Assessment of antioxidative and selenium status by seleno-dependent glutathione peroxidase activity in different blood fractions using a pig model: issues for clinical nutrition and research.

Blood seleno-dependent glutathione peroxidase (SeGPX) activity is widely used as a metabolic indicator of systemic antioxidative status despite inconsistent responses in the literature. This study aimed to compare SeGPX activity profiles in different blood fractions, expressed with different reference units, and assess their impact on interpretation of results. Two studies on selenium (Se) metabolism in gilts, including long-term and peri-oestrus SeGPX activity profiles, were submitted to analysis of variance with double repeated measures, after data set standardization. Differences between studies were experimental period (three post-pubertal oestrus or five post-pubertal oestrus +30 days of gestation) and sample type (whole blood or blood plasma). No difference was observed between whole-blood long-term profiles (three oestrus) for SeGPX activity/mg haemoglobin (SeGPXhb) vs. SeGPX activity/ml whole blood (SeGPXwb; p = 0.29). No long-term difference was observed in whole blood between profiles according to dietary Se provision (basal and dietary Se-supplemented groups; p ≥ 0.12). Blood plasma long-term profiles (five oestrus + 30 days gestation) for SeGPX/mg blood plasma protein (SeGPXpro) were different from SeGPX/ml blood plasma (SeGPXpla) according or not to Se provision (p ≤ 0.007 and p < 0.001 respectively). However, regardless of Se provision (p ≥ 0.80), when excluding gestation from the model, blood plasma profiles were similar. During the peri-oestrus period (day -4 to +3), regardless of Se provision, SeGPX activity profiles differed according to reference units in both studies (p < 0.001). However, considering Se provision, similar profiles were observed in whole blood and blood plasma (p ≥ 0.27) for basal Se groups, whereas in Se-supplemented groups they differed for both sample types (p ≤ 0.02). In conclusion, reference units influence interpretation of SeGPX activity according to physiological state. During oxidative stress periods, this effect depends upon dietary Se provision.

Gene expression of porcine blastocysts from gilts fed organic or inorganic selenium and pyridoxine.

In this study, we determined how maternal dietary supplementation with pyridoxine combined with different sources of selenium (Se) affected global gene expression of porcine expanded blastocysts (PEB) during pregnancy. Eighteen gilts were randomly assigned to one of the three experimental diets (n=6 per treatment): i) basal diet without supplemental Se or pyridoxine (CONT); ii) CONT+0.3 mg/kg of Na-selenite and 10 mg/kg of HCl-pyridoxine (MSeB610); and iii) CONT+0.3 mg/kg of Se-enriched yeast and 10 mg/kg of HCl-pyridoxine (OSeB610). All gilts were inseminated at their fifth post-pubertal estrus and killed 5 days later for embryo harvesting. A porcine embryo-specific microarray was used to detect differentially gene expression between MSeB610 vs CONT, OSeB610 vs CONT, and OSeB610 vs MSeB610. CONT gilts had lower whole blood Se and erythrocyte pyridoxal-5-P concentrations than supplemented gilts (P<0.05). No treatment effect was observed on blood plasma Se-glutathione peroxidase activity (P=0.57). There were 10, 247, and 96 differentially expressed genes for MSeB610 vs CONT, OSeB610 vs CONT, and OSeB610 vs MSeB610 respectively. No specific biological process was associated with MSeB610 vs CONT. However, for OSeB610 vs CONT, upregulated genes were related with global protein synthesis but not to selenoproteins. The stimulation of some genes related with monooxygenase and thioredoxin families was confirmed by quantitative real-time RT-PCR. In conclusion, OSeB610 affects PEB metabolism more markedly than MSeB610. Neither Se sources with pyridoxine influenced the Se-glutathione peroxidase metabolic pathway in the PEB, but OSeB610 selectively stimulated genes involved with antioxidant defense.

Effect of nutritional status on nutrient and gas utilization by the mammary gland of lactating sows.

