Fatty acid profile of phospholipids and sphingomyelin in milk and regulation of sphingomyelin synthesis of mammary glands in cows receiving increasing levels of crushed sunflower seeds.

The objective of this study was to investigate the effect of increasing dietary supplementation of crushed sunflower seed (CSS) in the diet of dairy cows on the fatty acid (FA) composition of phospholipids and sphingomyelin in milk, and on mammary transcription of genes that are important for sphingomyelin de novo synthesis. Four groups of 6 cows received diets supplemented with CSS at 0% (control), or 5, 10, or 15% of dry matter for a 5-wk experimental period. Milk samples and mammary biopsies were collected at the end of the experiment. Phospholipid concentration in milk fat decreased linearly with CSS supplementation. Sphingomyelin concentration in milk fat was unaffected by CSS supplementation. Daily yield of phospholipids decreased linearly with CSS supplementation. Daily yield of sphingomyelin was not significantly affected. The CSS supplementation linearly increased the proportion of monounsaturated FA in milk phospholipids. The major isomer incorporated into phospholipids was C18:1 (n-9 cis), which showed a linear increase with CSS supplementation. The C22:0 proportion in sphingomyelin increased linearly with CSS supplementation and constituted between 15.2 to 25.4% of total FA in sphingomyelin. However, CSS supplementation linearly decreased C23:0 sphingomyelin. Mammary transcription of serine palmitoyl transferase, long chain subunit 1 and subunit 2, the rate-limiting enzymes in ceramide synthesis, showed a linear decrease with increasing CSS supplementation. In conclusion, the data showed that dietary supplementation of CSS linearly increased the proportion of unsaturated FA and monounsaturated FA in milk phospholipids with no effect on phospholipid concentration. In addition, CSS supplementation linearly decreased n-3 polyunsaturated fatty acid proportion in sphingomyelin. The results further showed that mammary transcription of important genes for sphingomyelin de novo synthesis is regulated by lipid supplementation.

Transcriptional profiling of swine mammary gland during the transition from colostrogenesis to lactogenesis using RNA sequencing.

BACKGROUND: Colostrum and milk are essential sources of antibodies and nutrients for the neonate, playing a key role in their survival and growth. Slight abnormalities in the timing of colostrogenesis/lactogenesis potentially threaten piglet survival. To further delineate the genes and transcription regulators implicated in the control of the transition from colostrogenesis to lactogenesis, we applied RNA-seq analysis of swine mammary gland tissue from late-gestation to farrowing. Three 2nd parity sows were used for mammary tissue biopsies on days 14, 10, 6 and 2 before (-) parturition and on day 1 after (+) parturition. A total of 15 mRNA libraries were sequenced on a HiSeq2500 (Illumina Inc.). The Dynamic Impact Approach and the Ingenuity Pathway Analysis were used for pathway analysis and gene network analysis, respectively. RESULTS: A large number of differentially expressed genes were detected very close to parturition (-2d) and at farrowing (+ 1d). The results reflect the extraordinary metabolic changes in the swine mammary gland once it enters into the crucial phases of lactogenesis and underscore a strong transcriptional component in the control of colostrogenesis. There was marked upregulation of genes involved in synthesis of colostrum and main milk components (i.e. proteins, fat, lactose and antimicrobial factors) with a pivotal role of CSN1S2, LALBA, WAP, SAA2, and BTN1A1. The sustained activation of transcription regulators such as SREBP1 and XBP1 suggested they help coordinate these adaptations. CONCLUSIONS: Overall, the precise timing for the transition from colostrogenesis to lactogenesis in swine mammary gland remains uncharacterized. However, our transcriptomic data support the hypothesis that the transition occurs before parturition. This is likely attributable to upregulation of a wide array of genes including those involved in 'Protein and Carbohydrate Metabolism', 'Immune System', 'Lipid Metabolism', 'PPAR signaling pathway' and 'Prolactin signaling pathway' along with the activation of transcription regulators controlling lipid synthesis and endoplasmic reticulum biogenesis and stress response.

Effects of nitrogen supply on inter-organ fluxes of urea-N and renal urea-N kinetics in lactating Holstein cows.

