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Maternal nutritional plane during pregnancy may leave lifelong impacts on offspring in beef cattle. Given that, we hypothesized that the supplementation of one-carbon metabolites (OCM) during pregnancy would mitigate the negative effects of maternal rate of gain through brain gene expression regulation in early developing bovine fetuses. The objective of this study was to evaluate the effects of OCM supplementation and maternal rate of gain on gene expression in the developing fetal brain in gestating beef heifers. Herein, Angus crossbred heifers (n = 72, body weight = 406 ± 33 kg) were randomly assigned to 2 × 2 factorial arrangement of treatments with two levels of maternal rate of gain (CON, 0.60 kg/d; and RES, -0.23 kg/d) each with or without OCM supplementation (+OCM, and -OCM). Heifers were fed individually and were bred using female-sexed semen from a single sire. The OCM supplementation included rumen-protected choline (44.4 g/d) and methionine (7.4 g/d) in corn carrier fed daily and weekly injections of folate (320 mg) and vitamin B12 (20 mg). The -OCM heifers received the corn carrier and saline injections. The treatments started at the time of breeding and continued until d 63 of gestation when pregnant heifers (n = 29) were slaughtered to collect fetal brain tissues for RNA sequencing. The fetal brain phenotypic data were analyzed using MIXED procedures of SAS, indicating that restricted nutrition affected the fetal right hemisphere weight (P < 0.05), and there was a strong tendency for overall fetal brain weight (0.05 < P < 0.1) to be affected as well. Brain transcriptomics data were analyzed using STAR aligner and a total of 177 differentially expressed genes (DEGs, FDR < 0.05) were identified across all treatment comparisons using DESeq2. Most of the DEGs were either isoforms of spliceosomal non-coding RNAs or micro-RNAs (miRNAs) associated with biological processes (FDR < 0.1) like mRNA processing, cellular nitrogen metabolism, lipid metabolism, and post-transcriptional gene silencing. The key miRNAs upregulated in the brain tissue of restricted fetuses without OCM supplementation, bta-mir-2375, bta-mir-876, and bta-mir-196a-1, were found to be associated with early embryogenesis and essential cellular regulatory processes. Conversely, in restricted fetuses with OCM supplementation, upregulated miRNAs such as bta-mir-216b, bta-mir-2285dd, and btamir-1298 were linked to early embryonic development, muscular development, and lipid metabolism in cattle. Both phenotypic and transcriptomics data here revealed that the maternal rate of gain altered fetal brain development, with differences seemingly compensated by OCM supplementation. USDA is an equal opportunity provider and employer. [ABSTRACT FROM AUTHOR]

We hypothesized that yearling bulls selected for a 28‐d breeding season would have reduced sperm concentrations and morphology, and have increased seminal plasma concentrations of pro‐inflammatory cytokine interleukin‐8 (IL‐8). Yearling bulls were selected based on a breeding soundness examination (BSE) at approximately 415 d of age and contained at least 750 million sperm in the ejaculate, with 12 bulls randomly selected for breeding (BREEDERS) and 12 bulls not selected for breeding (NON‐BREEDERS). After a 28‐d breeding period, all bulls underwent a BSE. Plasma and seminal plasma were collected at each time point for analysis. Data were analysed utilizing either the MIXED or GLIMMIX procedures with repeated measures in SAS with breeding group, age and the interaction as fixed effects. Sperm concentration per ml of ejaculate was reduced (p <.05) in yearling bulls used for breeding compared with those not used for breeding at the end of the breeding season. Seminal plasma IL‐8 concentrations in yearling bulls used for breeding were increased (p <.05) after the breeding season compared with bulls not used for breeding. Taken together, yearling bulls selected for a 28‐d breeding season have reduced sperm production per ml of an ejaculate and increased inflammatory response in the seminal plasma that can lead to impaired breeding response if they are to be used for more than 30 d of breeding. [ABSTRACT FROM AUTHOR]

Hempseed cake is a byproduct of hempseed oil extraction and is potentially a useful source of protein and fiber for use in ruminant diets. However, data are lacking on the appearance and/or clearance of cannabinoids in tissues of animals fed hempseed cake. To this end, a rapid method for quantifying cannabinol (CBN), cannabidiol (CBD), cannabinolic acid (CBNA), cannabidiolic acid (CBDA), cannabigerolic acid (CBGA), cannabichromenic acid (CBCA), cannabidivarin (CBDV), cannabidivarinic acid (CBDVA), tetrahydrocannabinol (THC) and tetrahydrocannabinolic acid (THCA) in cattle tissues, plasma, and urine was developed using rapid screen electrospray ionization mass spectrometry (RS-ESI-MS). Regression coefficients of matrix-matched standard curves ranged from 0.9946 to >0.9999 and analyte recoveries averaged from 90.2 ± 15.5 to 108.7 ± 18.7% across all compounds. Limits of detection and quantification ranged from 0.05 to 2.79 ng · mL−1 and 0.17 to 9.30 ng · mL−1, respectively, while the inter-day relative standard deviation ranged from 5.1 to 15.1%. Rapid screening electrospray ionization mass spectrometry (RS-ESI-MS) returned no false positives for any cannabinoid in plasma, urine, and tissue (liver, skeletal muscle) samples from 6 non-dosed control animals (n = 90 samples; of which 72 samples were plasma or urine and 18 samples were tissues). Across-animal cannabinoid concentrations measured in 32 plasma samples of cattle dosed with ground hemp were quantified by RS-ESI-MS; analytical results correlated well (r2 = 0.963) with independent LC-MS/MS analysis of the same samples. [ABSTRACT FROM AUTHOR]

