Milk yield, milk composition, and hepatic lipid metabolism in transition dairy cows fed flaxseed or linola.

The response of transition dairy cows to dietary supplementation with fat sources of various fatty acid profiles could affect hepatic fat metabolism differently. Twenty-eight Holstein cows were blocked for similar calving date 4wk before expected parturition to compare the effects of feeding sources of n-3 and n-6 fatty acids on milk production and composition, plasma metabolites, and liver parameters. Cows within each block were assigned to 1 of 3 isonitrogenous and isoenergetic diets: control with a source of calcium salts of palm oil (MEG; 1.1 and 2.6% of the dry matter in prepartum and postpartum diets, respectively); n-3 fatty acids supplied as whole flaxseed (WFL; 4.8 and 7.7% of the dry matter in prepartum and postpartum diets, respectively); and n-6 fatty acids supplied as whole linola (WLO; 4.8 and 7.7% of the dry matter in prepartum and postpartum diets, respectively). Diets were fed until wk 14 of lactation. Contrasts of WFL versus WLO and polyunsaturated fatty acids versus MEG were compared. Cows fed polyunsaturated fatty acids increased dry matter intake over time at a greater extent than those fed MEG, which resulted in enhanced energy balance. Cows fed MEG produced more milk compared with those fed polyunsaturated fatty acids, and there was no difference between those fed WFL and WLO. We found no effect on body condition score and body weight. Plasma concentrations of glucose, fatty acids, and BHB were similar among diets. There was no effect of diet on concentration of glycogen and activities of superoxide dismutase and glutathione peroxidase in the liver. We observed higher concentrations of hepatic lipids and triacylglycerol in cows fed MEG compared with those fed polyunsaturated fatty acids, and no difference between WFL and WLO. Hepatic catalase activity tended to be higher on wk 4 after calving for cows supplemented with WFL compared with those fed WLO. Feeding linoleic and linolenic acids as unprotected oilseeds increased dry matter intake over time at a greater extent for cows fed MEG, improved the energy status, and lowered hepatic lipids and triacylglycerol contents, which may contribute to enhance the health status of transition dairy cows.

Oral administration of domperidone in the first or third week of lactation: Effects on prolactin concentrations and mammary gene expression in sows and piglet growth.

The scope of the present study is endocrine and metabolic control of sow lactation. This project aimed to determine the impact of increasing prolactin concentrations via oral administration of the dopamine receptor antagonist domperidone in the first or third week of lactation in sows. Effects on sow hormonal and metabolic status, lactational performance, and gene expression in mammary epithelial cells were determined. Primiparous sows were divided in 3 treatments: 1) 10 mL of vehicle (table syrup) per os twice daily during the first and third weeks of lactation (Control, CTL, n = 23), 2) 0.5 mg/kg of domperidone per os twice daily during the first week of lactation (LACT1, n = 23), or 3) 0.5 mg/kg of domperidone given per os twice daily during the third week of lactation (LACT3, n = 22). Treated sows also received 10 mL of the vehicle twice daily during the other treatment period. Litter size was standardized to 12 ± 1 and piglets were weighed at birth, 24 h, and on d 8, 15, 22 (weaning), 35, and 56. Sow feed intake was recorded daily. Representative milk samples were obtained on d 7 and 21 of lactation for compositional analyses, and milk fat globules were used to measure mRNA abundances of various genes. Jugular blood samples were obtained from sows on d 1, 7, 14, and 21 of lactation to measure concentrations of prolactin, IGF-1, insulin, urea, and FFA. Concentrations of prolactin were increased (P < 0.01) at the end of the 7-d treatment period with domperidone, whether imposed in the first (LACT1) or third (LACT 3) week of lactation. No other blood variables were affected by treatments and neither was milk composition (P > 0.10). Sow BW, backfat thickness, or feed intake were not altered by treatments (P > 0.10), but piglet BW tended to be greater in litters from LACT3 compared with CTL sows on d 22 and 35 (P ≤ 0.10). Gene expression of EGF in milk fat globules tended to be (LACT1, P < 0.10) or was increased (LACT3, P < 0.05) after treatment, and the effect in LACT1 sows was maintained until d 21 of lactation. The mRNA abundance of SPP1 was increased (P < 0.05) in LACT1 vs CTL sows on d 7, and that of 3 major milk proteins tended to be (CSN1S2 and WAP, P < 0.10) or was greater (LALBA, P < 0.05) in LACT3 vs CTL sows on d 21 of lactation. Oral administration of domperidone during the first or third week of lactation increased prolactin concentrations and altered mRNA abundances of selected genes in milk fat globules. Yet, only the LACT 3 treatment positively affected piglet performance.