The effects of decreasing ruminal urea infusion in lactating dairy cows fed a basal diet deficient in rumen degradable protein on inter-organ urea-N fluxes, epithelial urea-N extraction, and renal urea-N kinetics were investigated. Eight Danish Holstein cows fitted with a ruminal cannula and permanent indwelling catheters in the major splanchnic blood vessels and the gastrosplenic vein were used. The cows were randomly allocated to a triplicate incomplete 3 × 3 Latin square design with 14-d periods. Treatments were continuous ventral ruminal infusion of water, 4.1g of feed urea/kg of dry matter intake, and 8.5 g of feed urea/kg of dry matter intake. Dry matter intake and milk yield decreased linearly with decreasing urea infusion. Arterial blood urea-N and ruminal ammonia concentrations decreased linearly with decreasing urea infusion. In absolute amounts, the urea-N recycling did not increase when urea infusion was decreased. Arterial urea-N extraction across the portal-drained viscera and rumen wall increased linearly with decreasing urea infusion (2.46, 3.65, and 4.32 ± 0.31% and 7.5, 11.5, and 16.9 ± 0.9%, respectively), indicating that cows responded to the changes in N supply. The relative urea-N extraction across the ruminal wall increased compared with the total portal-drained viscera extraction. We observed a postprandial decrease in ruminal extraction of arterial urea-N that might reflect that the activity of the protein, presumably facilitating urea-N transport, is regulated by ruminal ammonia. The urea-N clearance by the kidneys decreased (35, 30, and 25 ± 2L/h) and the urea-N reabsorbed by the kidney increased (42, 51 and 56 ± 3%) with decreasing urea infusion, indicating that the kidneys salvaged urea-N with low-N supply. The urea transporter B mRNA abundance in rumen papillae (papillae harvested at sampling days) was not affected by dietary N supply. The study showed, that rumen wall extraction of arterial urea-N is subjected to both long- and short-term regulation. Extraction increases with decreasing N supply long-term; however, a short-term postprandial decrease in extraction was observed. No association between long-term adaptation of urea-N extraction across the rumen wall and urea transporter B mRNA abundance could be demonstrated.

Short communication: Effects of dietary nitrogen concentration on messenger RNA expression and protein abundance of urea transporter-B and aquaporins in ruminal papillae from lactating Holstein cows.

To test the hypothesis that dietary N concentrations affect gut epithelial urea transport by modifying the expression of urea transporter B (UT-B) and aquaporins (AQP), the mRNA expression and protein abundance of UT-B and AQP3, AQP7, AQP8, and AQP10 were investigated in ruminal papillae from 9 lactating dairy cows. Ruminal papillae were harvested from cows fed low N (12.9% crude protein) and high N (17.1% crude protein) diets in a crossover design with 21-d periods. The mRNA expression was determined by real-time reverse transcription-PCR and protein abundance by immunoblotting. The mRNA expression of UT-B was not affected by dietary treatment, whereas mRNA expression of AQP3, 7, and 10 were greater in the high N compared with the low N fed cows. Using peptide-derived rabbit antibodies to cow AQP3, 7, and 8, immunoblotting revealed bands of approximately 27, 27, and 24 kDa in ruminal papillae, respectively. A peptide-derived chicken antibody to cow UT-B detected a band of approximately 30 to 32 kDa in ruminal papillae. The abundance of UT-B and AQP3 and 7 were not affected by dietary treatment. In contrast, the abundance of AQP8 was greater in high N compared with low N diets. In conclusion, AQP3, 7, and 8 were found to be expressed in bovine rumen papillae. None of the investigated transcripts or proteins correlated to the increased rumen epithelial urea permeability observed with low dietary N concentration.

Impact of litter size, supplementary milk replacer and housing on the body composition of piglets from hyper-prolific sows at weaning.

The modern hyper-prolific sow gives birth to 17 live-born piglets on average. An alternative strategy to nurse sows and artificial rearing may be providing milk replacer while letting all the piglets stay with their dam. However, milk replacer is of lower nutritional quality than sow milk and may reduce the body fat content of piglets who use milk replacer to compensate for low suckling success due to competition at the udder. Therefore, the aim of this study was to investigate the body composition at weaning of two random sow-reared piglets per litter from 93 litters by using the deuterium oxide dilution technique. The piglets were part of large study with a 2×2×2 factorial design of either 14 or 17 piglets from day 1 (LS: LS14/LS17) with or without access to milk replacer (MILK: -MILK/+MILK) and reared by crated or loose-housed sows (HOUSING: CRATE/ LOOSE). From behavioral observations day 21 in +MILK, piglets were divided according to their frequency of drinking milk replacer and suckling (Nutrition Source). Increasing LS from 14 to 17 reduced the average daily gain from 258 to 228 g/d and body fat % from 14.4 to 12.7% (P<0.01). In a two-way interaction between LS and HOUSNG, the body fat percentage was lower (P=0.04) and the water percentage tended to be higher (P=0.07) in LS17 CRATE compared to the other treatments (i.e. LS17 LOOSE, LS14 CRATE and LOOSE). There was no effect of MILK on piglet composition day 28 (P>0.1). In +MILK, the Nutrition Source affected piglet body composition (P<0.05) as piglets with low suckling frequency (LOW) had lower body fat and higher water content compared to piglets who had high suckling frequency (SUCKLE). Unexpectedly, drinking milk replacer in addition to suckling (MIXED) did not increase piglet body fat content. Relying mainly on milk replacer (CUP) caused body fat and water contents to be intermediate to piglets with high (SUCKLE and MIXED) and low suckling frequency (LOW). In conclusion, LS had a clear impact on piglet growth and body composition at weaning. In contrast, supplementation of milk and housing had only negligible impact on litter performance. Some individual piglets that had low frequency of sow milk intake benefitted from milk supplementation. Loose housing appeared to benefit piglet body fat at weaning but this was due to a greater piglet mortality.