Simple Summary: Trace minerals play a critical role in several biological functions, being essential for fetal growth and development. Maternal nutrition influences the supply of trace minerals to the fetus, which can lead to long-term consequences on offspring performance and health. Our study shows that a maternal vitamin and mineral supplementation (VTM) pre-breeding and during the first trimester of gestation resulted in greater concentrations of Se, Cu, and Co in the liver of beef heifers and their fetuses. These minerals are an essential component of the antioxidant system, which may have positive impacts on the immune system of the future offspring. In addition, we reported that low rates of gain (LG, 0.28 kg/d) from breeding to d 83 of gestation resulted in greater concentrations of Mo and Co in fetal liver compared with moderate rates of gain (MG, 0.79 kg/d). Having a greater fetal trace mineral reserve may be beneficial for neonates, as the immune system of newborn calves is underdeveloped and minerals play a role in that development. Therefore, our study shows that liver weight and concentrations of hepatic trace minerals are affected by maternal dietary treatments, which may impact the performance and health of the future offspring. 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. [ABSTRACT FROM AUTHOR]

Simple Summary: Cow-calf operations rely mostly in forage-based systems. Different supplementation strategies are used by beef producers in order to overcome nutrient deficiencies and achieve targeted growth or reproductive performances. This study provides information on the impacts of feeding pregnant replacement heifers with vitamin/mineral and protein/energy supplements on heifer performance and fetal outcomes. Our study shows that moderate rates of gain (achieved via protein/energy supplementation) resulted in fetuses with heavier femurs and reduced liver mass relative to fetal body weight. Moreover, vitamin and mineral supplementation increased fetal liver mass, and vitamin and mineral supplementation combined with moderate gain treatments resulted in greater fetal intestinal weights. These findings indicate that replacement heifer nutrition during early gestation can alter the development of organs in the fetus that are relevant for future offspring performance. Liver and intestines are key organs related to energy metabolism; therefore, this study shows 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. 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. [ABSTRACT FROM AUTHOR]

As the hemp industry continues to develop in the United States, there is an increasing interest in feeding byproducts of industrial hemp production to livestock. A completely randomized design experiment using crossbred finishing heifers (initial body weight [ BW ] ± SE = 494 ± 10 kg) was conducted to determine the effects of feeding hempseed cake in a corn-based finishing diet (10% forage) formulated to meet or exceed ruminally degradable and metabolizable protein requirements on growth performance, carcass characteristics, feeding behavior, and plasma parameters. Dietary treatments were the inclusion of 20% (dry matter [ DM ] basis) of dried corn distillers grains plus solubles (DDGS , n  = 16) or hempseed cake (HEMP , n  = 15). Cattle were housed in two pens, had ad libitum access to feed and water, and individual intakes and feeding behavior were monitored using the Insentec feeding system. Cattle were fed treatment diets for 111 d, and every 14 d BW were measured and blood samples were collected. Blood plasma was analyzed for glucose, urea nitrogen, and individual amino acids, and results were analyzed using repeated-measures analysis in SAS. Final BW, average daily gain, gain:feed, and hot carcass weight decreased (P  ≤ 0.05) by 2.3%, 7.7%, 7.7%, and 2.6%, respectively, in heifers fed the HEMP diet than in heifers fed the DDGS diet. Net energy for maintenance and gain (Mcal/kg of feed, DM basis), estimated based on heifer intake and performance, were greater (P  = 0.02) for the DDGS diet than for the HEMP diet. All other performance and carcass characteristics were not different (P  ≥ 0.20) between treatments. Heifers fed the HEMP diet had greater (P  < 0.05) plasma urea nitrogen concentration in samples from each collection day compared with heifers fed the DDGS diet, although there was a treatment-by-day interaction (P  < 0.01) because of variability in the magnitude of treatment differences over time. Plasma glucose concentration was not influenced (P  = 0.17) by dietary treatment. Plasma concentrations of total amino acids, nonessential amino acids, and essential amino acids were not different between treatments (P  ≥ 0.09), although there were several interactions between treatment and day (P  ≤ 0.04) for individual amino acids. These data suggest that hempseed cake has a lower net energy for maintenance and gain relative to DDGS when adequate metabolizable protein is supplied, while still providing adequate nutrition to support the acceptable performance of finishing cattle. [ABSTRACT FROM AUTHOR]