Milk synthesis being a continuous process in lactating sows, the mammary gland has to adapt its metabolism in response to extreme short-term changes in nutrient availability in the arterial bloodstream, due to the feeding pattern. The objective of the present study was to better quantify and understand these adaptations. The effect of morning refeeding after an overnight 16-h feed withdrawal was measured on the uptake of energy-supplying nutrients, amino acids (AA), and some vitamins and minerals. After farrowing, catheters were fitted in the right anterior mammary vein and in the carotid artery of six sows. Blood samples were drawn on days 7, 14, and 21 of lactation, every 30 from 60 min before the morning meal to 300 min after the morning meal. Plasma concentrations of glucose, lactate, triglycerides (TG), non-esterified fatty acids (NEFA), glycerol, α-amino nitrogen (N), vitamins B12, and folates were determined on all samples. Riboflavin and AA concentrations were only measured 30 min before the meal and 120 min after the meal. Arterial and venous plasma concentrations of glucose, lactate, and α-amino N increased after the meal (P < 0.01), and concentrations of NEFA, glycerol, and TG decreased (P < 0.01). Mammary arteriovenous concentration difference increased after the meal for glucose, lactate, and α-amino N (P < 0.01), remained constant for TG, and decreased for NEFA (P < 0.01) and glycerol (P < 0.05). Arterial concentrations of all AA increased after the meal, but changes of arteriovenous difference with the meal differed among AA. Arteriovenous difference of energy (7.6 kJ/l plasma) concentration was similar in feed-deprived and fed sows, but the contribution of the various nutrients differed, and the respiratory quotient was lower (P < 0.01) before the meal (0.95) than after the meal (1.54). The relative contributions of glucose, lactate, TG, NEFA, and AA to arteriovenous difference in energy concentration were 50.2, 3.8, 25.1, 0, and 20.8% in fed and 24.6, 2.2, 24.9, 32.9, and 15.0% in feed-deprived sows, respectively. The daily mammary extraction of vitamin B12, estimated from arteriovenous differences was higher than the amount of this vitamin bioavailable from the diet, probably contributing to the 50% decrease in plasma concentration between day 7 and day 21 of lactation. For both riboflavin and folates, arteriovenous differences in plasma concentrations were small or not different from zero. These results indicate that the mammary gland has a great capacity to adapt nutrient uptake very rapidly and modify its metabolism according to the nutrients available in the bloodstream.

Effects of supplementary folic acid and vitamin B(12) on hepatic metabolism of dairy cows according to methionine supply.

The present experiment was undertaken to study the interactions between dietary supplements of rumen-protected methionine (RPM) and intramuscular injections of folic acid and vitamin B(12), given from 3 wk before calving to 16 wk of lactation, on hepatic metabolism of lactating dairy cows. Sixty multiparous Holstein cows were assigned to 10 blocks of 6 cows each according to their previous milk production. Within each block, 3 cows were fed a diet calculated to supply Met as 1.83% of metabolizable protein, whereas the 3 other cows were fed the same diet supplemented with 18g of RPM calculated to provide Met as 2.23% of metabolizable protein. Within each level of Met, the cows received no vitamin supplement or weekly intramuscular injections of 160mg of folic acid alone or combined with 10mg of vitamin B(12). Liver biopsies were taken at 2, 4, 8, and 16 wk of lactation. Liver concentrations of folates and vitamin B(12) were increased by their respective supplements but this response to vitamin supplements was altered by methionine supply. Concentrations of total lipids and triglycerides increased in livers of cows fed RPM, whereas concentrations of cholesterol ester, cholesterol, diglycerides, phosphatidylethanolamine, and phosphatidylcholine were not affected. Folic acid, alone or combined with vitamin B(12), tended to increase the ratio of phosphatidylcholine to phosphatidylethanolamine. Gene expression of 5,10-methylene-tetrahydrofolate reductase, microsomal transfer protein, and phosphatidylethanolamine methyltransferase were higher in liver of cows fed RPM supplements. The relative mRNA abundance of 5,10-methylene-tetrahydrofolate reductase and methylmalonyl-CoA mutase were increased by the combined injections of folic acid and vitamin B(12), whereas those of methionine synthase and methionine synthase reductase were not affected by treatments. These results suggest that increasing supply of methyl groups, as preformed labile methyl groups or through methylneogenesis, affected the methylation cycle but had a limited effect on dairy cow performance. The observed effects of the combined supplement of folic acid and vitamin B(12) on lactational performance of dairy cows probably result from an improvement of energy metabolism during early lactation.

Maternal perinatal transfer of vitamins and trace elements to piglets.

Nursing piglets are entirely dependent, for their micronutrient provisions, upon in utero, colostrum and milk transfers from the dam. An adequate maternal transfer of micronutrients is all the more important during these periods which, in fact, lasts for approximately half the life cycle (conception to slaughter) of modern pigs. The present study aimed to set up a simple approach to assess the maternal perinatal transfer of vitamins and trace elements in sows. Prenatal transfer (R-u) was estimated as limited, passive or active using the ratio between pre-colostral serum concentrations of a given micronutrient in newborn piglets and corresponding pre-farrowing values in sows. Efficiency of the postnatal transfer (R-c) was estimated from the ratio between serum concentrations of post- and pre-colostral micronutrients in piglets. Data from literature (12 studies) were used for vitamins A, D, E, C, folic acid and B12, whereas vitamins B2, B3, B6 and B8 as well as Zn, Fe, Cu and Se were generated from a trial where blood sera from 20 sows, and their litter were collected during the perinatal period. In sow trial, statistical t tests were used to determine if ratios differed from 1. Prenatal transfer was active and in favour of piglets (R-u > 1, P < 0.03) for Zn and vitamins B6 and B8 (sow trial) as well as for vitamins C and B12 (literature data). This transfer was limited (R-u < 1, P < 0.01) for vitamin B2, Fe, Cu and Se (sow trial) and for vitamins A, E, D and folic acid (literature data) whereas it was passive for vitamin B3 (R-u = 1, P > 0.37). After birth, the early postnatal transfer through colostrum was active towards piglets for most micronutrients but vitamins B6 and B8 (R-c < 1, P < 0.01). Globally, the perinatal transfer (combination of R-u and R-c) was favourable to the neonatal piglets for most micronutrients except for vitamins A and D as well as Fe, Cu and Se whereas there is apparently a barrier for prenatal transfer which is not compensated by the colostrum provision to neonatal piglets. Then, post-colostral concentrations of these micronutrients in piglets remain below prenatal levels of their dam. Neonatal strategies of micronutrient provision are known for Fe (intramuscular injection) and Se (sow milk enrichment). Further studies are needed to assess the importance of the unfavourable perinatal transfer for Cu and vitamins A and D for piglet robustness later in life.

