DOHaD: A MENAGERIE OF ADAPTATIONS AND PERSPECTIVES: Large animal models of developmental programming: sustenance, stress, and sex matter.

In brief: Developmental programming refers to the long-term programming of gene expression during fetal and postnatal development, resulting in altered organ function even into adulthood. This review describes how maternal and paternal sustenance and stress, as well as fetal sex, all matter in large animal models and affect developmental programming of the offspring. Abstract: Developmental programming is the concept that certain health outcomes throughout life can be linked to early fetal or postnatal development. Progress in understanding concepts and mechanisms surrounding developmental programming is heavily leveraged by the use of large animal models. Numerous large animal models have been developed that apply a host of different maternal stressors and, more recently, paternal stressors. Maternal nutrition is the most researched maternal stressor applied during gestation and includes both global nutrient supply and models that target specific macro- or micro- nutrients. The focus of this review is to provide an overview of the many large animal models of developmental programming and to discuss the importance of sex effects (including paternal contributions) in study design and data interpretation.

Effect of maternal overnutrition on predisposition to insulin resistance in the foal: Foal skeletal muscle development and insulin signaling.

Skeletal muscle plays an integral role in the ability of a horse to perform at high levels. Shifts in skeletal muscle development in response to maternal plane of nutrition may have substantial and lasting impacts on athletic performance and whole-body metabolism. Therefore, sixteen Quarter Horse mares were used in a completely randomized design and maintained at a body condition score (BCS) 6 until start of third trimester. On d 235 of gestation, mares were randomly assigned to receive one of two dietary treatments with a diet formulated to meet requirements during late gestation (CON; n = 8), and an overfed diet (HIGH; n = 8) where mares received an additional 40% above CON. Five h after parturition, foals were euthanized, and gluteus medius, triceps brachii, and semitendinosus were harvested for analyses. Gene expression was determined by qPCR and western immunoblotting was used to quantify total and phosphorylated forms of proteins involved in skeletal muscle metabolism with tubulin as the loading control. All data were analyzed using PROC MIXED of SAS. Foals from HIGH mares exhibited larger skeletal muscle fibers by area (P <0.05), and a shift in muscle fiber development towards type I slow twitch muscle fibers (P <0.05). Relative expression of glucose transporter 4 (GLUT4) was lower in HIGH foals compared to CON in gluteus medius (P = 0.05). Insulin receptor isoform B (INSR-B) and insulin-like growth factor 1 receptor (IGF1R) were greater in triceps brachii of HIGH foals compared to CON (P ≤ 0.03). Insulin receptor isoform A (INSR-A), however, tended to be lower in triceps brachii of HIGH compared to CON (P = 0.10). Ratios of phosphorylated to total extracellular signal-regulated protein kinase 1/2 (ERK1/2) and c-June N-terminal kinase (JNK) were higher in HIGH foals compared to CON (P ≤0.04) in gluteus medius. There were no differences observed for phosphorylated to total protein ratios in semitendinosus and triceps brachii muscles; however, total ERK1/2 tended to be elevated (P <0.10) in semitendinosus from CON foals compared to HIGH. There was no difference in phosphorylated or total protein kinase B (AKT) (P >0.14). These data indicate hypertrophy of skeletal muscle fibers and a shift towards type I slow twitch fibers in HIGH foals. Furthermore, this study identifies muscle specific changes in gene expression and downstream insulin receptor signaling, which may contribute to future metabolic abnormalities in response to maternal overnutrition.

Effect of maternal overnutrition on predisposition to insulin resistance in the foal: Maternal parameters and foal pancreas histoarchitecture.

Results from previous studies indicate that maternal overnutrition during late gestation predisposes foals to metabolic disease, however, specific mechanisms resulting in disease remain unknown. Quarter Horse mares (n = 16), were randomly assigned to dietary treatments, beginning on gestational day 235, and consisted of a control group (CON- diet meeting nutrient requirement; n = 8) or an overfed diet (HIGH; n = 8) where mares received an additional 40 % above CON. On gestational days 285 and 315, an intravenous glucose tolerance test (FSIGTT) was conducted. Following parturition, foals were separated from the mare, prohibited from nursing, and an FSIGTT was conducted at 2 h postpartum. Foals were immediately euthanized and tissues preserved for analyses. There was no effect of treatment on foal BW (P = 0.50), pancreas weight (P = 0.60), or FSIGTT area under the curve for glucose (P = 0.80) and insulin (P = 0.70). Colocalization of α-amylase to isolate pancreatic islets of Langerhans indicated increased islet number and size in foals from HIGH mares (P < 0.01). Immunofluoresent analysis of insulin, glucagon, and somatostatin indicate no difference in intensity of staining (P> 0.10). Foals exposed to overnutrition during peak fetal growth had altered pancreatic islet development that may lead to adult-onset metabolic disease.

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

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

HORSE SPECIES SYMPOSIUM: Nutritional programming and the impact on mare and foal performance.

