Milk yield loss in response to feed restriction is associated with mammary epithelial cell exfoliation in dairy cows.

In dairy cows, feed restriction is known to decrease milk yield by reducing the number of mammary epithelial cells (MEC) in the udder through a shift in the MEC proliferation-apoptosis balance, by reducing the metabolic activity of MEC, or both. The exfoliation of MEC from the mammary epithelium into milk is another process that may participate in regulating the number of MEC during feed restriction. The aim of the present study was to clarify the mechanisms that underlie the milk yield loss induced by feed restriction. Nineteen Holstein dairy cows producing 40.0 ± 0.7 kg/d at 77 ± 5 d in milk were divided into a control group (n = 9) and a feed-restricted group (n = 10). Ad libitum dry matter intake (DMI) was recorded during a pre-experimental period of 2 wk. For 29 d (period 1), cows were fed either 100 (control) or 80% (feed-restricted) of their ad libitum DMI measured during the pre-experimental period. Then, all cows were fed ad libitum for 35 d (period 2). Milk production and DMI were recorded daily. Blood and milk samples were collected once during the pre-experimental period; on d 5, 9, and 27 of period 1; and on d 5, 9, and 30 of period 2. Mammary epithelial cells were purified from milk using an immunomagnetic method to determine the rate of MEC exfoliation. Mammary tissue samples were collected by biopsy at the end of each period to analyze the rates of cell proliferation and apoptosis and the expression of genes involved in synthesizing constituents of milk. Feed restriction decreased milk yield by 3 kg/d but had no effect on rates of proliferation and apoptosis in the mammary tissue or on the expression of genes involved in milk synthesis. The daily MEC exfoliation rate was 65% greater in feed-restricted cows than in control cows. These effects in feed-restricted cows were associated with reduced insulin-like growth factor-1 and cortisol plasma concentrations. When all cows returned to ad libitum feeding, no significant difference on milk yield or MEC exfoliation rate was observed between feed-restricted and control cows, but refeeding increased prolactin release during milking. These results show that the exfoliation process may play a role in regulating the number of MEC in the udders of dairy cows during feed restriction without any carryover effect on their milk production.

Oxytocin Induces Mammary Epithelium Disruption and Could Stimulate Epithelial Cell Exfoliation.

Mammary epithelial cells (MEC) are exfoliated from the epithelium into milk, influencing the number of MEC present in the udder. This process is associated with epithelium integrity. The release of oxytocin (OT) induced by milking causes myoepithelial cell contraction, which, in turn, may stimulate MEC exfoliation through mechanical forces. To investigate the role of OT in MEC exfoliation, we inhibited or induced myoepithelial cell contraction by injecting the OT receptor antagonist atosiban (Ato) or a supraphysiological dose of OT, respectively. Eight cows were assigned to 2 treatments during 2 milkings according to a crossover experimental design: Control+OT (cows were first milked to collect standard milk and then received 5 IU of OT to collect residual milk through a second milking) and Ato + OT (cows were injected with Ato (50 µg/kg of body weight) and milked to collect cisternal milk, then received 5 IU of OT to collect alveolar milk through a second milking). Milk MEC were purified to determine their concentration and number in milk. Mammary epithelium integrity was assessed by measuring the kinetics of plasma lactose concentration. Inhibiting myoepithelial cell contraction by Ato injection decreased the number of exfoliated MEC in milk. In contrast, OT injection increased the concentration of MEC in the residual milk and the number of MEC in the alveolar milk. Ato injection reduced plasma lactose concentration, whereas, in both treatments, OT injections increased it. Our results suggested that myoepithelial cell contraction caused by OT could stimulate MEC exfoliation into milk and was associated with epithelium disruption.

Mammary epithelium disruption and mammary epithelial cell exfoliation during milking in dairy cows.

