Longitudinal Data to Assess Relationships among Plasma Folate, Vitamin B12, Non-esterified Fatty Acid, and ß-Hydroxybutyrate Concentrations of Holstein Cows during the Transition Period.

It is well established that the plasma metabolite profile changes during metabolic dysfunction, such as elevated non-esterified fatty acid (NEFA) release when body reserve mobilization is excessive in early lactation cows. Relationships between changes in plasma concentrations of metabolites caused by a metabolic impairment and the status of vitamins, such as folates and vitamin B12, have barely been studied in cattle. This study was undertaken to assess relationships between peripartum plasma concentrations of folates, vitamin B12, NEFA, and beta-hydroxybutyrate (BHB). Longitudinal data of 48 multiparous Holstein cows from 5 studies were taken from days -14 to 21 relative to calving. Blood samples were taken weekly before calving and either twice or thrice per week postpartum, and plasma was analyzed for folate, vitamin B12, NEFA, and BHB concentrations. Postpartum plasma NEFA and BHB concentrations were negatively related to plasma folate concentration at days -14 and -7 relative to parturition, whereas the opposite relationship was noted for the plasma vitamin B12:folate ratio. The plasma folate and NEFA areas under the curve from the whole studied period were negatively associated, and the opposite was observed with the association between the plasma vitamin B12:folate ratio and NEFA as well as the BHB areas under the curve. The results suggest that there is an increased use of folate for metabolic functions during elevated concentrations of plasma NEFA and BHB. Future research should focus on finding an optimal plasma vitamin B12:folate ratio to favor cow health during the challenging period of parturition.

Effect of dietary supplementation of 2 forms of a B vitamin and choline blend on the performance of Holstein calves during the transition and postweaning phase.

The transition from a liquid- to a solid-based diet involves several adaptations in calves. Development of ruminal function is likely to alter B vitamin and choline supply, although little is known about the extent of these changes relative to the calf's requirements and consequences for the calf around weaning. Moreover, literature data are equivocal concerning the need to supplement B vitamins and choline through weaning and transition phase of the dairy calf. To evaluate the effect of increasing B vitamin and choline supply on performance, 61 Holstein calves were individually housed and raised from birth to 13 wk of age. Calves were fed milk replacer (28% crude protein, 15% fat) up to 1.6 kg of dry matter (DM)/d at 15% solids (3 times/d) from birth to 4 wk of age. At that time, calves were randomly assigned to one of 3 treatments: a rumen-protected blend of B vitamins and choline (RPBV); a 30:70 mix of a nonprotected blend of B vitamins and choline and fat (UPBV); or fat only, used as control (CTRL). Calves were maintained on milk replacer and offered ad libitum quantities of a starter grain (25.5% crude protein) specifically formulated to supply all essential amino acids with no added B vitamins or choline. The supplements were provided in gel capsules and administered once a day to each calf in quantities corresponding to 0.39 and 0.28% of the previous day's starter DM intake for the vitamin blends and control, respectively. Calves were weaned gradually from d 49 to 63. Body weight and stature were measured, and blood was collected and analyzed for hematocrit, plasma urea nitrogen, ß-hydroxybutyrate, folates, and vitamin B12. Body weight and stature were similar among treatments. Overall gain (0.99 kg/d), DM intake (1.90 kg of DM/d), and feed efficiency (0.52) were not affected by vitamin supplementation. Plasma vitamin B12 concentrations were not different between RPBV and UPBV but tended to be higher at the end of weaning and were greater postweaning in RPBV and UPBV treatments compared with CTRL. Both forms of the vitamin blend effectively improved vitamin B12 status postweaning with no effect on folate status. No differences were observed in other blood measurements. Under conditions of this study, additional B vitamins and choline did not improve calf performance before, during, or after weaning.

Comparison of the Potential Abilities of Three Spectroscopy Methods: Near-Infrared, Mid-Infrared, and Molecular Fluorescence, to Predict Carotenoid, Vitamin and Fatty Acid Contents in Cow Milk.