Continuous lactation effects on mammary remodeling during late gestation and lactation in dairy goats.

The present study aimed to 1) elucidate whether continuous milking during late gestation in dairy goats negatively affects mammary remodeling and hence milk production in the subsequent lactation, and 2) identify the regulatory factors responsible for changes in cell turnover and angiogenesis in the continuously lactating mammary gland. Nine multiparous dairy goats were used. One udder half was dried off approximately 9 wk prepartum (normal lactation; NL), and the other udder half of the same goat was milked continuously (continuous lactation; CL) until parturition or until the half-udder milk yields had dropped to below 50 g/d. Mammary biopsies were obtained from each udder half just before the NL gland was dried off (before dry period), within the first 2 wk after drying-off (early dry period, samples available only for NL glands), in the mid dry period, within the last 2 wk before parturition (late dry period), and at d 1 (the day of parturition), 3, 10, 60, and 180 of lactation. Mammary morphology was characterized in biopsies by quantitative histology, and cell turnover was determined by immunohistochemistry (terminal deoxynucleotidyl transferase dUTP nick end labeling and Ki-67). Transcription of genes encoding factors involved in mammary epithelial cell (MEC) turnover and vascular function was quantified by quantitative reverse transcription PCR. Results demonstrated that omitting the dry period was possible in goats but was not as easy as claimed before. Renewal of MEC was suppressed in CL glands, which resulted in a smaller MEC population in the subsequent lactation. At the time of parturition (and throughout lactation), the mammary glands subjected to CL had smaller alveoli, more fully differentiated MEC, and a substantially larger capillary fraction compared with NL glands. The continuously lactating gland thus resembled a normally lactating gland in an advanced stage of lactation. None of the studied genomic factors could account for these treatment differences. The described characteristics in CL glands compared with NL glands indicated a gland maintained in lactation for a longer period.

Mammary remodeling in primiparous and multiparous dairy goats during lactation.

Milk production is generally lower but lactation persistency higher in primiparous (PP) than in multiparous (MP) goats. This may be related to differences in development and maintenance of mammary gland function, but the underlying mechanisms are not well understood. The present study aimed to elucidate whether differences in lactational performance between PP and MP mammary glands are related to the time course of development and maintenance, not only of the mammary epithelial cell (MEC) population, but also of the mammary vasculature that sustains synthetic activity. Mammary biopsies were obtained from both mammary glands of 3 PP and 6 MP (>or=2 parity) dairy goats at parturition (d 1), d 10, 60, and 180 of lactation. Gene transcription relating to MEC turnover and vascular function was quantified by real-time reverse transcription-PCR, mammary morphology was characterized (quantitative histology), and cell turnover was determined (terminal deoxynucleotidyl transferase dUTP nick end labeling assay and Ki-67). Primiparous glands showed higher expression for the genes involved in angiogenesis; namely, vascular endothelial growth factor receptor 2, and angiopoietin 1 and 2 and their receptor, a few days after parturition (d 10). Primiparous glands also had higher rates of MEC proliferation in early lactation. It therefore appears that initiation of lactation is associated with development and growth of the mammary gland into early lactation, which continues for a longer period in PP compared with MP glands. In addition, MEC survival was found to be higher in PP glands throughout lactation, and MEC in PP glands underwent more extensive differentiation. This could explain the reported flatter lactation curve and higher lactation persistency in PP glands. Although some of the genes included in this study were differentially expressed in PP and MP glands during the course of lactation, it was not possible to identify any specific genomic factor(s) that could account for the differences between PP and MP glands with respect to mammary development and MEC survival during lactation. It remains to be established why parity number affects MEC and vascular development and survival during lactation, and, in particular, which regulatory mechanisms are involved.

Impact of sow parity on yield and composition of colostrum and milk in Danish Landrace × Yorkshire crossbred sows.

