Effects of nutrient restriction and melatonin supplementation from mid-to-late gestation on maternal and fetal small intestinal carbohydrase activities in sheep.

The objective of this experiment was to evaluate the effects of nutrient restriction and melatonin supplementation during mid-to-late gestation on maternal and fetal small intestinal carbohydrase activities in sheep. Ewes were randomly assigned to one of 4 dietary treatments arranged in a 2 × 2 factorial design. Ewes were fed to provide 100% (adequate; ADQ) or 60% (restricted; RES) of nutrient recommendations, and diets were supplemented with either no melatonin (control; CON) or 5 mg melatonin/d (melatonin; MEL). This resulted in 4 treatment groups: CON-ADQ (n = 7), CON-RES (n = 8), MEL-ADQ (n = 8), MEL-RES (n = 8). Treatments began on day 50 of gestation, and ewes were euthanized on day 130 for tissue collection. The maternal and fetal small intestine were collected and assayed for small intestinal carbohydrase activities. Data were analyzed using the GLM procedure of SAS with fetal sex, melatonin, nutrition, and the melatonin by nutrition interaction included in the model statement. There were no melatonin by nutrition interactions for maternal or fetal small intestinal protein concentration or carbohydrase activities (P ≥ 0.11). Dietary melatonin supplementation decreased (P = 0.03) maternal small intestinal protein concentration by 22.7% and increased (P = 0.03) maternal small intestinal glucoamylase, isomaltase, and maltase activity per gram protein by 45.5%, 41.3%, and 40.6%, respectively. Nutrient restriction from mid-to-late gestation did not influence (P ≥ 0.46) maternal small intestinal protein concentration, or maltase, isomaltase, and lactase activity. Maternal glucoamylase activity per gram intestine increased (P = 0.05) with nutrient restriction by 49.1%. Melatonin supplementation and maternal nutrient restriction did not influence (P ≥ 0.15) fetal small intestinal protein concentration, or glucoamylase, isomaltase, and lactase activity. Maternal nutrient restriction from mid-to-late gestation decreased (P = 0.05) fetal maltase activity per gram intestine by 20.5% but did not influence fetal maltase activity per gram protein. These data indicate that some maternal and fetal carbohydrases are influenced by nutrient restriction and melatonin supplementation in sheep. More information is needed to understand how nutritional and hormonal factors regulate digestive enzyme activity in ruminants to design improved maternal nutrition programs to optimize fetal growth and development while maintaining maternal productivity.

PHYSIOLOGY AND ENDOCRINOLOGY SYMPOSIUM: Alterations in uteroplacental hemodynamics during melatonin supplementation in sheep and cattle.

Compromised placental function can result in fetal growth restriction which is associated with greater risk of neonatal morbidity and mortality. Large increases in transplacental nutrient and waste exchange, which support the exponential increase in fetal growth during the last half of gestation, are dependent primarily on the rapid growth and vascularization of the uteroplacenta. The amplitude of melatonin secretion has been associated with improved oxidative status and altered cardiovascular function in several mammalian species; however, melatonin mediated alterations of uteroplacental capacity in sheep and cattle are lacking. Therefore, our laboratories are examining uteroplacental blood flow and fetal development during maternal melatonin supplementation. Using a mid- to late-gestation ovine model of intrauterine growth restriction, we examined uteroplacental blood flow and fetal growth during supplementation with 5 mg/d of dietary melatonin. Maternal nutrient restriction decreased uterine arterial blood flow, while melatonin supplementation increased umbilical arterial blood flow compared with non-supplemented controls. Although melatonin treatment did not rescue fetal weight in nutrient restricted ewes; we observed disproportionate fetal size and fetal organ development. Elevated fetal concentrations of melatonin may result in altered blood flow distribution during important time points of development. These melatonin specific responses on umbilical arterial hemodynamics and fetal development may be partially mediated through vascular melatonin receptors. Recently, we examined the effects of supplementing Holstein heifers with 20 mg/d of dietary melatonin during the last third of gestation. Uterine arterial blood flow was increased by 25% and total serum antioxidant capacity was increased by 43% in melatonin supplemented heifers vs. non-supplemented controls. In addition, peripheral concentrations of progesterone were decreased in melatonin supplemented heifers vs. non-supplemented controls. Using an in vitro model, melatonin treatment increased the activity of cytochrome P450 2C, a progesterone inactivating enzyme, which was blocked by treatment with the melatonin receptor antagonist, luzindole. Elucidating the consequences of specific hormonal supplements on the continual plasticity of placental function will allow us to determine important endogenous mediators of offspring growth and development.

Supplementation of metabolizable protein during late gestation and fetal number impact ewe organ mass, maternal serum hormone and metabolite concentrations, and conceptus measurements.

To examine the effects of maternal metabolizable protein (MP) supplementation during late gestation on serum hormone and metabolites and organ masses, multiparous ewes (n = 45) carrying singletons or twins were allotted randomly (within pregnancy group) to 1 of 3 treatments: 60% (MP60), 80% (MP80), or 100% (MP100) of MP requirements. Blood samples were drawn before the initiation of diets (day 100) and before slaughter (day 130) for chemistry panel analysis and weekly for hormone analysis including progesterone (P4) and estradiol-17ß (E2). At day 130, ewe organ masses were recorded. Despite being fed isocaloric diets, MP60 ewes gained less weight throughout pregnancy compared with MP80 and MP100 ewes which were similar. Although diet did not impact E2 or P4 concentrations, ewes carrying twins had greater (P < 0.05) concentrations of both as gestation advanced. Albumin, aspartate aminotransferase, and total protein were reduced (P < 0.05) in MP60 compared with MP100 ewes near term. There was a diet by fetal number interaction (P = 0.03) for lactate dehydrogenase. Twin-carrying MP80 ewes had greater lactate dehydrogenase compared with all other groups on day 130 of gestation. Ewes that were fed MP80 had greater body weight on day 130 of gestation compared with MP60 ewes. Kidney and heart weights were lighter in MP60 ewes compared with MP80 ewes. There was a maternal diet by fetal number interaction (P = 0.05) on fetal weight per unit empty ewe body weight. In ewes carrying singletons, MP60 ewes supported less fetal weight compared with MP100. In contrast, MP60 ewes supported more fetal mass compared with MP100 ewes when carrying twins. The level of protein, and not just total energy, in the diet appears to impact some aspects of the maternal system. Moreover, it appears some measurements of mobilizing maternal body resources are enhanced in ewes carrying twins.