Hyperprolactinemia using domperidone in prepubertal gilts: Effects on hormonal status, mammary development and mammary and pituitary gene expression.

Objectives of this experiment were to determine if the domperidone protocol previously used for gestating gilts can also lead to hyperprolactinemia in growing gilts, and to assess the effects of such a protocol on hormonal status, mammary development and gene expression in mammary and pituitary tissue of gilts at puberty. The impact on future lactation performance was also determined. At 75 ± 3 kg body weight (BW), gilts were divided between: 1) controls (CTL), receiving daily intramuscular (IM) injections of canola oil (1.1 mL) for 29 d (n = 41), and 2) treated (DOMP), receiving daily IM injections with 0.5 mg/kg BW of the dopamine receptor antagonist domperidone for 29 d (n = 40). In addition to that daily injection, treated gilts also received twice daily IM injections with 0.5 mg/kg BW of domperidone over the first 3 d of treatment. Fifteen gilts per treatment were sacrificed at 210 ± 5 d of age to collect mammary glands (for compositional analysis and gene expression) and the anterior pituitary (for gene expression). Remaining gilts were bred and allowed to farrow. Blood was sampled at the onset of treatment and on days 14 and 30. Gilts that farrowed were also blood sampled on days 3 and 20 of lactation. Blood was assayed for prolactin (PRL), leptin, insulin-like growth factor 1 (IGF-1), urea, free fatty acids and glucose. Concentrations of PRL increased after 14 d and 30 d of treatment (P < 0.01) and were lesser on day 3 of lactation in DOMP than CTL gilts (P < 0.01). At puberty, there were tendencies (P < 0.10) for total parenchymal protein and DNA to be greater in DOMP than CTL gilts. Treatment did not affect mRNA abundance of PRL or the long form of the PRL receptor genes in the pituitary gland at puberty but expression level of the dopamine receptor D2 and PRL genes was much lower in pubertal than late-pregnant gilts (P < 0.001). Furthermore, many genes related with PRL had a much greater expression level in late pregnancy than at puberty. On day 20 of lactation, CTL sows had greater concentrations of urea than DOMP sows (P < 0.01). The growth rate of litters was not affected by treatment nor was milk composition (P > 0.10). Even though PRL concentrations were increased with treatment, the absence of effect on mammary development was either due to timing relative to developmental stage, whereby treatment was initiated when gilts were too young, or was because all PRL receptors may have been saturated thereby preventing biological action of additional PRL.

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.

Effects of sustained hyperprolactinemia in late gestation on mammary development of gilts.

