The role of altered uterine-embryo synchrony on conceptus growth in the pig

This study was conducted to determine whether inducing an embryo-uterine asynchrony during the preimplantation period would alter fetal and(or) placental size at term. Yorkshire gilts (n = 24) were checked twice daily for estrus and bred to a Yorkshire boar 24 h after the first exhibition of estrus. Embryos (1 to 4 cells) were flushed from the oviducts of each donor gilt on d 2.5 of gestation and transferred in equal numbers to the oviducts of a recipient gilt on d 1.5, 2.5, or 3.5 of the estrous cycle. Gilts were slaughtered on d 112 of gestation (calculated on the age of the conceptus) and fetal and placental weight, placental surface area, and implantation site lengths were determined. Although litter sizes were similar (9.1+/-0.9), conceptuses transferred to d 3.5 recipients became heavier fetuses (1.44+/-0.05 vs 1.23+/-0.04 kg, P < 0.001), with larger placental surface areas (1,793+/-60 vs 1,459+/-43 cm2, P < 0.01), and longer implantation sites (32.1+/-1.5 vs 24.9+/-0.6 cm, P < 0.001) than those transferred to recipients on d 2.5. These data demonstrate that oviductal transfer of embryos into a reproductive tract that is more advanced by as little as 24 h can result in alterations in placental growth and function during gestation.     

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.     

Overfeeding and underfeeding have detrimental effects on oocyte quality measured by in vitro fertilization and early embryonic development in sheep

To determine effects of maternal diet on in vitro fertilization (IVF) and early embryonic development, ewes (n = 48) were divided into control, overfed (ad libitum feeding), and underfed (60% of control) nutritional planes for 8 wk before oocyte collection. Follicular development was induced by twice-daily injections of FSH on days 13 and 14 of the estrous cycle, and ovaries and blood samples were collected on day 15 of the estrous cycle. During the 8-wk experiment, for control ewes BW and BCS did not change, but for overfed ewes mean (± SEM) BW and BCS increased (11.8 ± 1.1 kg and 2.0 ± 0.1, respectively) and for underfed ewes decreased (14.2 ± 0.9 kg and 0.7 ± 0.1, respectively). The number of follicles was determined; oocytes were collected and subjected to in vitro maturation and fertilization. After IVF, developing embryos were evaluated throughout the 8-d culture period. The proportion of cleaved oocytes after IVF and developing morula and blastocyst were less (P < 0.0001) in overfed and underfed ewes than in control ewes. However, number of visible follicles, total number of oocytes, number of healthy oocytes, and percentage of healthy oocytes were similar for control, overfed, and underfed ewes. Serum insulin concentration was greater (P < 0.05) in overfed ewes than in underfed ewes, estradiol 17-β (E₂) concentration was greater (P < 0.05) in underfed ewes than in overfed ewes, but triiodothyronine (T₃) and thyroxine (T₄) concentrations were similar in all treatment groups. These data show that inadequate feeding has a negative effect on oocyte quality which results in lower oocyte cleavage after IVF and morula and blastocyst formation; overfeeding increased serum insulin and underfeeding increased serum E₂ but not T₃ or T₄. These data emphasize the importance of diet for reproductive and metabolic functions. Furthermore, the mechanisms through which enhanced or decreased energy in diet affect oocyte quality and serum insulin and E₂ concentrations remain to be elucidated.     

Mammary gland growth and vascularity at parturition and during lactation in primiparous ewes fed differing levels of selenium and nutritional plane during gestation

