Ovine placental steroid synthesis and metabolism in late gestation.

Steroid synthesis is required for pregnancy maintenance and for parturition, but comparatively little is known about the major metabolic routes that influence circulating concentrations. Dietary intake changes progesterone and estradiol concentrations in pregnant ewes but whether this reflects placental synthesis is unknown. Progesterone metabolism by 5alpha-reduction is a major metabolic route in other species and can influence the onset of parturition. Therefore, studies were conducted to (1) determine placental enzyme activity, progesterone, and estradiol measured by immunoassay in late gestation ewes on low-, moderate-, and high-nutritional planes, (2) to assess the significance of 5alpha-reduction of progesterone in determining progesterone concentrations in late gestation ewes (gestation day 145) given finasteride to inhibit 5alpha-reductase metabolism. In the second experiment, steroid profiles were examined comprehensively in blood and tissues by liquid chromatography tandem mass spectrometry for the first time in this species. Dietary intake altered progesterone and estradiol serum concentrations but without correlated changes in placental 3beta-hydroxysteroid dehydrogenase, 17alpha-hydroxylase/17,20-lyase cytochrome P450 or aromatase activity. 5alpha-reduced pregnane metabolites were identified in ewes at 145 days of gestation, but concentrations were lower than those of progesterone. Finasteride inhibited 5alpha-reduced progesterone metabolism but did not impact serum progesterone concentrations in these ewes. We conclude that (1) diet-induced changes in serum progesterone and estradiol concentrations are not likely a result of altered placental synthesis of sex steroid but most likely by their metabolism, and (2) metabolism by 5α-reduction is not a major determinant of systemic progesterone concentrations in late gestation ewes.

Consumption of ground beef obtained from cattle that had received steroidal growth promotants does not trigger early onset of estrus in prepubertal pigs.

BACKGROUND: The earlier onset of puberty seen in young American girls has led researchers to question if a causal relation exists between dietary sources of estrogenic compounds and precocious puberty. OBJECTIVE: Using the prepubertal gilt (young female pig) as an animal model, our hypothesis is that feeding beef obtained from cattle receiving growth-promoting steroidal implants postweaning does not alter the onset of puberty or the peripubertal body composition of gilts compared with contemporaries fed nonimplanted "natural" beef or a common meat alternative, tofu. METHOD: The base diet was formulated using canola meal replacing soybean meal to reduce diet estrogenicity. Feed intake was monitored and controlled to ensure similar intake. Gilts were assigned to treatments based on dam and initial body weight (mean: 24.5 ± 3.20 kg) at 61 d of age. The negative control base diet was supplemented with daily feedings of a cooked patty from nonimplanted steers (natural), from steers that had been treated with growth promotants [100 mg trenbolone acetate and 14 mg estradiol (E2) benzoate; implanted], or cooked tofu patty. RESULTS: E2 equivalents (nanogram per kilogram, as fed as analyzed by E-Screen) of the tofu (a soy-based product) supplement were ∼570 times the natural and ∼170 times the implanted supplements. There were no observed differences across treatments in live weight gain (P = 0.90), longissimus muscle area developed at the 10th and 11th rib interface (P = 0.46), and subcutaneous fat deposition (P = 0.41) at the same location over time or in the number of days to reach estrus (P = 0.55). CONCLUSIONS: Consumption of beef from growth implanted or natural steers or tofu at levels similar to those typically consumed by humans did not impact growth or onset of estrus in these prepubertal gilts.

Evaluation of Improvest effects on production parameters of gilts from two different genetic sire lines.

The objective was to evaluate the effects of Improvest on the performance and carcass characteristics of gilts from two different genetic sire lines and the performance of Improvest gilts with castrated male pigs. It was hypothesized that performance parameters observed for Improvest gilts would be similar to barrows, thus narrowing the performance gaps between traditionally managed gilts and barrows. Pigs were from Large White/Landrace dams and either Duroc-Pietrain (DP) or Duroc (D) sires. Females within each genetic sire line were randomized by weight to receive the first dose of Improvest (IMP) on day 25 of the study or to serve as a nontreated control (DP IMP gilt (n = 6 pens; 19 pigs/pen), D IMP gilt (n = 6 pens; 19 pigs/pen), DP CON gilt (n = 6 pens; 19 pigs/pen), D CON gilt (n = 6 pens; 19 pigs/pen). The second dose of Improvest was administered 6 wk later (i.e., day 67). Barrows did not receive Improvest (DP barrow [n = 10 pens; 19 to 20 pigs/pen], D barrow [n = 10 pens; 19 to 20 pigs/pen]). Average daily gain (ADG), average daily feed intake (ADFI), and gain-to-feed ratio (G:F) were measured at 21 d intervals throughout the duration of the study. The targeted weight for pigs to be marketed was 133 ±â€…2.5 kg. Carcass characteristics and loin quality parameters were evaluated on a subset of pigs (n = 283). Improvest-treated gilts of both genetic lines had increased (P ≤ 0.05) ADG and ADFI compared to untreated gilts during the post-second dose intervals with values exceeding that of barrows from day 84 to marketing. Overall, DP IMP gilts had increased (P ≤ 0.05) G:F post-second dose compared to DP CON gilts and DP barrows, yet all other treatments were similar. As pigs were marketed at a similar weight, there was no difference in the final weight, however, DP IMP gilts and DP barrows reached market weight sooner (P ≤ 0.05) than DP CON gilts (109.9 and 111.8 vs. 114.3 ±â€…0.8 d). Backfat and loin weight were greater (P < 0.01) in IMP gilts versus CON gilts, while IMP gilts and barrows had similar values within each respective genetic sire line. There were no differences between treatments (P ≥ 0.08) for pH and instrumental color of the loins. When the pass rate of loins (Japanese color score ≥ 3.0 and marbling ≥ 2.0) was evaluated, IMP gilts were at intermediate values between CON gilts and barrows for each respective genetic sire line. Overall, Improvest within a genetic line improved gilt carcass measurements so that they were more similar to barrows.