The objective of this project was to determine the effects of sustained hyperprolactinemia for 7 or 20 d on mammary development in late-pregnant gilts. On day 90 of gestation, gilts were assigned to one of 3 groups to receive intramuscular (IM) injections of (1) canola oil (CTL, n = 18) until day 109 ± 1 of gestation; (2) a dopamine receptor antagonist, domperidone (0.5 mg/kg of body weight [BW]) until day 96 ± 1 of gestation (T7, n = 17); or (3) domperidone (0.5 mg/kg BW) until day 109 ± 1 of gestation (T20, n = 17). Domperidone-treated gilts also received 100 mg of domperidone per os twice daily from days 90 to 93 of gestation. Blood was sampled on days 89, 97, 104, and 110 for prolactin (PRL), insulin-like growth factor 1 (IGF1), lactose, urea, and glucose assays. Mammary glands were collected at necropsy, on day 110, for compositional and cell proliferation analyses. Abundance of mRNA for selected genes was also determined in the mammary gland and the pituitary gland. On day 97 of gestation, PRL concentrations were 3 times greater for T20 and T7 than CTL gilts and were also greater for T20 than T7 and CTL gilts on days 104 and 110 (P < 0.001). Concentrations of IGF1 in T20 and T7 gilts were elevated relative to controls on days 97 and 104 and were greater for T20 vs T7 and CTL gilts on day 110 (P < 0.05). There were no treatment effects (P > 0.1) on parenchymal or extraparenchymal tissue weights, or on epithelial proliferation as measured by immunohistochemistry for Ki-67. Treatments did not alter concentrations of dry matter (DM), fat, or DNA (P > 0.1) in parenchyma. Concentrations of RNA (P < 0.05) and protein (P < 0.10) as well as total parenchymal protein, RNA, and DNA (P < 0.05) were lower, or tended to be, in T20 than T7 or CTL gilts. Hyperprolactinemia for 20 d in late gestation increased mRNA abundance of the milk protein genes beta-casein (CSN2) and whey acidic protein (WAP) (P < 0.05) in mammary parenchyma and also decreased mRNA abundance of the long form of the prolactin receptor (PRLR-LF). Increasing PRL concentrations for 7 or 20 d in late gestation had no beneficial effects on the composition of the mammary gland, and sustained exposure to domperidone for 20 d reduced metabolic activity either by a lower expression of the long form of the PRL receptor in mammary parenchymal tissue or, most likely, by the early involution of parenchymal tissue. In conclusion, results do not support the hypothesis that a sustained hyperprolactinemia in late gestation could enhance mammary development of gilts.

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.

The 'beneficial' adipokines in reproduction and fertility.

The objective of this study was to review the available information on the signaling proteins produced by adipose tissue in the context of their role in regulating reproductive processes, including ovarian and uterine function. It is well known that both obesity and excessive leanness are associated with reproductive dysfunction. Adipokines are cytokines predominantely or exclusively expressed by adipose tissue that circulate and affect target tissues. Four known adipokines, adiponectin, visfatin/PBEF, omentin and vaspin, all increase tissue sensitivity to insulin, and are thus described as 'beneficial'. There is strong support for a role for adiponectin in the function of the ovary and placenta. There is evidence for direct effects of this adipokine on the late stages of folliculogenesis, and additive interactions of adiponectin with insulin and gonadotropins in inducing periovulatory changes in ovarian follicles. In addition, clinical and genomic studies associate hypoadiponectinemia with obesity-related reproductive disorders, including the polycystic ovarian syndrome. The roles for visfatin/PBEF, omentin and vaspin in reproduction remain to be established. The conclusion thus drawn is that the expression of insulin-sensitizing adipokines varies with adipose abundance. These adipokines have demonstrated both the potential effects on ovarian function and the possible effects on the formation of the placenta, acting through multiple mechanisms.

Evaluation of molecular methods used for establishing the interactions and functions of microorganisms in anaerobic bioreactors.

Molecular techniques have unveiled the complexity of the microbial consortium in anaerobic bioreactors and revealed the presence of several uncultivated species. This paper presents a review of the panoply of classical and recent molecular approaches and multivariate analyses that have been, or might be used to establish the interactions and functions of these anaerobic microorganisms. Most of the molecular approaches used so far are based on the analysis of small subunit ribosomal RNA but recent studies also use quantification of functional gene expressions. There are now several studies that have developed quantitative real-time PCR assays to investigate methanogens. With a view to improving the stability and performance of bioreactors, monitoring with molecular methods is also discussed. Advances in metagenomics and proteomics will lead to the development of promising lab-on chip technologies for cost-effective monitoring.

Characterization of swine adiponectin and adiponectin receptor polymorphisms and their association with reproductive traits.

