K. A. Vonnahme

Maternal Undernutrition from Early- to Mid-Gestation Leads to Growth Retardation, Cardiac Ventricular Hypertrophy, and Increased Liver Weight in the Fetal Sheep

Abstract Early gestation is critical for placentomal growth, differentiation, and vascularization, as well as fetal organogenesis. The fetal origins of adult disease hypothesis proposes that alterations in fetal nutrition and endocrine status result in developmental adaptations that permanently change structure, physiology, and metabolism, thereby predisposing individuals to cardiovascular, metabolic, and endocrine disease in adult life. Multiparous ewes were fed to 50% (nutrient restricted) or 100% (control fed) of total digestible nutrients from Days 28 to 78 of gestation. All ewes were weighed weekly and diets adjusted for individual weight loss or gain. Ewes were killed on Day 78 of gestation and gravid uteri recovered. Fetal body and organ weights were determined, and numbers, morphologies, diameters, and weights of all placentomes were obtained. From Day 28 to Day 78, restricted ewes lost 7.4% of body weight, while control ewes gained 7.5%. Maternal and fetal blood glucose concentrations were reduced in restricted versus control pregnancies. Fetuses were markedly smaller in the restricted group than in the control group. Further, restricted fetuses exhibited greater right- and left-ventricular and liver weights per unit fetal weight than control fetuses. No treatment differences were observed in any gross placentomal measurement. However, caruncular vascularity was enhanced in conceptuses from nutrient-restricted ewes but only in twin pregnancies. While these alterations in fetal/placental development may be beneficial to early fetal survival in the face of a nutrient restriction, their effects later in gestation as well as in postnatal life need further investigation.

Evidence for altered placental blood flow and vascularity in compromised pregnancies

The placenta is the organ that transports nutrients, respiratory gases, and wastes between the maternal and fetal systems. Consequently, placental blood flow and vascular development are essential components of normal placental function and are critical to fetal growth and development. Normal fetal growth and development are important to ensure optimum health of offspring throughout their subsequent life course. In numerous sheep models of compromised pregnancy, in which fetal or placental growth, or both, are impaired, utero‐placental blood flows are reduced. In the models that have been evaluated, placental vascular development also is altered. Recent studies found that treatments designed to increase placental blood flow can ‘rescue’ fetal growth that was reduced due to low maternal dietary intake. Placental blood flow and vascular development are thus potential therapeutic targets in compromised pregnancies.

Effects of maternal nutrition on conceptus growth and offspring performance: Implications for beef cattle production1

Developmental programming is the concept that a maternal stimulus or insult at a critical period in fetal development has long-term effects on the offspring. Historically, considerable effort has been made to understand how nutrition influences health and productivity during the postnatal period. Whereas maternal nutrition during pregnancy plays an essential role in proper fetal and placental development, less is known about how maternal nutrition affects the health and productivity of the offspring. Conceptus growth is sensitive to direct and indirect effects of maternal dietary intake. Even from the earliest stages of embryonic life, when nutrient requirements for conceptus growth are negligible, alterations in tissue composition can occur, influencing future growth of the compromised organ system. Not only is neonatal health compromised, but subsequent health may also be programmed because offspring from undernourished dams have exhibited poor growth and productivity and have developed significant diseases later in life. Although the literature is now evolving, with increasing evidence of how maternal nutrient restriction impairs several prenatal physiological variables, few studies have evaluated postnatal growth and development in livestock species, and fewer have evaluated it in beef cattle. In addition, very few studies have evaluated restriction of specific components of the diet during pregnancy (such as protein) on offspring growth and performance. This review focuses on how maternal nutrition affects conceptus growth and postnatal responses in beef cattle.