Supplementation of corn dried distillers' grains plus solubles to gestating beef cows fed low-quality forage: II. Impacts on uterine blood flow, circulating estradiol-17ß and progesterone, and hepatic steroid metabolizing enzyme activity.

The objective of this study was to investigate the effects of supplementing dried distillers' grains plus solubles (DDGS) during late gestation on uterine blood flow (BF), circulating steroid hormones and hepatic steroid metabolizing enzymes, and calf and placental weights. Multiparous beef cows were randomly divided into a control group (CON; = 15) consuming a diet containing 90% corn stover and 10% corn silage (DM basis) for ad libitum intake and a treatment group (SUP; = 12) consuming the same diet and DDGS (0.3% of BW). Corn silage inclusion was increased to 30% as gestation progressed to meet increasing caloric requirements. Ipsilateral and contralateral uterine BF and cross-sectional area (CSA) of each uterine artery were measured by Doppler ultrasonography on d 180, 216, and 246 of pregnancy. Contralateral BF and CSA increased ( < 0.01) as gestation advanced. Ipsilateral BF and CSA was affected by a treatment × day of gestation interaction ( < 0.05). A main effect of treatment ( = 0.02) and day ( < 0.01) was observed for total BF; BF increased over time and SUP cows had greater BF than CON cows. Circulating concentrations of both progesterone (P4) and estradiol-17ß (E2) were affected by an interaction of treatment and day ( < 0.01). Concentrations of circulating E2 steadily increased throughout the study and were greater in CON cows than in SUP cows by d 242. Concentrations of P4 also increased over time; P4 of CON cows was greater than that of SUP cows by d 242. Uridine 5'-diphospho-glucuronosyltransferase (UGT) and cytochrome P450 1A (CYP1A) activity increased with advancing gestation ( < 0.01). There was greater UGT activity ( < 0.05) and a trend for greater CYP1A activity ( = 0.06) in SUP cows than in CON cows. Activity of cytochrome P450 3A was greater ( < 0.01) in SUP cows and decreased ( < 0.05) with advancing gestation. Supplementing DDGS to cows fed low-quality forage during late gestation increased uterine BF but decreased circulating E2 and P4 concentrations and altered hepatic steroid metabolizing enzyme activity. It was anticipated that enzyme activity would reflect circulating hormone levels; however, our data suggests the observed increases in BF are not driven by alterations in hormone concentration. Therefore, further research is warranted to elucidate the underlying mechanisms.

Supplementation of corn dried distillers grains plus solubles to gestating beef cows fed low-quality forage: I. Altered intake behavior, body condition, and reproduction.

To investigate the effects of corn dried distillers grains plus solubles (DDGS) supplementation to cows fed corn stover and silage during late gestation, 27 multiparous beef cows (674 ± 17 kg; BCS, 5.6 ± 0.1) were divided randomly into 2 pens equipped with electronic feeders. For 10 wk, both groups were fed the basal diet for ad libitum intake while 1 group was supplemented (SUP; = 12) with DDGS at 0.3% of BW (DM basis). Following parturition, all cows received the same diet for an additional 8 wk. During gestation, SUP cows gained BW ( < 0.01), and there was no change in BCS ( 0.79). Nonsupplemented (CON) cows tended to lose BW ( 0.06) and lost BCS ( < 0.01) during gestation. Supplemented cows consumed more forage ( 0.01) and total feed than CON cows. An interaction of treatment and day was observed for time spent consuming forage ( < 0.01); SUP cows consumed forage faster than CON cows ( ≤ 0.01) early in gestation. Control cows ate more meals than SUP cows ( = 0.06) from d 201 to 218 of gestation. Supplemented cows tended ( = 0.09) to consume larger meals than CON cows and spent more ( < 0.01) time eating than CON cows around d 240 of gestation. Calves born to SUP cows tended ( = 0.06) to be heavier than calves born to CON cows. During lactation, both groups gained ( < 0.01) BW. Body condition score was less ( < 0.05) in CON cows than it was in SUP cows at the end of the study. Dry matter intake during lactation increased ( < 0.01) over time but was not influenced ( = 0.44) by treatment. Supplemented cows spent more time ( < 0.01) eating than CON cows after wk 4 of lactation, and they ate faster than CON cows until wk 3 of lactation whereas CON cows ate faster than SUP cows after wk 6 of lactation ( 0.01). The number of meals increased with advancing lactation ( < 0.01) and CON cows averaged more meals daily than SUP cows ( = 0.01). Conversely, meal size decreased as lactation advanced ( < 0.01), and SUP cows consumed larger meals than CON cows ( = 0.05). Supplementation with DDGS during gestation influenced intake behavior during gestation and lactation as well as the maintenance of maternal BW and BCS and calf birth BW.

Influence of nutrient restriction and melatonin supplementation of pregnant ewes on maternal and fetal pancreatic digestive enzymes and insulin-containing clusters.