Many environmental factors can alter the phenotype of offspring when applied during critical periods of early development. In most domestic species, maternal nutrition influences fetal development and the fetus is sensitive to the nutrition of the dam during pregnancy. Many experimental models have been explored including both under- and overnutrition of the dam. Both nutritional strategies have yielded potential consequences including altered glucose tolerance, pancreatic endocrine function, insulin sensitivity, body composition, and colostrum quality. Although the impact of maternal nutrition on fetal development in the equine has not been thoroughly investigated, overnutrition is a common occurrence in the industry. Work in our laboratory has focused on effects of maternal overnutrition on mare and foal performance, mare DMI, foaling parameters, colostrum quality and passive transfer of immunity, and glucose and insulin dynamics. Over several trials, mares were fed either 100 or 140% of NRC requirements for DE, and supplemental Se and arginine were added to diets in an attempt to mitigate potential intrauterine growth retardation resulting from dams overfed during the last third of pregnancy. As expected, when mares were overfed, BW, BCS, and rump fat values increased. Foal growth over 150 d was also not influenced. Maternal nutrition did not alter colostrum volume but influenced colostrum quality. Maternal overnutrition resulted in lower colostrum IgG concentrations but did not cause failure of passive transfer in foals. Supplemental Se and arginine were unable to mitigate this reduction in colostrum IgG. Additionally, mare and foal glucose and insulin dynamics were influenced by maternal nutrition. Mare glucose and insulin area under the curve (AUC) increased with increased concentrate supplementation. Foal insulin AUC and peak insulin concentrations were increased when mares were fed concentrate and, in a later trial, foal peak glucose values were reduced with arginine supplementation of the mare. This influence of maternal nutrition on glucose and insulin dynamics warrants further investigation because it may be related to athletic performance and metabolic disease in the adult. Further studies will be necessary to fully elucidate the influence of mare nutrition during pregnancy on development of the fetus as well as long-term consequences of developmental programming.

Maternal metabolizable protein restriction during late gestation on uterine and umbilical blood flows and maternal and fetal amino acid concentrations near term in sheep.

To examine the effects of maternal metabolizable protein (MP) restriction during late gestation on uterine and umbilical blood flows, conceptus size, and amino acid concentrations in the uterine and umbilical vessels, 11 ewes with singleton pregnancies were assigned to one of three isocaloric diets formulated to provide 60% of MP (MP60), 80% of MP (MP80), or 100% of MP (MP100) requirements from days 100 to 130 of gestation. On day 130 of gestation, intraoperative uterine and umbilical blood flows were obtained as well as serum samples from the uterine artery, uterine vein, umbilical artery, and umbilical vein. Ewes on the MP60 diet had lighter (P=0.04) and smaller (P≤0.05) fetuses, but increased (P=0.02) uterine blood flow relative to fetal weight compared with MP100 ewes, with MP80 being intermediate. Umbilical blood flow was similar (P=0.70) across treatments. Glutamine, glycine, isoleucine, leucine, ornithine, serine, and valine concentrations were impacted (P≤0.02) by maternal treatment. While uterine flux of total serum nitrites was greater (P=0.03) in MP60 and MP80 ewes compared with MP100 ewes, fetal flux did not differ. Decreased maternal protein intake resulted in less (P<0.01) maternal cytochrome P450 1A enzyme activity. There were minimal impacts of maternal diet on steroid concentrations. Maternal dietary protein may alter fetal growth by impacting placental vasculature function and nutrient absorptive capabilities.

Influence of maternal plane of nutrition on mares and their foals: determination of mare performance and voluntary dry matter intake during late pregnancy using a dual-marker system.

Thirty pregnant mares (538 to 695 kg BW; 4 to 19 yr of age) were used to evaluate the effects of plane of nutrition on DMI of hay and mare performance (BW, BCS, and rump fat) during the last third of pregnancy. Mares were divided into 4 blocks by their expected foaling date and randomly assigned within block to either a hay or concentrate plus hay diet (concentrate fed at 0.75% BW, as-fed basis) with 15 mares per treatment. Treatments began 110 d before expected foaling date (230 d of gestation) and terminated at parturition. Mares were housed by block and allowed ad libitum access to coastal Bermuda grass (C. dactylon) hay, and concentrate-supplemented mares were fed twice daily in individual stalls. Performance variables were recorded every 14 d, with the last measurements obtained before foaling being considered a prepartum measurement. To evaluate DMI of hay, a dual-marker system was used at 9, 10, and 11 mo of gestation. Titanium dioxide was dosed at 10 g for 14 d. Fecal grab samples were obtained on the last 4 d twice daily via rectal palpation at 12-h intervals with times advancing 3 h each day to account for diurnal variation and to ultimately represent a 24-h period. Fecal samples were analyzed for TiO2 using a colorimetric procedure. Fecal, concentrate, and hay samples were also analyzed for acid detergent insoluble ash. Treatment tended to influence prepartum BW (P = 0.09) and affected prepartum BCS (P < 0.01) and rump fat (P = 0.01), with hay-fed mares having decreased BW and BCS from d 0 (beginning of feeding trial or d 230 of gestation) until parturition, whereas mares fed concentrate gained BW and BCS (P < 0.01). Mares fed only hay consumed 2.3% BW of forage compared with 1.8% BW for concentrate-fed mares (P < 0.01). Regardless of treatment, month of gestation influenced forage intake (P < 0.06), with mares consuming less during the 10th month of gestation and more in the 11th month (1.9% and 2.2% BW, respectively). These data indicate that the altered plane of nutrition of mares in late gestation influenced mare performance. Furthermore, DMI of hay was influenced by both diet and month of gestation. Continued research investigating manipulation of maternal nutrition and its effects on DMI would be beneficial to completely understand the relationships of these observations.