Uterine infusion of melatonin or melatonin receptor antagonist alters ovine feto-placental hemodynamics during midgestation.

Dietary melatonin supplementation from mid- to late gestation increases umbilical artery blood flow and causes disproportionate fetal growth. Melatonin receptors have been described throughout the cardiovascular system; however, there is a paucity of data on the function of placental melatonin receptors. The objectives of the current experiment were to determine fetal descending aorta blood flow, umbilical artery blood flow, and placental and fetal development following a 4-wk uterine infusion of melatonin (MEL), melatonin receptor 1 and 2 antagonist (luzindole; LUZ), or vehicle (CON) from Day 62 to Day 90 of gestation. After 4 wk of infusion, umbilical artery blood flow and umbilical artery blood flow relative to placentome weight were increased (P < 0.05) in MEL- versus CON- and LUZ-infused dams. Fetal descending aorta blood flow was increased (P < 0.05) in MEL- versus CON- and LUZ-infused dams, while fetal descending aorta blood flow relative to fetal weight was increased in MEL- versus CON-infused dams and decreased in LUZ- versus CON-infused dams. Following the 4-wk infusion, we observed an increase in placental efficiency (fetal-placentome weight ratio) in MEL- versus LUZ-infused dams. The increase in umbilical artery blood flow due to chronic uterine melatonin infusion is potentiated by an increased fetal cardiac output through the descending aorta. Moreover, melatonin receptor antagonism decreased fetal descending aorta blood flow relative to fetal weight. Therefore, melatonin receptor activation may partially mediate the observed increase in fetal blood flow following dietary melatonin supplementation.

The effect of timing of Improvest administration on growth performance and carcass characteristics in gilts.

Improvest (IMP; Zoetis Inc., Parsippany, NJ) has been approved by the U.S. Food and Drug Administration for use in gilts. Improvest is administered twice: the first dose should be administered no earlier than 9 wk of age and the second dose (D2) at least 4 wk after the first dose. The aim of this study was to determine how the timing of IMP before harvest affects growth performance and carcass characteristics in gilts. A total of 1,632 gilts were allocated to four groups (12 pens/treatment; 34 gilts/pen): 1) a control group did not receive IMP; 2) T-early gilts received IMP on day 7 (day 0 = 10 wk postweaning), and D2 on day 40 (i.e., 35 d prior to first removal for harvest); 3) T-medium gilts received IMP on day 21 and D2 on day 56 (i.e., 19 d prior to first removal for harvest); 4) T-late gilts received IMP on day 35 and D2 on day 70 (i.e., 5 d before first removal for harvest). Pigs were selected for harvest by visual observation on days 75, 89, 103, and 117: 1) the heaviest 7 gilts/pen for each treatment on day 75; 2) the heaviest 10 gilts/pen of each treatment at day 89; 3) the heaviest 10 gilts/pen of each treatment on day 103; and 4) the remaining 7 gilts/pen on day 117. Weights and feed disappearance were recorded every 2 wk and during harvest dates to calculate average daily gain (ADG), average daily feed intake (ADFI), and feed efficiency (Gain:Feed; G:F). Generalized linear mixed models of SAS were used to analyze all variables. The increase in ADFI over Control gilts was observed 15 d post D2 and continued through 77 d post D2, with advantages in ADG occurring between 15 and 35 d post D2. Control and IMP treated gilts had similar G:F 15 to 33 d post D2. The overall ADG and ADFI from day 0 to market, final live weights, and hot carcass weights were significantly greater (P ≤ 0.05) in IMP gilts compared to Control. When G:F based on live weight was averaged across all groups (i.e., from day 0 to market), T-early had the lowest (P ≤ 0.05) G:F compared to Control, T-medium, and T-late gilts, which did not differ. Carcasses from IMP gilts had increased (P < 0.01) backfat, but similar (P = 0.5) Longissimus muscle depth, compared to Control. Within a cohort of similar aged gilts finishing during the summer, this study indicates that the trajectory of growth is enhanced within a similar window post D2 of IMP. Gilts treated with IMP had heavier carcasses with increased backfat and similar Longissimus muscle depth.

Timing of maternal nutrient restriction during mid- to late-gestation influences net umbilical uptake of glucose and amino acids in adolescent sheep.