The objectives of this study were to investigate the effects of feeding 60% dried corn distillers grains plus solubles (DDGS) or the equivalent sulfur as calcium sulfate (CaSO 4) on semen quality and performance characteristics in yearling bulls. Thirty-six half-sibling Angus bulls [291 ± 8.5 d; initial body weight (BW) = 320 ± 2.7 kg] were assigned to one of three diets: 1) 60% concentrate as corn (CON; S = 0.18%; n = 12); 2) 60% DDGS replacing corn (60DDGS; S = 0.55% DM; n = 12); 3) CON + equivalent sulfur of 60DDGS added as CaSO 4 (SULF; S = 0.54%; n = 12). Bulls were fed for 112 d to target an average daily gain (ADG) of 1.6 kg/d. Blood samples were collected on d 0, 56, and 112, and evaluated for testosterone, thyroxine, triiodothyronine (T3) and glutathione peroxidase (GP x) activity. Ruminal H 2 S was measured on d 0, 14, and 42. Scrotal circumference and semen were collected on d 0, 28, 56, 84, and 112 to evaluate sperm characteristics and GP x activity in seminal plasma. A computer assisted semen analysis was used to evaluate kinematic profiles in motile and progressive sperm throughout the study. Data were analyzed as repeated measures using MIXED procedures of SAS. No differences (P ≥ 0.14) were observed for final BW, ADG, or scrotal circumference; however, SULF tended (P = 0.07) to have reduced gain:feed compared with CON, with 60DDGS being intermediate. Concentrations of ruminal H 2 S on d 42 were greatest (P < 0.01) for SULF. Increased ejaculate volume was observed for 60DDGS and CON (P < 0.01) compared with SULF. For motile populations of sperm, velocity on an average path (VAP) and curvilinear velocity (VCL) were reduced (P ≤ 0.02) for SULF compared with CON, with 60DDGS being intermediate. In progressively motile sperm throughout the study, VAP and VSL were reduced (P ≤ 0.05) in 60DDGS and SULF compared to CON. For VCL, SULF was reduced (P ≤ 0.02) compared with CON, with 60DDGS being intermediate. In serum, concentrations of T3 were reduced (P = 0.009) in 60DDGS compared with CON or SULF. A treatment by day interaction (P = 0.03) was observed for seminal plasma GP x. At d 56, GP x activity was greater (P = 0.03) for 60DDGS compared with CON, with SULF intermediate; and at d 112, 60DDGS had the greatest (P ≤ 0.02) GP x activity. Therefore, feeding 60% DDGS to developing bulls altered semen kinematics, T3 concentrations, and GP x activity leading to the conclusion that these differences may not be solely dependent on concentrations of dietary sulfur. • Increased ruminal H 2 S was observed for bulls consuming diets with increased sulfur. • Feed efficiency tended to be reduced for diets with elevated percentages of sulfur. • Parameters in motile and progressively motile populations of sperm were altered. • Triiodothyronine and glutathione peroxidase in seminal plasma was altered in bulls. [ABSTRACT FROM AUTHOR]

The molecular basis underlying fetal programming in response to maternal nutrition remains unclear. Herein, we investigated the regulatory relationships between genes in fetal cerebrum, liver, and muscle tissues to shed light on the putative mechanisms that underlie the effects of early maternal nutrient restriction on bovine developmental programming. To this end, cerebrum, liver, and muscle gene expression were measured with RNA-Seq in 14 fetuses collected on day 50 of gestation from dams fed a diet initiated at breeding to either achieve 60% (RES, n = 7) or 100% (CON, n = 7) of energy requirements. To build a tissue-to-tissue gene network, we prioritized tissue-specific genes, transcription factors, and differentially expressed genes. Furthermore, we built condition-specific networks to identify differentially co-expressed or connected genes. Nutrient restriction led to differential tissue regulation between the treatments. Myogenic factors differentially regulated by ZBTB33 and ZNF131 may negatively affect myogenesis. Additionally, nutrient-sensing pathways, such as mTOR and PI3K/Akt, were affected by gene expression changes in response to nutrient restriction. By unveiling the network properties, we identified major regulators driving gene expression. However, further research is still needed to determine the impact of early maternal nutrition and strategic supplementation on pre- and post-natal performance. [ABSTRACT FROM AUTHOR]

The focus of this review is maternal nutrition during the periconceptual period and offspring developmental outcomes in beef cattle, with an emphasis on the first 50 d of gestation, which represents the embryonic period. Animal agriculture in general, and specifically the beef cattle industry, currently faces immense challenges. The world needs to significantly increase its output of animal food products by 2050 and beyond to meet the food security and agricultural sustainability needs of the rapidly growing human population. Consequently, efficient and sustainable approaches to livestock production are essential. Maternal nutritional status is a major factor that leads to developmental programming of offspring outcomes. Developmental programming refers to the influence of pre-and postnatal factors, such as inappropriate maternal nutrition, that affect growth and development and result in long-term consequences for health and productivity of the offspring. In this review, we discuss recent studies in which we and others have addressed the questions, "Is development programmed periconceptually?" and, if so, "Does it matter practically to the offspring in production settings?" The reviewed studies have demonstrated that the periconceptual period is important not only for pregnancy establishment but also may be a critical period during which fetal, placental, and potentially postnatal development and function are programmed. The evidence for fetal and placental programming during the periconceptual period is strong and implies that research efforts to mitigate the negative and foster the positive benefits of developmental programming need to include robust investigative efforts during the periconceptual period to better understand the implications for life-long health and productivity. [ABSTRACT FROM AUTHOR]

During early pregnancy offspring are directly exposed to nutrients consumed by their mother, and the development of their own reproductive tract is underway. The objective of this research was to determine how characteristics of offspring ovaries were affected by different maternal rates of gain during the first trimester of gestation in beef heifers. Before breeding antral follicle counts were determined via ultrasound. Beginning at breeding Angus heifers were managed to achieve one of two rates of gain: low (0.20 kg/d, n = 8; LG) or moderate (0.75 kg/d, n = 8; MG) for the first trimester of pregnancy, after which they were managed as a single group through F1 calving. Calves remained with their dams until weaning and were managed as a single group through breeding. These F1 heifer calves were then synchronized and bred to a single sire using female sexed semen. On the 84th day of gestation (approximate heifer age 17 months) ovaries were removed from the reproductive tract, weighed, and visible antral follicles were counted. Cross-sections from each ovary were post-fixed and embedded in paraffin, then each of 3 sections (5 μm, with 10 sections between to avoid counting follicles more than once) were stained with Hematoxylin and Eosin for histological evaluation. Number of primordial, primary, secondary, and antral follicles were determined for each section. Data were analyzed using the GLM procedure of SAS with significance declared at P ≤ 0.05. The CORR procedure of SAS was used to calculate correlations between prebreeding antral follicle counts, surface follicle counts, and histological follicle counts. Though no differences were observed in number of visible follicles (P = 0.45), the corpus luteum (CL) was heavier (P = 0.03) and average ovarian length was greater (P = 0.04) in offspring from LG dams compared with those from MG dams. No differences were observed in the number of primordial, primary, secondary, or antral follicles between treatments (P ≤ 0.18). There was a correlation between the number of histological follicles and surface follicles (r = 0.73; P = 0.002) and pre-breeding antral follicles and surface follicles (r = 0.60; P = 0.014), but there was no correlation between pre-breeding antral follicles and histological follicles (P = 0.10). Heavier CLs have a positive correlation with amount of progesterone released, which is essential for pregnancy maintenance. Longer lengths of ovaries suggest more area for follicles to develop, which is indicative of future reproductive success. Data corroborates previous reports regarding positive correlations between antral follicle count determined via ultrasound and counts of surface follicles on extracted ovaries. These findings demonstrate that nutrition during the first trimester of gestation impacts offspring reproductive tract development. [ABSTRACT FROM AUTHOR]