Background： Objectives were to examine the effects of selenium （Se） supply and maternal nutritional plane during gestation on mammary gland growth, cellular proliferation, and vascularity at parturition and d 20 of lactation. Rambouillet primiparous ewes （n = 84） were allocated to treatments in a 2 x 3 factorial. Factors were dietary Se （adequate Se [ASe, 11.5 μg/kg BW] or high Se [HSe, 77.0 μg/kg BVV]） and nutritional plane （60% IRES], 100% [CON], or 140% [EXC]）. At parturition, lambs were removed and 42 ewes （7/treatment） were necropsied. Remaining ewes were fed a common diet meeting requirements for lactation and mechanically milked twice daily until necropsy on d 20. At both necropsy periods, mammary glands were dissected and tissues harvested. Samples were analyzed for RNA, DNA, and protein content, cell proliferation, and vascularity. Where interactions were present （P 〈 0.05）, least squares means from the highest-order interaction are presented. Results： Final body weight of ewes was least （P 〈 0.002） in RES, intermediate for CON, and greatest for EXC, regardless of stage of the ewe at necropsy （parturition or d 20 of lactation）. In ewes necropsied at parturition, mammary glands were heavier （P = 0.02） in EXC compared to RES, with CON intermediate. Concentration of RNA （rag/g） was decreased （P= 0.01） in EXC compared to CON at parturition. There was a tendency （P= 0.07） for a Se by nutrition interaction in percentage of cells proliferating where ASe-EXC ewes had greater （P_〈 0.02） number of proliferating cells then all other treatments. Mammary vascular area tended （P = 0.08） to be affected by a Se by nutrition interaction where ASe-CON had less （P= 0.007） vascular area than HSe-CON ewes. In ewes necropsied at d 20 of lactation, the number of alveoli per area was decreased （P 〈- 0.05） in RES compared to CON and EXC-fed ewes. Conclusions： Results of this study indicate that proper maternal nutritional plane during gestation is important for mammary gland development, even out to d 20 of lactation.     

Effect of early gestational undernutrition on angiogenic factor expression and vascularity in the bovine placentome

The effect of early gestation maternal undernutrition followed by realimentation on placentomal vascular growth and angiogenic factor expression was determined in multiparous beef cows bred to the same bull. Cows gestating only female fetuses (n = 30) were fed in equal numbers to meet the NRC requirements (control) or were fed below the NRC requirements to lose BW (nutrient restricted; NR) from d 30 to 125 of gestation. After slaughter on d 125 of gestation, 10 control and 10 NR cows were necropsied. The remaining NR cows (n = 5) were then fed to achieve a BCS equal to their control group contemporaries (n = 5) by d 220 of gestation. All cows were fed the control diet from d 220 until 250 of gestation, when the remaining control and NR cows were slaughtered and necropsied. At necropsy, placentomes were fixed via perfusion of the caruncular and cotyledonary arteries to determine capillary vascular density. Cotyledonary (fetal placental) and caruncular (maternal placental) tissues also were snap-frozen in liquid nitrogen, and mRNA concentrations of vascular endothelial growth factor and its 2 specific receptors, fms-like tyrosine kinase and kinase insert domain containing receptor, as well as placental growth factor, were determined. There was no effect of diet or day of gestation on the percentage of proliferating caruncular cells. Although diet did not impact cotyledonary cellular proliferation, there was an increase (P < 0.05) in the percentage of proliferating cells on d 250 compared with d 125 of gestation. Nutrient restriction from d 30 to 125 increased (P < or = 0.10) placental mRNA concentrations of placental growth factor and fms-like tyrosine kinase; however, there was no alteration in vascularity. By d 250 of gestation, NR cows had increased (P < 0.05) caruncular capillary surface density and decreased (P < 0.05) cotyledonary capillary area density, capillary number density, and capillary surface density compared with control cows. Although nutrient restriction had little effect on placental vascularity by d 125, upon realimentation, alterations in vascularity became apparent by d 250 of gestation, suggesting a placental programming effect.     

Effect of post-insemination progesterone supplementation on pregnancy rate in dairy cows

Progesterone plays an important role in maintenance of pregnancy. It is hypothesized that insufficient progesterone early in pregnancy may result in embryonic loss, and that supplemental progesterone would decrease pregnancy loss in dairy cows. In Experiment 1, 84 cows and 16 heifers from a single dairy operation were selected randomly. Within each age category, controlled internal drug release (CIDR) devices were inserted into the vagina of every other female on Day 4 post-insemination and removed on Day 18 post-insemination. Transrectal ultrasonography was performed to determine pregnancy at 4 time periods [days 30 to 37 (week 5), days 44 to 51 (week 7), days 58 to 65 (week 9), and days 86 to 93 (week 13)]. Progesterone supplementation had no effect on pregnancy rate. In Experiment 2, there were no differences in progesterone concentrations between cows that did and did not receive a CIDR. Further, cows receiving CIDR devices did not have an increase in circulating progesterone concentrations 30 min or 1 h after CIDR insertion. It appears that progesterone supplementation does not increase circulating levels of progesterone in the early pregnant lactating dairy cow. Alterative methods to influence progesterone concentrations and/or early embryonic loss need to be investigated.     