Previous research demonstrated that maternal nutrient restriction during mid- to late-gestation influenced net umbilical uptakes of glucose and amino acids in sheep. However, it is unclear how the timing and duration of nutrient restriction during mid- to late-gestation influences net uterine, uteroplacental, and fetal flux of glucose and amino acids. On day 50 of gestation, 41 adolescent ewe lambs carrying singletons were randomly assigned to one of six dietary treatments: 1) 100% of nutrient requirements from days 50 to 90 of gestation (CON; n = 7); 2) 60% of nutrient requirements (RES; n = 7) from days 50 to 90 of gestation; 3) 100% of nutrient requirements from days 50 to 130 of gestation (CON-CON; n = 6); 4) 100% of nutrient requirements from days 50 to 90 of gestation and 60% of nutrient requirements from days 90 to 130 of gestation (CON-RES; n = 7); 5) 60% of nutrient requirements from days 50 to 90 of gestation and 100% of nutrient requirements from days 90 to 130 of gestation (RES-CON; n = 7); or 6) 60% of nutrient requirements from days 50 to 130 of gestation (RES-RES; n = 7). On day 90 (n = 14) and day 130 (n = 27), intraoperative procedures were performed to evaluate uteroplacental blood flows, collect blood samples, and then ewes were euthanized. Net uterine, uteroplacental, and umbilical fluxes of glucose and amino acids were calculated by multiplying blood flow by the arterial-venous concentration difference. Data from days 90 and 130 were analyzed separately using ANOVA in SAS. Maternal nutrient restriction during mid-gestation increased (P = 0.04) net umbilical glucose uptake but, maternal nutrient restriction during late-gestation decreased (P = 0.02) net umbilical glucose uptake. Net umbilical essential amino acid uptake decreased (P = 0.03) with nutrient restriction during mid-gestation; however, net umbilical uptakes of Phe (P = 0.02), Thr (P = 0.05), Met (P = 0.09), and His (P = 0.08) increased or tended to increase after nutrient restriction during late-gestation. These data demonstrate that net umbilical glucose and amino acid uptakes were influenced by the timing of nutrient restriction during mid- to late-gestation. Elevated net umbilical glucose uptake after mid-gestational nutrient restriction was sustained throughout late-gestation, independent of late-gestational feeding level. Long-term adaptations in umbilical glucose uptake may have implications for prenatal and postnatal growth and development of the offspring.

Maternal undernutrition during gestation can lead to decreased fetal growth, decreased uteroplacental blood flow, and changes in nutrient supply to the fetus. However, it is unclear how the timing (mid-gestation vs. late-gestation) and duration (40 d vs. 80 d) of nutrient restriction influence nutrient supply to the fetus during mid- to late-gestation. Pregnant ewe lambs fed a pelleted diet to meet 100% of nutritional requirements or 60% of nutritional requirements during mid-gestation alone (days 50 to 90) or during mid- and late-gestation (days 50 to 130). At the end of mid-gestation and late-gestation, the net nutrient supply between the maternal, placental, and fetal compartments was measured. The results indicated that the timing of nutrient restriction influenced the net nutrient supply to the fetus but, the duration of nutrient restriction did not. Nutrient restriction during mid-gestation increased glucose to the fetus but nutrient restriction during late-gestation decreased glucose to the fetus. The opposite response occurred for fetal essential amino acid supply where nutrient restriction during mid-gestation decreased essential amino acid supply to the fetus but increased for several essential amino acids during late-gestational nutrient restriction. The timing of maternal undernutrition during mid- to late-gestation can affect the amount of nutrients delivered to the fetus and thus, could potentially impact postnatal growth and development.

Melatonin supplementation alters uteroplacental hemodynamics and fetal development in an ovine model of intrauterine growth restriction.

Using a mid- to late-gestation ovine model of intrauterine growth restriction (IUGR), we examined uteroplacental blood flow and fetal growth during melatonin supplementation 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 until day 130 of gestation. Umbilical artery blood flow was increased from day 60 to day 110 of gestation in MEL vs. CON dams, while umbilical artery blood flow was decreased from day 80 to day 110 of gestation in RES vs. ADQ dams. At day 130 of gestation, uteroplacental hemodynamics, measured under general anesthesia, and fetal growth were evaluated. Uterine artery blood flow was decreased in RES vs. ADQ dams, while melatonin supplementation did not affect uterine artery blood flow. Total placentome weight and placentome number were not different between treatment groups. Fetal weight was decreased by nutrient restriction. Abdominal girth and ponderal index were increased in fetuses from MEL-ADQ dams vs. all other groups. Fetal biparietal distance was decreased in CON-RES vs. CON-ADQ dams, while melatonin supplementation rescued fetal biparietal distance. Fetal kidney length and width were increased by maternal melatonin treatment. Fetal cardiomyocyte area was altered by both maternal melatonin treatment and nutritional plane. In summary, melatonin may negate the consequences of IUGR during specific abnormalities in umbilical blood flow as long as sufficient uterine blood perfusion is maintained during pregnancy.

Neonatal hormone changes and growth in lambs born to dams receiving differing nutritional intakes and selenium supplementation during gestation.

