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Thirty-five crossbred Angus heifers (initial BW = 359.5 ± 7.1 kg) were randomly assigned to a 2 × 2 factorial design to evaluate effects of vitamin and mineral supplementation [VMSUP; supplemented (VTM) vs. unsupplemented (NoVTM)] and different rates of gain [GAIN; low gain (LG), 0.28 kg/d, vs. moderate gain (MG), 0.79 kg/d] during the first 83 d of gestation on dam hormone and metabolic status, fetal tissue and organ mass, and concentration of glucose and fructose in fetal fluids. The VMSUP was initiated 71 to 148 d before artificial insemination (AI), allowing time for mineral status of heifers to be altered in advance of breeding. At AI heifers were assigned their GAIN treatment. Heifers received treatments until the time of ovariohysterectomy (d 83 ± 0.27 after AI). Throughout the experiment, serum samples were collected and analyzed for non-esterified fatty acids (NEFA), progesterone (P4), insulin, and insulin-like growth factor 1 (IGF-1). At ovariohysterectomy, gravid reproductive tracts were collected, measurements were taken, samples of allantoic (ALF) and amniotic (AMF) fluids were collected, and fetuses were dissected. By design, MG had greater ADG compared to LG (0.85 vs. 0.34 ± 0.04 kg/d, respectively; p < 0.01). Concentrations of NEFA were greater for LG than MG (p = 0.04) and were affected by a VMSUP × day interaction (p < 0.01), with greater concentrations for NoVTM on d 83. Insulin was greater for NoVTM than VTM (p = 0.01). A GAIN × day interaction (p < 0.01) was observed for IGF-1, with greater concentrations for MG on d 83. At d 83, P4 concentrations were greater for MG than LG (GAIN × day, p < 0.01), and MG had greater (p < 0.01) corpus luteum weights versus LG. Even though fetal BW was not affected (p ≥ 0.27), MG fetuses had heavier (p = 0.01) femurs than LG, and VTM fetuses had heavier (p = 0.05) livers than those from NoVTM. Additionally, fetal liver as a percentage of BW was greater in fetuses from VTM (P = 0.05; 3.96 ± 0.06% BW) than NoVTM (3.79 ± 0.06% BW), and from LG (p = 0.04; 3.96 ± 0.06% BW) than MG (3.78 ± 0.06% BW). A VMSUP × GAIN interaction was observed for fetal small intestinal weight (p = 0.03), with VTM-MG being heavier than VTM-LG. Therefore, replacement heifer nutrition during early gestation can alter the development of organs that are relevant for future offspring performance. These data imply that compensatory mechanisms are in place in the developing conceptus that can alter the growth rate of key metabolic organs possibly in an attempt to increase or decrease energy utilization.