Tissue-specific profiling reveals modulation of cellular and mitochondrial oxidative stress in normal- and low-birthweight piglets throughout the peri-weaning period.

Weaning is known to induce important nutritional and energetic stress in piglets. Low-birthweight (LBW) piglets, now frequently observed in swine production, are more likely to be affected. The weaning period is also associated with dysfunctional immune responses, uncontrolled inflammation and oxidative stress conditions that are recognized risk factors for infections and diseases. Mounting evidence indicates that mitochondria, the main cellular sources of energy in the form of adenosine 5' triphosphate (ATP) and primary sites of reactive oxygen species production, are related to immunity, inflammation and bacterial pathogenesis. However, no information is currently available regarding the link between mitochondrial energy production and oxidative stress in weaned piglets. The objective of this study was to characterize markers of cellular and mitochondrial energy metabolism and oxidative status in both normal-birthweight (NBW) and LBW piglets throughout the peri-weaning period. To conduct the study, 30 multiparous sows were inseminated and litters were standardized to 12 piglets. All the piglets were weighted at day 1 and 120 piglets were selected and assigned to 1 of 2 experimental groups: NBW (n = 60, mean weight of 1.73 ± 0.01 kg) and LBW piglets weighing less than 1.2 kg (n = 60, 1.01 ± 0.01 kg). Then, 10 piglets from each group were selected at 14, 21 (weaning), 23, 25, 29 and 35 days of age to collect plasma and organ (liver, intestine and kidney) samples. Analysis revealed that ATP concentrations were lower in liver of piglets after weaning than during lactation (P < 0.05) thus suggesting a significant impact of weaning stress on mitochondrial energy production. Oxidative damage to DNA (8-hydroxy-2'-deoxyguanosine, 8-OHdG) and proteins (carbonyls) measured in plasma increased after weaning and this coincides with a rise in enzymatic antioxidant activity of glutathione peroxidase (GPx) and superoxide dismutase (SOD) (P < 0.05). Mitochondrial activities of both GPx and SOD are also significantly higher (P < 0.05) in kidney of piglets after weaning. Additionally, oxidative damage to macromolecules is more important in LBW piglets as measured concentrations of 8-OHdG and protein carbonyls are significantly higher (P < 0.05) in plasma and liver samples, respectively, than for NBW piglets. These results provide novel information about the nature, intensity and duration of weaning stress by revealing that weaning induces mitochondrial dysfunction and cellular oxidative stress conditions which last for at least 2 weeks and more severely impact smaller piglets.

Influence of methionine supply on the response of lactational performance of dairy cows to supplementary folic acid and vitamin B12.

The present experiment was undertaken to determine if the effects of supplementary folic acid on lactational performance were caused by improved methylneogenesis and if the supply in vitamin B(12) could affect this metabolic pathway. In this eventuality, supplementary Met, a major source of preformed methyl groups, should reduce the requirements for these vitamins. Sixty multiparous Holstein cows were assigned to 10 blocks of 6 cows each according to their previous milk production. Within each block, 3 cows were fed a diet estimated to supply Met as 1.83% metabolizable protein and 3 cows were fed the same diet supplemented with 18 g of rumen-protected methionine (RPM) to supply Met as 2.23% of metabolizable protein. Within each level of Met, cows received no vitamin supplement or weekly intramuscular injections of 160 mg of folic acid alone or combined with 10 mg of vitamin B(12) from 3 wk before to 16 wk after calving. There was no treatment effect on dry matter intake during pre- and postcalving periods: 13.4 +/- 0.4 and 21.8 +/- 0.4 kg/d, respectively. Milk production was not affected by RPM supplementation. Folic acid and vitamin B(12) given together tended to increase milk production during the 16 wk of lactation. This effect was more pronounced during the first 4 wk of lactation: 37.5, 37.7, and 40.3 +/- 0.9 kg/d for cows receiving no vitamin supplement, folic acid alone, or folic acid combined with vitamin B(12), respectively. Milk fat yield was not affected by treatments. Lactose, crude protein, and total solid yields were greater, in early lactation, in cows injected with folic acid and vitamin B(12) together but this effect diminished as lactation progressed. Intramuscular injections of folic acid alone or combined with vitamin B(12) tended to decrease plasma concentrations of homocysteine from 5.51 microM with no vitamin supplement to 4.54 and 4.77 +/- 0.37 microM, respectively. Results of the present experiment suggest that the effects of the combined supplement of folic acid and vitamin B(12) on lactational performance of dairy cows were not due to an improvement in methyl groups supply, because RPM supplement, a source of preformed methyl groups, did not alter the cow responsiveness to vitamin supplements.