In this study, polymorphisms in genes encoding porcine adiponectin (ADIPOQ) and its receptors (ADIPOR1 and ADIPOR2) were evaluated for associations with reproductive traits in a Landrace sow population. Sixteen SNPs were identified, and among these, associations were found between reproductive traits and five SNPs. Heterozygous multiparous females for SNP ADIPOQEF601160:c.178G>A had fewer stillborn piglets (P < 0.05) and shorter weaning-to-oestrus intervals (P < 0.05). Multiparous females bearing the mutant allele for SNP ADIPOQEF601160:c.*1094_1095insC gave birth to fewer stillborn piglets (P < 0.05). In addition, selection for the ADIPOQ [A;C] haplotype is expected to result in multiparous sows having the lowest number of stillborn piglets and shorter weaning-to-oestrus intervals. In second-parity sows, the polymorphism in ADIPOR1 (AY856513:c.*129A>C) showed significant associations with live-born (P < 0.01) and stillborn (P < 0.05) piglets. In multiparous sows, a significant association was observed for an ADIPOR2 polymorphism (AY856514:c.*112G>A), with the c.*112GA genotype associated with shorter weaning-to-oestrus intervals (P < 0.01). Haplotype analyses of ADIPOR2 SNPs revealed that selection in favour of the [A;C] haplotype and against the [G;G] haplotype may result in sows having an increased number of live-born piglets and shorter weaning-to-oestrus intervals. We have therefore described specific SNPs and haplotypes that are associated with large litter size, fewer stillborn and mummified piglets and shorter weaning-to-oestrus intervals. Selection for these SNPs and haplotypes is a strategy to improve reproductive success in pigs.

Hepatic lipid metabolism in transition dairy cows fed flaxseed.

Thirty-three Holstein cows averaging 687 kg of body weight were allotted 6 wk before the expected date of parturition to 11 groups of 3 cows blocked within parity for similar calving dates to determine the effects of feeding different sources of fatty acids on blood parameters related to fatty liver and profile of fatty acids in plasma and liver. Cows were fed lipid supplements from 6 wk before the expected date of parturition until d 28 of lactation. Cows within each block were assigned to 1 of 3 isonitrogenous and isoenergetic dietary supplements: control with no added lipids (CO); unsaturated lipids supplied as whole flaxseed (FL; 3.3 and 11.0% of the dry matter in prepartum and postpartum diets, respectively); and saturated lipids supplied as Energy Booster (EB; 1.7 and 3.5% of the DM in prepartum and postpartum diets, respectively). Diets EB and FL had similar ether extract concentrations. Multiparous cows fed EB had lower dry matter intake and milk production, higher concentrations of nonesterified fatty acids and beta-hydroxybutyrate in plasma and triglycerides (TG) and total lipids in liver, and lower concentrations of plasma glucose and liver glycogen than those fed FL and CO. Production of 4% fat-corrected milk was similar among treatments. Multiparous cows fed FL had the highest liver concentrations of glycogen on wk 2 and 4 after calving and lowest concentrations of TG on wk 4 after calving. Liver C16:0 relative percentages in multiparous cows increased after calving whereas those of C18:0 decreased. Relative percentages of liver C16:0 were higher in wk 2 and 4 postpartum for multiparous cows fed EB compared with those fed CO and FL; those of C18:0 were lower in wk 4 postpartum for cows fed EB compared with those fed CO and FL. Liver C18:1 relative percentages of multiparous cows increased after calving and were higher in wk 4 for cows fed EB compared with those fed CO and FL. The inverse was observed for liver C18:2 relative percentages. In general, diets had more significant effects on plasma concentrations of nonesterified fatty acids, beta-hydroxybutyrate, and glucose and liver profiles of fatty acids, TG, total lipids, and glycogen of multiparous than primiparous cows. These data suggest that feeding a source of saturated fatty acids increased the risk of fatty liver in the transition cow compared with feeding no lipids or whole flaxseed. Feeding flaxseed compared with no lipids or a source of saturated fatty acids from 6 wk before calving could be a useful strategy to increase liver concentrations of glycogen and decrease liver concentrations of TG after calving, which may prevent the development of fatty liver in the transition dairy cow.

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.

TRIENNIAL LACTATION SYMPOSIUM/BOLFA: Adipokines affect mammary growth and function in farm animals.

The essential role of mammary fat pads in mammary growth and morphogenesis was the first indication that biologically active molecules, secreted from adipocytes or other stromal cells, could regulate endocrine cues for growth and function of the mammary gland. The presence of leptin and adiponectin receptors in mammary tissues suggested that locally produced or circulating adipokines could affect mammary growth and function. Herein, we present the current knowledge on the role of adipokines in mammary cell proliferation and differentiation and in lactogenesis and galactopoiesis in farm animals. We also address the role of milk adipokines in the neonate. Accumulating evidence suggests that adipokines could act as metabolic sensors, regulating mammary growth and function in periods of metabolic adaptations such as late pregnancy and early lactation. Indeed, different experiments reported that adiponectin and leptin expression varies according to physiological stages and nutritional status of the animal. The current review also demonstrates that adipokines, such as leptin and adiponectin, are important regulators of the action of lactogenic hormones in the mammary gland. Findings also suggest important roles for adipokines in growth and intestinal maturation of the neonate.