The presence of mammary epithelial cells (MEC) in the milk of ruminants indicates that some MEC are shed from the mammary epithelium; however, the mechanisms that regulate the MEC exfoliation process are not known. Through the release of oxytocin, prolactin, and cortisol and through oxytocin-induced mechanical forces on the mammary epithelium, milking could participate in regulating the MEC exfoliation process. The aims of the present study were to determine the rate of MEC exfoliation throughout milking and to investigate its relationship to mammary epithelium integrity and milking-induced hormone release. Milk samples from 9 Holstein dairy cows producing 40.6 ± 1.36 kg of milk/d were collected at the beginning (after 1 and 2 min), in the middle, and at the end of milking. Milk MEC were purified using an immunomagnetic method. Blood samples were collected before, during, and after milking, and the oxytocin, prolactin, and cortisol concentrations in the samples were measured. Tight junction opening was assessed by plasma lactose concentration and the Na+:K+ ratio in milk. The somatic cell count in milk varied during the course of milking; it decreased at the beginning of milking and then increased, reaching the highest values at the end of milking. Exfoliated MEC were present in all milk samples collected. The presence of MEC in the milk sample collected during min 1 of milking, likely corresponding to the cisternal milk fraction, suggests that MEC were exfoliated between milkings. The observed increase in the Na+:K+ ratio in milk and in the plasma concentration of lactose indicated that disruption of mammary epithelium integrity occurred during milking. The MEC exfoliation rate at milking was not correlated with the variables describing milking-induced prolactin release but was negatively correlated with cortisol release, suggesting that cortisol may play a role in limiting exfoliation. In conclusion, milking induced a disruption of the mammary epithelial barrier. Mammary epithelial cells may be continuously exfoliated between milkings or exfoliated during milking as a consequence of the oxytocin-induced mechanical forces and the disruption of mammary epithelium integrity.

Regulation of cell number in the mammary gland by controlling the exfoliation process in milk in ruminants.

Milk yield is partly influenced by the number of mammary epithelial cells (MEC) in the mammary gland. It is well known that variations in MEC number are due to cell proliferation and apoptosis. The exfoliation of MEC from the mammary epithelium into milk is another process that might influence MEC number in the mammary tissue. The rate of MEC exfoliation can be assessed by measuring the milk MEC content through light microscopy, flow cytometry analysis, or an immuno-magnetic method for MEC purification. Various experimental models have been used to affect milk yield and study the rate of MEC exfoliation. Reducing milking frequency from twice to once daily did not seem to have any effect on MEC loss in goat and cow milk after 7 d, but increased MEC loss per day in goats when applied for a longer period. An increase in MEC exfoliation was also observed during short days as compared with long days, or in response to an endotoxin-induced mastitis in cows. Other animal models were designed to investigate the endocrine control of the exfoliation process and its link with milk production. Suppression of ovarian steroids by ovariectomy resulted in a greater persistency of lactation and a decrease in MEC exfoliation. Administering prolactin inhibitors during lactation or at dry-off enhanced MEC exfoliation, whereas exogenous prolactin during lactation tended to prevent the negative effect of prolactin inhibitors. These findings suggest that prolactin could regulate MEC exfoliation. In most of these studies, variations of MEC exfoliation were associated with variations in milk yield and changes in mammary epithelium integrity. Exfoliation of MEC could thus influence milk yield by regulating MEC number in mammary tissue.

Physiological traits of newborn piglets associated with colostrum intake, neonatal survival and preweaning growth.

Colostrum intake, which is critical for piglet survival after birth and growth up to weaning, greatly depends on piglet weight and vitality at birth. Our aim was to identify a set of biological variables explaining individual variations in colostrum intake, preweaning growth and risk of dying. Farrowing traits, morphological traits and colostrum intake were determined for 504 piglets born alive from 37 Landrace × Large White sows. A subset of 203 of these piglets was used to measure plasma neonatal concentrations of metabolites and hormones in blood collected from the umbilical cord at birth. From univariate analyses, we established that colostrum intake was positively associated with plasma neonatal concentrations of IGF-I, albumin, thyroid hormones (P < 0.001), and non-esterified fatty acids (P < 0.05), and was negatively associated with concentrations of lactate (P < 0.001). In a multivariable analysis, the variables explaining the variation in colostrum intake were piglet birth weight and rectal temperature 1 h after birth (positive effect, P < 0.001), time of birth after the onset of parturition, and fructose plasma concentrations at birth (negative effects, P < 0.001 and P < 0.05, respectively). Piglets that died within 3 days after birth had lower neonatal concentrations of albumin (P < 0.001), IGF-I and thyroxine (P < 0.01) than surviving piglets. Preweaning growth was positively associated with neonatal concentrations of IGF-I, thyroxine (P < 0.001), albumin and insulin (P < 0.05). Cortisol and glucose concentrations at birth were not related to colostrum intake, neonatal survival or preweaning growth. Multivariable analyses confirmed that colostrum intake was the predominant factor influencing piglet survival within 3 days after birth and preweaning growth. These results provide physiological indicators of piglet colostrum intake, besides birth weight. They also confirm the impact of time of birth during farrowing on colostrum intake and the crucial importance of physiological maturity at birth for postnatal adaptation.