Effects of supplements of folic acid, vitamin B12, and rumen-protected methionine on whole body metabolism of methionine and glucose in lactating dairy cows.

The present experiment was undertaken to determine the effects of dietary supplements of rumen-protected methionine and intramuscular injections of folic acid and vitamin B(12), given 3 wk before to 16 wk after calving, on glucose and methionine metabolism of lactating dairy cows. Twenty-four multiparous Holstein cows were assigned to 6 blocks of 4 cows each according to their previous milk production. Within each block, 2 cows were fed a diet estimated to supply methionine as 1.83% metabolizable protein, equivalent to 76% of methionine requirement, whereas the 2 other cows were fed the same diet supplemented daily with 18 g of rumen-protected methionine. Within each diet, the cows were administrated either no vitamin supplement or weekly intramuscular injections of 160 mg of folic acid plus 10 mg of vitamin B(12.) To investigate metabolic changes at 12 wk of lactation, glucose and methionine kinetics were measured by isotope dilution using infusions of 3[U-(13)C]glucose, [(13)C]NaHCO(3) and 3[1-(13)C,(2)H(3)] methionine. Milk and plasma concentrations of folic acid and vitamin B(12) increased with vitamin injections. Supplementary B-vitamins increased milk production from 34.7 to 38.9 +/- 1.0 kg/d and increased milk lactose, protein, and total solids yields. Whole-body glucose flux tended to increase with vitamin supplementation with a similar quantitative magnitude as the milk lactose yield increase. Vitamin supplementation increased methionine utilization for protein synthesis through increased protein turnover when methionine was deficient and through decreased methionine oxidation when rumen-protected methionine was fed. Vitamin supplementation decreased plasma concentrations of homocysteine independently of rumen-protected methionine feeding, although no effect of vitamin supplementation was measured on methionine remethylation, but this could be due to the limitation of the technique used. Therefore, the effects of these B-vitamins on lactation performance were not mainly explained by methionine economy because of a more efficient methylneogenesis but were rather related to increased glucose availability and changes in methionine metabolism.

Effects of dietary vitamin supplementation and semen collection frequency on hormonal profile during ejaculation in the boar.

To evaluate the effect of dietary and management factors on boar hormonal status during ejaculation, 39 boars were canulated to determine the profiles of luteinizing hormone (LH), follicle-stimulating hormone (FSH), 17beta-estradiol (E2), and testosterone (T) in blood plasma and seminal fluid. Prior to canulation, 18 boars were fed a basal diet (control), whereas the remainder (n=21) were fed a basal diet supplemented with extra vitamins (supplemented). Within each dietary treatment, two regimens of semen collection were used over the 3mo preceding the hormonal evaluation: three times per 2wk (3/2) or three times per wk (3/1). Plasma E2 was lower (P<0.01) before ejaculation (232.5+/-22.6pg/mL) than at the onset of ejaculation (255.2+/-27.1ng/mL). Plasma T increased from 5.14+/-0.72, before ejaculation to 5.87+/-0.86ng/mL at the onset of ejaculation in supplemented boars, whereas it decreased from 5.15+/-0.65 to 4.87+/-0.70ng/mL in controls (diet by time, P<0.05). At the onset of ejaculation, plasma FSH was higher in 3/2 boars (0.436+/-0.06ng/mL) than in 3/1 boars (0.266+/-0.04ng/mL; P<0.05). During ejaculation, plasma LH increased linearly (P<0.01) from 0.59+/-0.07 to 0.97+/-0.10ng/mL, and plasma E2 and T concentrations were correlated (r=0.62, P<0.01). Plasma FSH before and during ejaculation was negatively correlated with sperm production (r=-0.60, P<0.01) and testicular weight (r=-0.50, P<0.01). In conclusion, dietary and management factors had few impacts on hormonal profiles during ejaculation, but homeostasis of some hormones was related to some criteria of reproductive performance in boars.

Effects of dietary supplements of folic acid and vitamin B12 on metabolism of dairy cows in early lactation.