Body condition of gilts at the end of gestation affects their mammary development.

The impact of body condition at 110 d of gestation on mammary gland development, mammary gene expression, and hormonal and metabolite status of gilts was studied. Thirty-nine gilts were equally divided into 3 groups based on their backfat thickness at the end of gestation: 1) low backfat (LBF; 12-15 mm), 2) medium backfat (MBF; 17-19 mm), or 3) high backfat (HBF; 21-26 mm). Gilts had similar BW (138.1 ± 8.2 kg) and backfat thicknesses (16.4 ± 1.0 mm) at mating and the 3 groups were achieved via ingestion of varying amounts of feed throughout gestation. Jugular blood samples were obtained from all gilts at mating and at 109 d of gestation to assess hormonal and metabolic statuses, and animals were slaughtered on d 110 to collect mammary glands for compositional analyses and for measure of gene expression. The LBF gilts had less extraparenchymal tissue ( < 0.01) and parenchymal tissue ( < 0.05) than HBF gilts. Mammary parenchyma from LBF gilts also tended to contain less DM ( < 0.10), contained more protein ( < 0.05), and had greater RNA concentrations ( < 0.01) than that from HBF gilts. None of the 15 genes studied in mammary parenchymal tissue differed in terms of expression level, and the rate of mammary cell proliferation was similar among treatments ( > 0.10). There was a tendency for circulating leptin concentrations on d 109 of gestation to be lower in LBF gilts than in MBF gilts ( < 0.10), whereas values for HBF gilts did not differ from those of the other treatments ( > 0.10). Current results demonstrate that being too thin at the end of gestation (12-15 mm backfat) has a negative impact on mammary development in gilts, whereas having backfats varying from 17 to 26 mm seems to have no detrimental effects on mammogenesis. Backfat thickness in late pregnancy must therefore be considered to achieve optimal sow lactation performance.

Differences in body condition of gilts that are maintained from mating to the end of gestation affect mammary development.

The goal of this project was to determine if different body conditions in late gestation that are due to varying body conditions at mating affect mammary development and mammary gene expression of gilts. Gilts that were fed ad libitum in the growing period were selected based on their backfat depths to form 3 groups at mating, namely, low backfat (LBF; 12-15 mm; = 14), medium backfat (MBF; 17-19 mm; = 15), and high backfat (HBF; 22-26 mm; = 16). During gestation, LBF, MBF, and HBF gilts were fed approximately 1.25, 1.43, and 1.63 times maintenance requirements to maintain their differences in body condition. Feed intake was increased by 1 kg in the last 10 d of gestation. Backfat depths of gilts were ultrasonically measured at mating and on d 30, 50, 70, 100, and 109 of gestation. Blood samples were obtained at mating and on d 109 of gestation to measure concentrations of IGF-1, glucose, insulin, estradiol, urea, free fatty acids, leptin, and adiponectin. Gilts were slaughtered on d 110 of gestation to collect mammary glands for compositional analyses. Mammary extraparenchymal tissue weight was lesser in LBF and MBF gilts than in HBF gilts (1,259.3, 1,402.7, and 1,951.5 ± 70.4 g, respectively; < 0.01). The weight of parenchymal tissue was not affected by treatment ( > 0.10), but its composition was altered. Concentrations of DNA and RNA decreased as backfat depth increased ( < 0.05), whereas percent fat and DM increased ( < 0.05). Circulating concentrations of leptin tended to be lower at mating ( < 0.10) and were lower on d 109 of gestation ( < 0.05) in LBF gilts than in HBF gilts. On d 109 of gestation, concentrations of insulin ( < 0.01) and IGF-1 ( < 0.05) were lower in LBF and MBF gilts than in HBF gilts, whereas those of urea were greater ( < 0.05). The mRNA abundance in parenchymal tissue for all genes studied was not affected by treatments ( > 0.10) with the exception of , which had a greater expression level in LBF gilts than in MFB or HBF gilts ( < 0.05). Percent of Ki-67-positive cells, used to assess mammary cell proliferation rate, was greater in HBF gilts than in LBF gilts ( < 0.05). When differences in body conditions of gilts that were present at mating were maintained throughout gestation, it had an impact on mammary development. Extraparenchymal tissue mass was affected and, more importantly, composition of parenchymal tissue was altered, indicating a beneficial effect of gilts being in the thinner treatment groups at mating.