The present study aims to characterize the colostrum, milk yield and composition and to determine whether sow parity would influence yield and composition of colostrum and milk in Danish Landrace × Yorkshire crossbred sows. The data were collected from sow parity numbers 1 (n = 27), 2-4 (n = 48) and 5-6 (n = 30) from Danish Landrace × Yorkshire crossbred sows reared in a commercial swine herd in Thailand. The piglets were weighed on day 0 (<1 h), 1 (24 h), 3, 10 and 17 after birth to determine the colostrum and milk yields of the sows using a prediction equation. Milk samples were collected manually within 1 h of the onset of parturition and on days 3, 10 and 17 after farrowing to evaluate milk composition. A general linear model procedure was used to analyze the effects of sow parity numbers on colostrum yield and composition and a general linear mixed model procedure was used to analyze the effects of sow parity numbers on yield and composition of milk. The model included the fixed effects of sow parity number and time (day after parturition). The sow parity numbers 2-4 (7.0 kg) had a higher colostrum yield than 1st parity sows (5.4 kg, P = 0.002) and parity 5-6 sows (5.9 kg, P = 0.025). No evidence of parity differences was observed on milk yield (P = 0.306). No effect of sow parity numbers on fat, protein and lactose in milk was observed. The dry matter in sow parity numbers 2-4 (19.8 g/100 g) had a tendency to be higher than sow parity number 1 (18.6 g/100 g, P = 0.107) and 5-6 (18.4 g/ 100 g, P = 0.053). In conclusion, sow parity number had an impact on colostrum yield in Danish Landrace × Yorkshire crossbred sows in a tropical climate but did not influence colostrum, milk composition and milk yield. Colostrum yield in Danish Landrace × Yorkshire crossbred sows was the highest in sow parity numbers 2-4.

Effect of litter size, milk replacer and housing on production results of hyper-prolific sows.

The modern hyper-prolific sow gives birth to more piglets than she has functional teats (in the following called supernumerary piglets). The aim of the present study was (1) to investigate the production consequences of hyper-prolific sows rearing supernumerary piglets equal to the mean live-born litter size, and (2) investigate whether potential negative effects on survival and growth could be alleviated by providing access to milk replacer and/or providing easier access to the udder (by loose housing). At day 1 (D1) postpartum (pp), 93 litters were standardised to 14 or 17 piglets (LS14/LS17) after which no piglets were moved between sows leading to decreased litter size if piglets died. Litters were provided with or without milk replacer in milk cups (+MILK/-MILK), and sows were either crated or loose housed (CRATE/LOOSE) in a 2 × 2 × 2 factorial design. Piglet mortality was higher in LS17 compared to LS14 (P < 0.01; OR = 2.0), higher in -MILK compared to +MILK (P = 0.01; OR = 1.2) and higher in LOOSE compared to CRATE (P = 0.02; OR = 1.8). This study showed that sow rearing of supernumerary piglets while supplying with milk replacer can increase piglet survival. It also showed that early mortality before piglets learned to drink milk replacer posed a challenge using this automatic milk replacer system. An interaction between access to milk replacer and the standardised litter size D1 affected litter weight (P < 0.01) and piglet weight day 28 (D28) (P = 0.03). The highest litter weight D28 was found in LS17 +MILK (P < 0.01) but with a lower individual piglet weight than in LS14 -MILK. Piglet weight D28 was higher in LS14 -MILK compared to LS17 regardless of access to milk replacer. Heterogeneity in piglet weight within litters D28 was larger in LS17 (P = 0.03) but could be reduced with +MILK in CRATE (P < 0.01). No effects were found on sow weight loss and feed intake (P > 0.05). In conclusion, the results showed that sows cannot rear the supernumerary piglets without further management interventions to reduce mortality. Supplying supernumerary piglets equal to the mean live-born litter size of hyper-prolific sows with milk replacer can from results of this study be an alternative strategy to the use of nurse sows.

Increased dietary protein for lactating sows affects body composition, blood metabolites and milk production.

Hyper-prolific sows nurse more piglets than less productive sows, putting a high demand on the nutrient supply for milk production. In addition, the high production level can increase mobilization from body tissues. The effect of increased dietary protein (104, 113, 121, 129, 139 and 150 g standardized ileal digestible (SID) CP/kg) on sow body composition, milk production and plasma metabolite concentrations was investigated from litter standardization (day 2) until weaning (day 24). Sow body composition was determined using the deuterium oxide dilution technique on days 3 and 24 postpartum. Blood samples were collected weekly, and milk samples were obtained on days 3, 10 and 17 of lactation. Litter average daily gain (ADG) peaked at 135 g SID CP/kg (P < 0.001). Sow BW and back fat loss reached a breakpoint at 143 and 127 g SID CP/kg (P < 0.001). Milk fat increased linearly with increasing dietary SID CP (P < 0.05), and milk lactose decreased until a breakpoint at 124 g SID CP/kg and 5.3% (P < 0.001) on day 17. The concentration of milk protein on day 17 increased until a breakpoint at 136 g SID CP/kg (5.0%; P < 0.001). The loss of body protein from day 3 until weaning decreased with increased dietary SID CP until it reached a breakpoint at 128 g SID CP/kg (P < 0.001). The body ash loss declined linearly with increasing dietary SID CP (P < 0.01), and the change in body fat was unaffected by dietary treatment (P=0.41). In early lactation (day 3 + day 10), plasma urea N (PUN) increased linearly after the breakpoint at 139 g SID CP/kg at a concentration of 3.8 mmol/l, and in late lactation (day 17 + day 24), PUN increased linearly after a breakpoint at 133 g SID CP/kg (P < 0.001) at a concentration of 4.5 mmol/l. In conclusion, the SID CP requirement for sows was estimated to 135 g/kg based on litter ADG, and this was supported by the breakpoints of other response variables within the interval 124 to 143 g/kg.