A static model to analyze carbon and nitrogen partitioning in the mammary gland of lactating sows.

Quantitative estimates of mammary nutrient inputs, outputs and metabolism in sows are scarce, despite being critical elements to identify parameters controlling milk synthesis central for the feeding of lactating sows. The objective of this study was to quantify the mammary gland input and output of nutrients as well as the intramammary partitioning of carbon and nitrogen with the purpose to identify mechanisms controlling mammary nutrient inputs, metabolism and milk production in lactating sows. A data set was assembled by integration of results from four studies. The data set included data on litter performance, mammary arterial-venous concentration differences (AV-difference) of energy metabolites and amino acids, and the contents of lactose, fat and amino acids in milk. Milk yield was estimated based on average litter size and litter gain, and mammary plasma flow (MPF) was estimated using the sum of phenylalanine and tyrosine as internal flow markers. The yield and composition of milk were used to estimate mammary nutrient output in milk, and MPF and AV-difference were used to estimate net mammary input of carbon and nitrogen and output of CO2. Carbon and nitrogen used for the synthesis of lactose, fat and protein in milk and CO2-yielding processes were represented in a static nutrient partitioning model. The origin of mammary CO2 output was calculated using theoretical estimates of carbon released in processes supporting mammary synthesis of de novo fat, protein and lactose in milk, mammary tissue protein turnover and transport of glucose and amino acids. Results indicated that total input of carbon from glucose and lactate was partitioned into lactose (36%), fat (31%) and CO2-yielding processes (34%). Theoretical CO2 estimates indicated that de novo fat synthesis, milk protein synthesis and mammary tissue protein turnover were the main processes related to mammary CO2 production. More than 90% of mammary gland amino acid input was used for milk protein. The quadratic relationship between AV-difference and mammary input of essential amino acids indicated that both changes in AV-difference and MPF contributed to the regulation of mammary input of essential amino acids. The impact of the arterial supply of amino acids on mammary input may be greater for the branched-chain amino acids, arginine and phenylalanine than for other essential amino acids. In conclusion, relationships between input and output parameters indicate that AV-difference and MPF regulate mammary nutrient input to match the supply and demand of nutrients for the mammary gland.

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.

Review: nutritional and endocrine control of colostrogenesis in swine.

Colostrum plays an essential role in ensuring the survival, growth and health of piglets by providing energy, nutrients, immunoglobulins, growth factors and many other bioactive components and cells. Both colostrum yield and composition are highly variable among sows, yet mechanisms and factors that regulate colostrogenesis are not fully known. Unlike sow milk yield, sow colostrum yield is not highly determined by litter size and suckling intensity but is largely driven by sow-related factors. Colostrum synthesis is under hormonal control, with prolactin and progesterone concentrations prepartum having, respectively, positive and negative influences on colostrum yield. Less is known about the endocrine control of the end of colostrogenesis in swine, which is characterized by the closure of tight junctions in the mammary epithelium and the cessation of transfer of immunoglobulin G (IgG) into lacteal secretions. Recent studies indicate that exogenous hormones may influence colostrogenesis. Inducing parturition by injecting prostaglandin F2α on day 114 of gestation in combination with an oxytocin-like molecule reduced colostrum yield, and injection of prostaglandin F2α alone either reduced colostrum yield or had no effect. Injecting a supraphysiological dose of oxytocin to sows in the early postpartum period delayed the tightening of mammary tight junctions, thereby prolonging the colostral phase and increasing concentrations of IGF-I and IgG and IgA in early milk. The development of strategies to improve colostrum composition in swine through maternal feeding has been largely explored but very few attempts were made to increase colostrum yield. This is most likely because of the difficulty in measuring colostrum yield in swine. The fatty acid content of colostrum greatly depends on the amount of lipids provided in the sow diet during late gestation, whereas the fatty acid profile is largely influenced by the type of lipid being fed to the pregnant sow. Moreover, various ingredients that presumably have immuno-modulating effects (such as fish oil, prebiotics and probiotics) increased concentrations of IgG, IgA and/or IgM in sow colostrum when they were provided during the last weeks of gestation. Finally, there is some evidence that sow nutrition during late gestation may influence colostrum yield but this clearly warrants more research. This review emphasizes that although progress has been made in understanding the control of colostrogenesis in swine, and that strategies exist to manipulate fat and immunoglobulin contents of colostrum, ways to increase colostrum yield are still lacking.