The cotyledon and caruncle tissues provide a functional bridge between the fetus and the dam. However, the relationship between these tissues and the transcriptomic profile that underlies the tissue functions remains elusive. Herein we investigate the expression profile of cotyledon and caruncle from nulliparous beef heifers carrying female fetuses at day 83 of pregnancy to identify changes occurring across tissues that contribute to placental function and their tissue-specific roles. We identified 2654 differentially expressed genes [ padj ≤ 0.05, abs(log2FC) ≥ 1], including nutrient transporters and paternally imprinted genes. We found key regulators of tissue function and differentiation, including FOXO4 , GATA2 , GATA3 , and HAND1, rewired between the tissues. Finally, we shed light on the over-represented pathways related to immune tolerance, tissue differentiation and remodeling. Our findings highlighted the intricate and coordinated cross-talk between fetal-maternal tissues. They provided evidence of a fine-tuned gene regulatory network underlying pregnancy and tissue-specific function in the bovine placenta. • Genes involved with nutrient transport and tissue differentiation and remodeling were differentially expressed between the fetal and maternal tissues. • The Wnt pathway is likely repressed in cotyledon at day 83 of pregnancy. • Downregulated differentially expressed genes are involved with immune tolerance in the fetal placental tissue. • Key transcription factors play a key role in modulating gene expression between cotyledon and caruncle. • Paternally imprinted genes were upregulated in the cotyledon tissue. [ABSTRACT FROM AUTHOR]

Abstract: We determined whether source of trace mineral supplementation prior to embryo collection affected embryo production and quality. Angus half-sibling heifers (n =20) originating from a common herd were assigned to three treatment groups using a 3×3 latin square design replicated in time (3×) and space (6× complete and 1× incomplete): (1) heifers received no added mineral to their diet (control; n =53); (2) heifers received a commercially available organic mineral supplement (organic; n =52); or (3) heifers received an all inorganic mineral supplement (inorganic; n =55). All heifers had ad libitum access to hay and were fed a supplement containing corn and soybean meal. Treatments were initiated 23 days prior to embryo recovery. Heifers were given a 45-day adaptation period of no mineral supplementation before initiating a new treatment. Ovarian structures were evaluated using transrectal ultrasonography to determine the presence and number of follicles and CL on each ovary. The mean number of recovered ova/embryos was similar among treatments (4.1±0.7, 3.8±0.7, and 3.3±0.7 for control, inorganic, and organic treatments, respectively), the number of unfertilized oocytes was greater (P <0.05) for inorganic (2.3±0.5) and control (1.6±0.5) treated heifers than organic (0.4±0.4) treated heifers. No differences among treatments existed for the number of degenerate or transferable embryos, but individual heifer influenced the total number of embryos/ova, unfertilized ova, and transferable embryos recovered. We conclude that heifer accounted for the greatest differences in embryo production and quality. Source of trace mineral supplementation did not significantly alter embryo number or quality in superovulated purebred Angus heifers fed a well-balanced diet, meeting all trace mineral requirements. [Copyright &y& Elsevier]

• Infrared thermography technology was investigated for beef cattle temperament evaluation. • Eye temperature and percentage of the eye white area were extracted for beef cattle temperament prediction. • Both popular subjective and objective temperament measurements were compared to the infrared technology. Beef cattle temperament can impact animal performance and meat quality in cattle. The aim of this study was to evaluate the use of infrared thermography (IRT) maximum, average, and minimum eye temperature (IRT MIN , IRT AVG , IRT MAX) and percentage of the eye with the sclera visible in a digital image (eye white percentage, EW) to predict beef cattle temperament by (1) assessing the relationship between IRT and EW traits with 4 established subjective and objective temperament scoring methods (n = 16 traits total) and (2) identify behavioral characteristics that IRT and EW traits are predicting. Traits were measured on Angus- and Hereford-influenced steers (n = 203; age 183.00 ± 15.80 days; BW 264.50 ± 30.84 kg) and heifers (n = 200; age 186.60 ± 15.18 days; BW 251.90 ± 28.77 kg). Computer vision and image processing technologies were used to extract IRT MIN , IRT AVG , IRT MAX , and EW features. Temperament Scores (TS), Docility Scores (DS), Four-platform Standing Scale (FPSS), Qualitative Behavioral Assessment (QBA) attributes were measured to compare with the imagery methods. Results showed there were no statistically significant relationships of IRT MIN , IRT AVG IRT MAX for TS, DS, FPSS and QBA. The correlations value between EW and DS and between EW and TS were also low but indicated correlated at certain level (0.258 and 0.179 at P < 0.0001). This study showed to prove using thermal and eye white area to objectively predict beef cattle temperament, more cattle sample numbers should be further investigated to validate the hypothesis. [ABSTRACT FROM AUTHOR]