Primiparous ewes (n=32) were assigned to dietary treatments in a 2×2 factorial arrangement to determine effects of nutrient restriction and melatonin supplementation on maternal and fetal pancreatic weight, digestive enzyme activity, concentration of insulin-containing clusters and plasma insulin concentrations. Treatments consisted of nutrient intake with 60% (RES) or 100% (ADQ) of requirements and melatonin supplementation at 0 (CON) or 5 mg/day (MEL). Treatments began on day 50 of gestation and continued until day 130. On day 130, blood was collected under general anesthesia from the uterine artery, uterine vein, umbilical artery and umbilical vein for plasma insulin analysis. Ewes were then euthanized and the pancreas removed from the ewe and fetus, trimmed of mesentery and fat, weighed and snap-frozen until enzyme analysis. In addition, samples of pancreatic tissue were fixed in 10% formalin solution for histological examination including quantitative characterization of size and distribution of insulin-containing cell clusters. Nutrient restriction decreased (P⩽0.001) maternal pancreatic mass (g) and α-amylase activity (U/g, kU/pancreas, U/kg BW). Ewes supplemented with melatonin had increased pancreatic mass (P=0.03) and α-amylase content (kU/pancreas and U/kg BW). Melatonin supplementation decreased (P=0.002) maternal pancreatic insulin-positive tissue area (relative to section of tissue), and size of the largest insulin-containing cell cluster (P=0.04). Nutrient restriction decreased pancreatic insulin-positive tissue area (P=0.03) and percent of large (32 001 to 512 000 µm2) and giant (⩾512 001 µm2) insulin-containing cell clusters (P=0.04) in the fetus. Insulin concentrations in plasma from the uterine vein, umbilical artery and umbilical vein were greater (P⩽0.01) in animals receiving 100% requirements. When comparing ewes to fetuses, ewes had a greater percentage of medium insulin-containing cell clusters (2001 to 32 000 µm2) while fetuses had more (P<0.001) pancreatic insulin-positive area (relative to section of tissue) and a greater percent of small, large and giant insulin-containing cell clusters (P⩽0.02). Larger insulin-containing clusters were observed in fetuses (P<0.001) compared with ewes. In summary, the maternal pancreas responded to nutrient restriction by decreasing pancreatic weight and activity of digestive enzymes while melatonin supplementation increased α-amylase content. Nutrient restriction decreased the number of pancreatic insulin-containing clusters in fetuses while melatonin supplementation did not influence insulin concentration. This indicated using melatonin as a therapeutic agent to mitigate reduced pancreatic function in the fetus due to maternal nutrient restriction may not be beneficial.

Supplementing metabolizable protein to ewes during late gestation: II. Effects on ewe lamb performance and reproductive efficiency.

The objective of the current study was to determine the effects of maternal MP intake in isocaloric diets during late gestation on female offspring growth from birth to breeding and measure reproductive performance of those offspring in their first breeding season. In yr 1, maternal dietary treatments were applied at d 100 of gestation, were similar in total energy, and contained 60MP1, 60% of MP requirements; 80MP1, 80% of MP requirements; and 100MP1, 100% of the MP requirements on a DM basis during late gestation. In yr 2, maternal dietary treatments were similar in total energy and contained 60MP2, 60% of MP requirements; 100MP2, 100% of the MP requirements; and 140MP2, 140% of MP requirements on a DM basis during late gestation. While there was a quadratic effect ( = 0.003) for ewe lamb birth weight with the ewe lambs from 80MP1 ewes having increased birth weights compared with ewe lambs from 60MP1 and 100MP1 ewes in yr 1, there was no effect ( ≥ 0.22) of maternal diet on growth of ewe lamb offspring thereafter. A quadratic effect ( = 0.02) was observed for the percentage of ewe lambs bred during the first 17 d of the breeding season, with more ewe lambs born to ewes fed 80MP1 bred compared with ewe lambs born to ewes fed 60MP1 and 100MP1. Ewe lambs giving birth within the first 17 d of lambing season increased ( = 0.001) linearly as MP intake increased in the maternal diet. In yr 2, there was no effect ( ≥ 0.07) of maternal MP treatment during late gestation on growth of ewe lambs and reproductive performance except ADG from birth to weaning and lamb birth weight. There was a quadratic effect ( = 0.01) for ADG from birth to weaning of ewe lambs from ewes consuming 100MP2 being increased compared with ewe lambs from ewes fed 60MP2 and 140MP2. There was a linear ( = 0.04) reduction in birth weight of lambs born to ewe lambs as the dam's maternal dietary MP intake increased. The data from the current study suggest that feeding 80% or 100% of MP requirements during late gestation may have the greatest positive impacts on female offspring reproductive performance.

Arginine supplementation between 41 and 146 days of pregnancy reduces uterine blood flow in dairy heifers.