We evaluated the effects of vitamin and mineral supplementation (from pre-breeding to day 83 of gestation) and two rates of gain (from breeding to day 83 of gestation) on trace mineral concentrations in maternal and fetal liver, fetal muscle, and allantoic (ALF) and amniotic (AMF) fluids. Crossbred Angus heifers (n = 35; BW = 359.5 ± 7.1 kg) were randomly assigned to one of two vitamin and mineral supplementation treatments (VMSUP; supplemented (VTM) vs. unsupplemented (NoVTM)). The VMSUP factor was initiated 71 to 148 d before artificial insemination (AI), allowing time for the mineral status of heifers to be altered in advance of breeding. The VTM supplement (113 g·heifer−1·d−1) provided macro and trace minerals and vitamins A, D, and E to meet 110% of the requirements specified by the NASEM, and the NoVTM supplement was a pelleted product fed at a 0.45 kg·heifer−1·day−1 with no added vitamin and mineral supplement. At AI, heifers were assigned to one of two rates of gain treatments (GAIN; low gain (LG) 0.28 kg/d or moderate gain (MG) 0.79 kg/d) within their respective VMSUP groups. On d 83 of gestation fetal liver, fetal muscle, ALF, and AMF were collected. Liver biopsies were performed prior to VMSUP factor initiation, at the time of AI, and at the time of ovariohysterectomy. Samples were analyzed for concentrations of Se, Cu, Zn, Mo, Mn, and Co. A VMSUP × GAIN × day interaction was present for Se and Cu (p < 0.01 and p = 0.02, respectively), with concentrations for heifers receiving VTM being greater at AI and tissue collection compared with heifers not receiving VTM (p < 0.01). A VMSUP × day interaction (p = 0.01) was present for Co, with greater (p < 0.01) concentrations for VTM than NoVTM at the time of breeding. VTM-MG heifers had greater concentrations of Mn than all other treatments (VMSUP × GAIN, p < 0.01). Mo was greater (p = 0.04) for MG than LG, while Zn concentrations decreased throughout the experiment (p < 0.01). Concentrations of Se (p < 0.01), Cu (p = 0.01), Mn (p = 0.04), and Co (p = 0.01) were greater in fetal liver from VTM than NoVTM. Mo (p ≤ 0.04) and Co (p < 0.01) were affected by GAIN, with greater concentrations in fetal liver from LG than MG. In fetal muscle, Se (p = 0.02) and Zn (p < 0.01) were greater for VTM than NoVTM. Additionally, Zn in fetal muscle was affected by GAIN (p < 0.01), with greater concentrations in LG than MG. The ALF in VTM heifers (p < 0.01) had greater Se and Co than NoVTM. In AMF, trace mineral concentrations were not affected (p ≥ 0.13) by VMSUP, GAIN, or their interaction. Collectively, these data suggest that maternal nutrition pre-breeding and in the first trimester of gestation affects fetal reserves of some trace minerals, which may have long-lasting impacts on offspring performance and health.

During pregnancy, the fetus relies on the dam for its nutrient supply. Nutritional stimuli during fetal organ development can program hepatic metabolism and function. Herein, we investigated the role of vitamin and mineral supplementation (VTM or NoVTM—at least 71 days pre-breeding to day 83 of gestation) and rate of weight gain (low (LG) or moderate (MG)—from breeding to day 83) on the fetal liver transcriptome and the underlying biological pathways. Crossbred Angus beef heifers (n = 35) were randomly assigned to one of four treatments in a 2 × 2 factorial design (VTM_LG, VTM_MG, NoVTM_LG, and NoVTM_MG). Gene expression was measured with RNA-Seq in fetal livers collected on day 83 ± 0.27 of gestation. Our results show that vitamin and mineral supplementation and rate of weight gain led to the differential expression of hepatic genes in all treatments. We identified 591 unique differentially expressed genes across all six VTM-gain contrasts (FDR ≤ 0.1). Over-represented pathways were related to energy metabolism, including PPAR and PI3K-Akt signaling pathways, as well as lipid metabolism, mineral transport, and amino acid transport. Our findings suggest that periconceptual maternal nutrition affects fetal hepatic function through altered expression of energy- and lipid-related genes.