The objective of this work is to compare the ability of three spectroscopy techniques: molecular fluorescence, near-infrared (NIR), and mid-infrared with attenuated total reflectance (MIR-ATR) spectroscopy to predict the concentrations of 8 carotenoids, 6 vitamins and 22 fatty acids (FA) in cow's milk. A dataset was built through the analysis of 242 frozen milk samples from different experiments. The milk compounds were analysed using reference methods and by NIR, MIR-ATR, and fluorescence to establish different predictive models. NIR spectroscopy allowed for better prediction of cis9-ß-carotene, ß-cryptoxanthin and the sum of carotenoids than the other techniques, with a coefficient of cross-validation in calibration (R2CV) > 0.60 and a coefficient of determination in validation (R2V) > 0.50. Their standard errors of prediction (SEP) were equal to 0.01, except for the sum of carotenoids (SEP = 0.15). However, MIR-ATR and fluorescence seem usable for the prediction of lutein and all-trans-ß-carotene, respectively. These three spectroscopy methods did not allow us to predict (R2CV < 0.30) vitamin contents except, for vitamin A (the best R²CV = 0.65 with NIR and SEP = 0.15) and α-tocopherol (the best R²CV = 0.56 with MIR-ATR and SEP = 0.41), but all R²V were <0.30. NIR spectroscopy yielded the best prediction of the selected milk FA.

Correlations between the Composition of the Bovine Microbiota and Vitamin B12 Abundance.

Vitamin B12 is synthesized by prokaryotes in the rumens of dairy cows-and this has implications in human nutrition since humans rely on consumption of dairy products for vitamin B12 acquisition. However, the concentration of vitamin B12 in milk is highly variable, and there is interest in determining what causes vitamin B12 variability. We collected 92 temporally linked rumen, fecal, blood, and milk sample sets from Holstein cows at various stages of lactation fitted with rumen cannula and attempted to define which bacterial genera correlated well with vitamin B12 abundance. The level of vitamin B12 present in each sample was measured, and the bacterial population of each rumen, fecal, and milk sample (n = 263) was analyzed by 16S rRNA-targeted amplicon sequencing of the V4 region. The bacterial populations present in the rumen, small intestine, and milk were highly dissimilar. Combined diet and lactation status had significant effects on the composition of the microbiota in the rumen and in the feces. A high ruminal concentration of vitamin B12 was correlated with the increased abundance of Prevotella, while a low ruminal concentration of vitamin B12 was correlated with increased abundance of Bacteroidetes, Ruminiclostridium, and Butyrivibrio The ultimate concentration of vitamin B12 is controlled by the complex interaction of several factors, including the composition of the microbiota. Bacterial consumption of vitamin B12 in the rumen may be more important in determining overall levels than bacterial production.IMPORTANCE In this paper, we examined the microbiome of the bovine rumen, feces, and milk and attempted to understand how the bacterial communities at each site affected the production and movement of vitamin B12 around the animal's body. It was determined that the composition of the bovine rumen microbiome correlates well with vitamin B12 concentration, indicating that the rumen microbiota may be a good target for manipulation to improve production of this important vitamin.

FGF21 underlies a hormetic response to metabolic stress in methylmalonic acidemia.

Methylmalonic acidemia (MMA), an organic acidemia characterized by metabolic instability and multiorgan complications, is most frequently caused by mutations in methylmalonyl-CoA mutase (MUT). To define the metabolic adaptations in MMA in acute and chronic settings, we studied a mouse model generated by transgenic expression of Mut in the muscle. Mut-/-;TgINS-MCK-Mut mice accurately replicate the hepatorenal mitochondriopathy and growth failure seen in severely affected patients and were used to characterize the response to fasting. The hepatic transcriptome in MMA mice was characterized by the chronic activation of stress-related pathways and an aberrant fasting response when compared with controls. A key metabolic regulator, Fgf21, emerged as a significantly dysregulated transcript in mice and was subsequently studied in a large patient cohort. The concentration of plasma FGF21 in MMA patients correlated with disease subtype, growth indices, and markers of mitochondrial dysfunction but was not affected by renal disease. Restoration of liver Mut activity, by transgenesis and liver-directed gene therapy in mice or liver transplantation in patients, drastically reduced plasma FGF21 and was associated with improved outcomes. Our studies identify mitocellular hormesis as a hepatic adaptation to metabolic stress in MMA and define FGF21 as a highly predictive disease biomarker.

Bioavailability of Vitamin B12 from Dairy Products Using a Pig Model.