We hypothesized that arginine (Arg) supplementation during early pregnancy could foster placental vascularization. The objective of this study was to evaluate the effects of Arg supplementation on uterine artery hemodynamics between 40 and 140 days of gestation. At 41 days of gestation, 17 Holstein heifers (448 ± 73.9 kg) were surgically fitted with a catheter in the peritoneal cavity. After surgery, nine heifers received a daily intraperitoneal infusion of 40 mg of Arg/kg of body weight (BW; ARG), whereas the remaining eight heifers received a daily intraperitoneal infusion of saline solution at equivalent volumes as ARG heifers relative to BW (CTRL). Daily infusions took place every 12 hours until heifers reached 146 days of pregnancy. At 41, 62, 83, 104, 125, and 146 days of pregnancy, all heifers were body weighed, bled, and uterine blood flow volume (FV) and other hemodynamics were determined using Doppler ultrasonography. The measurements included heart rate, FV, pulsatility index, and resistance index. Plasma concentrations of amino acids (AAs), nitric oxide, glucose, insulin growth factor, progesterone (P4), growth hormone, and prolactin were analyzed. In a second experiment, we evaluated Arg metabolism when infused either intravenously or intraperitoneally. Overall, FV did not differ between the treatments, but it increased (P < 0.001) with pregnancy day. Uterine blood FV evolved differently between the treatments, with CTRL heifers having a greater (P < 0.05) FV (1104 ± 65.42 mL/min) at 146 days of pregnancy than in ARG heifers (806 ± 65.32 mL/min). Pulsatility index and resistance index decreased throughout pregnancy but did not differ between the treatments. Heart rate was decreased (P < 0.05) in ARG (74 ± 1.4 beats/min) compared with CTRL heifers (81 ± 1.5 beats/min). Plasma concentrations of Arg tended (P < 0.09) to be greater in ARG than those in CTRL heifers, but carnitine, valine, isoleucine, leucine, phenylalanine, and tryptophan were lower (P < 0.05) in ARG than those in CTRL heifers. The proportion of essential AAs (with respect to total AAs) in plasma was greater (P < 0.05) in CTRL (73 ± 2.4%) than that in ARG heifers (65 ± 2.3%). No differences between the treatments were found in plasma prolactin (P = 0.43), insulin growth factor (P = 0.97), glucose (P = 0.45), growth hormone (P = 0.39), or nitric oxide (P = 0.89) concentrations. However, plasma P4 concentration was greater (P < 0.05) in ARG than in CTRL heifers. Contrary to our hypothesis, Arg supplementation did not increase blood flow to the uterus but did change other parameters that could influence placental and fetal growth such as heart rate, maternal concentration of plasma AAs, or P4 synthesis. Furthermore, infusion sites (blood vs. peritoneum) of Arg render different metabolic responses.

Effects of nutrient restriction and melatonin supplementation on maternal and foetal hepatic and small intestinal energy utilization.

To determine how nutrient restriction and melatonin supplementation influence ewe and foetal hepatic and small intestinal energy use, 32 primiparous ewes on d 50 of gestation were fed 60% (RES) or 100% (ADQ) of NRC recommendations with 0 (CON) or 5 mg/d (MEL) of dietary melatonin. On d 130 of gestation, small intestine and liver were weighed and collected. Data were analysed as a completely randomized design with a 2 × 2 factorial arrangement of treatments. Liver weight (g/kg EBW) decreased (p = 0.02) in RES ewes. Jejunum weight (g/kg BW) increased (interaction p = 0.04) in ADQ-MEL ewes compared with all other treatments. Total in vitro O2 consumption (mol/min/tissue) and total citrate synthase activity (mol/min/tissue and mol/min/kg EBW) in liver decreased (p ≤ 0.03) in RES ewes. Oxygen consumption (mol/min/kg EBW) increased (interaction p = 0.02) in jejunum of ADQ-CON versus RES-MEL and ADQ-CON. Citrate synthase activity (mol/min/kg of EBW) increased (interaction p = 0.03) in jejunum of ADQ-MEL compared with RES-MEL and ADQ-CON. Foetal liver weight (g/kg BW) decreased (p = 0.02) in RES versus ADQ. Foetal small intestine weight (g/kg BW) decreased (interaction p = 0.05) in RES-MEL versus ADQ-MEL. Total O2 consumption (mol/min/tissue) and total citrate synthase activity (mol/min/kg of BW) in foetal liver decreased (p ≤ 0.05) in RES versus ADQ. Foetal small intestinal O2 consumption (mol/min/kg of BW) was greater (interaction p = 0.03) in RES-CON and ADQ-MEL than RES-MEL and ADQ-CON. Maternal nutrient restriction had a greater effect than melatonin supplementation on liver and jejunum mass and energy utilization in dams and foetuses. Because intestinal mass and energy utilization were more responsive to melatonin supplementation in ewes fed adequate nutrition compared with restricted ewes, melatonin may have limited use as a therapeutic supplement to help overcome potential negative effects of nutrient restriction.

Effect of maternal nutrient restriction and melatonin supplementation from mid to late gestation on vascular reactivity of maternal and fetal placental arteries.

INTRODUCTION: Maternal nutrient restriction and decreased scotophase concentrations of melatonin have been associated with severely compromised pregnancies. We hypothesized that melatonin supplementation in a compromised pregnancy enhances the bradykinin (BK)-induced relaxations of placental arteries thereby ensuring sufficient umbilical blood flow to the developing fetus. METHODS: Pregnant ewes (n = 31) were fed an adequate (ADQ) or nutrient restricted (RES) diet supplemented with 5 mg of melatonin (MEL) or without melatonin (CON) from day 50 to 130 of gestation. On day 130 of gestation, the maternal (caruncular; CAR) and fetal (cotyledonary; COT) placental arteries were suspended in organ chambers for isometric tension recording. RESULTS: There were no treatment or dietary effects on CAR arteries for any vasoactive agent. However, in COT arteries, MEL ewes were more sensitive (P < 0.01) to bradykinin-induced relaxation than CON ewes. There was a melatonin by nutritional level interaction (P < 0.01) with sodium nitroprusside-induced relaxation of COT arteries where CON-RES were more sensitive to sodium nitroprusside compared to CON-ADQ, which was in contrast to when ewes were fed MEL. There was a significant melatonin by nutritional interaction (P = 0.04) for responsiveness to norepinephrine. The sensitivity of the COT arteries to norepinephrine in CON-RES ewes was decreased compared to CON-ADQ. Melatonin supplementation, regardless of maternal dietary intake, resulted in COT arteries having similar responsiveness to CON-RES ewes. CONCLUSION: An increase in placental vessel sensitivity to bradykinin-induced relaxation may contribute to melatonin-induced increases in umbilical artery blood flow.