To investigate the effects of maternal selenium (Se) supplementation and nutritional intake during gestation on hormone changes, percentage body weight (BW) change, and organ mass in neonatal lambs, ewes were allocated to differing Se levels (adequate Se (ASe, 11.5 µg/kg BW) or high Se (HSe, 77.0 µg/kg BW)) initiated at breeding and nutritional intake (60% (RES), 100% (CON), or 140% (HIGH) of NRC requirements) initiated at day 40 of gestation. At parturition, all lambs were removed from dams, fed common diets, and BW and blood samples were collected until day 19. There was a Se × nutritional intake × day interaction for percentage BW change from birth. Lambs born to ASe-HIGH ewes tended to have decreased BW change compared with ASe-CON and ASe-RES groups on day 7. Lambs from HSe-HIGH ewes tended to have increased BW change compared with HSe-RES and HSe-CON groups from days 7 to 19. At birth, there was a Se × sex of offspring interaction, in which male lambs from HSe ewes had decreased cortisol concentrations compared with all other lambs. By 24 h, lambs from RES ewes had decreased cortisol compared with those from HIGH ewes, with lambs from CON ewes being intermediate. Lambs from RES- and CON-fed ewes had greater thyroxine than HIGH ewes at 24 h. Organ masses on day 19 were mainly impacted by maternal nutritional intake and sex of the offspring. Birth weight alone did not predict growth performance during neonatal life. Moreover, despite a similar postnatal diet, maternal nutritional plane and Se status did impact neonatal endocrine profiles. Exact mechanisms of how neonatal endocrine status can influence later growth and development need to be determined.

Barbed compared with standard suture: effects on cellular composition and proliferation of the healing wound in the ovine uterus.

OBJECTIVE: To compare cellular composition (fibroblasts vs. smooth muscle cells) and proliferation in uterine healing wounds after application of barbed compared with standard suture in a sheep model. DESIGN: Randomized trial (Canadian Task Force classification I) using each animal as its own control. SETTING: Certified animal research facility. Population or sample. 23 non-pregnant ewes. METHODS: A myometrial incision was created with the harmonic scalpel in each horn of the bicornuate uterus. The incisions were randomly allocated to be closed using either polyglactin 210 (Vicryl®) or barbed suture. Three months later, uterine tissues were collected, fixed and used for determination of cellular composition and proliferation using histochemistry (Masson trichrome staining) and immunohistochemistry (staining of smooth muscle cell actin and Ki67, a marker of proliferating cells) followed by image analysis. MAIN OUTCOME MEASURES: Evaluation and comparison of the cellular composition and proliferation of uterine wounds after application of barbed vs. standard suture. RESULTS: The ratio between connective tissue elements and smooth muscle cells, expression of smooth muscle cell actin and labeling index were similar in wounds after application of barbed compared with standard suture, but were different (p < 0.0001-0.05) in wounds than in non-wounded areas in uterus. CONCLUSION: Both barbed and standard sutures had similar effects on cellular composition and proliferation of uterine wounds in an animal model.

Maternal metabolizable protein restriction during gestation affects the vascular function of maternal and fetal placental arteries in sheep.

We hypothesized isocaloric diets low in protein would decrease the sensitivity of caruncular (CAR) and cotyledonary (COT) arteries compared to placental arteries from ewes receiving adequate metabolizable protein (MP) requirements. Pregnant ewes were fed one of three isocaloric dietary treatments that provided 60% (MP60), 80% (MP80), or 100% (MP100) of the MP requirements. Diets were fed from day 100-130 of gestation. In vitro dose response curves to bradykinin (BK), sodium nitroprusside (SNP), potassium chloride (KCl), and phenylephrine (PE) in CAR and COT arteries were performed. As MP decreased, the sensitivity to a low dose of KCl increased (P = 0.05) in the COT arteries. There was an overall treatment effect in the CAR and COT arteries for the BK dose response curve, where CAR arteries of MP80 ewes were more sensitive (P = 0.05) to BK compared with MP60 and MP100 ewes, and COT arteries of MP60 and MP80 ewes were more sensitive (P = 0.01) to BK compared with MP100 ewes. There were no treatment effects (P ≥ 0.09) on the SNP or PE dose response curves in CAR or COT arteries. The mechanism of the BK induced vasodilation needs to be elucidated. Moreover, MP restriction appears to alter placental vascular function, which could help explain the differences in nutrient flux previously reported.

Programming the offspring through altered uteroplacental hemodynamics: how maternal environment impacts uterine and umbilical blood flow in cattle, sheep and pigs.

As placental growth and vascularity precedes exponential fetal growth, not only is proper establishment of the placenta important, but also a continual plasticity of placental function throughout gestation. Inadequate maternal environment, such as nutritional plane, has been documented to alter fetal organogenesis and growth, thus leading to improper postnatal growth and performance in many livestock species. The timing and duration of maternal nutritional restriction appears to influence the capillary vascularity, angiogenic profile and vascular function of the placenta in cattle and sheep. In environments where fetal growth and/or fetal organogenesis are compromised, potential therapeutics may augment placental nutrient transport capacity and improve offspring performance. Supplementation of specific nutrients, including protein, as well as hormone supplements, such as indolamines, during times of nutrient restriction may assist placental function. Current use of Doppler ultrasonography has allowed for repeated measurements of uterine and umbilical blood flow including assessment of uteroplacental hemodynamics in cattle, sheep and swine. Moreover, these variables can be monitored in conjugation with placental capacity and fetal growth at specific time points of gestation. Elucidating the consequences of inadequate maternal intake on the continual plasticity of placental function will allow us to determine the proper timing and duration for intervention.