Yearling Angus bulls (n = 36) were assigned one of three diets: 1) 60 % concentrate as corn (CON, 0.2 % S, 13.4 % CP; n = 12); 2) 60 % dried corn distiller's grains plus solubles (60DDGS 0.5 % S, 22.0 % CP; n = 12); 3) CON diet + equivalent sulfur of 60DDGS as CaSO 4 (SULF, 0.5 % S, 13.9 % CP; n = 12) to evaluate effects of feeding 60 % DDGS or sulfur as CaSO 4 on mineral and metabolite concentrations in serum and seminal plasma. Treatment × day interactions (P < 0.03) were observed for serum Cu, Se, and Mo. For Cu at d 112, lesser (P < 0.01) concentrations were observed in bulls fed the 60DDGS compared to SULF and CON diets. There were greater (P < 0.01) concentrations of Se at d 112 in bulls fed 60DDGS than CON and SULF diets. Concentrations of Mo were greater at d 56 and 112 (P < 0.01) in bulls fed CON compared to SULF and 60DDGS diets. In seminal plasma, there were treatment × day interactions (P < 0.02) for Cu and Mo. For Cu, at d 112, there was a lesser (P < 0.01) concentration in the bulls fed SULF compared to CON and 60DDGS diets. For Mo, there was a greater (P < 0.01) concentration in bulls fed the CON than 60DDGS and SULF diets at d 56 and 112. Changes in mineral and metabolite concentrations may have effects on bull reproductive performance when there is a relatively greater dietary sulfur content. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Rambouillet rams were managed on either a positive (POS; gain 12% body weight [BW]; n = 8), maintenance (MAINT; maintain BW; n = 8), or negative (NEG; lose 12% BW; n = 8) plane of nutrition before breeding. Rams were bred to ewes (n = 10 per ram) that were managed similarly throughout gestation, and lambs were fed a common diet postnatally. Two ewe lambs (7.6 ± 0.02 months of age, BW = 47.1 ± 1.17 kg) from each sire were selected and within pair, randomly assigned to be managed for a moderate (MOD, 0.11 kg/d; n = 23) or accelerated (ACC, 0.20 kg/d; n = 22) rate of gain for 56 d. Ewe lamb BW was recorded on a weekly basis and blood was collected on d 0, 28, and 56 for analysis of insulin-like growth factor 1 (IGF-1), triiodothyronine (T3), thyroxine (T4), glucose, blood urea nitrogen (BUN), and non-esterified fatty acids (NEFA). Intravenous glucose tolerance tests (IVGTT) were conducted from d -7 to -4 and d 57 to 64. A unilateral ovariectomy was performed and ovarian follicles were staged and counted macro and microscopically. Sire treatment × day and ewe treatment × day interactions were present for BW (P ≤ 0.05), where POS had slower growth than MAINT and NEG, and tended (P = 0.10) to have reduced average daily gain (ADG) when managed at an accelerated rate of gain.By design, ACC had greater BW and ADG than MOD (P < 0.05). Concentrations of IGF-1 and T4 were greater in ACC than MOD (P ≤ 0.05), and NEG tended to have greater concentrations of IGF-1 than POS and MAINT (P = 0.08). At the first IVGTT, concentration of insulin was influenced by a sire treatment × time interaction (P ≤ 0.05), suggesting impaired secretion in NEG-sires ewes, but no differences in area under the curve (AUC) for glucose, insulin, or their ratio (P ≥ 0.11). No interactive effects of sire and ewe treatment (P ≥ 0.52) were observed at the second IVGTT, but insulin and insulin:glucose ratio were influenced by sire treatment × time (P ≤ 0.02), as NEG had greater insulin concentration at 60 min than MAINT (P = 0.03) and greater AUC than POS and MAINT (P ≤ 0.04). No differences in ovary size, weight, or total counts of macro and microscopic follicles were observed (P ≥ 0.23). Ewes fed ACC had a greater number of small surface follicles (P = 0.02), whereas MOD tended to have a greater number of large surface follicles and tertiary follicles (P < 0.06). These findings suggest that paternal plane of nutrition influences female offspring physiology, particularly at varying growth rates., (© The Author(s) 2024. Published by Oxford University Press on behalf of the American Society of Animal Science. All rights reserved. For commercial re-use, please contact reprints@oup.com for reprints and translation rights for reprints. All other permissions can be obtained through our RightsLink service via the Permissions link on the article page on our site—for further information please contact journals.permissions@oup.com.)

Background: Maternal diet quality and quantity have significant impacts on both maternal and fetal health and development. The composition and function of the maternal gut microbiome is also significantly influenced by diet; however, little is known about the impact of gestational nutrient restriction on the bovine maternal microbiome during early gestation, which is a critical stage for maternal microbiome-mediated fetal programming to take place. The objective of the present study was to evaluate the impacts of diet restriction and one-carbon metabolite (OCM) supplementation during early gestation on maternal ruminal, vaginal, and blood microbiota in cattle. Thirty-three beef heifers (approx. 14 months old) were used in a 2 × 2 factorial experiment with main factors of target gain (control [CON]; targeted 0.45 kg/d gain vs restricted [RES]; targeted - 0.23 kg/d gain), and OCM supplementation (+ OCM vs - OCM; n = 8/treatment; except n = 9 for RES-OCM). Heifers were individually fed, starting treatment at breeding (d 0) and concluding at d 63 of gestation. Ruminal fluid and vaginal swabs were collected on d - 2, d 35, and d 63 (at necropsy) and whole blood was collected on d 63 (necropsy). Bacterial microbiota was assessed using 16S rRNA gene (V3-V4) sequencing., Results: Overall ruminal microbiota structure was affected by gain, OCM, time, and their interactions. The RES heifers had greater microbial richness (observed ASVs) but neither Shannon nor Inverse Simpson diversity was significantly influenced by gain or OCM supplementation; however, on d 63, 34 bacterial genera showed differential abundance in the ruminal fluid, with 25 genera enriched in RES heifers as compared to CON heifers. In addition, the overall interaction network structure of the ruminal microbiota changed due to diet restriction. The vaginal microbiota community structure was influenced by gain and time. Overall microbial richness and diversity of the vaginal microbiota steadily increased as pregnancy progressed. The vaginal ecological network structure was distinctive between RES and CON heifers with genera-genera interactions being intensified in RES heifers. A relatively diverse bacterial community was detected in blood samples, and the composition of the blood microbiota differed from that of ruminal and vaginal microbiota., Conclusion: Restricted dietary intake during early gestation induced significant alterations in the ruminal microbiota which also extended to the vaginal microbiota. The composition of these two microbial communities was largely unaffected by OCM supplementation. Blood associated microbiota was largely distinctive from the ruminal and vaginal microbiota., (© 2024. The Author(s).)