Effects of short-term oilseed supplementation on plasma fatty acid composition, progesterone and prostaglandin F metabolite in lactating beef cows.

Twenty-four 3-year-old Angus cows (512.2±21.6 kg) and six ruminally cannulated beef heifers (523.1±16.9 kg) were used to determine the impact of feeding oilseeds starting at the beginning of estrous synchronization until maternal recognition of pregnancy on plasma fatty acid composition. Starting ~60 days postpartum cows were synchronized with the Select Synch+controlled internal drug-release (CIDR) device and timed artificial insemination (AI) protocol. The day CIDR was inserted; cattle were randomly assigned to one of the three treatments being grazing only (CON) or a supplement containing whole soybeans (SOY); or whole flaxseed (FLX). Cattle continued to receive these diets for 28 days. Blood was collected every 3 days until 10 days after insemination and then every day until 18 days after insemination. All cattle grazed a common pasture and supplemented cattle were individually fed their respective supplements once daily. Ruminally cannulated heifers were used to evaluate the impact supplements had on forage intake, which was reduced (P=0.05) with oilseed supplementation. Feeding oilseeds increased total fatty acid intake (P<0.001) across treatments with SOY having greater (P<0.001) 18:2n-6 intake than either CON or FLX. Likewise, cattle fed FLX had greater (P<0.001) 18:3n-3 intake than either CON or SOY. There was a treatment×time interaction (P≤0.05) for all fatty acids identified except for 20:5n-3 (P=0.99). Within 3 days after the start of supplementation, plasma concentrations of 18:2n-6 increased (P<0.001) for cattle fed SOY compared with CON or FLX, whereas flax-fed cattle did not exhibit an increase (P=0.02) until day 15 of supplementation over that of CON. Plasma concentrations for 18:3n-3 was greater (P<0.013) for FLX than both CON and SOY by day 12. Feeding flaxseed tended to (P=0.07) increase and increased (P=0.01) plasma concentrations of 20:4n-6 by day 18 over CON and SOY, respectively. Overall, treatment did not affect serum concentration of progesterone (P=0.18) or prostaglandin F metabolite (P=0.89). However, day after breeding had an effect on serum progesterone (P=0.01) with day 16 after timed AI being lower compared with other days. Feeding oilseeds during the time of estrous synchronization will not only increase the energy density of the diet but will provide key fatty acids around the time of maternal recognition of pregnancy.

Dietary selenium and nutritional plane alter specific aspects of maternal endocrine status during pregnancy and lactation.

Objectives were to examine effects of selenium (Se) supply and maternal nutritional plane during gestation on placental size at term and maternal endocrine profiles throughout gestation and early lactation. Ewe lambs (n = 84) were allocated to treatments that included Se supply of adequate Se (ASe; 11.5 µg/kg BW) or high Se (HSe; 77 µg/kg BW) initiated at breeding and nutritional plane of 60% (RES), 100% (CON), or 140% (EXC) of requirements beginning on day 40 of gestation. At parturition, lambs were removed from their dams, and ewes were transitioned to a common diet that met requirements of lactation. Blood samples were taken from a subset of ewes (n = 42) throughout gestation, during parturition, and throughout lactation to determine hormone concentrations. Cotyledon number was reduced (P = 0.03) in RES and EXC ewes compared with CON ewes. Placental delivery time tended (P = 0.08) to be shorter in HSe ewes than in ASe ewes, whereas placental delivery time was longer (P = 0.02) in RES ewes than in CON and EXC ewes. During gestation, maternal progesterone, estradiol-17ß, and GH were increased (P < 0.05) in RES ewes and decreased (P < 0.05) in EXC ewes compared with CON ewes. In contrast, maternal cortisol, IGF-I, prolactin, triiodothyronine, and thyroxine were decreased in RES ewes and increased in EXC ewes compared with CON ewes during gestation. Selenium supply did not alter maternal hormone profiles during gestation. During parturition and lactation, maternal hormone concentrations were influenced by both Se and maternal nutritional plane. During the parturient process, HSe ewes tended to have greater (P = 0.06) concentrations of estradiol-17ß than ASe ewes. Three hours after parturition a surge of GH was observed in ASe-RES ewes that was muted in HSe-RES ewes and not apparent in other ewes. Growth hormone area under the curve during the parturient process was increased (P < 0.05) in ASe-RES vs HSe-RES ewes. Ewes that were overfed during gestation had reduced (P < 0.05) estradiol-17ß but greater IGF-I, triiodothyronine, and thyroxine (P < 0.05) compared with RES ewes. Even though ewes were transitioned to a common diet after parturition, endocrine status continued to be affected into lactation. Moreover, it appears that gestational diet may partially affect lactational performance through altered endocrine status.

Supplementing metabolizable protein to ewes during late gestation: I. Effects on ewe performance and offspring performance from birth to weaning.