Cellular mechanisms in regulating mammary cell turnover during lactation and dry period in dairy cows.

The mechanisms involved in regulating mammary cell turnover during the pregnancy-lactation cycle in dairy cows are unclear. The objective of present experiment was to describe expression of genes encoding proteins known to be involved in pathways regulating mammary cell proliferation, apoptosis, differentiation, cell survival, and tissue remodeling. Mammary gland biopsies were taken 7 times during the pregnancy-lactation cycle of 10 dairy cows, and samples were analyzed by immunohistochemistry and real-time PCR. Cell proliferation was greatest during the dry period and apoptosis was high in early dry period and early lactation. Based on Fas (tumor necrosis factor receptor superfamily member 6), Fas ligand, and caspase-3, caspase-8, and caspase-9 gene expression, no indication was found of a stage-dependent shift between the extrinsic and intrinsic pathways leading to apoptosis. Gene expression of microsomal glutathione S-transferase (mGST) did not vary significantly, whereas B-cell leukemia/lymphoma 2 (Bcl-2) and BCL2-associated X protein (Bax) gene expression was greatest during the dry period and early lactation and coincided with high cell turnover. Gene expression of early response genes c-Fos, c-Jun, and c-Myc correlated to neither rate of cell proliferation nor plasma concentration of insulin-like growth factor (IGF)-I and insulin. Gene expression of nuclear factor of kappa light chain gene enhancer in B-cells (NFkappaB) and NFkappaB inhibitor alpha was greatest in the periparturient period, and NFkappaB gene expression coincided with an anticipated need for cell survival factors. Expression of transforming growth factor beta (TGF-beta) receptor 1 and 2 mRNA was greatest in early lactation, whereas TGF-beta1 did not vary significant during the pregnancy-lactation cycle. Even though our results on the TGF-beta system did not comply with other studies, the gene expression pattern of the TGF-beta receptors indicates a role in regulating apoptosis in early lactation. Signal transducer and activator of transcription 5 (STAT5) gene expression was high in the periparturient period, which suggests a role for STAT5 in regulation of mammary cell proliferation and differentiation in dairy cows. Expression of tissue-plasminogen activator, plasminogen activator inhibitor-1, and IGF binding protein 5 genes was greatest in early lactation, suggesting a role for IGF binding protein 5 in coordinating regulation of apoptosis and tissue remodeling.

Temporal changes in glycogenolytic enzyme mRNAs during myogenesis of primary porcine satellite cells.

The objective was to study the regulation of glycogenolytic enzyme mRNAs in porcine satellite cells during proliferation and differentiation. Beyond 80% confluence, cells were grown in absence or presence of 1µM insulin. The observed increases in abundance of mRNA for glycogenin, glycogen synthase, phosphorylase kinase, phosphorylase and glycogen debranching enzyme, and no alterations of the transporter molecule GLUT4, clearly indicate that glycogenolytic enzymes of potential importance to meat quality development are regulated at the gene level during myogenesis, and are heavily involved in muscle cell and muscle fibre development. The genes, however, are not influenced by insulin, and the lack of response to insulin of expression of gene-encoding enzymes involved in the formation and degradation of glycogen may question the applicability of porcine cell culture systems, like the one applied, as a model to study the regulation and regulatory mechanism of energy metabolism in muscles.

In vitro and in vivo studies of creatine monohydrate supplementation to Duroc and Landrace pigs.

Duroc and Landrace pigs as well as primary myotubes from these breeds were used to investigate mechanisms behind differences in their response to creatine monohydrate (CMH). Pigs were supplemented with 0, 12.5, 25 or 50g CMH/d for 5 days (n=10 per treatment and breed). Plasma levels of creatine increased dose-dependently in both breeds, while muscle-creatine phosphate content increased only in the Duroc pigs. (1)H NMR metabolic profiling showed a tendency towards clustering according to CMH supplementation only among Duroc pigs, revealing a stronger response compared to Landrace pigs. The abundance of insulin-like growth factor I and myostatin mRNA was decreased by CMH supplementation while that of type 1 IGF-receptor and creatine transporter was unaffected. Protein synthesis, increased in the myotubes from both breeds, indicating protein accretion, but no effect was observed on the mRNA abundance of IGF-I, type 1 IGF-receptor, myostatin or the creatine transporter in myotubes.