Maternal nutrition during pregnancy influences offspring wool production and wool follicle development

The effects of maternal nutrition on offspring wool production (quality and quantity) were evaluated. Primiparous Rambouillet ewes (n = 84) were randomly allocated to 1 of 6 treatments in a 2 × 3 factorial design. Selenium treatment [adequate Se (ASe, 9.5 μg/kg of BW) vs. high Se (HSe, 81.8 μg/kg of BW)] was initiated at breeding, and maternal nutritional intake [control (CON, 100% of requirements) vs. restricted (60% of CON) vs. overfed (140% of CON)] was initiated at d 50 of gestation. Lamb birth weight was recorded at delivery, and all lambs were placed on the same diet immediately after birth to determine the effects of prenatal nutrition on postnatal wool production and follicle development. At 180 ± 2.2 d of age, lambs were necropsied and pelt weights were recorded. Wool samples were collected from the side and britch areas, whereas skin samples were collected from the side of each lamb only. Although Se status did not influence side staple length in males, female lambs born from ewes on the ASe treatment had a shorter staple length (P < 0.05) when compared with females from ewes on the HSe treatment. Maternal nutritional intake and Se status did not influence (P ≥ 0.23) wool characteristics on the britch. However, at the britch, wool from female lambs had a reduced comfort factor (P = 0.01) and a greater (P = 0.02) fiber diameter compared with wool from male lambs. Maternal Se supplementation, maternal nutritional plane, sex of the offspring, or their interactions had no effect (P > 0.13) on primary (29.10 ± 1.40/100 μm(2)) and secondary (529.84 ± 21.57/100 μm(2)) wool follicle numbers. Lambs from ASe ewes had a greater (P = 0.03) secondary:primary wool follicle ratio compared with lambs from HSe ewes (20.93 vs. 18.01 ± 1.00). Despite similar postnatal diets, wool quality was affected by maternal Se status and the maternal nutritional plane.     

Effects of maternal selenium supply and plane of nutrition during gestation on passive transfer of immunity and health in neonatal lambs

To investigate the influence of maternal Se supply and plane of nutrition on lamb morbidity, mortality, and passive transfer of IgG, pregnant ewe lambs were used in 2 experiments with 2 × 3 factorial treatment arrangements. Supplementation of Se began at breeding and was either adequate Se (ASe, 9.5 μg/kg of BW) or high Se (HSe, 81.8 μg/kg of BW) in Exp. 1 or ASe (11.5 μg/kg of BW) or HSe (77.0 μg/kg of BW) in Exp. 2. On d 50 or 40 of gestation for Exp. 1 or 2, respectively, ewes were assigned randomly to 1 of 3 nutritional planes: 60% (RES), 100% (control, CON), or 140% (HI) of NRC requirements. This resulted in the following treatments: ASe-RES, ASe-CON, ASe-HI, HSe-RES, HSe-CON, and HSe-HI. Upon parturition, lambs were separated from their dams and serum samples obtained. Lambs were fed artificial colostrum for the first 20 h and then placed on milk replacer and grain pellets until completion of the study (Exp. 1, 57 d; Exp. 2, 21 d). Twenty-four hours after parturition, lamb serum samples were collected for IgG analysis. All lambs were reared similarly and morbidity and mortality assessed. Main effects were considered significant when P ≤ 0.05. In Exp. 1, there was a Se × plane of nutrition interaction (P ≤ 0.01) for lamb morbidity from birth to weaning and for 24-h IgG concentration. Lambs from ASe-RES and HSe-HI ewes were treated more frequently (P < 0.01) for respiratory and gastrointestinal disease, and lambs from HSe-HI ewes had the smallest (P < 0.01) 24-h serum IgG concentration. In Exp. 1, lambs from HI ewes also had the greatest (P < 0.01) mortality rates from birth to weaning compared with lambs from CON and RES ewes. In Exp. 2, there was an effect (P < 0.01) of maternal plane of nutrition with lambs from RES ewes having increased 24-h IgG compared with lambs from CON and HI ewes. There was no effect of maternal Se supplementation on lamb 24-h IgG in Exp. 2; however, there was a Se × plane of nutrition interaction (P < 0.01) for morbidity. From birth to 21 d of age, lambs from ASe-CON ewes had fewer (P < 0.01) treatment days compared with lambs from any of the other treatment groups. There also tended (P = 0.08) to be an effect of maternal Se supplementation on lamb mortality with increased mortality observed in lambs from HSe ewes. Results from the studies show a restricted maternal plane of nutrition can increase lamb serum IgG concentration. Selenium results were not consistent between the 2 experiments and may be due to differences in maternal Se.     