We hypothesized that a maternal diet greater in MP during the last third of gestation would yield improved offspring growth due to enhanced placental or mammary gland function. Our objective was to determine how MP intake during late gestation affected ewe and offspring performance from birth to weaning in ewes fed diets similar in total energy. In yr 1, maternal dietary treatments were applied at d 100 of gestation, were similar in total energy, and contained 60% of MP requirements (60MP1), 80% of MP requirements (80MP1), and 100% of the MP requirements (100MP1) on a DM basis during late gestation. In yr 2, maternal dietary treatments were similar in total energy and contained 60% of MP requirements (60MP2), 100% of the MP requirements (100MP2), and 140% of MP requirements (140MP2) on a DM basis during late gestation. In yr 1, there was no effect (P ≥ 0.30) of maternal MP restriction on gestation length, lamb birth weight, or milk production of the dam. At lambing, ewe BW (P = 0.02) and BCS (P = 0.01) increased linearly as MP in the diet increased. While there were no differences in lamb BW at birth, lamb weaning BW (P = 0.08) and ADG from birth to weaning (P = 0.10) tended to increase linearly as maternal MP intake increased. In yr 2, maternal BW increased with increased dietary MP. However, there were no significant effects (P ≥ 0.45) of maternal dietary treatment on maternal BCS or lamb birth weight. Moreover, lamb weaning weight and ADG were not influenced by maternal MP levels. These results suggest that ewes maintain and gain BW and BCS when consuming diets similar in total energy with increased MP during late gestation, but the increased MP had minimal effects on lamb performance through weaning.

Maternal dietary intake alters organ mass and endocrine and metabolic profiles in pregnant ewe lambs.

To determine the impacts of Se supply and maternal dietary intake on ewe organ mass and endocrine and metabolic changes throughout gestation, pregnant first parity ewes (n=77) were allocated to 6 treatments in a 2×3 factorial array. Factors included Se [adequate Se (ASe; 9.5µg/kg BW) vs. high Se (HSe; 81.8µg/kg BW)] initiated at breeding and dietary intake [60% (RES), 100% (CON), or 140% (EXC) of requirements] initiated on d 50 of gestation. Ewes were individually fed and blood samples from the jugular vein were obtained approximately every 14 d from d 50 until parturition. Maternal Se supply did not impact endocrine or metabolic status. There was a nutritional intake by day interaction for NEFA, blood urea nitrogen (BUN), insulin, triiodothyronine (T3), thyroxine (T4), progesterone (P4), and estradiol-17ß (E2). As expected, with increased maternal intake, NEFA concentrations were reduced. During the last weeks of gestation, BUN and insulin were elevated in EXC compared with RES ewes. Although the pattern of T3 and T4 differed throughout gestation within a treatment group, as maternal intake increased, circulating T3 and T4 were increased. For P4 and E2, as maternal dietary intake increased, there was a reduction in the steroid concentrations in jugular blood. There was only a main effect of maternal nutrition (P<0.001) for cortisol concentrations with EXC ewes having greater concentrations than RES and CON ewes, which did not differ. Although Se is known to influence thyroid hormone metabolism, supranutritional levels during pregnancy did not alter circulating T3 and T4 concentrations. Alterations in maternal endocrine status may have influenced placental transport of nutrients to the developing fetus, which we have shown previously is affected by maternal dietary Se and intake. In addition, the alterations in mammary gland weight that we observed may explain the impact of maternal nutrition on mammary gland function and colostrum production, thereby further impairing growth of developing neonates.

Dietary melatonin supplementation alters uteroplacental amino acid flux during intrauterine growth restriction in ewes.

Dietary melatonin supplementation during mid- to late-gestation increased umbilical artery blood flow and caused disproportionate fetal growth. This melatonin-induced increase in umbilical artery blood flow may alter nutrient availability to the fetus, which may lead to alterations in fetal size. The objectives of the current experiment were to determine amino acid (AA) and glucose concentrations as well as AA and glucose flux across the uteroplacenta using a mid- to late-gestation model of intrauterine growth restriction supplemented with dietary melatonin as a 2 × 2 factorial design. At day 50 of gestation, 32 ewes were supplemented with 5 mg of melatonin (MEL) or no melatonin (CON) and were allocated to receive 100% (adequate; ADQ) or 60% (restricted; RES) of nutrient requirements. On day 130 of gestation, uterine and umbilical blood flows were determined via Doppler ultrasonography during a non-survival surgery. Blood samples were collected under general anesthesia from the maternal saphenous artery, gravid uterine vein, umbilical artery, and umbilical vein for AA analysis and glucose. Total α-AA concentrations in maternal artery and gravid uterine vein were decreased (P < 0.05) in RES v. ADQ fed ewes. Maternal arterial - venous difference in total α-AA was increased (P ⩽ 0.01) in RES v. ADQ fed ewes, while total uterine α-AA flux was not different (P > 0.40) across all treatment groups. Fetal venous - arterial difference in total α-AA as well as uteroplacental flux of total α-AA were decreased (P < 0.05) in CON-RES v. CON-ADQ, and similar (P > 0.20) in MEL-RES v. CON-ADQ. Maternal concentrations and uterine flux of branched-chain AA (BCAA) were not different across all treatment groups; however, fetal uptake of BCAA was decreased (P < 0.05) in CON-RES v. CON-ADQ, and similar (P > 0.20) in MEL-RES v. CON-ADQ. Uterine uptake of glucose was not different (P ⩾ 0.08) across all treatment groups, while uteroplacental uptake of glucose was increased (P ⩽ 0.05) in RES v. ADQ ewes. In conclusion, maternal nutrient restriction increased maternal arterial - venous difference in total α-AA, while total uterine α-AA flux was unaffected by maternal nutrient restriction. Melatonin supplementation did not impact maternal serum concentrations or uterine flux of glucose or AA; however, melatonin did improve fetal BCAA uptake during maternal nutrient restriction.

Maternal nutritional plane and selenium supply during gestation impact visceral organ mass and intestinal growth and vascularity of neonatal lamb offspring.