Herein, we evaluated the hepatic lipid metabolic profiles of bovine fetuses in response to maternal vitamin and mineral supplementation (VMSUP; supplemented (VTM) or not (NoVTM)) and two different rates of gain (GAIN; low gain (LG), 0.28 kg/d, or moderate gain (MG), 0.79 kg/d). Crossbred Angus heifers (n = 35; initial BW = 359.5 ± 7.1 kg) were randomly assigned to a 2 × 2 factorial arrangement, resulting in the following treatment combinations: NoVTM-LG (n = 9), NoVTM-MG (n = 9), VTM-LG (n = 9), and VTM-MG (n = 8). Heifers received their treatments until d 83 of gestation, when they were ovariohysterectomized. Fetuses were harvested and liver samples were analyzed via ultrahigh-performance liquid chromatography–tandem mass spectroscopy to characterize lipid profiles and abundances. We identified 374 biochemicals/metabolites belonging to 57 sub-pathways of the lipid metabolism super-pathway. The majority of the biochemicals/metabolites (n = 152) were significantly affected by the main effect of GAIN. Maternal moderate rates of gain resulted in greater abundances (p ≤ 0.0001) of ω-3 fatty acids (eicosapentaenoate, docosapentaenoate, and docosahexaenoate) and lower abundances (p ≤ 0.0001) of ω-6 fatty acids. Further, MG resulted in the accumulation of several diacylglycerols and depletion of the majority of the monoacylglycerols. Concentrations of nearly all acylcarnitines (p ≤ 0.03) were decreased in VTM-LG fetal livers compared to all other treatment combinations, indicating a greater rate of complete oxidation of fatty acids. Levels of secondary bile acids were impacted by VMSUP, being greater (p ≤ 0.0048) in NoVTM than in VTM fetal livers. Moreover, NoVTM combined with lower rate of gain resulted in greater concentrations of most secondary bile acid biochemicals/metabolites. These data indicate that maternal diet influenced and altered fetal hepatic lipid composition in the first trimester of gestation. Maternal body weight gain exerted a greater influence on fetal lipid profiles than vitamin and mineral supplementation. Specifically, lower rate of gain (0.28 kg/d) resulted in an increased abundance of the majority of the biochemicals/metabolites identified in this study.

Maternal nutrients are essential for proper fetal and placental development and function. However, the effects of vitamin and mineral supplementation under two rates of maternal weight gain on placental genome-wide gene expression have not been investigated so far. Furthermore, biological processes and pathways in the placenta that act in response to early maternal nutrition are yet to be elucidated. Herein, we examined the impact of maternal vitamin and mineral supplementation (from pre-breeding to day 83 post-breeding) and two rates of gain during the first 83 days of pregnancy on the gene expression of placental caruncles (CAR; maternal placenta) and cotyledons (COT; fetal placenta) of crossbred Angus beef heifers. We identified 267 unique differentially expressed genes (DEG). Among the DEGs from CAR, we identified ACAT2, SREBF2, and HMGCCS1 that underlie the cholesterol biosynthesis pathway. Furthermore, the transcription factors PAX2 and PAX8 were over-represented in biological processes related to kidney organogenesis. The DEGs from COT included SLC2A1, SLC2A3, SLC27A4, and INSIG1. Our over-representation analysis retrieved biological processes related to nutrient transport and ion homeostasis, whereas the pathways included insulin secretion, PPAR signaling, and biosynthesis of amino acids. Vitamin and mineral supplementation and rate of gain were associated with changes in gene expression, biological processes, and KEGG pathways in beef cattle placental tissues.

Thirty-six half-sibling yearling Angus bulls [256 ± 8 d; initial BW = 320 ± 2 kg] were assigned one of three diets: 1) 60% concentrate as corn (CON; n = 12); 2) 60% DDGS (60DDGS; n = 12); 3) CON diet + equivalent sulfur of 60DDGS as CaSO4 (SULF; n = 12) to evaluate effects of feeding 60% DDGS or sulfur as CaSO4 on trace mineral (TM) concentrations in serum and seminal plasma. Bulls were fed in Calan gates for 112 days and targeted to gain 1.6 kg/d. Data were analyzed as repeated measures using PROC MIXED in SAS. In serum, treatment × day interactions were observed (P ≤ 0.03) for Cu, Se, and Mo. For Cu, no differences (P > 0.15) were observed at d 0 or 56, but at d 112, 60DDGS was reduced (P < 0.01) compared with SULF and CON. At d 0, no differences (P > 0.09) were observed for Se; however, at d 56 and 112, 60DDGS was greater (P < 0.01) than CON and SULF. For Mo, at d 0, 60DDGS was greater (P = 0.03) than CON, whereas SULF was intermediate, but at d 56 and 112, CON was greater (P < 0.01) than SULF and 60DDGS. In seminal plasma, treatment × day interactions were observed (P ≤ 0.02) for Cu and Mo. For Cu, no differences (P ≥ 0.09) were observed at d 0 or 56, but at d 112, CON and 60DDGS were greater (P < 0.01) compared with SULF. For Mo, at d 0, 60DDGS was greater (P = 0.03) than SULF, whereas CON was intermediate, but on d 56 and 112, CON was greater (P < 0.01) than 60DDGS and SULF. Differences observed for TM may have influenced enzyme activity and semen kinematics which were previously reported.