Delayed muscle development in small pig fetuses around birth cannot be rectified by maternal early feed restriction and subsequent overfeeding during gestation.

Intrauterine variations in nutrient allowance can alter body composition and tissue features of the porcine offspring around birth. This study aimed to determine the effects of fetal weight variations between littermates and of maternal dietary regimen during gestation on fetal muscle traits just before birth. Fourteen pregnant gilts were reared under a conventional (control, CTL; n=7) or an experimental (treatment, TRT; n=7) dietary regimen during gestation. The dietary treatment provided 70% of the protein and digestible energy contents of the CTL diet during the first 70 days of gestation and then, 115% of the protein and digestible energy contents up to farrowing. At 110 days of gestation, sows were sacrificed and one fetus having a low (824±140 g) and one having a normal (1218±192 g) BW per litter were sampled. Irrespective of maternal dietary regimen, the longissimus muscle of the small fetuses exhibited higher expression levels of DLK1/Pref1 and NCAM1/CD56, two genes known to be downregulated during normal skeletal muscle development. Expression levels of the embryonic isoform of the myosin heavy chain (MyHC), both at the mRNA and at the protein levels, were also higher in small fetuses. In addition, the ratios of perinatal to embryonic and of adult fast to developmental MyHC isoforms were generally lower in light fetuses compared with their medium-weight littermates. These modifications suggest a delayed myofiber development in spontaneous growth-retarded fetuses. Finally, GLUT1 was expressed to a lesser extent in the muscle of small v. normal fetuses, suggesting decreased ability for glucose uptake in muscle. Initial feed restriction and subsequent overfeeding of sows during gestation led to a lower expression of the myogenic factor MYOD1, a prerequisite for myogenic initiation in skeletal muscle. This maternal strategy was also associated with a lower expression level of insulin-like growth factor 1 receptor (IGFR) but an upregulation of IGF2. This suggests an altered susceptibility of muscle cells to IGFs' signal in fetuses from treated sows. Altogether, intrauterine growth restriction impaired fetal muscle development, and restricted feeding followed by overfeeding of gestating sows did not allow small fetuses to recover normal contractile and metabolic characteristics.

Mammary gland development and hormone levels in pregnant Upton-Meishan and large white gilts.

Genetic differences between Upton-Meishan (UM, n = 13) and Large White (LW, n = 14) gilts were studied with regard to mammary gland development and concentrations of hormones. Gilts were weighed and their backfat measured at mating, and at d 70 and 109 of gestation. Jugular blood samples were also collected at these times and assayed for prolactin, cortisol, IGF-I, insulin, glucose, progesterone, and estradiol. Gilts were slaughtered on d 110 of gestation. One row of mammary glands was used for dissection and biochemical analyses. The other row was used for determination of prolactin receptor number and affinity. UM gilts weighed less (P<0.05) and had more backfat (P<0.01) than LW gilts at all times. Parenchymal tissue weight was less (P<0.05) in UM gilts. Percent fat (P<0.001) and dry matter (P<0.001) in parenchymal tissue were greater in UM gilts while that of protein (P<0.001) was lower. Total protein weight in parenchyma was also lower in parenchyma was also lower in UM gilts (P = 0.01). Both DNA (P<0.001) and RNA (P<0.001) contents were lower in UM gilts while RNA/DNA remained similar (P>0.1). Number of prolactin receptors were lower (P = 0.06) and affinity greater (P<0.05) in UM gilts. Cortisol levels were greater (P<0.01) in UM gilts while other hormones were not affected (P> 0.1). Results clearly demonstrate genetic differences with regard to mammogenesis in gilts and suggest that the less mammary gland development in Upton-Meishan compared with Large White breed of gilts may be related to lower number of prolactin receptors.