To investigate effects of nutritional plane and Se supply during gestation on neonatal offspring visceral organ mass and intestinal growth and vascularity, 84 nulliparous Rambouillet ewes (age = 240 ± 17 d, BW = 52.1 ± 6.2 kg) were allocated to a 2 × 3 factorial design. Ewes were fed 1 of 2 Se diets [adequate Se (ASe, 11.5 µg/kg BW) or high Se (HSe, 77.0 µg/kg BW)], initiated at breeding, and 1 of 3 nutritional planes [60% (restricted; RES), 100% (control; CON), or 140% (high; HIH) of NRC requirements], initiated at d 40 of gestation. Ewes were fed individually and remained on treatments through parturition. All lambs were removed from their dams at birth and fed milk replacer. At 20.6 ± 0.9 d of age, lambs were necropsied, visceral organs dissected, and jejunal samples collected. Lambs born to ewes fed CON and HIH had greater (P < 0.05) BW, gastrointestinal tract, stomach complex, and liver masses at necropsy than RES. Large intestinal and pancreatic masses, as well as stomach complex, large intestinal, and liver proportional masses, demonstrated (P ≤ 0.08) a nutritional plane × Se supply interaction. Proportional pancreatic mass was greater (P = 0.03) for lambs born to RES ewes than HIH. Although small intestinal mass was not affected (P ≥ 0.18) by gestational treatments, lambs born to HIH-fed ewes had greater (P ≤ 0.09) jejunal DNA concentration than RES and CON, and greater (P = 0.01) total DNA than RES. Nutritional plane and Se supply interacted to affect (P ≤ 0.003) jejunal percent proliferation and total proliferating small intestinal cells, although jejunal crypt depth and villus length were not affected by gestational treatment (P ≥ 0.17). Jejunal glucagon-like peptide-2 mRNA expression was greater (P ≤ 0.07) in lambs born to ewes fed RES compared with CON and HIH. Jejunal capillary size was affected (P = 0.09) by the interaction of nutritional plane × Se supply. Lambs from CON ewes had greater (P ≤ 0.04) jejunal capillary surface density than RES. Nutritional plane and Se supply interacted to affect (P = 0.07) jejunal soluble guanylate cyclase mRNA expression in a manner opposite of capillary size. In conclusion, neonatal lamb visceral organ mass was affected by gestational nutrition, even when lambs had ad libitum intake and similar management postnatally. Despite similar small intestinal mass at 20 d of age, jejunal growth, vascularity, and gene expression were altered by maternal nutrition during gestation.

Impacts of maternal selenium supply and nutritional plane on visceral tissues and intestinal biology in 180-day-old offspring in sheep.

Objectives were to investigate the effects of maternal Se supply and nutritional plane during gestation on offspring visceral tissues and indices of intestinal growth, vascularity, and function at 180 d of age. Rambouillet ewe lambs (n = 82, approximately 240 d of age; 52 ± 0.8 kg BW at breeding) were allocated to a 2 × 3 factorial arrangement of treatments. Treatments included dietary Se [adequate Se (ASe, 9.5 µg/kg BW) or high Se (HSe, 81.8 µg/kg BW)] initiated at breeding and nutritional plane [60% (restricted, RES), 100% (control, CON), and 140% (high, HI) of requirements] initiated at d 50 of gestation. Ewes were fed pelleted diets and housed individually indoors. At parturition, lambs were immediately removed and fed artificial colostrum for the first 20 h followed by ad libitum access to milk replacer. At 180 ± 2 d of age, lambs were euthanized and tissues were harvested. Birth weight was affected by nutritional treatments (P < 0.001), with decreased birth weight in RES and HI compared with CON. Offspring from RES and HI ewes had decreased (P = 0.07) blood volume compared with CON, and those born to HSe ewes had increased (P < 0.04) total visceral adiposity. Within offspring from CON ewes, those from HSe ewes had greater (P < 0.02) intestinal mass compared with ASe ewes. Within offspring from HSe ewes, both RES and HI had reduced (P ≤ 0.05) intestinal mass compared with CON. Jejunal capillary area density was greater (P = 0.08) in offspring from ewes fed HSe compared with ASe. In addition, area per capillary was greater (P ≤ 0.09) in CON compared with RES. Maternal nutritional plane tended (P ≤ 0.11) to alter total small intestinal vascularity, with lambs from CON being greater than RES. Expression of most mRNA for measured angiogenic factors and receptors was not altered (P ≤ 0.13) by maternal treatments; however, expression of glucagon-like peptide-2 (GLP-2) was decreased (P = 0.07) in offspring from RES compared with CON ewes. Offspring from ewes fed HI diets had increased (P = 0.08) jejunal mucosal maltase activity. In conclusion, maternal Se supply and nutritional plane during gestation resulted in measurable changes in offspring visceral tissues and intestinal biology, including perirenal fat, blood volume, intestinal mass, total jejunal crypt cell proliferation, area per capillary in jejunal villi, GLP-2 mRNA expression, and maltase activity at 180 d. Additional work is needed to determine impacts on intestinal function and nutrient uptake.

Influence of nitrogen and sulfur intake on bovine uterine pH throughout the luteal phase.