The field of Developmental Origins of Health and Disease has primarily focused on maternal programming of offspring health. However, emerging evidence suggests that paternal factors, including the seminal microbiome, could potentially play important roles in shaping the developmental trajectory and long-term offspring health outcomes. Historically, the microbes present in the semen were regarded as inherently pathogenic agents. However, this dogma has recently been challenged by the discovery of a diverse commensal microbial community within the semen of healthy males. In addition, recent studies suggest that the transmission of semen-associated microbes into the female reproductive tract during mating has potentials to not only influence female fertility and embryo development but could also contribute to paternal programming in the offspring. In this review, we summarize the current knowledge on the seminal microbiota in both humans and animals followed by discussing their potential involvement in paternal programming of offspring health. We also propose and discuss potential mechanisms through which paternal influences are transmitted to offspring via the seminal microbiome. Overall, this review provides insights into the seminal microbiome-based paternal programing, which will expand our understanding of the potential paternal programming mechanisms which are currently focused primarily on the epigenetic modifications, oxidative stresses, and cytokines., (© The Author(s) 2024. Published by Oxford University Press behalf of Society for the Study of Reproduction.)

To investigate the effects of nutrient restriction and one-carbon metabolite (OCM) supplementation (folate, vitamin B 12 , methionine, and choline) on fetal small intestine weight, vascularity, and cell proliferation, 29 ( n = 7 ± 1 per treatment) crossbred Angus beef heifers (436 ± 42 kg) were estrous synchronized and conceived by artificial insemination with female sexed semen from a single sire. Then, they were allotted randomly to one of four treatments in a 2 × 2 factorial arrangement with the main factors of nutritional plane [control (CON) vs. restricted feed intake (RES)] and OCM supplementation [without OCM (-OCM) or with OCM (+OCM)]. Heifers receiving the CON level of intake were fed to target an average daily gain of 0.45 kg/day, which would allow them to reach 80% of mature BW by calving. Heifers receiving the RES level of intake were fed to lose 0.23 kg/heifer daily, which mimics observed production responses in heifers that experience a diet and environment change during early gestation. Targeted heifer gain and OCM treatments were administered from d 0 to 63 of gestation, and then all heifers were fed a common diet targeting 0.45 kg/d gain until d 161 of gestation, when heifers were slaughtered, and fetal jejunum was collected. Gain had no effect ( p = 0.17) on the fetal small intestinal weight. However, OCM treatments ( p = 0.02) displayed less weight compared to the -OCM groups. Capillary area density was increased in fetal jejunal villi of RES - OCM ( p = 0.02). Vascular endothelial growth factor receptor 2 (VEGFR2) positivity ratio tended to be greater ( p = 0.08) in villi and was less in the crypts ( p = 0.02) of the RES + OCM group. Cell proliferation decreased ( p = 0.02) in villi and crypts of fetal jejunal tissue from heifers fed the RES + OCM treatment compared with all groups and CON - OCM, respectively. Spatial cell density increased in RES - OCM compared with CON + OCM ( p = 0.05). Combined, these data show OCM supplementation can increase expression of VEGFR2 in jejunal villi, which will promote maintenance of the microvascular beds, while at the same time decreasing small intestine weight and crypt cell proliferation.

The effect of vitamins and minerals supplementation (VTM) and/or two rates of body weight gain (GAIN) on bovine placental vascular development and angiogenic factors gene expression were evaluated in two experiments: In Exp. 1, crossbred Angus heifers ( n = 34) were assigned to VTM/NoVTM treatments at least 71 days before breeding to allow changes in the mineral status. At breeding, through artificial insemination (AI), heifers were assigned to low-gain (LG) 0.28 kg/d or moderate-gain (MG) 0.79 kg/d treatments, resulting in NoVTM-LG (Control; n = 8), NoVTM-MG ( n = 8), VTM-LG ( n = 9), and VTM-MG ( n = 9) until day 83 of gestation; In Exp. 2, crossbred angus heifers ( n = 28), were assigned to control (CON; n = 12), receiving a basal total mixed ration (TMR) or TMR + VTM (VTM; n = 16) from breeding until parturition. Placentomes from Exp. 1 and cotyledons (COT) from Exp. 2 were evaluated by immunohistochemistry for COT vascular density area. COTs from Exp. 1 were evaluated for angiogenic factor ( ANGPT -1, ANGPT -2, eNOS 2, eNOS 3, FLT 1, KDR , TEK , VEGFA ) gene expression. In Exp. 1, COT vascularity was not affected by the interaction of VTM and GAIN ( p = 0.67) or the main effects of VTM ( p = 0.50) and GAIN ( p = 0.55). Likewise, angiogenic factors were not differentially expressed between treatments ( p < 0.05). In Exp. 2, COT vascularity was greater in VTM vs. CON ( p = 0.07). In conclusion, there is a suggested later-stage influence of vitamin and mineral supplementation on placental vascularity, emphasizing the importance of supplementation beyond early pregnancy.