Placental angiogenesis in sheep models of compromised pregnancy

Because the placenta is the organ that transports nutrients, respiratory gases and wastes between the maternal and fetal systems, development of its vascular beds is essential to normal placental function, and thus in supporting normal fetal growth. Compromised fetal growth and development have adverse health consequences during the neonatal period and throughout adult life. To establish the role of placental angiogenesis in compromised pregnancies, we first evaluated the pattern of placental angiogenesis and expression of angiogenic factors throughout normal pregnancy. In addition, we and others have established a variety of sheep models to evaluate the effects on fetal growth of various factors including maternal nutrient excess or deprivation and specific nutrients, maternal age, maternal and fetal genotype, increased numbers of fetuses, environmental thermal stress, and high altitude (hypobaric) conditions. Although placental angiogenesis is altered in each of these models in which fetal growth is adversely affected, the specific effect on placental angiogenesis depends on the type of ‘stress’ to which the pregnancy is subjected, and also differs between the fetal and maternal systems and between genotypes. We believe that the models of compromised pregnancy and the methods described in this review will enable us to develop a much better understanding of the mechanisms responsible for alterations in placental vascular development.

Developmental programming: The concept, large animal models, and the key role of uteroplacental vascular development

Developmental programming refers to the programming of various bodily systems and processes by a stressor of the maternal system during pregnancy or during the neonatal period. Such stressors include nutritional stress, multiple pregnancy (i.e., increased numbers of fetuses in the gravid uterus), environmental stress (e.g., high environmental temperature, high altitude, prenatal steroid exposure), gynecological immaturity, and maternal or fetal genotype. Programming refers to impaired function of numerous bodily systems or processes, leading to poor growth, altered body composition, metabolic dysfunction, and poor productivity (e.g., poor growth, reproductive dysfunction) of the offspring throughout their lifespan and even across generations. A key component of developmental programming seems to be placental dysfunction, leading to altered fetal growth and development. We discuss various large animal models of developmental programming and how they have and will continue to contribute to our understanding of the mechanisms underlying altered placental function and developmental programming, and, further, how large animal models also will be critical to the identification and application of therapeutic strategies that will alleviate the negative consequences of developmental programming to improve offspring performance in livestock production and human medicine.

Effects of gestational plane of nutrition and selenium supplementation on mammary development and colostrum quality in pregnant ewe lambs

To examine effects of nutritional plane and Se supplementation on colostrum quality and mammary development, individually fed, pregnant Rambouillet ewe lambs were allotted randomly to 1 of 6 treatments in a 2 x 3 factorial arrangement. Main effects included dietary Se level, which began at breeding (d = 0) [adequate Se (9.5 mug/kg of BW) vs. high Se (81.8 mug/kg of BW)], and plane of nutrition, which began at d 50 of gestation [60% (RES), 100% (CON), and 140% (HIGH) of requirements]. Upon parturition, lambs were immediately separated from dams and weighed. Three hours after lambing, colostrum yield was determined, and samples were obtained for components and immunoglobulin G (IgG) analysis. Ewes were slaughtered within 24 h of parturition, and mammary tissues were collected for determination of alveolar secretory epithelial cell proliferation index and luminal area. Gestation length was reduced (P < 0.01) in HIGH ewes compared with RES and CON ewes. Although birth weights were reduced (P < 0.01) in RES and HIGH compared with CON ewes, there was little effect of diet on placental size. Mammary gland weight was reduced (P </= 0.05) in RES compared with CON and HIGH, which were similar. However, when expressed as grams per kilogram of BW, mammary gland weight in HIGH ewes was less (P = 0.03) compared with RES and CON. Colostrum weight and volume were reduced (P < 0.01) in RES and HIGH ewes compared with CON. Although colostrum IgG concentration was greater in RES ewes compared with CON and HIGH, total IgG was lower (P </= 0.06) in RES and HIGH compared with CON ewes. The percentage of alveolar cells proliferating was increased (P < 0.04) in HIGH compared with RES ewes, with CON being intermediate. Percentage of alveoli luminal area per unit tissue area was increased (P = 0.04) in RES compared with HIGH and CON ewes, which did not differ. Selenium had no effect (P >/= 0.15) on mammary gland weight, colostrum quantity, or IgG concentration in pregnant ewe lambs. Improper nutrition from mid to late pregnancy in ewe lambs altered colostrum quality and quantity and reduced offspring birth weight, which may have negative implications for lamb health and survival during the early postnatal period.