Impact of fat source and dietary fibers on feed intake, plasma metabolites, litter gain and the yield and composition of milk in sows.

Sow lactation diets often include fat sources without considering the impact on digestion, metabolism and performance. Fiber ingredients may reduce feed intake and are often completely excluded from lactation diets, although locally available ingredients may be cost-efficient alternatives to partly replace cereals in lactation diets. Thus, a standard lactation diet low in dietary fiber, and two high-fiber diets based on sugar beet pulp (SBP) or alfalfa meal (ALF) were formulated. The SBP diet was high in soluble non-starch polysaccharides (NSP), whereas ALF being high in insoluble NSP. Each diet was divided in three portions and combined with 3% soybean oil (SOYO), palm fatty acid distillate (PFAD), or glycerol trioctanoate (C8TG) as the dietary fat source. Equal amounts of metabolizable energy were fed to 36 second parity sows from day 105 of gestation and throughout lactation to study the impact on feed intake, plasma metabolites, milk production and litter performance. Backfat thickness and BW of sows were recorded on days 3, 17 and 28 of lactation; blood was sampled on days 3 and 17; milk samples were obtained on days 3, 10, 17 and 24 of lactation; and piglets were weighed on days 2, 7, 14, 21 and 28 of lactation. Litter gain and milk yield during late lactation were greater in sows fed C8TG or SOYO than in sows fed PFAD (P=0.05), whereas loss of BW (P=0.60) and backfat (P=0.70) was unaffected by fat source. Milk protein on days 3 and 10 of lactation were lower in C8TG and SOYO sows, than in PFAD sows (P<0.05). The lowest concentration of plasma lactate on day 3 (P<0.05) and plasma acetate on day 17 (P<0.05) was observed in C8TG sows. Milk yield was unaffected by fiber treatment (P=0.43), whereas milk protein concentration was lowest in ALF sows (P<0.05). Feed intake tended to be lower (P=0.09), and litter gain during the 3rd week of lactation was decreased (P<0.05) in SBP sows. In conclusion, performance was enhanced in SOYO and C8TG compared with PFAD sows, possibly associated with reduced energy intake in PFAD-fed sows. Furthermore, the SBP diet seemed to impair feed intake and litter gain at peak lactation, suggesting that effects of the dietary fiber fraction on energy intake determines the potential inclusion level of fiber-rich ingredients.

Mammary nutrient uptake in multiparous sows fed supplementary arginine during gestation and lactation.

Arginine is the precursor for the synthesis of nitric oxide and may increase mammary plasma flow (MPF), which may in turn increase mammary nutrient uptake. Quantifying mammary nutrient uptake improves our understanding of mammary nutrient metabolism and may potentially allow identification of limiting nutrients for colostrum and milk production. Thus, the objectives of the present study were 1) to study the impact of 25 g/d of crystalline Arg (ARG) on MPF and uptake of nutrients by the mammary glands compared with an isonitrogenous supply of Ala (51 g/d; control [CON]) fed to a total of 8 sows from d 30 of gestation until weaning on d 28 of lactation and 2) to quantify mammary nutrient uptake in late gestation and in early and at peak lactation. Sows were surgically fitted with indwelling catheters on d 76 ± 2 SEM of gestation. -amino hippuric acid (AH) was infused (3.0 mmol/h) in the infusion catheter inserted in the mammary vein, initiated 1 h before the first blood sample at -10, -3, 3, and 17 d in milk (DIM). Blood samples were simultaneously drawn from catheters inserted in the femoral artery and the mammary vein, and the samples were collected in hourly intervals from 0.5 h before to 6.5 h after feeding. Sow milk production was assessed at 3 and 17 DIM. Arterial plasma concentrations of Arg and Ala were increased in ARG and CON sows, respectively ( < 0.01), whereas we did not succeed in detecting a greater MPF in ARG sows ( = 0.30). Arterial-venous differences ( = 0.03) and net mammary flux ( = 0.01) of Ala were increased in CON sows, while the net flux of most other metabolites ( > 0.05) was unaffected by treatment. The mammary extraction of all essential AA was below 13% in late gestation. The average mammary extraction of essential AA at peak lactation was greatest for Leu (51%), while the preprandial extraction was greatest for Lys (57%). The mammary carbon balance (input-output) was negative (-39 ± 12 mol C/d) in early lactation but almost balanced at peak lactation (-13 ± 14 mol C/d), suggesting that mammary fat depots contributed to milk synthesis. In conclusion, we failed to observe an increased MPF and mammary uptake of AA and energy metabolites in ARG-supplemented sows. The mammary extraction rate of essential AA indicated that AA were not limiting for the mammary glands in late gestation, while Lys and Leu appeared to be the 2 most limiting essential AA for milk production at peak lactation.