Barbed vs standard suture: randomized single-blinded comparison of adhesion formation and ease of use in an animal model.

STUDY OBJECTIVE: To compare adhesion formation and ease of use of barbed vs traditional suture during myometrial closure in a sheep model. DESIGN: Randomized trial using each subject as its own control (Canadian Task Force classification I). SETTING: Certified animal research facility. SUBJECTS: Twenty-three nonpregnant ewes. INTERVENTIONS: The Harmonic scalpel was used to create a myometrial incision in each uterine horn, and the incisions were randomly allocated to be closed using either polyglactin 210 (Vicryl) or barbed suture. Each animal served as its own control, with 1 horn sutured using barbed suture and the other horn sutured using Vicryl suture. Ease of use was evaluated by comparing closure times. Adhesion formation was compared at necropsy 3 months later using a standardized adhesion-formation scoring system. The evaluator of the adhesion score was blinded to the exposure at surgery. MEASUREMENTS AND MAIN RESULTS: Mean total procedure time was 13.3 minutes. Myometrial closure was significantly faster using barbed vs traditional suture (126.5 seconds vs 272.6 seconds; p <.001). At necropsy 3 months later, adhesions were observed in 12 uterine horns (52.2%) in the barbed suture group vs 10 uterine horns (43.5%) in the Vicryl group (p = .62). The mean (SD) adhesion score was not significantly different between the barbed suture group (3.78 [3.92]) vs the Vicryl group (3.04 [3.75]) ( p = .16). CONCLUSION: Barbed suture significantly facilitates myometrial closure and is associated with adhesion formation and adhesion severity that is not different from that using Vicryl in an animal model.

Mid- to late-gestational maternal nutrient restriction followed by realimentation alters development and lipid composition of liver and skeletal muscles in ovine fetuses.

Maternal nutrient restriction during gestation adversely affects offspring growth and development of liver and skeletal muscle tissues. Realimentation following nutrient restriction may alleviate these negative impacts on development but may alter metabolism and tissue composition. Forty-eight ewes, pregnant with singletons, were fed to meet 100% National Research Council (NRC) recommendations starting at the beginning of gestation. On day 50 of gestation, seven ewes were euthanized (BASE), and fetal liver, skeletal muscles, and blood samples were collected. The remaining animals were fed either 100% of NRC recommendations (CON) or 60% NRC recommendations (RES), a subset were euthanized at day 90 of gestation (n = 7/treatment), and fetal samples were collected. Remaining ewes were maintained on the current diet (CON-CON, n = 6; RES-RES, n = 7) or switched to the alternate diet (CON-RES, RES-CON; n = 7/treatment). On day 130 of gestation, the remaining ewes were euthanized, and fetal samples were collected. At day 130 of gestation, maternal nutrient restriction during late-gestation (RES-RES and CON-RES) decreased fetal liver weight (P < 0.01) and cross-sectional area in triceps brachii (P = 0.01; TB), longissimus dorsi (P = 0.02; LM), and semitendinosus (P = 0.05; STN) muscles. Maternal nutrient restriction during mid-gestation increased hepatocyte vacuole size at day 130 of gestation. Late-gestational maternal nutrient restriction increased mRNA expression of insulin-like growth factor (IGF) binding protein-1 (P < 0.01), glycogen synthase 2 (P = 0.01; GYS2), and pyruvate dehydrogenase kinase 1 (P < 0.01; PDHK1) in the liver and IGF receptor 1 (P = 0.05) in the LM. Lipid concentration in the LM was decreased by late-gestational nutrient restriction (P = 0.01) and increased by mid-gestational nutrient restriction in STN (P = 0.03) and TB (P < 0.01). Principal component analysis of lipidomics data demonstrated clustering of principal components by day of gestation and elastic net regression identified 50, 44, and 29 lipids that classified the treatments in the fetal liver, LM, and blood, respectively. In conclusion, restricting maternal nutrition impacts fetal liver and muscle morphology, gene expression, and lipid metabolism, whereas realimentation attenuated some of these effects. Therefore, realimentation may be a viable strategy to reduce the impacts of nutrient restriction, but can lead to alterations in lipid metabolism in sheep.

Water intake of pigs consuming tiamulin during the nursery phase.