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

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

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

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

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.

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.

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.

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.

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

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

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.

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 /= 0.15) 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.

Characterization of a colostrum replacer and a colostrum supplement containing IgG concentrate and growth factors.

The objective of this study was to characterize absorption of colostrum replacer (CR) or supplement (CS) containing fractions of bovine plasma. Immunoglobulin concentrate (IGC) was prepared from bovine blood to a final purity of approximately 90% IgG. Blood was also processed to produce a fraction containing elevated concentrations of IGF-I and TGF-beta (GF). Both IGC and GF were spray-dried and blended with other ingredients to produce CR (30% IgG) or CS (15% IgG) containing 0 or 5% GF. Holstein bull calves were assigned to one of five treatments: 1) Pooled colostrum (MC)--1.9 L of pooled colostrum at 1 and 8 h of age to provide over 150 g of IgG; 2) Low supplement (LC)--1.9 L of CS at 1 and 8 h of age to provide 150 g of IgG; 3) Low supplement + GF (LG)--1.9 L of a CS with GF at 1 and 8 h of age to provide 150 g of IgG; 4) High supplement (HC)--1.9 L of CR at 1 h of age to provide 150 g of IgG and 1.9 L of a commercial milk replacer (MR) at 8 h of age; and 5) High supplement + GF (HG)--1.9 L of a CR with GF at 1 h of age to provide 150 g of IgG and 1.9 L of MR at 8 h of age. Apparent efficiency of IgG absorption was higher for calves fed HC and HG compared with those fed LC and LG and was lower for calves fed LG and HG compared with those fed LC and HC. IgG concentrations at 24 h were highest in calves fed MC compared with other calves and were higher in calves fed HC and HG compared to LC and LG. Calves fed LG and HG had lower IgG concentrations at 24 h of age compared with those fed LC and HC. These results indicate that 150 g of IgG provided in one dose soon after birth is superior to 150 g of IgG fed in two doses 7 h apart. Also, addition of a plasma fraction containing elevated concentrations of IGF-I and TGF-beta decreased IgG absorption in the neonatal calf.

Compromised development of calves (Bos gaurus) derived from in vitro-generated embryos and transferred interspecifically into domestic cattle (Bos taurus).

Advanced reproductive technologies, incuding IVF and interspecies embryo transfer, are becoming increasingly important for the preservation of endangered species. Previous attempts at interspecies transfers between Bos gaurus and Bos taurus have yielded compromised offspring. The goal of this investigation was to characterize the effects of interspecies transfer of IVF-derived embryos on subsequent neonatal outcome. To achieve this goal, fresh Bos gaurus IVF-derived embryos were transferred into Holstein (Bos taurus) recipients. Four fetuses were carried to term. Calf weight, temperature, heart rate, and respiration rate were recorded after birth. Blood samples also were obtained for determination of blood glucose, pH, packed cell volume (PCV), total hemoglobin (tHB), PO2, and PCO2. After parturition, milk production and health status of the recipients were recorded. Two calves were alive at birth, and two calves were stillborn. One of the calves that was born alive died within minutes after birth, while the other lived until approximately 26 h of age. Blood samples obtained from the calf that lived for 26 h showed it to be extremely acidotic and hypoglycemic; this calf also had marked difficulty thermoregulating. At necropsy, all calves showed evidence of in utero gasping and hypoxia, suggestive of premature placental separation. None of the recipient cows showed typical signs of impending parturition. After parturition, lactogenesis in all recipient cows was markedly decreased. On gross examination, placentae resulting from the interspecies transfers had fewer cotyledons that were also much larger in size compared to cotyledons from normal gaur placentae. Calves in this study had abnormalities consistent with those noted from previous interspecies transfers and with IVF and nuclear transfer (cloned) calves. Due to the design of this study, it is not possible to differentiate between problems resulting from the IVF process and those resulting from potential interspecies incompatibilities. However, interspecies transfers of in vitro-produced gaur embryos into Bos taurus are strongly discouraged.