Sow environment during gestation: part II. Influence on piglet physiology and tissue maturity at birth.

Sow environment during gestation can generate maternal stress which could alter foetal development. The effects of two group-housing systems for gestating sows on piglet morphological and physiological traits at birth were investigated. During gestation, sows were reared in a conventional system on a slatted floor (C, 18 sows), demonstrated as being stressful for sows or in an enriched system in larger pens and on deep straw bedding (E, 19 sows). On gestation day 105, sows were transferred into identical individual farrowing crates on a slatted floor. Farrowing was supervised to allow sampling from piglets at birth. In each litter, one male piglet of average birth weight was euthanized immediately after birth to study organ development and tissue traits. Blood samples were collected from 6 or 7 piglets per litter at birth and 2 piglets per litter at 4 days of lactation (DL4). At birth, mean piglet BW did not differ between groups (P > 0.10); however, the percentage of light ( 0.10) between C and E piglets, but the insulin to glucose ratio was greater (P = 0.02) in C than in E piglets. Compared with E piglets, C piglets had a lighter gut at birth (P = 0.01) and their glycogen content in longissimus muscle was lower (P < 0.01). In this muscle, messenger RNA levels of PAX7, a marker of satellite cells and of PPARGC1A, a transcriptional coactivator involved in mitochondriogenesis and mitochondrial energy metabolism, were greater (P < 0.05), whereas the expression level of PRDX6, a gene playing a role in antioxidant pathway, was lower (P = 0.03) in C than in E piglets. Other studied genes involved in myogenesis did not differ between C and E piglets. No system effect was observed on target genes in liver and subcutaneous adipose tissue. On DL4, C piglets exhibited a lower plasma antioxidant capacity than E piglets (P = 0.002). In conclusion, exposure of sows to a stressful environment during gestation had mild negative effects on the maturity of piglets at birth.

Sow environment during gestation: part I. Influence on maternal physiology and lacteal secretions in relation with neonatal survival.

In pig husbandry, pregnant females are often exposed to stressful conditions, and their outcomes on maternal and offspring health have not been well evaluated. The present study aimed at testing whether improving the welfare of gestating sows could be associated with a better maternal health during gestation, changes in the composition of lacteal secretions and improvement in piglet survival. Two contrasted group-housing systems for gestating sows were used, that is, a French conventional system on slatted floor (C, 49 sows) and an enriched system using larger pens on deep straw (E, 57 sows). On the 105th days of gestation (DG105), sows were transferred into identical farrowing crates on slatted floor. Saliva was collected from all sows on DG35, DG105 and DG107. Blood samples were collected on DG105 from all sows and on the 1st day of lactation (DL1) from a subset of them (C, n=18; E, n=19). Colostrum and milk samples were collected from this subset of sows at farrowing (DL0) and DL4. Saliva concentration of cortisol was greater in C than in E sows at DG35 and DG105, and dropped to concentrations comparable to E sows after transfer into farrowing crates (DG107). On DG105, plasma concentrations of haptoglobin, immunoglobulins G (IgG) and A (IgA), blood lymphocyte counts and plasma antioxidant potential did not differ between groups (P > 0.10), whereas blood granulocyte count, and plasma hydroperoxide concentration were lower in E than in C sows (P < 0.05). Concentrations of IgG and IgA in colostrum and milk did not differ between the two groups. The number of cells did not differ in colostrum but was greater in milk from E than C sows (P < 0.05). Pre-weaning mortality rates were lower in E than C piglets (16.7% v. 25.8%, P < 0.001), and especially between 12 and 72 h postpartum (P < 0.001). Plasma concentration of IgG was similar in E and C piglets on DL4. In conclusion, differences in salivary cortisol, blood granulocyte count and oxidative stress markers between groups suggested improved welfare and reduced immune solicitation during late gestation in sows of the E compared with the C system. However, the better survival observed for neonates in the E environment could not be explained by variations in colostrum composition.