Mass medication to manage population health can be achieved by providing therapeutics in the drinking water. Young nursery pigs are highly sensitive to the flavor and smell of water. Medications that reduce water palatability often lead to an interruption in water and feed intake. With the availability of several generic water-soluble antimicrobials for pigs, questions have arisen about their palatability compared with the original product. In this study, we compared the intake of water containing tiamulin hydrogen fumarate from two different manufacturers with the intake of unmedicated water. The hypothesis was that the intake of tiamulin-containing water would be similar to unmedicated water. Water intake was monitored upon entry into the nursery and just prior to leaving the nursery. Also, average daily gain (ADG) and feed efficiency (FE) were determined. A total of 300 pigs were individually weighed (4.2-10.9 kg; avg = 6.8 kg) for randomization to pen (n = 30 pens). The experiment had two time points: 1) early nursery (periods 1-3) and 2) late nursery (period 4). Pens were randomly assigned to a sequence (period 1-3) in a crossover experimental design containing three 10-d periods, with 5 d for the resetting of baseline where unmedicated water was provided followed by 5 d on tiamulin source addition [i.e., TriamuloxTM (Zoetis, Parsippany, NJ); Denagard (Elanco Animal Health, Greenfield, IN)] or unmedicated water. After period 3 was concluded, all pens were given unmedicated water (via nipple waterers) and the number of pigs per pen was reduced to six pigs to maintain adequate space per pig. Ten days prior to pigs leaving the nursery, a fourth period was performed. After a 5-d water baseline was achieved, pens were treated with either unmedicated water or Triamulox- or Denagard-containing water. Pigs had ad libitum access to water and feed. During the testing periods, daily water intake was measured by a cup water system in each pen. Feed intake was measured every 5 d. There was no effect of treatment on initial body weights or weights at the beginning or end of each period (P ≥ 0.51). Therefore, there was no effect of treatment on ADG (P ≥ 0.23). Water intake (P ≥ 0.16) and FE (P ≥ 0.35) were not affected by treatment. Water consumption was similar among all treatments in each of the four periods. There appears to be no aversion to water intake when tiamulin hydrogen fumarate is added to the drinking water.

Effects of administering exogenous bovine somatotropin to beef heifers during the first trimester on conceptus development as well as steroid- and eicosanoid-metabolizing enzymes.

The aim of this study was to evaluate the effect of bovine somatotropin (bST) on fetal and placental development during the first third of gestation in beef heifers. Angus heifers (n = 97) were randomly assigned to either receive a 500-mg injection of bST (BST) biweekly on days 0, 15, 29, 43, and 57 of gestation or not receive bST (CTL) throughout the experiment. Body weight (BW) was assessed on days -9, -3, 0, 15, 22, 29, 43, 50, 57, 64, and 77, while blood samples were collected on days 0, 22, 50, and 64. Pregnancy status was determined via transrectal ultrasonography on days 29 and 64. A subset of pregnant heifers (BST, n = 7; CTL, n = 5) were harvested on day 84, and complete gravid reproductive tracts and liver tissue were collected for analysis. Cytochrome P450 1A (CYP1A), 2C (CYP2C), 3A (CYP3A), and uridine 5'-diphospho-glucuronosyltransferase (UGT) activities were determined. Mean change in BW and average daily gain of heifers between fixed-time artificial insemination (day 0) and day 77 did not differ between treatments (P ≥ 0.05). Mean concentrations of insulin-like growth factor 1 (IGF-1) were greater (P < 0.001) in BST (347 ± 27.7 ng/mL) compared with CTL (135 ± 32.8 ng/mL) heifers. Mean placental weight, fetal membrane weight, uterine weight, and ovarian and corpus luteum (CL) weights, as well as fetal morphometric data, did not differ (P ≥ 0.05) between treatments. However, BST heifers had greater (P = 0.03) quantities of combined fetal fluid compared with CTL (521.6 ± 22.9 vs. 429.6 ± 27.14 g, respectively). Tendencies were observed for BST heifers to have reproductive tracts with fewer placentomes (P = 0.08) and fetuses with greater umbilical diameters (P = 0.09) compared with CTL. The activity of CYP1A did not differ (P ≥ 0.05) within the maternal and fetal liver, caruncle, cotyledon, or CL tissue samples between treatments. Furthermore, CYP3A activity was only observed in maternal liver samples and was not different between treatments (P ≥ 0.05). Interestingly, CYP2C activity was greater (P = 0.01) in the liver of BST vs. CTL heifers, and UGT activity was greater (P = 0.02) in the CL from BST heifers compared with CTL. In conclusion, the administration of bST during the first third of gestation increased plasma concentrations of IGF-1, which resulted in an increase in fetal fluid, decrease in placentome number, and greater umbilical diameter, but failed to alter fetal development.

Effects of Nutrient Restriction During Midgestation to Late Gestation on Maternal and Fetal Postruminal Carbohydrase Activities in Sheep.