The present experiment was undertaken to determine the effects of dietary supplements of folic acid and vitamin B12 given from 3 wk before to 8 wk after calving on lactational performance and metabolism of 24 multiparous Holstein cows assigned to 6 blocks of 4 cows each according to their previous milk production. Supplementary folic acid at 0 or 2.6 g/d and vitamin B12 at 0 or 0.5 g/d were used in a 2 x 2 factorial arrangement. Supplementary folic acid increased milk production from 38.0 +/- 0.9 to 41.4 +/- 1.0 kg/d and milk crude protein yield from 1.17 +/- 0.02 to 1.25 +/- 0.03 kg/d. It also increased plasma Gly, Ser, Thr, and total sulfur AA, decreased Asp, and tended to increase plasma Met. Supplementary B12 decreased milk urea N, plasma Ile, and Leu and tended to decrease Val but increased homocysteine, Cys, and total sulfur AA. Liver concentration of phospholipids was higher in cows fed supplementary B12. Plasma and liver concentrations of folates and B12 were increased by their respective supplements, but the increase in plasma folates and plasma and liver B12 was smaller for cows fed the 2 vitamins together. In cows fed folic acid supplements, supplementary B12 increased plasma glucose and alanine, tended to decrease plasma biotin, and decreased Km of the methylmalonyl-coenzyme A mutase in hepatic tissues following addition of deoxyadenosylcobalamin, whereas it had no effect when cows were not fed folic acid supplements. There was no treatment effect on plasma nonesterified fatty acids as well as specific activity and gene expression of Met synthase and methylmalonyl-coenzyme A mutase in the liver. Ingestion of folic acid supplements by cows fed no supplementary B12 increased total lipid and triacylglycerols in liver, whereas these supplements had no effect in cows supplemented with B12. The increases in milk and milk protein yields due to folic acid supplements did not seem to be dependent on the vitamin B12 supply. However, when vitamin B12 was given in combination with folic acid, utilization of the 2 vitamins seems to be increased, probably more so in extrahepatic tissues. Metabolic efficiency seems also to be improved as suggested by similar lactational performance and dry matter intake for cows fed supplementary folic acid but increased plasma glucose and decreased hepatic lipids in cows fed folic acid and vitamin B12 together.

Comparison of plasma tryptophan-related metabolites in crossbred Piétrain and Duroc pigs.

Besides being incorporated into proteins, Trp, an indispensable AA, is involved in numerous metabolic pathways. Previous data showed that Trp conversion into kynurenine (Kyn) and nicotinamide (Nam) differs among studies, and such differences cannot be explained by different dietary niacin supplies. We hypothesized that pig genotype influences Trp metabolism and thus the conversion of Trp into its metabolites. The objective of this study was to compare plasma appearance of Trp and related metabolites in 12 Duroc and 12 Piétrain crossbred postweaning pigs fed 2 contrasting dietary Trp levels. Within each genotype, 6 pigs were fed a basal (B-Trp: 17% and 15% standardized ileal digestible [SID] Trp:Lys for starter and prestarter diets) or supplemented (S-Trp: 24% and 23% SID Trp:Lys for starter and prestarter diets) Trp diet. Growth was monitored, and plasma fasted concentrations were measured over 4 wk, and then pigs were fitted with a jugular catheter for frequent blood samplings. After overnight fasting, 350 g of the experimental diets were offered to each pig, and plasma concentrations of Trp, Kyn, Nam, and 5-hydroxytryptamine (5-HT) were measured for 6 h. The activities of Trp-degrading enzymes were measured in different tissues collected after pig slaughtering. Plasma Trp fasted concentrations did not differ between B-Trp and S-Trp diets and increased from weaning to 2 and 4 wk after weaning for Piétrain but not for Duroc crossbred pigs (time × genotype, = 0.001). Plasma Kyn concentrations were greater 4 wk after weaning ( = 0.002) than at weaning and for Piétrain compared to Duroc genetics ( = 0.008). Plasma Nam concentrations were greater for pigs fed the S-Trp diet than for those fed the B-Trp diet ( = 0.0001) and for Duroc than for Piétrain genetic lines ( = 0.001); this difference tends to be greater at weaning than after ( = 0.055). Our data showed an increase in plasma concentrations of Trp, Kyn, Nam, and 5-HT according to time after a meal and to the dietary Trp content. However, postprandial plasma concentrations of Trp metabolites and enzyme activities were not significantly different between Duroc and Piétrain crossbred pigs. In conclusion, our results suggest that Nam endogenous synthesis capacity from Trp is greater in Duroc than in Piétrain crossbred pigs, but this was apparent only at weaning.

Effect of a post-weaning diet supplemented with functional feed additives on ileal transcriptome activity and serum cytokines in piglets challenged with lipopolysaccharide.