The present study compares the bioavailability of vitamin B12 (B12) of dairy products or synthetic B12, using the pig as an experimental model for humans. Eleven pigs were used in a cross-over design to assess the net portal drained viscera (PDV) flux of blood plasma B12 after ingestion of tofu (TF; devoid of B12), Swiss cheese (SC), Cheddar cheese (CC), yogurt (YG), and synthetic B12 (TB12; TF supplemented with cyanocobalamin), providing a total of 25 µg of B12 each. PDV blood plasma flow for SC and CC were higher than for TF and TB12 (p ≤ 0.04) whereas YG was higher than TF (p = 0.05). Porto-arterial difference of blood plasma B12 concentrations were higher for CC and TB12 than for TF and YG (p ≤ 0.04) but not different from SC (p ≥ 0.15). Net PDV flux of B12 was only different from zero for CC. However, the net PDV flux of B12 for CC was not different from SC or TB12. Cumulative net PDV flux of B12 for SC, TB12, and CC were 2.9, 4.4, and 8.3 µg 23 h post-meal, corresponding to a bioavailability of 11.6%, 17.5%, and 33.0%, respectively. In conclusion, CC had the best bioavailability of B12 among the tested dairy products or compared to synthetic B12.

Intestinal fate of dietary zinc and copper: Postprandial net fluxes of these trace elements in portal vein of pigs.

In pig, the assessment of bioavailability of dietary trace minerals with classical approaches such as relative bioavailability estimates or digestive tract balances have often generated inconsistent responses. In the present study, net portal-drained-viscera fluxes were monitored after a meal to assess intestinal absorption of zinc (Zn) or copper (Cu) according to dietary sources and levels of these trace minerals. Twelve pigs were surgically equipped with portal and carotid catheters and a portal ultrasonic flow probe for 12-h postprandial measurements. In a cross-over design, pigs received boluses of inorganic (I) or organic (O) dietary Cu and Zn at adequate (A, 20 and 200mg, respectively) or high (H, 40 and 400mg, respectively) level just before a 0.8-kg meal (semi-purified diet). Whatever treatments, arterial Zn increased by 72% at 45min postprandial and gradually declined thereafter (P<0.01). Arterial Zn were greater by 11% after O than I (P=0.02) and by 19% after H than A (P<0.01) meals. Net portal-drained-viscera fluxes of Zn during the first 240min postprandial were greater by 44% after O than I (P=0.10) and by 51% after H than A (P=0.07) meals. For Cu, portal-drained-viscera fluxes of Cu up to 240min postprandial were greater (P=0.03) after A than H meals. Those results suggest that Zn is absorbed rapidly, likely in the upper digestive tract of pigs and, whatever dietary levels, more efficiently after O meals. It appears that H levels of both Zn and Cu interfered with intestinal absorption of Cu and/or stimulate post-absorption enterocyte sequestration of this mineral.

Supplements of vitamins B9 and B12 affect hepatic and mammary gland gene expression profiles in lactating dairy cows.

BACKGROUND: A combined supplement of vitamins B9 and B12 was reported to increase milk and milk component yields of dairy cows without effect on feed intake. The present study was undertaken to verify whether this supplementation positively modifies the pathways involved in milk and milk component synthesis. Thus, by studying the transcriptome activity in these tissues, the effect of supplements of both vitamins on the metabolism of both liver and mammary gland, was investigated. For this study, 24 multiparous Holstein dairy cows were assigned to 6 blocks of 4 animals each according to previous 305-day milk production. Within each block, cows were randomly assigned to weekly intramuscular injections of 5 mL of either saline 0.9 % NaCl, 320 mg of vitamin B9, 10 mg of vitamin B12 or a combination of both vitamins (B9 + B12). The experimental period began 3 weeks before the expected calving date and lasted 9 weeks of lactation. Liver and mammary biopsies were performed on lactating dairy cows 64 ± 3 days after calving. Samples from both tissues were analyzed by microarray and qPCR to identify genes differentially expressed in hepatic and mammary tissues. RESULTS: Microarray analysis identified 47 genes in hepatic tissue and 16 genes in the mammary gland whose expression was modified by the vitamin supplements. Gene ontology (GO) categorizes genes in non-overlapping domains of molecular biology. Panther is one of the online GO resources used for gene function classification. It classifies the 63 genes according to Molecular Function, Biological Process and Protein Class. Most of the biological processes modulated by the vitamin supplements were associated to developmental process, protein metabolic process, transport and response to inflammation. In the liver, most of the genes modulated by the vitamin treatments involved protein metabolic process while developmental process appeared to be more affected by the treatments in mammary gland. Out of 25 genes analysed by qPCR, 7 were validated. CONCLUSION: The results indicate that several metabolic processes were modulated by the supplementation of vitamins in early-lactating dairy cows. In addition, the results suggest that the vitamin supplements promoted liver regeneration and reduced catabolism of lipids in early lactation.