The objectives were to investigate the effects of feeding 60% dried corn distillers grains plus solubles (DCD) or equivalent sulfur as calcium sulfate (CaSO4) on performance, hormone concentrations, and glutathione peroxidase (GPx) activity in yearling bulls. Thirty-six half-sibling Angus bulls [256 ± 8 d; initial BW = 320 ± 2 kg] were assigned one of three treatments: 1) corn-based diet containing 60% concentrate (CON; S = 0.18%; n = 12); 2) diet containing 60% DCD as a replacement for corn (DDGS; S = 0.53% DM; n = 12); 3) CON diet + equivalent sulfur of the DDGS diet added as CaSO4 (SULF; S = 0.51%; n = 12). Bulls were fed in Calan gates to target an average daily gain (ADG) of 1.6 kg/d. Data were analyzed as repeated measures using PROC MIXED in SAS. Blood and semen samples were collected on d 0, 56, and 112 then evaluated for concentrations of testosterone, thyroxine, triiodothyronine in serum, and GPx activity in seminal plasma. By design, no differences (P ³ 0.14) were observed among treatments for final BW, ADG or dry matter intake. However, DDGS tended (P = 0.07) to have reduced gain: feed when compared with CON, whereas SULF was intermediate. No differences (P ³ 0.13) were observed among treatments for serum concentrations of testosterone and thyroxine; however, DDGS had reduced (P = 0.009) triiodothyronine concentrations compared with CON and SULF. A treatment ′ d interaction (P = 0.03) was observed for seminal plasma GPx. On d 56, GPx activity was greater (P = 0.03) for DDGS compared with CON, whereas SULF was intermediate, but on d 112, DDGS had greater (P £ 0.02) GPx activity compared with CON and SULF. Therefore, alterations in triiodothyronine concentrations and GPx activity among treatments indicates that DDGS may influence characteristics of semen quality.

During pregnancy, the placenta plays a pivotal role in fetal-maternal communication. Despite the known involvement of trophoblast mononuclear (MNC) and binuclear (BNC) cells in placentation and fetal development, the regulatory mechanisms underlying the gene expression in these cells and their role in pregnancy have not been fully elucidated. Thus, a genome-wide expression profile analysis of MNC and BNC from ovine placentomes on day 90 and 130 of pregnancy was performed to identify differentially expressed genes (DEG) and regulatory pathways. After tissue separation, enzymatic digestion, and elutriation, MNC and BNC were obtained, and the total RNA was isolated and sequenced (n = 2 ewes/day). Differential expression analysis was carried out with DESeq2 after library quality control and read mapping using FastQC and STAR, respectively. Among six pairwise contrasts possible, herein, we focused on the one between MNC and BNC on day 130. Accordingly, we identified 514 genes upregulated and 161 downregulated in BNC (adj.Pval < 0.05). Interestingly, we found nine DE long intergenic noncoding RNAs (lincRNA). Although the function of lincRNAs remains under investigation, its transcription has been associated with gene expression regulation in utero. The DEGs functional over-representation analysis included Rap1 and PI3k-Akt signaling pathways as up-regulated in BNC (adj.Pval. < 0.05). These pathways are activated by extracellular signals and are involved in cell proliferation and differentiation, morphogenesis, and energy metabolism. Furthermore, we identified PLA2G10 gene that codes a phospholipase protein involved in the production of prostaglandins, which have been associated with placental blood flow. These findings suggest an intricate and complex network underlying gene expression between cells and time points that are essential for the fetal-maternal placenta relationship. This project was supported by Agriculture and Food Research Initiative Competitive Grant n. 2016-67016-24884 from the USDA National Institute of Food and Agriculture.