This study evaluated the potential of a weanling diet supplemented with trace minerals, vitamins, prebiotics, essential oils, antioxidants and bovine colostrum (BC) to modulate the inflammatory response of low-weight (LW) and high-weight (HW) piglets challenged with lipopolysaccharide (LPS). At weaning (20±1 d), litters from 32 sows were assigned to four groups: control diet (CTL), CTL plus dietary supplements (DS) or the antibiotic chlortetracycline (ATB), or DS plus BC in place of plasma proteins in the weanling diet (DS+BC). At 37 d (T0), two LW and two HW piglets were bled to evaluate ex vivo cytokine production by LPS activated peripheral blood mononuclear cells (PBMCs). In parallel, LW and HW piglets received intraperitoneal LPS and were bled at slaughter at 4h (T4) or 18h (T18) post-injection. Ileal tissues from these piglets and two unchallenged medium weight (MW) piglets per treatment were excised and analyzed by microarray. At T0, cytokine production of LPS-activated PBMCs was not affected by dietary treatments. At T4 after LPS challenge, serum concentrations of TNF-α, IL-6, IL-8, and IL-10 were increased in all piglets (P<0.01). Interestingly, the LW piglets had a higher TNF-α level than the HW piglets did (P=0.05). Dietary treatments had no effect on the piglet serum concentration of these cytokines neither at T4 nor at T18. Microarray data and QPCR analysis reveal that several genes were differentially expressed in the LPS-challenged piglets in comparison with the two control MW piglets (P<0.001). However, the dietary treatments had a slight effect on the ileal gene expression of the T4 and T18 LPS-challenged piglets when all piglets were included in the analysis. But when body weight (LW and HW) was considered as a fixed effect, the microarray analysis showed that the expression of 54 genes was differentially modulated by the dietary treatments in the T4 and T18 LPS-challenged LW piglets (P<0.05) while in HW piglets no difference was observed. QPCR analyses confirm that the level expression of several genes was reduced in LW piglets fed DS or DS+BC diet compared with ATB piglets. In conclusion, LPS challenge induced a transitional inflammation in weanling piglets that was characterized by increased blood-circulating cytokines and gut transcriptome activity. Results also suggest that the weanling diet supplemented with feed additives attenuated the ileal gene response to the LPS challenge, an effect that was more pronounced in the LW piglets.

Fate of supplementary B-vitamins in the gastrointestinal tract of dairy cows.

Four lactating Holstein cows equipped with ruminal, duodenal, and ileal cannulas were used in 2 studies to evaluate the disappearance of supplementary B-vitamins before and from the small intestine. The cows were fed a total mixed ration with chromic oxide in 12 daily meals. Each study consisted of a control (no vitamin supplementation) and a treatment period (with vitamin supplementation). Amounts of vitamins (mg/d) supplemented in studies 1 and 2, respectively, were: thiamin: 300 and 10; riboflavin: 1600 and 2.0; niacin: 12,000 and 600; vitamin B6: 800 and 34; biotin: 20 and 0.02; folic acid: 2600 and 111; vitamin B12: 500 and 0.4. In study 1, vitamins were added to the feed 5 d before and during the 4-d collection period. In study 2, vitamins were infused postruminally 1 d before and during the 4-d collection period. Substantial disappearance before the duodenal cannula was noted in study 1 (67.8% thiamin, 99.3% riboflavin, 98.5% nicotinamide, 41.0% pyridoxine, 45.2% biotin, 97.0% folic acid, and 62.9% vitamin B12). Except for nicotinamide and folate, there was almost no disappearance of postruminally infused vitamins before the duodenal cannula (study 2), suggesting extensive ruminal destruction or use. Apparent intestinal absorption values differed greatly among vitamins, but the proportion of vitamins disappearing from the small intestine was not negatively influenced by supplementation. Except for riboflavin and niacin, absolute amounts disappearing from the small intestine were greater during the treatment than the control periods, suggesting that B-vitamin supply in dairy cows is increased by supplementation, although losses in the rumen are extensive.

Homocysteine metabolism, growth performance, and immune responses in suckling and weanling piglets.

Homocysteine (Hcy), an intermediary sulfur AA, is recognized as a powerful prooxidant with deleterious effects on physiological and immune functions. In piglets, there is an acute 10-fold increase of plasma concentrations of homocysteine (pHcy) during the first 2 wk of life. This project aimed to maximize pHcy variations within physiological ranges using typical supplies of folates and vitamin B12 (B12) to sows and piglets. Growth, immune response, and Hcy metabolism of piglets were studied until piglets reached 56 d of age. Third-parity sows were randomly assigned to a 2 × 2 split-plot design with 2 dietary treatments during gestation and lactation, S(-) (1 mg/kg folates and 20 µg/kg B12, n = 15) and S(+) (10-fold S(-) levels, n = 16), and 2 treatments to piglets within each half litter, intramuscular injections (150 µg) of B12 (P(+)) at d 1 and 21 (weaning) and saline (P(-)). Within each litter of 12 piglets, 3 P(+) and 3 P(-) piglets were studied for growth and Hcy metabolism, and the others were studied for immune responses. During lactation, plasma B12 decreased and was transiently greater in S(+) vs. S(-) piglets on d 1 and P(+) vs. P(-) piglets on d 7 (sow treatment × age and piglet treatment × age; P < 0.05). From 14 to 21 d of age, pHcy was 33% lower in S(+)P(+) vs. S(-)P(-) piglets (sow treatment × piglet treatment interaction; P < 0.05). At 56 d of age, hepatic B12 was greater and pHcy was lower for P(+) vs. P(-) piglets (P < 0.05). No treatment effect was observed on growth except for a lower postweaning G:F in S(+)P(-) piglets than in others (sow treatment × piglet treatment interaction; P < 0.05). Positive correlations were observed between pHcy and growth (r > 0.29, P < 0.05) before and after weaning. Antibody responses to ovalbumin and serum tumor necrosis factor-α were not affected by treatments, but postweaning serum IL-8 peaked earlier in S(-)P(-) vs. S(+)P(+) piglets (piglet treatment × age; sow treatment × piglet treatment interaction, P < 0.05). Proliferation of lymphocytes in response to the mitogen concanavalin A tended to be lower in culture media supplemented with sera from S(-) vs. S(+) piglets (P = 0.081) and P(-) vs. P(+) piglets (P = 0.098), and the reduction of response was more marked (P < 0.05) with high (>21 µM) compared to medium (17 to 21 µM) or low (<17 µM) pHcy. In conclusion, the present vitamin supplements to sows and/or piglets produced variations of pHcy that were not apparently harmful for growth performance of piglets. The greater pHcy, particularly prevalent in S(-) and/or P(-) piglets, had negative effects on some indicators of immune responses, suggesting that these young animals may be immunologically more fragile.