Effects of plane of nutrition and selenium supply during gestation on ewe and neonatal offspring performance, body composition, and serum selenium

To investigate the effects of nutritional plane and Se supply during gestation on ewe and offspring performance and body composition, 84 Rambouillet ewe lambs (age = 240 +/- 17 d, BW = 52.1 +/- 6.2 kg) were allocated to a 2 x 3 x 2 factorial arrangement of treatments. Factors included Se [adequate Se (ASe, 11.5 microg/kg of BW) or high Se (HSe, 77.0 microg/kg of BW)] initiated at breeding, nutritional plane [60% (restricted, RES), 100% (control, CON), or 140% (high, HIH) of NRC requirements] initiated at d 40 of gestation, and physiological stage at necropsy [3 to 24 h postpartum or d 20 of lactation]. Ewes were fed and housed individually in a temperature-controlled facility. At parturition, all lambs were removed and artificially reared until necropsy on d 20.6 +/- 0.9 of age. Ewes assigned to the treatment at d 20 of lactation were transitioned to a common diet meeting lactation requirements and were mechanically milked. From d 95 of gestation through parturition and d 20 of lactation, ewe BW and BCS were least (P <or= 0.01) in the RES treatment, intermediate in the CON treatment, and greatest in the HIH treatment. Ewes fed HSe had a greater (P <or= 0.05) BCS increase than those fed ASe during mid- and late gestation. During gestation, ewes in the HIH treatment had the greatest (P < 0.001) ADG and G:F, those in the CON treatment were intermediate, and those in the RES treatment were least, whereas ewes fed HSe had greater (P < 0.001) ADG and G:F than those fed ASe during midgestation. Ewe backfat and LM area on d 135 of gestation were least (P < 0.001) in the RES treatment, intermediate in the CON treatment, and greatest in the HIH treatment, with ewes fed HSe having greater (P <or= 0.03) backfat than those fed ASe. During the first 20 d of lactation, ewes fed the RES diet had greater (P < 0.09) G:F than those fed the CON and HIH diets. Physiological stage had no effect on ewe omental and mesenteric fat or perirenal fat weights, although both were greater (P < 0.001) for ewes fed the HIH diet than for those fed the RES and CON diets. At birth, lambs born to ewes in the RES group weighed less and had decreased curved crown rump lengths (P = 0.08) compared with those born to ewes in the CON and HIH groups, and lambs from ewes in the ASe-RES treatment were lighter (P < 0.08) than those from ewes in the HSe-RES, ASe-CON, and ASe-HIH treatments. Lambs from dams in the RES group had less (P < 0.05) BW from d 7 to 19 and decreased (P < 0.07) overall ADG compared with lambs from dams in the CON and HIH groups. Additionally, lambs from dams in the RES group had less (P <or= 0.08) perirenal fat than their counterparts, and lambs from dams in the HIH group had greater (P = 0.01) omental and mesenteric fat than lambs from dams in the RES group. Postpartum serum Se of ewes and lambs (birth and d 19) was increased (P < 0.001) by HSe feeding during gestation. Results indicate that BW differences in pregnant ewes attributable to nutritional plane are accompanied by changes in body composition and offspring BW, both of which may be affected by Se supply.

Effects of early gestational undernutrition on fetal growth, organ development, and placentomal composition in the bovine

