C. J. Hammer

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.

Nutritional plane and selenium supply during gestation affect yield and nutrient composition of colostrum and milk in primiparous ewes.

The objectives were to investigate effects of nutritional plane and Se supply during gestation on yield and nutrient composition of colostrum and milk in first parity ewes. Rambouillet ewe lambs (n = 84, age = 240 ± 17 d, BW = 52.1 ± 6.2 kg) were allocated to 6 treatments in a 2 × 3 factorial array. 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 [60 (RES), 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. Colostrum was milked from all ewes at 3 h postpartum, and one-half of the ewes (n = 42) were transitioned to a common diet meeting lactation requirements and mechanically milked for 20 d. Colostrum yield was greater (P = 0.02) for HSe ewes than ASe, whereas CON had greater (P < 0.05) colostrum yield than RES and HIH. Colostrum Se (%) was greater (P < 0.01) for HSe than ASe. Colostrum from ewes fed HSe had less (P = 0.03) butterfat (%), but greater (P ≤ 0.05) total butterfat, solids-not-fat, lactose, protein, milk urea N, and Se than ASe. Colostrum from HIH ewes had greater (P ≤ 0.02) solids-not-fat (%) than RES, whereas RES had greater (P ≤ 0.04) butterfat (%) than CON and HIH. Colostrum from ewes fed the CON diet had greater (P = 0.01) total butterfat than HIH. Total solids-not-fat, lactose, and protein were greater (P < 0.05) in colostrum from CON than RES and HIH. Ewes fed HSe had greater (P < 0.01) milk yield (g/d and mL/d) than ASe, and CON and HIH had greater (P < 0.01) yield than RES. Milk protein (%) was greater (P ≤ 0.01) in RES compared with CON or HIH. Ewes fed HSe had greater (P < 0.01) milk Se (µg/g and mg/d) than ASe on each sampling day. Milk from CON and HIH ewes had greater (P < 0.01) total solids-not-fat, lactose, protein, and milk urea N than RES. Total Se was greater (P = 0.02) in milk from ewes fed the CON diet compared with RES. Somatic cell count and total somatic cells were greater (P ≤ 0.05) in milk from CON than RES. A cubic effect of day (P ≥ 0.01) was observed for milk yield (g and mL). Butterfat, solids-not-fat, lactose, milk urea N, and Se concentration responded quadratically (P ≤ 0.01) to day. Protein (%), total butterfat, and total Se, and somatic cells (cells/mL and cells/d) decreased linearly (P < 0.01) with day. Results indicate that gestational nutrition affects colostrum and milk yield and nutrient content, even when lactational nutrient requirements are met.

Ovine offspring growth and diet digestibility are influenced by maternal selenium supplementation and nutritional intake during pregnancy despite a common postnatal diet