Review: the cellular mechanisms underlying mammary tissue plasticity during lactation in ruminants.

The mammary tissue is characterized by its capacity to adapt in response to a wide variety of changing conditions. This adaptation capacity is referred to as the plasticity of mammary tissue. In dairy ruminants, lactation is challenged by modifications that can either be induced on purpose, such as by modifying management practices, or occur involuntarily, when adverse environmental constraints arise. These modifications can elicit both immediate changes in milk yield and composition and carryover effects that persist after the end of the challenge. This review focuses on the current knowledge concerning the cellular mechanisms underlying mammary tissue plasticity. The main mechanisms contributing to this phenomenon are changes in the activity and number of mammary epithelial cells (MECs). Changes in the number of these cells result from variations in the rates of cell proliferation and death as well as changes in the rate MEC exfoliation. The number of MECs also depends on the number of resident adult mammary stem cells and their progenitors, which can regenerate the pools of the various mammary cells. Several challenges, including changes in milking frequency, changes in level of feed supply and hormonal manipulations, have been shown to modulate milk yield together with changes in mammary cell activity, turnover and exfoliation. Epigenetic changes may be an additional mechanism of adaptation. Indeed, changes in DNA methylation and reductions in milk yield have been observed during once-daily milking and during mastitis in dairy cows and may affect cell activity persistently. In contrast to what has been assumed for a long time, no carryover effect on milk yield were observed after feed supply challenges in dairy cows and modification of milking frequency in dairy goats, even though the number of mammary cells was affected. In addition, mammary tissue plasticity has been shown to be influenced by the stage of lactation, health status and genetic factors. In conclusion, the cellular mechanisms underlying mammary tissue plasticity are diverse, and the mammary tissue either does or does not show elastic properties (with no permanent deformation), in response to environmental changes.

In vivo gene expression in granulosa cells during pig terminal follicular development.

Ovarian antral follicular development is clearly dependent on pituitary gonadotrophins FSH and LH. Although the endocrine mechanism that controls ovarian folliculogenesis leading to ovulation is quite well understood, the detailed mechanisms and molecular determinants in the different follicular compartments remain to be clarified. The aim of this study was to identify the genes differentially expressed in pig granulosa cells along the terminal ovarian follicle growth, to gain a comprehensive view of these molecular mechanisms. First, we developed a specific micro-array using cDNAs from suppression subtractive hybridization libraries (345 contigs) obtained by comparison of three follicle size classes: small, medium and large antral healthy follicles. In a second step, a transcriptomic analysis using cDNA probes from these three follicle classes identified 79 differentially expressed transcripts along the terminal follicular growth and 26 predictive genes of size classes. The differential expression of 18 genes has been controlled using real-time PCR experiments validating the micro-array analysis. Finally, the integration of the data using Ingenuity Pathways Analysis identified five gene networks providing descriptive elements of the terminal follicular development. Specifically, we observed: (1) the down-expression of ribosomal protein genes, (2) the genes involved in lipid metabolism and (3) the down-expression of cell morphology and ion-binding genes. In conclusion, this study gives new insight into the gene expression during pig terminal follicular growth in vivo and suggested, in particular, a morphological change in pig granulosa cells accompanying terminal follicular growth.

Porcine luteal function in relation to IGF-1 levels following ovulation during lactation or after weaning.

This study presents relationships between peripheral progesterone and Insulin-like Growth Factor-1 (IGF-1) concentrations during the early luteal phase in sows. Data were derived from three experiments, one with primiparous weaned sows (n = 21) and two with multiparous sows that either ovulated during lactation (n = 23) or after weaning (n = 12). The sows that ovulated during lactation did so due to an intermittent suckling regime (inhibition of suckling for 12 h each day from day 14 of lactation) or due to treatment with PG600. IGF-1 concentrations varied considerably among experiments, and were the lowest in the multiparous sows, regardless of whether they were weaned or lactating: 68 +/- 5 and 85 +/- 8 ng/ml in the two experiments with multiparous sows vs 188 +/- 15 ng/ml in the primiparous sows. Progesterone concentrations were lowest for the lactating sows. Overall, the increase in progesterone during the early luteal phase was strongly correlated with IGF-1 concentrations (r = 0.7). However, the correlation was low in multiparous lactating sows (r = 0.28; p < 0.10) and nonsignificant in multiparous weaned sows (r = 0). The weaned multiparous sows had IGF-1 levels comparable to lactating multiparous sows, but higher progesterone levels. In conclusion, these data show a positive relationship between peripheral IGF-1 and progesterone concentrations in vivo during the early luteal phase. In lactating sows, IGF-1 concentrations are probably a limiting factor for progesterone secretion, although other factors may be involved.