To examine the effects of nutrient restriction during midgestation to late gestation on maternal and fetal digestive enzyme activities, 41 singleton ewes (48.3 ±â€…0.6 kg of BW) were randomly assigned to dietary treatments: 100% (control; CON; n = 20) or 60% of nutrient requirements (restricted; RES; n = 21) from day 50 until day 90 (midgestation). At day 90, 14 ewes (CON, n = 7; RES, n = 7) were euthanized. The remaining ewes were subjected to treatments of nutrient restriction or remained on a control diet from day 90 until day 130 (late gestation): CON-CON (n = 6), CON-RES (n = 7), RES-CON (n = 7), and RES-RES (n = 7) and were euthanized on day 130. The fetal and maternal pancreas and small intestines were weighed, subsampled, and assayed for digestive enzyme activity. One unit (U) of enzyme activity is equal to 1 µmol of product produced per minute for amylase, glucoamylase, lactase, and trypsin and 0.5 µmol of product produced per minute for maltase and isomaltase. Nutrient restriction during midgestation and late gestation decreased (P < 0.05) maternal pancreatic and small intestinal mass but did not affect fetal pancreatic or small intestinal mass. Maternal nutrient restriction during late gestation decreased (P = 0.03) fetal pancreatic trypsin content (U/pancreas) and tended to decrease (P < 0.08) fetal pancreatic trypsin concentration (U/g), specific activity (U/g protein), and content relative to BW (U/kg of BW). Nutrient restriction of gestating ewes decreased the total content of α-amylase (P = 0.04) and tended to decrease total content of trypsin (P = 0.06) and protein (P = 0.06) in the maternal pancreas on day 90. Nutrient restriction during midgestation on day 90 and during late gestation on day 130 decreased (P = 0.04) maternal pancreatic α-amylase-specific activity. Sucrase activity was undetected in the fetal and maternal small intestine. Nutrient restriction during late gestation increased (P = 0.01) maternal small intestinal maltase and lactase concentration and tended to increase (P = 0.06) isomaltase concentration. Realimentation during late gestation after nutrient restriction during midgestation increased lactase concentration (P = 0.04) and specific activity (P = 0.05) in the fetal small intestine. Fetal small intestinal maltase, isomaltase, and glucoamylase did not respond to maternal nutrient restriction. These data indicate that some maternal and fetal digestive enzyme activities may change in response to maternal nutrient restriction.

Fetal expression of genes related to metabolic function is impacted by supplementation of ground beef and sucrose during gestation in a swine model.

To determine the effects of maternal supplementation on the mRNA abundance of genes associated with metabolic function in fetal muscle and liver, pregnant sows (Landrace × Yorkshire; initial body weight (BW) 221.58 ± 33.26 kg; n = 21) fed a complete gestation diet (corn-soybean meal based diet, CSM) were randomly assigned to 1 of 4 isocaloric supplementation treatments: control (CON, 378 g/d CSM, n = 5), sucrose (SUGAR, 255 g/d crystalized sugar, n = 5), cooked ground beef (BEEF, 330 g/d n = 6), or BEEF + SUGAR (B+S, 165 g/d cooked ground beef and 129 g/d crystalized sugar, n = 5), from days 40 to 110 of gestation. Sows were euthanized on day 111 of gestation. Two male and 2 female fetuses of median BW were selected from each litter, and samples of the longissimus dorsi muscle and liver were collected. Relative transcript level was quantified via qPCR with HPRT1 as the reference gene for both muscle and liver samples. The following genes were selected and analyzed in the muscle: IGF1R, IGF2, IGF2R, GYS-1, IRS-1, INSR, SREBP-1C, and LEPR; while the following were analyzed in the liver: IGF2, IGF2R, FBFase, G6PC, PC, PCK1, FGF21, and LIPC. No effect of fetal sex by maternal treatment interaction was observed in mRNA abundance of any of the genes evaluated (P > 0.11). In muscle, the maternal nutritional treatment influenced (P = 0.02) IGF2 mRNA abundance, with B+S and SUGAR fetuses having lower abundance than CON, which was not different from BEEF. Additionally, SREBP-1 mRNA abundance was greater (P < 0.01) for B+S compared with CON, BEEF, or SUGAR fetuses; and females tended (P = 0.06) to have an increased abundance of SREBP-1 than males. In fetal liver, IGF2R mRNA abundance was greater (P = 0.01) for CON and BEEF than SUGAR and B+S; while FBPase mRNA abundance was greater (P = 0.03) for B+S compared with the other groups. In addition, maternal nutritional tended (P = 0.06) to influence LIPC mRNA abundance, with increased abundance in CON compared with SUGAR and B+S. These data indicate limited changes in transcript abundance due to substitution of supplemental sugar by ground beef during mid to late gestation. However, the differential expression of FBPase and SREBP-1c in response to the simultaneous supplementation of sucrose and ground beef warrants further investigations, since these genes may play important roles in determining the offspring susceptibility to metabolic diseases.

Nuclear and membrane progesterone receptors expression in placenta from early to late pregnancy in sheep: Effects of restricted nutrition and realimentation.