Ontogeny of epidermal growth factor (EGF), EGF receptor (EGFR) and basic fibroblast growth factor (bFGF) mRNA levels in pancreas, liver, kidney, and skeletal muscle of pig.

Epidermal growth factor (EGF), EGF receptor (EGFR), and basic fibroblast growth factor (bFGF) messenger RNA (mRNA) levels were examined by Northern blot analysis in four tissues (pancreas, liver, kidney, and skeletal muscle) of pig from fetal 90 d to postnatal 180 d of age. The present study shows for the first time that EGF mRNA increased with advancing age in the kidney and skeletal muscle of pig. A high level of EGF mRNA was observed in the kidney compared with the liver and skeletal muscle. In the pancreas, high levels of EGF mRNA were found in fetuses and newborns and were low in older pigs. Pancreatic EGFR mRNA level parallelled its EGF mRNA, whereas in the kidney and skeletal muscle, patterns of EGFR mRNA were reversed to their EGF mRNA levels. In the liver, EGFR mRNA was abundant but EGF mRNA was undetected. In the pancreas and skeletal muscle, the highest levels of bFGF mRNA were found in fetuses of 90 d of age and then decreased with advancing age. In the liver and kidney, there were no major changes in bFGF mRNA levels during the examined developmental periods. These results show that EGF, EGFR, and bFGF mRNA levels are developmentally and tissue specifically regulated in pig. In the pancreas, mRNA levels of EGF, EGFR and bFGF were high in fetal and neonatal life and low thereafter. In the kidney and skeletal muscle, EGF mRNA increased with advancing age. EGF may play a role in muscle growth and maintenance in growing pigs during the later stage of development.

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.

Uterine immune reaction and reproductive performance of sows inseminated with extended semen and infused with pooled whole dead semen.

The objective of this study was to investigate the effect of infusing whole dead semen (WDS) after AI with diluted commercial semen on uterine inflammatory reaction and embryonic survival rate in gilts. Sixty Yorkshire-Landrace gilts were assigned at their second estrus to one of the following AI treatments: 1) commercial semen adjusted to 1 x 10(9) sperm cells (S1) per dose, followed by an infusion of 80 mL of WDS (S1-WDS); 2) S1 followed by an infusion of 80 mL of Beltsville Thawing Solution (S1-BTS); 3) commercial semen adjusted to 3 x 10(9) sperm cells (S3) per dose, followed by an infusion of 80 mL of BTS (S3-BTS); and 4) a negative control group, in which gilts received two infusions of 80 mL of BTS (BTS). Two days after the first AI, eight gilts from Groups 1, 2, and 4 were slaughtered and reproductive tracts were collected. One horn was cut open longitudinally along the antimesometrial aspect and endometrial samples were taken and immediately frozen for analysis of messenger RNA (mRNA) abundance for inflammatory cytokines and growth factors. The other horn was flushed with 20 mL of PBS, and the contents of interferon-gamma (IFN-gamma), tumor necrosis factor-alpha (TNF-alpha) and transforming growth factor-beta1 (TGF-beta1) were determined by ELISA. On d 25 after AI, gilts from Groups 1, 2, and 3 were slaughtered and their reproductive tracts were collected to evaluate the number of fetuses and corpora lutea. On d 2 after the first AI, only TGF-beta1 was detected in the flush of all gilts, and no difference was observed between S1-WDS, S1-BTS, and BTS gilts. Endometrial levels of IFN-gamma and interleukin (IL)-6 mRNA were marked in all gilts, but they were not affected by the AI treatments, whereas the mRNA abundances for IL-1 and IL-2 were negligible. Infusions of WDS or BTS after a fertile AI did not affect IGF-I, IGF-I receptor, or IGF-II mRNA levels compared with gilts infused with BTS only, whereas the mRNA abundance for the IGF-II receptor was decreased (P < 0.05) in WDS-infused gilts. In gilts inseminated with S1 doses, infusion of WDS did not affect the number of live embryos. Although infusions of WDS did not affect the mRNA level and secretion of the cytokines measured and did not improve embryonic survival rates, further studies are needed to better understand the influence of semen composition on the uterine response after mating.