Proteomic analysis of adipose tissue during the last weeks of gestation in pure and crossbred Large White or Meishan fetuses gestated by sows of either breed.

BACKGROUND: The degree of adipose tissue development at birth may influence neonatal survival and subsequent health outcomes. Despite their lower birth weights, piglets from Meishan sows (a fat breed with excellent maternal ability) have a higher survival rate than piglets from Large White sows (a lean breed). To identify the main pathways involved in subcutaneous adipose tissue maturation during the last month of gestation, we compared the proteome and the expression levels of some genes at d 90 and d 110 of gestation in purebred and crossbred Large White or Meishan fetuses gestated by sows of either breed. RESULTS: A total of 52 proteins in fetal subcutaneous adipose tissue were identified as differentially expressed over the course of gestation. Many proteins involved in energy metabolism were more abundant, whereas some proteins participating in cytoskeleton organization were reduced in abundance on d 110 compared with d 90. Irrespective of age, 24 proteins differed in abundance between fetal genotypes, and an interaction effect between fetal age and genotype was observed for 13 proteins. The abundance levels of proteins known to be responsive to nutrient levels such as aldolase and fatty acid binding proteins, as well as the expression levels of FASN, a key lipogenic enzyme, and MLXIPL, a pivotal transcriptional mediator of glucose-related stimulation of lipogenic genes, were elevated in the adipose tissue of pure and crossbred fetuses from Meishan sows. These data suggested that the adipose tissue of these fetuses had superior metabolic functionality, whatever their paternal genes. Conversely, proteins participating in redox homeostasis and apoptotic cell clearance had a lower abundance in Meishan than in Large White fetuses. Time-course differences in adipose tissue protein abundance were revealed between fetal genotypes for a few secreted proteins participating in responses to organic substances, such as alpha-2-HS-glycoprotein, transferrin and albumin. CONCLUSIONS: These results underline the importance of not only fetal age but also maternal intrauterine environment in the regulation of several proteins in subcutaneous adipose tissue. These proteins may be used to estimate the maturity grade of piglet neonates.

Oxytocin injections in the postpartal period affect mammary tight junctions in sows.

The potential impacts of injecting oxytocin (OXY) to sows in the early postpartum period on the quality of mammary tight junctions, milk composition, and immune status of sows and piglets were studied. Postparturient sows received i.m. injections of either saline (control [CTL]; = 10) or 75 IU of OXY ( = 10). Injections were given twice daily (0800 and 1630 h) starting on d 2 of lactation (i.e., between 12 and 20 h after birth of the last piglet), totaling 4 injections. Milk samples were obtained before the first injection (d 2 morning [AM]), before the second injection (d 2 afternoon [PM]), and on d 4 PM and d 5 PM. Blood samples were obtained from sows before milking on d 2 AM, d 2 PM, and d 5 PM. On d 5 of lactation, a blood sample was obtained from 3 piglets per litter. Circulating concentrations of prolactin, IGF-I, lactose, and IgA in sows did not differ between treatments at any time ( > 0.10), but OXY sows had less IgG than CTL sows ( < 0.01) on d 2 PM before the second OXY injection. There were differences in milk composition on d 2 PM, with OXY sows having more IGF-I ( < 0.01), solids ( < 0.05), protein ( < 0.01), energy ( < 0.05), and IgA ( < 0.01) and a greater Na:K ratio ( < 0.01) than CTL sows. These differences were not seen in the next 2 milk samples, except for protein and IgA that still tended ( < 0.10) to be greater in OXY vs. CTL sows on d 4 PM (for protein) and on d 5 PM (for IgA) after the last injection. Milk lactose content was lower in OXY vs. CTL sows on d 5 PM ( < 0.01). Values for immunoglobulin immunocrit, IgG, IgA, and IGF-I in piglet blood did not differ between treatments ( > 0.10). Injecting OXY to sows in the early postpartum period increased leakiness of the mammary tight junctions, improved composition of early milk, and may potentially affect immune status of neonatal piglets.