Oral supplementations of betaine, choline, creatine and vitamin B6 and their influence on the development of homocysteinaemia in neonatal piglets.

Homocysteine (Hcy) is an intermediary sulphur amino acid recognised for pro-oxidative properties in several species which may weaken immune competence in piglets. In this species, there is an acute 10-fold increase of concentrations of plasma Hcy (pHcy) during the first 2 weeks of life. The present experiment aimed to determine if pHcy in piglets can be regulated by oral supplementations of betaine as a methyl group supplier, creatine for reducing the demand for methyl groups, choline with both previous functions and vitamin B6 as enzymic co-factor for Hcy catabolism. A total of seventeen sows (second parity) were fed gestation and lactation diets supplemented with folic acid (10 mg/kg) and vitamin B12 (150 µg/kg). Eight piglets in each litter received daily one of the eight following oral treatments (mg/kg body weight): (1) control (saline); (2) betaine (50); (3) choline (70); (4) creatine (300); (5) pyridoxine (0·2); (6) treatments 2 and 5; (7) treatments 3 and 4; and (8) treatments 2, 3, 4 and 5. According to age, pHcy increased sharply from 2·48 µm at birth to 17·96 µm at 21 d of age (P < 0·01). Concentrations of pHcy tended to be lower (P = 0·09) in treated than in control piglets but the highest and sole pairwise significant decrease (23 %) was observed between treatments 1 and 8 (P = 0·03). Growth from birth to 21 d of age was not influenced by treatments (P > 0·70). Therefore, it appears possible to reduce pHcy concentrations in suckling piglets but a combination of all chosen nutrients is required.

Optimization and validation of a reversed-phase high performance liquid chromatography method for the measurement of bovine liver methylmalonyl-coenzyme a mutase activity.

BACKGROUND: Methylmalonyl-CoA mutase (MCM) is an adenosylcobalamin-dependent enzyme that catalyses the interconversion of (2R)-methylmalonyl-CoA to succinyl-CoA. In humans, a deficit in activity of MCM, due to an impairment of intracellular formation of adenosylcobalamin and methylcobalamin results in a wide spectrum of clinical manifestations ranging from moderate to fatal. Consequently, MCM is the subject of abundant literature. However, there is a lack of consensus on the reliable method to monitor its activity. This metabolic pathway is highly solicited in ruminants because it is essential for the utilization of propionate formed during ruminal fermentation. In lactating dairy cows, propionate is the major substrate for glucose formation. In present study, a reversed-phase high performance liquid chromatography (RP-HPLC) was optimized and validated to evaluate MCM activity in bovine liver. The major aim of the study was to describe the conditions to optimize reproducibility of the method and to determine stability of the enzyme and its product during storage and processing of samples. RESULTS: Specificity of the method was good, as there was no interfering peak from liver extract at the retention times corresponding to methylmalonyl-CoA or succinyl-CoA. Repeatability of the method was improved as compared to previous RP-HPLC published data. Using 66 µg of protein, intra-assay coefficient of variation (CV) of specific activities, ranged from 0.90 to 8.05% and the CV inter-day was 7.40%. Storage and processing conditions (frozen homogenate of fresh tissue vs. fresh homogenate of tissue snapped in liquid nitrogen) did not alter the enzyme activity. The analyte was also stable in liver crude extract for three frozen/thawed cycles when stored at -20°C and thawed to room temperature. CONCLUSIONS: The improved method provides a way for studying the effects of stages of lactation, diet composition, and physiology in cattle on MCM activity over long periods of time, such as a complete lactation period. Interestingly, this sensitive and accurate method could benefit the study of the cobalamin status in experimental studies and clinical cases.

Bioavailability of vitamin B12 in cows' milk.