Impacts on conceptus survival in a commercial swine herd

An estimated 30 to 40% of potential piglets are lost before farrowing in U.S. or European pig breeds. Because these studies were conducted in limited numbers of university research herds, we decided to characterize the timing, pattern, and extent of conceptus loss in a commercial swine herd in Iowa (Pig Improvement Company; Camborough Line). Sows (parities 2 to 14) were slaughtered on d 25 (n = 83), 36 (n = 78), or 44 (n = 83) of gestation. These days coincide with periods before, during, and after uterine capacity becomes limiting to conceptus survival. At slaughter, numbers of corpora lutea (CL) and uterine horn length were determined, and conceptuses were removed and evaluated. Uterine horn length and CL number did not differ among these days of gestation, averaging 217 cm and 26.6 CL, respectively. In contrast, numbers of conceptuses decreased (P < 0.05) from 15.8 on d 25 to 12.9 on d 36, then remained relatively constant through d 44 (12.1). Thus, conceptus survival averaged 60.2% on d 25, 50.1% on d 36, and 46.3% on d 44, based on numbers of CL present. There was a positive correlation (P < 0.001; r = 0.50) between numbers of viable conceptuses on d 25 and ovulation rate, but this association was completely lost by d 36 (P > 0.10) when uterine capacity becomes limiting. In agreement with this premise, uterine horn length and conceptus number were not associated on d 25 but exhibited positive correlations (P < 0.05) on d 36 (r = 0.36) and d 44 (r = 0.40). On all 3 d examined, the numbers of viable conceptuses were not associated with fetal weight but were negatively correlated (P < 0.05) with placental weight. Compared with the commonly reported values for ovulation rate and percentage conceptus loss in university research herds, values from these production animals were extremely high. Data suggest that throughout this period, larger litters were associated with conceptuses exhibiting small placentae. These data lend support to the concept that increased placental efficiency (fetal weight/placental weight) may contribute to increased litter size in the pig.     

Conceptus competition for uterine space: different strategies exhibited by the Meishan and Yorkshire pig

Our laboratory has demonstrated that Yorkshire placentae increase in size and surface area during the final third of gestation. In contrast, Meishan placental size remains constant during late gestation, but the density of blood vessels at the placental-endometrial interface increases markedly. Preliminary observations from our laboratory suggest that if one of two adjacent Meishan fetuses dies, the placenta of the remaining Meishan conceptus fails to increase its length of implantation or its placental weight or surface area. In contrast, if one of two adjacent Yorkshire fetuses dies, the adjacent conceptus accelerates its placental growth. The objective of this experiment was to document that Yorkshire, but not Meishan, conceptuses accelerate placental growth when adjacent fetuses are experimentally destroyed on d 40 of gestation. Straightbred Meishan (n = 5) and Yorkshire (n = 5) females were laparotomized and one uterine horn was randomly assigned to receive fetal crushing (treated horn); the other uterine horn served as a within-animal control. In the treated horn, every other fetus was then crushed through the uterine wall and the animals were allowed to recover. On d 111, animals were killed, uteri were recovered, and fetal weight, crown-rump length (CRL), placental weight, implantation site length, and placental surface area were recorded. Although there were no statistically significant differences in fetal weight or CRL observed between treated or control horns of females of either breed, there was a tendency for the fetuses in the treated uterine horn to be longer and heavier in both breeds. There were no differences in placental weight, placental surface area, or implantation site length between conceptuses in Meishan treated and control horns, which averaged 173.8+/-6.4 g, 1,162.7+/-35.9 cm2, and 19.0+/-0.4 cm, respectively. In contrast, placental weight, placental surface area, and implantation site length were increased (P < 0.05) in Yorkshire treated horns compared to Yorkshire control horns (306.1+/-26.0 g, 1,835+/-93.9 cm2, and 33.4+/-1.5 cm vs 253.7+/-13.4 g, 1,474.3+/-50.4 cm2, and 27.2+/-0.8 cm; respectively). These data confirm that Yorkshire conceptuses, but not Meishan conceptuses, accelerate placental growth when adjacent littermates perish as late as d 40 of gestation. These data indicate that differences exist in the strategies employed by Meishan and Yorkshire conceptuses in the competition for nutrients during gestation.