The objective of this study was to determine the dose-dependent response of one-carbon metabolite (OCM: methionine, choline, folate, and vitamin B12) supplementation on heifer dry matter intake on fixed gain, organ mass, hematology, cytokine concentration, pancreatic and jejunal enzyme activity, and muscle hydrogen peroxide production. Angus heifers (n = 30; body weight [BW] = 392.6 ± 12.6 kg) were individually fed and assigned to one of five treatments: 0XNEG: total mixed ration (TMR) and saline injections at days 0 and 7 of the estrous cycle, 0XPOS: TMR, rumen-protected methionine (MET) fed at 0.08% of the diet dry matter, rumen-protected choline (CHOL) fed at 60 g/d, and saline injections at days 0 and 7, 0.5X: TMR, MET, CHOL, 5-mg B12, and 80-mg folate injections at days 0 and 7, 1X: TMR, MET CHOL, 10-mg vitamin B12, and 160-mg folate at days 0 and 7, and 2X: TMR, MET, CHOL, 20-mg vitamin B12, and 320-mg folate at days 0 and 7. All heifers were estrus synchronized but not bred, and blood samples were collected on days 0, 7, and at slaughter (day 14) during which tissues were collected. By design, heifer ADG did not differ (P = 0.96). Spleen weight and uterine weight were affected cubically (P = 0.03) decreasing from 0XPOS to 0.5X. Ovarian weight decreased linearly (P < 0.01) with increasing folate and B12 injection. Hemoglobin and hematocrit percentage were decreased (P < 0.01) in the 0.5X treatment compared with all other treatments. Plasma glucose, histotroph protein, and pancreatic α-amylase were decreased (P ≤ 0.04) in the 0.5X treatment. Heifers on the 2X treatment had greater pancreatic α-amylase compared with 0XNEG and 0.5X treatment. Interleukin-6 in plasma tended (P = 0.08) to be greater in the 0XPOS heifers compared with all other treatments. Lastly, 0XPOS-treated heifers had reduced (P ≤ 0.07) hydrogen peroxide production in muscle compared with 0XNEG heifers. These data imply that while certain doses of OCM do not improve whole animal physiology, OCM supplementation doses that disrupt one-carbon metabolism, such as that of the 0.5X treatment, can induce a negative systemic response that results in negative effects in both the dam and the conceptus during early gestation. Therefore, it is necessary to simultaneously establish an optimal OCM dose that increases circulating concentrations for use by the dam and the conceptus, while avoiding potential negative side effects of a disruptive OCM, to evaluate the long-term impacts of OCM supplementation of offspring programming., (© The Author(s) 2024. Published by Oxford University Press on behalf of the American Society of Animal Science.)

We evaluated the effects of feeding a vitamin and mineral supplement to nulliparous beef heifers throughout gestation on the mineral status of the dam, calf, placenta, and colostrum; offspring growth performance; and physiological responses of offspring raised as replacement heifers. Angus-based heifers (n = 31, initial body weight [BW] = 412.5 ± 53.68 kg) were adapted to an individual feeding system for 14 d, estrus synchronized and bred with female-sexed semen. Heifers were ranked by BW and randomly assigned to receive either a basal diet (CON; n = 14) or the basal diet plus 113 g heifer-1 d-1 of the vitamin and mineral supplement (VTM; n = 17). Targeted BW gains for both treatments was 0.45 kg heifer-1 d-1. Liver biopsies were obtained from dams at breeding, days 84 and 180 of gestation. At calving, liver biopsies were taken from dams and calves; colostrum, placenta, and blood samples were collected; and calf body measurements were recorded. After calving, all cow-calf pairs received a common diet through weaning, and F1 heifer calves were managed similarly after weaning. Offspring growth performance, feeding behavior, blood metabolites, and hormones were evaluated from birth through 15 mo of age. Data were analyzed using the MIXED procedure in SAS with repeated measures where appropriate. Hepatic concentrations of Se decreased in VTM dams (P ≤ 0.05) from day 84 to calving, while concentrations of Cu decreased in VTM and CON (P ≤ 0.05) from day 84 to calving. Calf liver concentrations of Se, Cu, Zn, and Co at birth were greater for VTM than CON (P ≤ 0.05), but calf birth BW and body measurements were not different (P = 0.45). Placental Se, colostrum quantity, total Se, Cu, Zn, and Mn in colostrum were greater (P ≤ 0.04) in VTM dams than CON. Finally, offspring from VTM dams were heavier than CON (P < 0.0001) from weaning through 15 mo of age. These results were coupled with greater (P ≤ 0.04) blood glucose at birth, decreased (P ≤ 0.05) blood urea nitrogen at pasture turn out and weaning, and altered feeding behaviors in VTM offspring compared with CON. Maternal gestational vitamin and mineral supplementation enhanced mineral status in dams and F1 progeny, augmented postnatal offspring growth and blood metabolites. Consequently, in utero vitamin and mineral supplementation may exert programming outcomes on the performance and productivity of females raised as herd replacements and should be considered when developing diets for gestating cows and heifers., (© The Author(s) 2024. Published by Oxford University Press on behalf of the American Society of Animal Science. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Importance: Emerging evidence suggests that microbiome-targeted approaches may provide a novel opportunity to reduce the incidence of reproductive failures in cattle. To develop such microbiome-based strategies, one of the first logical steps is to identify reproductive microbiome features related to fertility and to isolate the fertility-associated microbial species for developing a future bacterial consortium that could be administered before breeding to enhance pregnancy outcomes. Here, we characterized the vaginal and uterine microbiota in beef cattle that became pregnant or remained open via artificial insemination and identified microbiota features associated with fertility. We compared similarities between vaginal and uterine microbiota and between heifers and cows. Using culturing, we provided new insights into the culturable fraction of the vaginal and uterine microbiota and their antimicrobial resistance. Overall, our findings will serve as an important basis for future research aimed at harnessing the vaginal and uterine microbiome for improved cattle fertility., Competing Interests: The authors declare no conflict of interest.