Nutrient restriction and/or realimentation may affect several placental functions, such as expression of selected regulatory factors, blood flow and other processes in sheep and other species. To determine the effects of the plane of nutrition, nulliparous white face ewes (6-8 months) carrying singletons on day 50 of gestation were randomly assigned to two dietary treatments receiving 100% of National Research Council recommendations (control; C) or 60% of C (restricted; R). Two groups remained on C or R diets from day 50 until day 130. From day 90-130 another group of C fed ewes was switched to the R diet, and another group of R fed ewes was switched to the C diet. This resulted in 7 groups (n = 5-6 ewes/group): C (day 50, 90 and 130), R (day 90 and 130), CR (day 130) and RC (day 130). At these time points, placental tissues were collected for the evaluation of progesterone receptor (PGR) protein expression (whole tissue), and mRNA expression in maternal (caruncular, CAR) and fetal (cotyledon, COT) (separated tissues). Data were statistically analyzed using analysis of variance (SAS 9.4). Protein for PGRAB and PGRB isoforms was detected using immunohistochemistry in all placental tissues, but the pattern of expression differed depending on pregnancy stage and placental compartment (e.g., CAR vs COT). PGRAB protein expression, quantified using image analysis, was greater (P < 0.04) on day 50 than 90 or 130, and was not affected by plane of nutrition. In CAR and COT, PGRAB mRNA expression was greater (P < 0.05) on day 50 than 90 or 130. PGRB mRNA expression was greater (P < 0.03) in CAR on day 50 than 90 and 130, and was greatest (P < 0.02) in COT on day 50, less on day 130, and least on day 90. For the membrane progesterone receptors, PAQR7 (membrane PGR alpha) mRNA expression was greater (P < 0.05) on days 50 and 90 than 130 in CAR, and greater (P < 0.01) on days 50 than 90 and 130 in COT; PAQR8 (membrane PGR beta) was similar throughout pregnancy in CAR and COT, and PAQR5 (membrane PGR gamma) was greatest (P < 0.0001) on day 130 in COT, but similar throughout pregnancy in CAR. Plane of nutrition affected (P < 0.05) mRNA expression for all genes in CAR and COT throughout pregnancy. These data indicate that expression of PGR in ovine placenta is dependent on stage of pregnancy and plane of nutrition in sheep. The mechanisms of how diet and stage of pregnancy influences placental PGR expression and function remains to be elucidated.

Cellular proliferation and vascularization in ovine fetal ovaries: effects of undernutrition and selenium in maternal diet.

Sheep were fed a maintenance (M) diet with adequate (A) Se or high (H) Se concentration from 21 days before breeding to day 135 of pregnancy. From day 50 to day 135 of pregnancy (tissue collection day), a portion of the ewes from ASe and HSe groups were fed restricted (R; 60% of M) diet. Fetal ovarian sections were stained for: 1) the presence of proliferating cell nuclear antigen (a marker of proliferating cells) to determine the proportion of proliferating primordial follicles, or the labeling index (LI; percentage of proliferating cells) for primordial, primary, secondary and antral follicles, stromal tissues, and blood vessels; 2) factor VIII (a marker of endothelial cells) or 3) a presence of apoptotic cells/bodies. The number of proliferating primordial follicles and the LI of primordial follicles was decreased by R and/or HSe diets. The LI was similar for theca and granulosa cells, and for secondary or antral follicles, but was greater in secondary and antral than in primordial and primary follicles. R diet and/or Se affected the LI in all follicle types, in stromal tissues and blood vessels. A dense network of blood vessels was detected in the areas containing secondary to antral follicles, medulla, and hilus, but areas containing primordial follicles were poorly vascularized. The number of apoptotic cells was minimal. These results demonstrate that nutrient restriction and/or Se level in the maternal diet affected cellular proliferation in follicles, blood vessels, and stromal tissues in fetal ovaries. Thus, plane of nutrition and Se in the maternal diet may impact fetal ovarian development and function.

Late gestation supplementation of corn dried distiller's grains plus solubles to beef cows fed a low-quality forage: III. effects on mammary gland blood flow, colostrum and milk production, and calf body weights.

Objectives were to investigate the effects of supplementation with corn dried distiller's grains plus solubles (DDGS) to late gestating beef cows on arterial blood flow to the mammary glands during late gestation and early lactation; colostrum and milk production; dystocia and immunity; and calf BW. Cows were fed a control (CON; n = 15; 5.1% CP; 36.2% ADF) diet consisting of 90% corn stover and 10% corn silage on a dry basis offered ad libitum or CON diet with supplementation of DDGS (0.30% of BW; SUP n = 12). Mammary gland blood flow was assessed on day 245 of gestation. At parturition, maternal and calving parameters were assessed; colostrum and jugular blood was sampled; and dams were weighed. Mammary gland blood flow and milk production was measured on day 44 of lactation. Calves were weighed fortnightly for 8 wk and at weaning. Colostrum production tended to be greater in SUP dams than in CON dams (837 vs. 614 ± 95 g, P = 0.10). Calves of SUP dams were heavier at birth and 24 h (0 h, 43.2 vs. 39.8 ± 1.0 kg, P = 0.02; 24 h, 44.0 vs. 40.4 ± 1.1 kg, P = 0.02). At birth and 24 h, blood pCO2 was greater in calves born to SUP dams (6.82 vs. 6.00 ± 0.41 kPa, P = 0.04). Serum IgG did not differ (P = 0.21) at 24 h. Ipsilateral mammary gland blood flow of SUP cows was greater than CON cows (2.76 vs. 1.76 ± 0.30 L/min; P = 0.03); however, when summed with contralateral, total blood flow was similar (P = 0.33). Hemodynamic measures on day 44 of lactation were similar (P ≥ 0.32). Milk production tended to be increased (13.5 vs. 10.2 ± 1.2 kg/d, P = 0.07) in SUP vs. CON cows. Despite similar BW through 56 d, calves from SUP cows were heavier (P = 0.04) at weaning (309.7 vs. 292.0 ± 6.0 kg). In conclusion, we accept our hypothesis that DDGS supplementation during gestation influenced mammary blood flow, milk production and calf weights. These findings implicate maternal nutrition's leverage on both nutrient and passive immunity delivery to the calf early in life as well as potential advantages on long-term performance.