Fetal intrauterine growth restriction (IUGR) is known to negatively affect offspring health postnatally. This study evaluated the impacts of early gestational undernutrition followed by realimentation on bovine fetal and placental growth. Thirty multiparous beef cows bred to a single sire and gestating female fetuses were fed to meet NRC recommendations (control; n = 15) or fed below NRC recommendations (68.1% of NE(m) and 86.7% of MP recommendations; nutrient restricted, NR; n = 15) from d 30 to 125 of gestation. On d 125 of gestation, 10 control and 10 NR cows were necropsied. The remaining 5 NR cows were realimented to achieve similar BW and BCS with the remaining 5 control cows by d 190 of gestation; both groups were necropsied at d 245 of gestation. Fetal weight at d 125 of gestation was 948 +/- 14 g (n = 10) for control cows; however, fetal weights of NR cows fell into 2 distinct groups: NR non-IUGR cows had fetal weights similar to control cows (974 +/- 20 g, n = 6), whereas fetal weights of NR IUGR cows were reduced (773 +/- 23 g, n = 4; P < 0.01). Fetal brain weight as a percentage of fetal weight was increased (approximately 11%; P < 0.01) in the NR IUGR fetuses compared with fetuses from the other 2 groups, which were similar. Fetal heart weight as a percentage of fetal weight also tended to be increased (approximately 10%; P = 0.08) in NR IUGR fetuses compared with control fetuses. Nutrient-restricted IUGR cows exhibited reduced (P < 0.01) cotyledonary weights compared with NR non-IUGR and control cows, which were similar (192 +/- 27 vs. 309 +/- 22, and 337 +/- 17 g, respectively). Total placentome surface area also tended to be reduced (P = 0.07) in NR IUGR cows compared with NR non-IUGR and control cows, which again were similar (685.0 +/- 45.6 vs. 828.7 +/- 37.2 and 790.7 +/- 28.9 mm(2), respectively). On d 245 of gestation, fetal weights and caruncle weight were similar for NR and control cows; cotyledonary weights, however, were reduced in NR vs. control cows (1,430 +/- 133 vs. 2,137 +/- 133 g, P < 0.01). Decreased fetal growth in NR IUGR cows on d 125 of gestation was associated with decreased cotyledonary weights and reduced placentomal surface areas. The return of NR cows to a BW and BCS similar to that of control cows through realimentation beginning on d 126 resulted in similar fetal weights of NR and control cows by d 245 of gestation. Thus, a bout of fetal IUGR may go undetected if cows undernourished during early gestation receive feed supplementation in the second half of gestation to assure normal birth weight.

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.

Circulating levels of nitric oxide and vascular endothelial growth factor throughout ovine pregnancy

Nitric oxide (NO) production has been shown to increase uterine blood flow and be elevated in ewes carrying multiple fetuses during late gestation. Vascular endothelial growth factor (VEGF) has been reported to increase eNOS expression and NO production in endothelial cell cultures. As angiogenesis and vasodilatation of the uterine and placental vascular beds are important at all stages of pregnancy, it is important to understand how VEGF and NO change throughout gestation in circulation. Therefore the objectives of the current study were to evaluate the systemic levels of VEGF and NO metabolite (NOx) throughout ovine gestation and to determine if there was an effect of sheep carrying singletons versus multiple fetuses. NOx and VEGF concentrations were analysed in systemic blood from pregnant ewes starting on day 27 of pregnancy and at multiple intermittent intervals throughout pregnancy until term. Blood samples from non‐pregnant and postpartum ewes were also analysed. NOx concentrations in maternal blood expressed a biphasic pattern with NOx concentrations increasing (P < 0.05) over non‐pregnant values on days 40–69 of gestation, returning to non‐pregnant concentrations from days 70–100, and again increasing (P < 0.05) until term. Postpartum NOx concentrations were similar to non‐pregnant values. While ewes carrying multiple fetuses had increased (P < 0.05) concentrations of NOx on days 60–69, there were no differences in NOx concentrations in ewes carrying singletons or multiples from day 70–99 of gestation. Starting on day 100 and continuing throughout the duration of pregnancy, ewes carrying multiple fetuses had increased (P < 0.05) concentrations of NOx compared to ewes carrying singletons. Concentrations of VEGF showed a different pattern from NOx with VEGF decreasing (P < 0.05) from day 20–69 of pregnancy compared to non‐pregnant ewes. Concentrations of VEGF returned to non‐pregnant levels by day 70 and remained constant throughout the duration of pregnancy. On days 20–39, ewes carrying singleton fetuses had an increased VEGF concentration (P < 0.05), whereas ewes carrying multiple fetuses demonstrated elevated VEGF concentrations from day 90–109 of gestation. Concentrations from non‐pregnant and postpartum ewes did not differ (P > 0.1). While there was no effect of fetal number on circulating VEGF concentrations, circulating levels of NOx were substantially increased (P < 0.05) in ewes carrying multiple fetuses, compared to ewes carrying singletons. The pattern of the rise in NOx in circulating plasma was not directly associated with changes in VEGF regardless of the number of fetuses present. However, circulating concentrations of NOx and VEGF appear to, respectively, follow patterns of uterine blood flow and angiogenesis of the uterus. An understanding of these circulatory patterns may have important implications for fetal size, birth weight and fetal/developmental origins of adult disease.