The natural source of vitamin B12 in human diets comes from animal products. For example, one glass (250 ml) of milk provides approximately 50 % of the RDA (2·4 µg/d). It was hypothesised that the provision of vitamin B12 from milk is more efficiently absorbed than the synthetic form used in vitamin supplements. Pigs (n 10) were used as a model for intestinal absorption of vitamin B12 in humans to compare the net fluxes of vitamin B12 across the portal-drained viscera (PDV; an indicator of intestinal absorption) after ingestion of meals complemented with conventional and vitamin B12-enriched (via injections to cows) milk (raw, pasteurised or microfiltrated) or with equivalent amounts of cyanocobalamin, the synthetic form used in supplements or unsupplemented. Net flux of vitamin B12 across PDV after the ingestion of milk was positive, though not influenced by milk enrichment (P>0·3) or technological processes (P = 0·8) and was greater than after ingestion of equivalent amounts of cyanocobalamin (cyanocobalamin v. all milk, P ≤ 0·003). In fact, net fluxes of this vitamin were not different from 0 after either cyanocobalamin or the meal devoid of vitamin B12 (unsupplemented v. cyanocobalamin, P = 0·7). The cumulative PDV fluxes during the 24 h following ingestion of meals complemented with milk varied from 5·5 to 6·8 µg. These values correspond to an efficiency of intestinal absorption of vitamin B12 from milk varying between 8 and 10 %. Therefore, vitamin B12, which is abundant in cows' milk, is also substantially more available than the most commonly used synthetic form of this vitamin.

Identification of cobalamin and cobalamin analogues along the gastrointestinal tract of dairy cows.

Cobalamin analogues in samples of rumen fluid, duodenal and ileal digesta and faeces of lactating dairy cows collected during the course of two nutritional studies were identified using a recently developed liquid chromatography-mass spectrometry method in order to determine if changes in diet composition could alter their proportions. Only cobalamin and cobinamine were detected in feedstuffs; six supplementary analogues were detected in the gastrointestinal tract content. The proportion of analogues produced by the gastrointestinal microflora was higher than the vitamin itself. The mode of conservation of forage had no effect on the proportion of analogues in rumen fluid, duodenal digesta and faeces, whereas increasing metabolisable protein supply raised the proportion of analogues reaching the duodenum (p = 0.008). The proportion of analogues was higher in ileal than duodenal digesta, this observation probably indicates a preferential absorption of cobalamin in the small intestine. The present results showed that changes in diet composition could alter cobalamin synthesis in rumen.

Methionine, folic acid and vitamin B12 in growing-finishing pigs: impact on growth performance and meat quality.

Growth performance, metabolic variables, and meat quality were measured in 78 growing-finishing pigs using supplements of 0 (C), or 0.2% of DL-methionine (M), and three combinations of folic acid [mg/kg] and cyanocobalamin [microg/kg], respectively 0 and 0 (V0), 10 and 25 (V1), and 10 and 150 (V2) in a 2 x 3 factorial arrangement. Feed conversion was lower (p = 0.05) in M than in C pigs during the growing period (0-4 weeks). Both V1 and V2 treatments increased plasma vitamin B12 (p < 0.01) and decreased plasma homocysteine (p < 0.01). Plasma 5-methyl-tetrahydrofolates were the lowest, highest and intermediate in V0, V1 and V2 pigs (p < 0.04), respectively. In V2 meat, folates were 32% higher, vitamin B12, 55% higher and homocysteine, 28% lower than in V0 (p < 0.01). Oxidative stability of the fresh meat was similar among treatments during a storage period of 42 days. Therefore, methionine supplements improved growth performance during the growing period. Vitamin supplements interacted with the methionine cycle pathway, increased vitamin content of pork meat but did not improve oxidative stability of the fresh meat during storage.

Calculations of apparent ruminal synthesis and intestinal absorption of biotin in dairy cows as influenced by the extraction method.