Lambs born from feed-restricted or overfed ewes can be lighter at birth, whereas maternal Se supplementation can increase fetal size near term. We hypothesized that birth weight would be inversely related to feed efficiency and growth rates during postnatal development. To examine the effects of maternal dietary Se and nutrient restriction or excess on postnatal lamb growth, diet digestibility, and N retention, 82 ewe lambs (52.2 ± 0.8 kg) were allotted randomly to 1 of 6 treatments in a 2 × 3 factorial arrangement. Factors were dietary Se [adequate Se (9.5 μg/kg of BW; ASe) vs. high Se (Se-enriched yeast; 81.8 μg/kg of BW; HSe)] and maternal nutritional intake [60% (restricted, RES), 100% (control, CON), or 140% (high, HI) of NRC requirements]. Selenium treatments began at breeding. Nutritional treatments began on d 50 of gestation. Lambs were immediately removed from their dams at parturition, provided artificial colostrum, and fed milk replacer until weaning. After weaning, lambs were maintained using common management and on common diets until necropsy at 180 d. Male and female lambs from RES-fed ewes were lighter (P ≤ 0.03) at birth than lambs from CON-fed ewes, with lambs from HI-fed ewes being intermediate. Although maternal nutritional intake influenced (P < 0.06) BW gain before weaning on d 57, both maternal nutritional intake and sex of offspring influenced (P ≤ 0.09) BW gain from d 57 to 180. Although maternal nutritional intake did not influence (P ≥ 0.35) female lamb BW gain, male lambs from RES-fed ewes were lighter (P ≤ 0.09) than those from CON-fed ewes until d 162. By d 180, male lambs from RES- and HI-fed ewes were lighter (P ≤ 0.09) than those from CON-fed ewes. In a subset of lambs used in a feed efficiency study, namely, those born to ASe ewes, HI maternal nutritional intake decreased (P ≤ 0.09) ADG and G:F compared with lambs born to RES- and CON-fed ewes, which did not differ (P ≥ 0.60). Conversely, when lambs were born to HSe ewes, HI maternal nutritional intake increased (P ≤ 0.01) ADG and G:F compared with CON, with RES being intermediate. Moreover, lambs born to ASe-HI ewes had decreased (P < 0.01) ADG and G:F compared with lambs born to HSe-HI ewes. Furthermore, male lambs had a greater (P < 0.01) G:F than female lambs. Maternal diet did not affect (P ≥ 0.11) N retention in male lambs. These data indicate that maternal nutrition during gestation and sex of the offspring alter postnatal growth and efficiency of growth in offspring despite similar postnatal management.

Effect of selenium supplementation and plane of nutrition on mares and their foals: foaling data.

To investigate the maternal plane of nutrition and role of Se yeast on foaling variables and passive transfer of IgG, 28 Quarter Horse mares were used in a study with a randomized complete block design. Mares were blocked by expected foaling date and assigned randomly within block to dietary treatments. Dietary treatments were arranged as a 2 x 2 factorial with 2 planes of nutrition, pasture or pasture + grain mix (fed at 0.75% of BW on an as-fed basis) and 2 concentrations of Se yeast (0 or 0.3 mg/kg of DMI). This resulted in 4 treatments: pasture (PA), pasture + Se (PS), pasture + grain mix (PG), and pasture + grain mix + Se (PGS). Assuming DMI at 2% of BW, the mares fed PA and PS received approximately 100% of the calculated NRC (2007) DE requirements, whereas PG and PGS received 120%. Selenium supplementation began 110 d before the estimated foaling date, and all dietary treatments were terminated at parturition. At parturition, foaling variables were recorded. Additionally, placental weight was recorded and 2 samples from each placenta were collected for analysis of DNA, RNA, and protein. Colostrum was obtained for fat, protein, milk urea N, somatic cell count, and IgG analyses. Foal blood samples were collected at 0, 6, 12, 18, and 24 h after parturition for IgG analysis. There was no effect (P >or= 0.21) of Se or plane of nutrition on foaling variables; however, foal BW as a percentage of mare BW tended (P = 0.10) to be reduced in foals from mares on grain mix (PG and PGS; 7.6%) compared with mares not fed grain mix (PA and PS; 8.0%). There was also no effect (P >or= 0.20) of Se or plane of nutrition on placental cell number (mg of DNA/g), potential cellular activity (RNA:DNA), expulsion time, or weight. However, mares fed supplemental Se (PS and PGS) had decreased (P = 0.02) placental cell size (24.1 mg of protein/mg of DNA) compared with mares not fed supplemental Se (PA and PG; 32.5 mg of protein/mg of DNA). There was also no effect (P >or= 0.18) of Se or plane of nutrition on colostral fat, protein, milk urea N, or somatic cell count. However, mares fed grain mix (PG and PGS) had less (P = 0.03) colostral IgG (76.5 g/L) compared with mares not fed grain mix (PA and PS; 126.6 g/L). Foals from mares fed grain (PG and PGS) tended (P = 0.06) to have less overall serum IgG (13.6 g/L) compared with foals from mares not fed grain (PA and PS; 15.3 g/L). These data indicate that the maternal diet during the last one-third of gestation affects placental efficiency and colostral IgG.