During pregnancy, the placenta plays a pivotal role in fetal-maternal communication. Despite the known involvement of trophoblast mononuclear (MNC) and binuclear (BNC) cells in placentation and fetal development, the regulatory mechanisms underlying the gene expression in these cells and their role in pregnancy have not been fully elucidated. Thus, a genome-wide expression profile analysis of MNC and BNC from ovine placentomes on day 90 and 130 of pregnancy was performed to identify differentially expressed genes (DEG) and regulatory pathways. After tissue separation, enzymatic digestion, and elutriation, MNC and BNC were obtained, and the total RNA was isolated and sequenced (n = 2 ewes/day). Differential expression analysis was carried out with DESeq2 after library quality control and read mapping using FastQC and STAR, respectively. Among six pairwise contrasts possible, herein, we focused on the one between MNC and BNC on day 130. Accordingly, we identified 514 genes upregulated and 161 downregulated in BNC (adj.Pval < 0.05). Interestingly, we found nine DE long intergenic noncoding RNAs (lincRNA). Although the function of lincRNAs remains under investigation, its transcription has been associated with gene expression regulation in utero. The DEGs functional over-representation analysis included Rap1 and PI3k-Akt signaling pathways as up-regulated in BNC (adj.Pval. < 0.05). These pathways are activated by extracellular signals and are involved in cell proliferation and differentiation, morphogenesis, and energy metabolism. Furthermore, we identified PLA2G10 gene that codes a phospholipase protein involved in the production of prostaglandins, which have been associated with placental blood flow. These findings suggest an intricate and complex network underlying gene expression between cells and time points that are essential for the fetal-maternal placenta relationship. This project was supported by Agriculture and Food Research Initiative Competitive Grant n. 2016-67016-24884 from the USDA National Institute of Food and Agriculture.

The objectives were to investigate the effects of feeding 60% dried corn distillers grains plus solubles (DCD) or equivalent sulfur as calcium sulfate (CaSO4) on performance, hormone concentrations, and glutathione peroxidase (GPx) activity in yearling bulls. Thirty-six half-sibling Angus bulls [256 ± 8 d; initial BW = 320 ± 2 kg] were assigned one of three treatments: 1) cornbased diet containing 60% concentrate (CON; S = 0.18%; n = 12); 2) diet containing 60% DCD as a replacement for corn (DDGS; S = 0.53% DM; n = 12); 3) CON diet + equivalent sulfur of the DDGS diet added as CaSO4 (SULF; S = 0.51%; n = 12). Bulls were fed in Calan gates to target an average daily gain (ADG) of 1.6 kg/d. Data were analyzed as repeated measures using PROC MIXED in SAS. Blood and semen samples were collected on d 0, 56, and 112 then evaluated for concentrations of testosterone, thyroxine, triiodothyronine in serum, and GPx activity in seminal plasma. By design, no differences (P 3 0.14) were observed among treatments for final BW, ADG or dry matter intake. However, DDGS tended (P = 0.07) to have reduced gain: feed when compared with CON, whereas SULF was intermediate. No differences (P з 0.13) were observed among treatments for serum concentrations of testosterone and thyroxine; however, DDGS had reduced (P = 0.009) triiodothyronine concentrations compared with CON and SULF. A treatment ' d interaction (P = 0.03) was observed for seminal plasma GPx. On d 56, GPx activity was greater (P = 0.03) for DDGS compared with CON, whereas SULF was intermediate, but on d 112, DDGS had greater (P £ 0.02) GPx activity compared with CON and SULF. Therefore, alterations in triiodothyronine concentrations and GPx activity among treatments indicates that DDGS may influence characteristics of semen quality.