Biotin is present in nature either free or as biocytin, which is only degraded under the action of a specific enzyme: biotinidase. This enzyme is not included in analytical assays generally used. A method for sample preparation using biotinidase was developed in our laboratory before analysis by ELISA. Three cows equipped with duodenal and ileal cannulae were used to compare the effects of methods of sample preparation on calculations of apparent ruminal synthesis and intestinal absorption of biotin. There was no apparent ruminal synthesis of biotin, no matter whether free or total biotin was measured (p = 0.84). Results also suggested that rumen microbes cannot utilize nor degrade biocytin present in the feed. Estimates of apparent intestinal absorption were influenced by the sample preparation method (p = 0.002). Analysis of free biotin caused an artefact, suggesting intestinal synthesis of this vitamin; whereas determination of total biotin concentrations showed that absorption was taking place in the small intestine.

Influence of maturation and cooking treatments on the nutritional value of bovine meats: Water losses and vitamin B12.

The aim of the study was to determine the influence of maturation and of cooking processes on water losses and on the vitamin B12 content of meat. Three types of muscle (Longissimus lumborum, Longissimus thoracis and Triceps brachii) were sampled from a total of 16 animals, representative of animals raised for meat production in France. Three durations of maturation were compared: 1, 3 and 14 days. Different cooking processes were applied: Longissimus lumborum was deep-fat fried or roasted, Longissimus thoracis was pan fried or grilled and Triceps brachii was braised. The cooking yield averaged 55-56% for Triceps brachii, 73-77% for Longissimus lumborum and 85-87% for Longissimus thoracis. Vitamin B12 concentration in raw meat was significantly higher in Triceps brachii than in Longissimus lumborum and Longissimus thoracis (20.86, 11.53 and 9.21ng/g wet tissue, in the same respective order). When expressed on a wet weight basis, all concentrations were significantly increased by cooking. When expressed on a lipid-free dry basis, significant losses in vitamin B12 were measured only in the braised Triceps brachii (-25%) and in the deep-fat fried Longissimus lumborum (-5.5%) as a result of long duration and high temperature of cooking, respectively. Maturation did not affect the vitamin B12 content of meat, whether raw or cooked.

Nutritional value of meat: the influence of nutrition and physical activity on vitamin B12 concentrations in ruminant tissues.

An important nutritional characteristic of ruminant meat is its high content in vitamin B12. The variability of these contents is not known. Three studies were been set up in order to test the influence of the animal species (2 studies on Charolais steers slaughtered at 30-32 months of age, n = 24 and n = 30 and a third one on lambs slaughtered at 4.5 months of age, n = 21), of the nature of the diet (grass vs. maize silage, lucerne or concentrate diets) and of physical activity (without or with walking) on the vitamin B12 contents of different muscle types (rather oxidative (Rectus Abdominis, RA), intermediate (Longissimus Dorsi, LD), or glycolytic (Semi Tendinosus, ST)) and on the liver. The animals were supplemented in macro and trace minerals according to usual feeding practices in France in order to theoretically avoid any risk of deficiency. For this reason, cobalt allowances, which are necessary for the ruminal synthesis of vitamin B12, could differ among treatments. The results indicate the following: (1) cobalt allowances varied widely among treatments, from (sub-)deficient to plethoric allowances, influencing vitamin B12 contents of the liver, and muscles (only in case of deficiency), (2) the effects of dietary treatments or of physical exercise were essentially related to differences in cobalt allowances, (3) the oxidative type muscle (RA) showed contents which were double those in glycolytic type muscle (RA 10.8 vs. ST 5.0 ng.g(-1)) and (4) vitamin B12 contents of raw muscles were lower than the values indicated in tables of feed composition for humans for cooked meat (0.5 to 1 vs. 2 to 3 microg.100 g(-1)).

Effect of folic acid plus glycine supplement on uterine prostaglandin and endometrial granulocyte-macrophage colony-stimulating factor expression during early pregnancy in pigs.

