transition cows.

This study evaluated the effects of increasing the dose of a 6-phytase from Buttiauxella on phytate degradation, mineral, energy, and AA digestibility in weaned pigs fed complex diets based on wheat, corn, soybean meal, barley, and rapeseed meal. A negative control (NC) diet containing no added inorganic phosphorus (P) and a reduction of 0.1% calcium (Ca) and 36 kcal/kg ME was supplemented with Buttiauxella phytase at 0, 250, 500, 1,000, or 2,000 FTU/kg diet and tested against a nutritionally adequate, positive control (PC) diet. One phytase units (FTU) is the amount of enzyme that liberates 1 micromole of inorganic phosphate per minute from a sodium phytate substrate at pH 5.5 and 37 °C. Barrows (Topigs × Pietrian; initial mean body weight 19.3 kg) were housed individually in metabolic crates and fed the test diets in mash form via 2 equal meals per day for 9 d (fed at 2.5 times the maintenance energy requirement), with 8 replicate pigs per treatment, in 2 experimental runs (total n = 48). After a 3-d adaptation period, urine and feces were collected over 5 d for measurements of apparent total tract digestibility (ATTD) and retention of nutrients. On day 9, pigs were euthanized and ileal digesta collected for measurements of apparent ileal digestibility (AID) of nutrients. Phytase improved (P < 0.05) digestibility of all measured AA except Trp (P < 0.1), and AID P, nitrogen, phytate, ATTD P, Ca versus NC. Increasing phytase dose from 0 (NC) to 2,000 FTU/kg increased AID Lys, Cys, Thr, Val, Ile, Leu, mean AA, P, N, phytate, ATTD P, N, Na, energy, ME, P retention (g/d), and reduced P excretion (g/d) in a linear or exponential manner (P < 0.05). Phytase at 2,000 FTU/kg improved AA digestibility by between +3.1 percentage points (Trp) and +8.8 percentage points (Cys) versus NC (average +6.3 percentage points) (P < 0.05). Phytase inclusion at 2,000 FTU/kg reduced P excretion (g/d) by 57% versus PC (P < 0.05). In conclusion, increasing Buttiauxella phytase in the range of 0 to 2,000 FTU/kg increased phytate degradation, improved AA and P digestibility, and reduced P excretion in weaned pigs fed complex diets.

This study aimed to investigate the differences in the metabolic profiles in serum of dairy cows that were normal or overconditioned when dried off for elucidating the pathophysiological reasons for the increased health disturbances commonly associated with overconditioning. Fifteen weeks antepartum, 38 multiparous Holstein cows were allocated to either a high body condition (HBCS; n = 19) group or a normal body condition (NBCS; n = 19) group and were fed different diets until dry-off to amplify the difference. The groups were also stratified for comparable milk yields (NBCS: 10,361 ± 302 kg; HBCS: 10,315 ± 437 kg; mean ± standard deviation). At dry-off, the cows in the NBCS group (parity: 2.42 ± 1.84; body weight: 665 ± 64 kg) had a body condition score (BCS) <3.5 and backfat thickness (BFT) <1.2 cm, whereas the HBCS cows (parity: 3.37 ± 1.67; body weight: 720 ± 57 kg) had BCS >3.75 and BFT >1.4 cm. During the dry period and the subsequent lactation, both groups were fed identical diets but maintained the BCS and BFT differences. A targeted metabolomics (AbsoluteIDQ p180 kit, Biocrates Life Sciences AG, Innsbruck, Austria) approach was performed in serum samples collected on d -49, +3, +21, and +84 relative to calving for identifying and quantifying up to 188 metabolites from 6 different compound classes (acylcarnitines, AA, biogenic amines, glycerophospholipids, sphingolipids, and hexoses). The concentrations of 170 metabolites were above the limit of detection and could thus be used in this study. We used various machine learning (ML) algorithms (e.g., sequential minimal optimization, random forest, alternating decision tree, and naïve Bayes-updatable) to analyze the metabolome data sets. The performance of each algorithm was evaluated by a leave-one-out cross-validation method. The accuracy of classification by the ML algorithms was lowest on d 3 compared with the other time points. Various ML methods (partial least squares discriminant analysis, random forest, information gain ranking) were then performed to identify those metabolites that were contributing most significantly to discriminating the groups. On d 21 after parturition, 12 metabolites (acetylcarnitine, hexadecanoyl-carnitine, hydroxyhexadecenoyl-carnitine, octadecanoyl-carnitine, octadecenoyl-carnitine, hydroxybutyryl-carnitine, glycine, leucine, phosphatidylcholine-diacyl-C40:3, trans-4-hydroxyproline, carnosine, and creatinine) were identified in this way. Pathway enrichment analysis showed that branched-chain AA degradation (before calving) and mitochondrial β-oxidation of long-chain fatty acids along with fatty acid metabolism, purine metabolism, and alanine metabolism (after calving) were significantly enriched in HBCS compared with NBCS cows. Our results deepen the insights into the phenotype related to overconditioning from the preceding lactation and the pathophysiological sequelae such as increased lipolysis and ketogenesis and decreased feed intake.

Long noncoding RNAs (lncRNAs) emerged as important regulatory component of mechanisms involved in gene expression, chromatin modification and epigenetic processes, but they are rarely annotated in the bovine genome. Our study monitored the jejunum transcriptome of German Holstein calves fed two different milk diets using transcriptome sequencing (RNA-seq). To identify potential lncRNAs within the pool of unknown transcripts, four bioinformatic lncRNA prediction tools were applied. The intersection of the alignment-free lncRNA prediction tools (CNCI, PLEK and FEELnc) predicted 1,812 lncRNA transcripts concordantly comprising a catalogue of 1,042 putative lncRNA loci expressed in the calves' intestinal mucosa. Nine lncRNA loci were differentially expressed (DE lncRNAs) between both calf groups. To elucidate their biological function, we applied a systems biology approach that combines weighted gene co-expression network analysis with functional enrichment and biological pathway analysis. Four DE lncRNAs were found to be strongly correlated with a gene network module (GNM) enriched for genes from canonical pathways of remodeling of epithelial adherens junction, tight junction and integrin signaling. Another DE lncRNA was strongly correlated with a GNM enriched for genes associated with energy metabolism and maintaining of cellular homeostasis with a focus on mitochondrial processes. Our data suggest that these DE lncRNAs may play potential regulatory roles in modulating biological processes associated with energy metabolism pathways and cellular signaling processes affecting the barrier function of intestinal epithelial cells of calves in response to different feeding regimens in the pre-weaning period.