Forestomach motility and behavior of bull calves according to changes in regimen.

Six 23-week-old bull calves were randomly assigned to a crossover design to study the effects of hay and concentrates given separately (A), silage and concentrates given in complete mixed ration (B), or given separately (C) on reticulo-ruminal motility and eating and resting behaviors after an adaptation period of 14 days (day 14). The effects of a change from one treatment to another on the same variables were also investigated (day 1). The differences observed between day 1 and day 14 indicate that a change in regimen, even not drastic, seems to affect reticulo-ruminal motility, rumination pattern, ruminal inactivity, and resting behavior. As expected, the number of triphasic and biphasic reticular spike bursts, as well as eating and resting behaviors, were affected by the type of forage ingested (A vs. C) on day 14 (p < or = 0.05). Moreover, the mode of distribution of concentrates affected also the variables measured (p < or = 0.05). Although the length of particles in complete mixed ration (B) was the same than in silage and concentrates given separately (C), the effect of regimen B on many variables was similar to that of regimen A or intermediate between A and C. The effect of regimens B and C was similar only for the rumination pattern, even though the number of boluses regurgitated for rumination differed (p < or = 0.05).

Age-related changes in secretion rate and post-secretory metabolism of growth hormone in swine.

The effect of age on growth hormone (GH) metabolism and GH-releasing factor (GRF)-induced GH concentrations were studied in 7 young (3 mo, 39 kg) and 7 old (30 mo, 156 kg) Yorkshire x Landrace female pigs. Jugular catheters were surgically inserted and 60 hr later total serum volume was determined. The following day, all animals were infused for 3 hr with GH (30.3 ng.min/kg B.W.) in order to calculate GH metabolic clearance rate (MCR), secretion rate (SR) and half-life (t 1/2). Two days later, 15 micrograms/kg of GRF was injected i.v. into all pigs. On a per animal basis, aging increased (P < .01) MCR (299 vs 132 ml/min), SR (714 vs 422 ng/min) and serum volume (6.6 vs 2.01), whereas t1/2 was unaltered (P > .1). Basal GH concentrations were lower in older pigs (P < .10) but the GRF-induced GH concentrations (measured as GH peak or area under the curve, AUC) were not affected by age (P > .1). Yet, when induced total GH secretion (AUC x MCR) and average total serum GH (mean GH post-injection x serum volume) were calculated per pig, these variables significantly increased between 3 and 30 mo of age. Basal IGF-I concentrations were lower in older pigs (P < .01), yet, there was a tendency (P = .10) for these pigs to show a greater IGF-I response to GH infusion. The present data therefore indicate that age alters both SR and post-secretory metabolism of GH.(ABSTRACT TRUNCATED AT 250 WORDS)

Effect of oestrous cycle and early pregnancy on uterine production and expression of immune regulatory factors in gilts.

This study was undertaken to characterize uterine immune factors involved in the establishment of pregnancy in gilts. Thirty crossbred Yorkshire-Landrace gilts of similar age and weight were observed twice a day for oestrous behaviour with intact boars. On the day of first standing oestrus (Day 0) and 12h later, 15 gilts were inseminated with pooled semen from Duroc boars of proven fertility. Pregnant gilts were slaughtered either on Days 10, 15 or 25 of gestation (n=5 per day). The other 15 gilts were not inseminated and were slaughtered on either Days 0, 10 or 15 of the oestrous cycle (n=5 per day). Immediately after slaughter, endometrial tissue samples from the mesometrial side were removed for gene expression using RNase protection assay and in situ hybridization methodologies. The other uterine horn was flushed with 20 ml of PBS to collect the uterine fluid. In pregnant gilts, endometrial interleukin (IL)-6 mRNA expression was higher on Day 15 than on Days 10 and 25 (P<0.01 and P<0.1, respectively). On Day 15, IL-6 expression was also significantly higher (P<0.01) in pregnant gilts than in cyclic gilts. In both pregnant and cyclic gilts, transforming growth factor (TGF)-beta2 in uterine fluid was significantly higher (P<0.0001) on Day 15 than on Day 10. At the gene expression level, TGF-beta2 also increased between Days 10 and 15 in both cyclic and pregnant gilts but differences were not significant. On Day 15, concentrations of interferon-gamma and prostaglandin E(2) (PGE(2)) in uterine fluid were markedly higher (P<0.001) in pregnant gilts than in cyclic gilts, whereas the total amount of TGF-beta2 in uterine fluid and its endometrial expression were approximately 70% higher although this increase was not significant. Finally, tumour-necrosis factor-alpha and granulocyte-macrophage/colony-stimulating factor mRNA expressions were undetectable in all endometrial samples. In conclusion, production and/or expression of uterine TGF-beta2, IL-6 and PGE(2) increased during the embryonic attachment period and are coincidental with embryonic interferon-gamma production.