Impacts of maternal selenium and nutritional level on growth, adiposity, and glucose tolerance in female offspring in sheep

To examine effects of maternal nutrition and Se intake on adiposity and insulin sensitivity in female offspring, treatments were imposed during gestation on 82 pregnant primiparous Rambouillet ewe lambs (52.2 ± 0.8 kg) allotted randomly to 1 of 6 treatments in a 2 × 3 factorial arrangement. Factors were adequate (9.5 μg Se·kg BW(-1)·d(-1); ASe) or high (81.8 μg Se·kg BW(-1)·d(-1); HSe) levels of dietary Se (Se-enriched yeast) and maternal nutritional intake (100% of metabolizable energy [ME] requirement [MOD], 60% of MOD [LOW], and 140% of MOD [HIGH]). Selenium treatments were initiated at breeding and global nutritional treatments at day 50 of gestation. At parturition, lambs were removed from ewes before nursing and managed similarly. Glucose tolerance tests were performed at 107 and 148 d of age. Necropsies were performed at 180 d of age. Although there was no effect of Se on maternal body condition or weight during gestation, both maternal nutritional intake and selenium treatment influenced (P ≤ 0.04) offspring growth and response to a glucose tolerance test. Female lambs from HSe ewes were heavier (P = 0.04) at birth. There were nutritional intake and Se interactions (P ≤ 0.05) on the growth rate of the lambs and their insulin response to a glucose bolus at 2 different times during growth. By 180 d, ewe lambs from HSe ewes had more (P ≤ 0.07) internal fat stores than lambs from ASe ewes. It appears that both maternal nutritional level and Se intake can influence insulin sensitivity, and maternal Se intake alone can enhance fat deposition in female offspring.

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.

Effect of selenium supplementation and plane of nutrition on mares and their foals: Selenium concentrations and glutathione peroxidase

To investigate the maternal plane of nutrition and role of Se yeast on muscle Se concentration, plasma glutathione peroxidase (Gsh-Px) activity, and colostrum Se concentration in mares and their foals, 28 Quarter Horse mares (465 to 612 kg of BW, and 6 to 19 yr of age) were used in a study with a randomized complete block design. Mares were blocked by expected foaling date and randomly assigned to dietary treatments within blocks. Dietary treatments were arranged as a 2 x 2 factorial with 2 planes of nutrition, pasture or pasture + grain mix (fed at 0.75% of BW on an as-fed basis) and 2 concentrations of Se yeast supplementation (0 or 0.3 mg/kg of DMI), resulting in 4 treatments: pasture, pasture + grain mix, pasture + grain mix + Se, or pasture + Se. Mares fed diets of pasture and pasture + Se received approximately 100% of the calculated NRC (2007) DE requirements, whereas mares fed diets of pasture + grain mix and pasture + grain mix + Se received 120%. Selenium supplementation began 110 d before the estimated foaling date and treatments were terminated at parturition. Blood and muscle (biopsy) samples were collected on d 0 and then every 14 or 28 d, respectively, thereafter until parturition. Additionally, BW, BCS, and rump fat (RF) were recorded every 14 d. At parturition, colostrum, foal plasma, and foal muscle samples were collected and sampling continued every 14 d for plasma and every 28 d for muscle until d 56. Mare BW, BCS, and RF were affected by plane of nutrition (P <or= 0.02), but not by Se supplementation. Mares fed the grain mix had greater (P < 0.05) BW, BCS, and RF measurements throughout the experiment. Mare plasma, muscle, and colostrum Se concentrations were greater (P < 0.01) in mares fed Se. Mares fed the grain mix had greater plasma Se (P = 0.02) than mares on pasture alone. Mare and foal plasma Gsh-Px concentrations were not affected by treatment. Foal plasma and muscle Se concentrations were greater when dams were fed the supplemental grain mix (P = 0.04 and 0.02, respectively) and supplemental Se (P < 0.001). Results indicated that maternal plane of nutrition and Se supplementation affected mare and foal plasma, muscle, and colostrum Se concentrations, but not Gsh-Px activity.

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.