Relative prolactin-to-progesterone concentrations around farrowing influence colostrum yield in primiparous sows.

In swine, colostrum production is induced by the drop of progesterone (P4) concentrations which leads to the prepartum peak of prolactin (PRL). PRL regulates mammary cell turnover and stimulates lacteal nutrient synthesis. P4 inhibits PRL secretion and downregulates the PRL receptor in the mammary gland. The aim of the present study was to determine if the relative prepartum concentrations of P4 and PRL (PRL/P4 ratio) influence sow colostrum production. The performance of 29 Landrace × Large White primiparous sows was analyzed. Colostrum yield was estimated during 24 h starting at the onset of parturition (T0) using litter weight gains. Colostrum was collected at T0 and 24 h later (T24). Repeated jugular blood samples were collected during the peripartum period, that is, from -72 to +24 h related to farrowing and were assayed for P4 and PRL. Sows were retrospectively categorized in 2 groups according to their PRL/P4 ratio 24 h before farrowing being either <2 (low PRL/P4, n = 16) or >3 (high PRL/P4, n = 13). During the peripartum period, the circulating concentrations of P4 were lower (P < 0.05) and those of PRL tended to be greater (P < 0.10) in high PRL/P4 compared with low PRL/P4 sows. Colostrum yield was greater in high PRL/P4 compared with low PRL/P4 sows (4.11 vs 3.48 kg [root mean square error = 0.69], P < 0.05). Colostrum composition (dry matter, energy, protein, lipid, and lactose contents) and IgG and IgA concentrations did not differ between the 2 groups of sows (P > 0.10). The Na/K ratio in colostrum 24 h after the onset of farrowing was lower in high PRL/P4 compared with low PRL/P4 sows (P < 0.05). Piglet mortality between birth and T24 averaged 10.0% in low PRL/P4 litters and 7.0% in high PRL/P4 litters (P = 0.29). In conclusion, a greater PRL/P4 ratio 24 h prepartum, characterized by lower P4 concentrations and a trend for greater PRL concentrations peripartum, led to increased colostrum yield in primiparous sows.

Localization of binding sites for IGF-I, insulin and GH in the sow ovary.

Binding sites for IGF-I, insulin, and GH were localized by in situ binding of (125)I-labelled hormones to the different compartments of the sow ovary. Binding sites for IGF-I were detected in oocytes, granulosa and thecal cells of healthy and atretic follicles as well as in the antrum and the stroma. Competition of (125)I-labelled IGF-I with IGF-I, insulin and an analogue of IGF-I (Long R(3)IGF-I), which allowed discrimination between binding to binding proteins from binding to type-I receptors, suggested that type-I receptors were present in granulosa cells of healthy follicles, whilst binding in other compartments was mainly due to binding proteins. Binding of insulin was revealed in oocytes, granulosa and theca interna cells of healthy preantral and antral follicles, and, to a lesser extent, in theca externa and stromal cells, and was still observed in granulosa cells of atretic follicles. Labelling with (125)I-labelled bovine GH was demonstrated in oocytes, granulosa cells, theca interna cells, and, although less intense, in theca externa and stromal cells. It disappeared in granulosa cells during atresia. Binding sites for GH were detected at all follicular stages, from preantral to preovulatory stages, but the intensity of labelling in granulosa cells was more intense in preantral than in large follicles. These data support the participation of insulin, GH and IGF-I in oocyte maturation, follicular growth and stromal cell function in swine.

Influence of the nutritional status on ovarian development in female pigs.