Previous research has reported that diets high in protein and sulfur decreased uterine pH in cattle. The objective of this study was to determine the effect of high N and high S intake on uterine pH. Holstein (n = 15) and Angus-cross (n = 5) heifers (337.5 ± 8.4 kg of BW) were randomly assigned to 1 of 4 diets: control (CON; 13.4% CP and 0.17% S); high nitrogen (HN; CON plus urea supplement); high sulfur (HS; CON plus calcium sulfate); or both high nitrogen and sulfur (HNS). Diets were individually fed at 2.6% of BW on a DM basis using Calan gates and estrus was synchronized to occur on d 13 (d 0 = start of dietary treatment). Blood samples were collected on d -2 and daily (d 1 to 28) at 1400 h to determine concentrations of plasma urea nitrogen (PUN), sulfate (d 1, 4, 8, 12, 16, 20, 24, and 28), and progesterone. Uterine pH was measured on d 16, 20, 24, and 28 (d 3, 7, 11, and 15 of the estrous cycle). There was a treatment, time, and treatment by time interaction (P < 0.01) on concentrations of PUN. There was an effect of treatment (P < 0.01) on concentrations of sulfate, with concentrations being increased in HS compared with CON, HN, and HNS (P < 0.01), and HNS increased compared with CON (P < 0.01) and HN (P < 0.01). Uterine pH was increased in HN and HNS compared with CON (P < 0.02), whereas HS was not different from any treatment (P > 0.11). There was no effect of time (P = 0.26) or treatment by time interaction (P = 0.71) on uterine pH. In summary, uterine pH was increased in HN and HNS compared with CON, whereas HS was intermediate and was associated with increased concentrations of PUN.

Effects of nutritional plane and selenium supply during gestation on visceral organ mass and indices of intestinal growth and vascularity in primiparous ewes at parturition and during early lactation.

Objectives were to investigate effects of nutritional plane and Se supply during gestation on visceral organ mass and intestinal growth and vascularization in ewes at parturition and during early lactation. Primiparous Rambouillet ewes (n = 84) were allocated to 2 × 3 × 2 factorial arrangement of treatments. Factors included dietary Se [adequate Se (ASe, 11.5 µg/kg BW) or high Se (HSe, 77.0 µg/kg BW)], nutritional plane [60% (restricted; RES), 100% (control; CON), or 140% (high; HIH)], and physiological stage at necropsy (parturition or d 20 of lactation). At parturition, lambs were removed and 42 ewes (7 per treatment) were necropsied. Remaining ewes were transitioned to a common diet which met lactation requirements and mechanically milked for 20 d. In the absence of interactions (P > 0.10), main effects are reported. At parturition, stomach complex and liver masses were greatest for HIH, intermediate for CON, and least for RES (P < 0.02). Small intestinal mass was greater (P ≤ 0.002) for HIH than RES and CON, and greater (P < 0.01) for ASe than HSe. During early lactation, RES and CON gastrointestinal masses increased disproportionally to BW (P < 0.05). At parturition, jejunal mucosal density was less (P ≤ 0.01) for RES than CON and HIH, whereas CON had greater (P < 0.003) jejunal mucosal RNA concentration and RNA:DNA than RES and HIH. Although there were no differences (P > 0.17) at parturition, jejunal cell percent proliferation was greatest in RES, intermediate in CON, and least in HIH (P ≤ 0.09) at d 20 lactation. At both stages, RES had less (P = 0.01) jejunal capillary area density than HIH and less (P ≤ 0.03) capillary surface density than CON and HIH. During lactation, jejunal capillary size was greater (P = 0.04) for ewes previously fed HSe compared with ASe. At parturition, ASe-HIH had greater (P < 0.02) jejunal mucosal endothelial nitric oxide synthase 3 mRNA than all other treatments and greater (P = 0.10) vascular endothelial growth factor (VEGF) than all treatments, except ASe-RES. In addition, CON had less (P ≤ 0.08) jejunal VEGF receptor-1 (FLT1) mRNA compared with RES and HIH, and ASe had greater (P = 0.003) FLT1 than HSe at parturition. Ewes fed HIH had greater (P = 0.04) jejunal VEGF receptor-2 mRNA compared with RES. Results indicate that maternal intestinal growth and vascularization are responsive to nutritional plane and dietary Se during gestation and undergo changes postpartum when under similar lactational management.

Effects of maternal plane of nutrition and increased dietary selenium in first-parity ewes on inflammatory response in the ovine neonatal gut.

Many areas of the western United States have soils that have increased Se content, and ruminants grazing these rangelands may ingest increased quantities of Se. In addition, high-energy diets or increased Se intake may induce gut inflammation. The objective of this study was to evaluate the effects of maternal plane of nutrition and increased dietary Se during gestation on inflammatory responses in neonatal lamb ileal tissue, a major immune organ. Rambouillet ewes (age = 240 ± 17 d; initial BW = 52.1 ± 6.2 kg) were allocated to 4 treatments arranged in a 2 × 2 factorial. Factors included Se [adequate Se (ASe, 11.5 µg/kg of BW) or high Se (HSe, 77.0 µg/kg of BW)] initiated at breeding, and nutritional plane [100% (CON) or 140% (HIH) of requirements] initiated at d 40 of gestation. Ewes were fed individually from d 40, and lambs were removed at parturition and fed artificial colostrum and milk replacer. Lambs were necropsied at 20 d of age, and ileal tissues were sampled for immunoblotting and real-time quantitative reverse-transcription PCR analyses. The ASe-HIH and HSe-CON treatments had no effect (P = 0.179) on inflammatory signaling compared with ASe-CON. However, greater inflammatory signaling was detected in the HSe-HIH group, as shown by increased (P < 0.05) mRNA expression of tumor necrosis factor-α and chemotaxis IL-8. Consistently, phosphorylation of c-Jun N-terminal kinase, a primary inflammatory signaling mediator, was greater (P < 0.05) in the HSe-HIH group compared with other treatments. Consistent with cytokine expression, mast cell density was less in the HSe-CON group than in other treatments. The expression of transforming growth factor ß mRNA was greater (P < 0.05) in the HSe-HIH group; consistently, collagen content was increased in the HSe-HIH group compared with the ASe-CON group (P < 0.05). In conclusion, independently, neither HSe nor HIH had major effects on inflammation, but in combination, these maternal treatments induced an inflammatory response in the neonatal intestine.