The objective was to determine the effects of folic acid+glycine supplement on uterine metabolism of prostaglandin and mRNA expression of endometrial granulocyte-macrophage colony-stimulating factor (GM-CSF) in nulliparous (NYL) and multiparous Yorkshire-Landrace (YL) sows, and in multiparous Meishan-Landrace sows (ML). In each of these three groups, sows were randomly assigned to two treatments: 15 ppm folic acid+0.6% glycine or no supplement. The dietary supplement was given from the estrus before mating to slaughter on Day 25 of pregnancy. At slaughter, endometrial tissue was collected to determine endometrial expression levels of GM-CSF mRNA, cyclooxygenase-1 (COX1) and -2 (COX2) and to evaluate in vitro endometrial secretion of prostaglandin E2 (PGE2) secretion. Allantoic fluid samples were also collected to determine the concentration of PGE2, prostaglandin F2alpha (PGF2alpha), estradiol-17beta (E2), progesterone (P4), and transforming-growth factor-beta2 (TGF-beta2). The allantoic contents of PGF2alpha, E2 and P4, and endometrial in vitro secretion of PGE2 were not significantly influenced by the folic acid+glycine supplement. The folic acid+glycine supplement tended (P<0.07) to increase allantoic content of PGE2 and TGF-beta2 in all sows and increased (P<0.05) endometrial expression of COX2, especially in NYL sows. The endometrial expression of COX1 was decreased (P<0.05) by folic acid+glycine supplement, especially in multiparous YL sows. The allantoic contents of PGE2 and PGF2alpha were not significantly affected by sow type. However, NYL sows had higher (P<0.05) endometrial in vitro secretion of PGE2 and allantoic content of P4 than multiparous YL and ML sows. The allantoic content of E2 was also higher (P<0.05) in NYL sows than in multiparous ML sows only. The allantoic content of TGF-beta2 was lower (P<0.05) in multiparous ML than in multiparous YL only sows. Finally, in YL and NYL sows, folic acid+glycine supplement decreased (P<0.05) the endometrial expression of GM-CSF but not in ML sows. In summary, folic acid+glycine supplement altered endometrial expression of GM-CSF and uterine metabolism of prostaglandins during the post-attachment period of porcine embryos but some of these effects were manifest only in Meishan and nulliparous sows.

Effects of folic acid and vitamin B12 supplements on folate and homocysteine metabolism in pigs during early pregnancy.

The present experiment aimed to determine the effects of supplements of folic acid (FA) alone or in combination with vitamin B12 on folate and homocysteine metabolism in gestating nulliparous Yorkshire-Landrace (YL) and multiparous Landrace (LD) occidental sows and multiparous Chinese Meishan-Landrace (ML) sows. LD sows were randomly assigned to two treatments: 0 or 15 mg FA/kg diet while YL and ML sows were assigned to three treatments: 0 mg FA/kg diet, 15 mg FA/kg or 15 mg vitamin B12/kg diet. Supplements were given from the oestrus preceding insemination up to slaughter on day 15 of gestation. At slaughter, a uterine flush was collected to determine uterine contents of homocysteine, methionine, tetrahydrofolate (THF), 5-methyl-THF, pyridoxal 5-phosphate (P5P) and vitamin B12. Blood samples were taken at first oestrus, at insemination and on days 5, 10 and 15 of gestation to determine plasma concentrations of homocysteine, methionine, THF, 5-methyl-THF, P5P, vitamin B12 and relative total folate-binding capacity. In occidental sows (YL and LD), the FA supplement tended to decrease uterine flush content of homocysteine (P=0.06) and concentrations of plasma homocysteine (P=0.09). Nulliparous YL sows had lower concentrations of plasma homocysteine, methionine, THF and 5-methyl-THF (P<0.05) than multiparous LD sows. Multiparous ML and LD sows had similar concentrations of plasma THF, 5-methyl-THF, methionine and vitamin B12, but ML sows had lower concentrations of plasma homocysteine (P<0.05). The vitamin B12 supplement increased concentrations of plasma vitamin B12 (P<0.05) both in multiparous ML and nulliparous YL sows, but had no effect on the composition of either uterine flush or plasma. The present results showed also that sows had a low vitamin B12 status (<200 pg/ml) and high circulating homocysteine levels (>15 microm) during the first 15 d of gestation. Furthermore, the vitamin B12 content in uterine secretions represented between 180 and 300 % of the total content in plasma. The low plasma concentrations of homocysteine in multiparous ML sows suggest a more efficient remethylation pathway which may not be dependent upon dietary supply of FA or vitamin B12. In nulliparous YL sows, low concentrations of both homocysteine and methionine suggest that the methionine requirement for protein deposition might have reduced the amount of methionine available for the methylation pathway. The results of the present experiment suggest that the reduction of uterine homocysteine may be an important aspect of the role of FA supplement on the uterine environment in occidental sows. The presence of high levels of vitamin B12 in uterine secretions merits further investigation in relation to embryonic development.