In female pigs, undernutrition may influence growth of antral follicles from various size classes, decrease ovulation rate, delay puberty and return to oestrus after weaning. It could also affect the oocyte maturation and hence the number of viable embryos per litter. Inhibition of the gonadotrophin release due to undernutrition is presumably involved in these phenomena. Presence of receptors, as well as in vitro and in vivo studies suggest that insulin and hormones from the somatotrophic axis are able to alter folliculogenesis directly at the ovarian level. They should act as hormones controlling nutrition, proliferation, growth and differentiation of the cells and/or as amplifiers of the action of gonadotrophins. Information are needed to determine whether their availability at the ovarian level may become insufficient or excessive in case of nutritional deficit. Increase in plasma concentrations of progesterone due to lower hepatic metabolic rate in underfed females probably contributes to inhibit folliculogenesis.

Effect of insulin administration before weaning on reproductive performance in feed restricted primiparous sows.

The objective of this study was to determine whether exogenous insulin administration in late can improve reproductive performance of feed restricted primiparous sows. A total of 24 pure Large White gilts were assigned at farrowing to one of the three following experimental groups. Control sows (CTRL) were fed on a plane of nutrition close to ad libitum and received 6 kg food day-1. Restricted saline-treated (RS) and insulin-treated (RI) sows received only 4 kg food day-1. Sows from RI group received 0.4 IU insulin per kg of live weight once daily during the 5-day weaning period, whereas CTRL and RS sows received saline injections as a sham treatment. Litters (8-10 piglets) were weaned at 23.5 +/- 1 days post-partum. Blood samples were collected 2 days before (Day W - 2) and after weaning (Day W + 2). Restricted sows RS and RI lost significantly more weight than CTRL sows (25 kg and 32 kg, respectively, vs. 10 kg) and more fat (5.3 mm and 4.7 mm, respectively, vs. 2.2 mm). During insulin treatment, daily litter weight gain was lower in RI than in RS and CTRL groups (P < 0.05). On Day W - 2, insulin concentration was higher, while that of glucose was lower in RI than in RS and CTRL sows (P < 0.05). Concentrations of NEFA and IGF-I were similar in the three groups of sows. On Day W + 2, concentrations of FSH and oestradiol-17 beta did not differ significantly between females of the different groups. Weaning-to-oestrus interval and ovulation rate were similar in CTRL, RS and RI females (5.7, 5.9 and 5.6 days; 19.2, 20.7 and 22.1 corpora lutea, respectively). We concluded that insulin administration during late lactation to feed-restricted primiparous sows did not improve weaning-to-oestrus interval and post-weaning ovulation rate, but was likely to decrease milk production.

Influence of insulin treatment and feed restriction on follicular development in cyclic gilts.

Crossbred gilts were used to investigate whether exogenous insulin can restore normal follicular growth in feed-restricted gilts. After an 18-day altrenogest treatment, the first day of oestrous behaviour was designed as day 0. From day 0 to 13, all gilts received the same amount of feed, calculated to meet 200% of the energy requirements for maintenance. On day 14, luteolysis was induced by injection of an analogue of prostaglandin F2alpha. All gilts were slaughtered on day 19 and their ovaries removed. In Experiment 1, gilts received a high (240% of maintenance) or low (80%) level of feeding (n=10/group) from day 14 to 18. The number of large follicles (> or = 5 mm) on day 19 was reduced in feed-restricted gilts (16.9 versus 20.6, P<0.05). The same protocol of feed restriction was used in Experiment 2 (240% versus 80% of maintenance from day 14 to 18), and some gilts received daily injections of insulin (0.6 IU live weight kg(-1)). The three experimental groups were H: 240% and no insulin (n=8); H-I: 240%+insulin (n=8) and L-I: 80%+insulin (n=7). On day 18, 4 h after insulin injection, plasma insulin was higher in insulin-treated than in untreated gilts and glucose concentrations were reduced more dramatically in L-I than in H-I gilts (P<0.05). Concentrations of IGF-I were lower in L-I than in other gilts (P<0.05) and plasma IGFBPs were not significantly affected by treatments. On day 19, the number of large follicles (> or = 5 mm) was not significantly influenced by treatments (19.4, 17.6 and 15.3 for H, H-I and L-I gilts, respectively). Insulin, IGF-I and IGFBP-2 levels in follicular fluids from large follicles did not differ between females whereas IGFBP-3 levels were lower in L-I than in H gilts (P<0.05) and intermediate in H-I gilts. Intrafollicular levels of glucose were higher in feed-restricted than in well-fed gilts (P<0.05). These results suggest that exogenous insulin does not restore final follicular growth impaired by acute undernutrition.