Effects of intramuscular injections of folic acid during lactation on folates in serum and milk and performance of sows and piglets.

In order to determine the effect of folic acid on serum and milk folates in lactating sows as well as on serum folates and growth rate of the piglets, sows (n = 25) received either saline or 15 mg folic acid i.m. each week from d 2 after parturition to weaning, 26 d later. Blood samples were drawn from all sows at 110 d of gestation and every week during lactation. Milk samples were taken at d 7 and 21 of lactation. Piglets were weighed and blood samples were collected weekly during lactation. Serum folates of sows increased during lactation. The rate of increase was more pronounced (P less than .0002) after folic acid injections. Milk folates concentrations decreased (P less than .0007) from d 7 to 21 of lactation but were higher (P less than .0001) in treated sows (11.8 +/- .7 ng/ml) than in control sows (7.9 +/- .4 ng/ml). Serum folates of piglets in control litters increased from 55.0 +/- 2.2 ng/ml at 2 d of age to a peak value of 86.3 +/- 3.1 ng/ml 2 wk later, and then gradually decreased. In piglets from treated dams, the time response curve was similar to that of the controls, but values were about 15% higher (P less than .01). The growth rate of piglets until 8 wk of age was not changed (P greater than .47) by folic acid injection of sows. More studies are needed to evaluate the practical importance of changes in folates status in establishing the folic acid requirements of lactating sows.

An estimation of the requirement for folic acid in gestating sows: the metabolic utilization of folates as a criterion of measurement.

Sows at their second parity were randomly distributed in five groups of seven animals each to determine the dietary concentration of folic acid that optimizes the metabolic utilization of the vitamin during gestation. The groups differed by dietary supplement of folic acid: 0, 5, 10, 15, or 20 ppm. Sows were fed 2.5 kg of diet each day. The response of serum folates and folate binding capacity to treatments and the excretion of urinary folates after an i.v. injection of folic acid were measured. The total daily excretion of urinary folates was corrected according to the response to one i.v. injection of saline on the day preceding the i.v. injection of folic acid. The decrease of total serum folates throughout gestation was less pronounced in the groups fed 15 and 20 ppm of dietary folic acid (supplement x period interaction, P<.06) than it was in the other three treatments. The proportion of i.v. folic acid not recovered in sow urine (injected - excreted) decreased as the amount of dietary folic acid increased to reach a minimum, which differed according to the period (supplement x period interaction, P<.02); it was 15 ppm during wk 1 of gestation and 10 ppm for the other periods studied. The unrecovered folates increased over a dietary concentration of 15 ppm. These minimum values correspond to the most appropriate feed concentration that covered the whole body utilization (tissue and cell metabolism, catabolism, and storage) of folates by the sows and could be interpreted as a reliable index of the requirement.

Responses of serum folates of preruminant and ruminant calves to a dietary supplement of folic acid.

The effect of dietary supplemental folic acid on serum folates of preruminant and ruminant calves was studied. In Trial 1, doses of 0, .07, .14, .28, and .56 mg of folic acid per kilogram of BW were added to the milk of preruminant calves. In Trial 2, doses of 0, .5, 1, 2, and 4 mg of folic acid per kilogram of BW were incorporated into the concentrates of ruminant heifers. In the first part of each trial, serum folates were determined in blood samples taken 0, 1, 2, 4, 8, 16 (both trials), and 32 h (Trial 2) after a single meal supplemented with folic acid. In the second part of the two trials, the supplement of folic acid was given in feed during seven consecutive days. Blood samples were taken the day before the trial and subsequently every day during 7 d. In preruminant and ruminant calves, the area under the curve and the peak of concentration of serum folates after a meal increased with the dose ingested (P less than or equal to .05, linear and quadratic effect of doses, respectively) but the amount of folic acid needed to obtain a similar response was lower for preruminant than for ruminant calves. In preruminants, the time to reach the maximal concentration was 3 to 4 h after the meal, whatever the dose ingested (P less than or equal to .05), whereas in ruminants this time decreased with the dose ingested (quadratic effect of treatment, P less than or equal to .05).(ABSTRACT TRUNCATED AT 250 WORDS)