Effect of experimentally increased protein supply to postpartum dairy cows on plasma protein synthesis, rumen tissue proliferation, and immune homeostasis.

The effect of experimentally increasing the postpartum protein supply on plasma protein synthesis, rumen tissue proliferation, and immune homeostasis was studied using 8 periparturient Holstein cows in a complete randomized design. At calving, cows were assigned to abomasal infusion of water (CTRL) or casein (CAS) in addition to a lactation diet. Casein infusion was gradually decreased from 696 ± 1 g/d at +2 d relative to calving (DRTC) to 212 ± 10 g/d at +29 DRTC to avoid excessive supply. Synthesis rate of plasma proteins was measured at -14, +4, +15, and +29 DRTC by measuring [C]Phe isotopic enrichment in arterial plasma free Phe, total plasma proteins, and albumin after 3, 5, and 7 h of jugular ring[C]Phe infusion. Plasma volume was determined at +4 and +29 DRTC by dilution of a [I]BSA dose. Synthesis rate of tissue protein in biopsied rumen papillae was determined by measuring [C]Phe isotopic enrichment, and mRNA expression of selected genes was measured by real-time qPCR. Total and differential leukocyte counts were performed and immune responsiveness of monocytes was evaluated by tumor necrosis factor ɑ (TNFɑ) concentration on ex vivo whole blood stimulation with Escherichia coli lipopolysaccharide (LPS) and responsiveness of T-lymphocytes by interferon γ (IFNγ) concentration on stimulation with Staphylococcus aureus enterotoxin ß (SEB). Further, ELISA plasma concentrations of IgM, IgA, and IgG were determined. The DRTC affected the majority of investigated parameters as expected. The CAS treatment increased milk protein yield (P = 0.04), and tended to lower TNFɑ (P = 0.06), and lowered IFNγ (P = 0.03) responsiveness per monocyte and lymphocyte, respectively, compared with CTRL. Further, fractional synthesis rate of albumin was greater at +4 DRTC for CAS compared with CTRL but did not differ by +29 DRTC (interaction: P = 0.01). In rumen papillae, synthesis rate of tissue protein was greater for CAS compared with CTRL (P < 0.01) and mRNA expression of genes for cell proliferation tended to be or were greater for CAS compared with CTRL (P ≤ 0.07). In conclusion, increased postpartum protein supply seem to enhance vital body functions as interpreted from increased liver synthesis of albumin, increased rumen papillae proliferation, and stabilized the ex vivo inflammatory responsiveness of leukocytes. Further studies are needed to enlighten the importance of increased postpartum protein supply in periparturient cows.

Dietary supplement rich in fiber fed to late gestating sows during transition reduces rate of stillborn piglets.

The beneficial effects of dietary fiber (DF) from a behavioral and welfare perspective have been thoroughly studied. However, data on the effects of DF on reproductive performance are scarce. Therefore, the aim of this study was to investigate the impact of increased DF supply during the last 2 wk of gestation on stillbirth rate, preweaning mortality, and total piglet mortality. A total of 644 sows were selected for the experiment from a commercial farm, and the sows were inseminated in weekly batches. Sows in the control group ( = 310) were fed according to the normal feeding strategy of the farm with a gestation diet until 1 wk before expected farrowing, then a transition diet until d 5 of lactation, and then a lactation diet until weaning. Sows in the treatment group ( = 334) were fed as the control group except that 280 g/d of the gestation diet (from d 102 to 108 of gestation) and 570 g/d of the transition diet (from d 109 of gestation until farrowing) was daily replaced with 350 and 700 g/d, respectively, of a DF-rich supplement. Both groups received isocaloric diets on a NE basis. The numbers of live-born and stillborn piglets as well as mortality of live-born piglets with presumed causes of death were recorded. The supplemented DF reduced the proportion of stillborn piglets from 8.8 to 6.6% ( < 0.001) and mortality of total born piglets from 22.3 to 19.9% ( = 0.004) but had no impact on preweaning mortality of the piglets ( = 0.21). Moreover, supplemented DF reduced the proportion of death due to poor viability ( < 0.001; 2.8 vs. 1.5% in the control and treatment groups, respectively) and prevalence of piglet diarrhea ( = 0.004; 0.7 vs. 0.3% in the control and treatment groups, respectively). Crushing, low birth weight, and poor viability were the top 3 contributors to preweaning mortality of live-born piglets, in descending order. In conclusion, the supplemented DF reduced the proportion of stillborn piglets and total piglet mortality as well as mortality due to poor viability and piglet diarrhea in lactation.

Cell turnover and activity in mammary tissue during lactation and the dry period in dairy cows.

Milk yield of the dairy cow follows a pattern termed the lactation curve. We have investigated the cellular background for this pattern. Seven mammary biopsies were obtained from each of 10 cows: at the end of lactation (d 347, equal to d 77 before next parturition); during the dry period at d 48 (4 d after dry off); 16 d before parturition; and during lactation at d 14, 42, 88, and 172. The fraction of proliferating (staining positive for Ki-67) alveolar cells was higher during the dry period (8.6%) than during lactation (0.5%). The fraction of apoptotic (staining positive by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling) alveolar cells was higher immediately after dry off (0.37%) and in early lactation (0.76%) than during other periods (0.15%). The enzyme activities of fatty acid synthetase, acetyl CoA-carboxylase, and galactosyl transferase were approximately 12-, 11-, and 4-fold higher, respectively, during lactation than during the dry period. In conclusion, mammary cell proliferation is substantial in a period near parturition but otherwise low, and apoptosis is elevated at dry off and in early lactation. The increase in apoptosis in early lactation may be due to discarding nonfunctional or senescent cells or to removal of a surplus of newly synthesized cells. The activity of selected enzymes central for milk synthesis is probably not limiting for milk production.

Changes in proteolytic enzyme mRNAs relevant for meat quality during myogenesis of primary porcine satellite cells.

The objective was to study the regulation of proteolytic enzyme mRNA's in porcine satellite cells during proliferation and differentiation. Beyond 80% confluence, cells were grown in absence or presence of 1µM insulin. The temporal changes in transcription of micro molar-, milli molar- and muscle specific calpains (p94), calpastatin and caspase 3 in response to insulin was evaluated and myogenin transcription and creatine kinase activity was determined to indicate differentiation. The housekeeping genes (GAPDH and ß-actin) were slightly affected by developmental stage and transiently by the insulin treatment but this did not affect the conclusions. The mRNA abundance of micro molar calpain, p94 and calpastatin increased from proliferation to differentiation. Milli molar calpain- and caspase 3-transcriptions were up-regulated in two steps, suggesting these two enzymes are involved in two distinct processes. Insulin stimulated differentiation as indicated by elevated creatine kinase activity but did not affect myogenin transcription. Insulin down-regulated milli molar calpain and calpastatin transcription and tended to down-regulate caspase 3 transcription but did not affect p94 or micro molar calpain. In conclusion, proteolytic enzymes relevant for post-mortem tenderisation are regulated at the gene level during myogenesis, indicating they are involved in muscle cell and muscle fibre development. Thus, a porcine satellite cell culture may be a model system to study regulation and relative contribution to proteolysis by the calpains.

Technical note: Measurement of mammary plasma flow in sows by downstream dilution of mammary vein infused -aminohippuric acid.

The objectives of the present study were to design a method to estimate mammary plasma flow (MPF) in lactating sows using downstream dilution of -aminohippuric acid (AH) and to compare these estimates with MPF estimates based on specific AA as internal markers (MPF-AA). A permanent indwelling catheter was surgically implanted in the femoral artery, and another 2 were inserted in the right cranial mammary vein of 8 second- and third-parity sows on d 76 ± 2 SEM of gestation. On the 3rd and 17th days in milk, arterial and venous blood samples were drawn in hourly intervals from 0.5 h before until 6.5 h after feeding. The MPF in the right cranial mammary vein was measured by downstream dilution of infused AH (3.0 mmol/h). Total MPF-AH was calculated assuming that the measured flow constituted the flow from 5 out of 14 suckled glands on the basis of the anatomical structure of the mammary vascular system. Total MPF-AA was estimated on the basis of the output of the specific AA marker in milk and the arteriovenous differences of the marker as free AA in plasma, assuming a direct transfer of AA from plasma to milk protein. Total MPF-AH was 6,860 L/d in early lactation and increased to 8,953 L/d at peak lactation ( = 0.003). In early lactation, MPF-AA estimates were greater or tended to be greater (132% to 175%; < 0.10) than MPF-AH estimates for all internal markers, except Met (119%). Moreover, MPF-AH was correlated with MPF-AA only for MET as an internal marker ( = 0.74; = 0.03) in early lactation. In contrast, MPF-AH and MPF-AA estimates did not differ and were well correlated at peak lactation with the strongest correlation observed when Met ( = 0.84; = 0.009) and Phe + Tyr ( = 0.82; = 0.01) were used as the internal AA markers. Litter gain increased from d 3 to 17 of lactation (2.13 vs. 3.46 g/d; = 0.001) and was correlated with MPF-AH during lactation ( = 0.74; < 0.001), whereas no correlation between litter gain and MPF-AA was observed ( > 0.10). These results suggest that downstream dilution of infused AH and the AA methods are applicable methods to estimate MPF at peak lactation. The reason for the observed discrepancy in early lactation between MPF- AH and MPF-AA is not obvious but might be related to the rapid metabolic changes observed in early lactation. In conclusion, MPF measured by downstream dilution of mammary infused AH was higher at peak compared to early lactation, which the internal AA marker approach failed to show.