Despite the extensive research conducted on ruminal methanogens and anti-methanogenic intervention strategies over the last 50 years, most of the currently researched enteric methane (CH 4 ) abatement approaches have shown limited efficacy. This is largely because of the complex nature of animal production and the ruminal environment, host genetic variability of CH 4 production, and an incomplete understanding of the role of the ruminal microbiome in enteric CH 4 emissions. Recent sequencing-based studies suggest the presence of methanogenic archaea in extra-gastrointestinal tract tissues, including respiratory and reproductive tracts of cattle. While these sequencing data require further verification via culture-dependent methods, the consistent identification of methanogens with relatively greater frequency in the airway and urogenital tract of cattle, as well as increasing appreciation of the microbiome-gut-organ axis together highlight the potential interactions between ruminal and extra-gastrointestinal methanogenic communities. Thus, a traditional singular focus on ruminal methanogens may not be sufficient, and a holistic approach which takes into consideration of the transfer of methanogens between ruminal, extra-gastrointestinal, and environmental microbial communities is of necessity to develop more efficient and long-term ruminal CH 4 mitigation strategies. In the present review, we provide a holistic survey of the methanogenic archaea present in different anatomical sites of cattle and discuss potential seeding sources of the ruminal methanogens.

Our objectives were to evaluate the impacts of providing vitamin and mineral (VTM) supplements to cow-calf pairs during the summer grazing period on cow and calf performance and liver concentrations of minerals. During a two-year period, 727 crossbred cows and their calves (initial cow BW = 601.7 ± 48.1 kg; calf BW = 87.8 ± 5.0 kg; n = 381 in year 1, n = 346 in year 2) from the Central Grasslands Research Extension Center (Streeter, N.D.) were blocked by parity ( young [parity 1 to 3], and old [parity 4+]) and randomly assigned to pastures at the beginning of the grazing season (16 in year 1 and 14 in year 2). Pastures were assigned to receive a free-choice VTM supplement ( SUPP ) or no VTM supplement ( CON ) from pasture turnout to pasture removal (158 and 156 days in year 1 and 2, respectively). Consecutive day weights were taken from cows and calves at pasture turnout and removal and liver biopsies were collected from a subset of cows at both timepoints and from calves at weaning. Cows were bred via AI 37 to 41 d after pasture turnout and by natural service cleanup bulls for a 70 to 80 d breeding season. Calving and weaning data were collected from the calf conceived and gestated during treatments. Data were analyzed for the effect of VTM treatment (SUPP vs. CON), block of parity, and their interaction using the GLM procedure of SAS with pasture as the experimental unit. Year was considered a random effect in the final analysis. Cow pregnancy success was evaluated using the GLIMMIX procedure in SAS with model terms of VTM treatment, parity, and their interaction with year as a random effect. In year 2, cows in differing days postpartum (DPP) groups at pasture turnout (66.1, 48.8, and 34.5 ± 1.04 DPP for EARLY, MID, and LATE groups, respectively) were selected for liver biopsies with cow as the experimental unit. Cow and calf BW and BW change were not impacted ( P ≥ 0.20) by VTM access. Pregnancy rate to AI, overall pregnancy rate, gestating calf birth BW and calving distribution were not affected ( P ≥ 0.11) by treatment. Liver concentrations of Se, Cu, and Co were greater ( P ≤ 0.002) at pasture removal and weaning for cows and suckling calves that had access to VTM. Cows considered EARLY calving had greater ( P = 0.05) concentrations of liver Se compared with LATE calving cows. Although VTM supplementation enhanced concentrations of key minerals in the liver of cow-calf pairs, reproductive and growth performance was not affected., (© The Author(s) 2023. Published by Oxford University Press on behalf of the American Society of Animal Science.)

Early life microbial colonization and factors affecting colonization patterns are gaining interest due to recent developments suggesting that early life microbiome may play a role in Developmental Origins of Health and Disease. In cattle, limited information exists on the early microbial colonization of anatomical sites involved in bovine health beyond the gastrointestinal tract. Here, we investigated 1) the initial microbial colonization of seven different anatomical locations in newborn calves and 2) whether these early life microbial communities and 3) serum cytokine profiles are influenced by prenatal vitamin and mineral (VTM) supplementation. Samples were collected from the hoof, liver, lung, nasal cavity, eye, rumen (tissue and fluid), and vagina of beef calves that were born from dams that either received or did not receive VTM supplementation throughout gestation ( n  = 7/group). Calves were separated from dams immediately after birth and fed commercial colostrum and milk replacer until euthanasia at 30 h post-initial colostrum feeding. The microbiota of all samples was assessed using 16S rRNA gene sequencing and qPCR. Calf serum was subjected to multiplex quantification of 15 bovine cytokines and chemokines. Our results indicated that the hoof, eye, liver, lung, nasal cavity, and vagina of newborn calves were colonized by site-specific microbiota, whose community structure differed from the ruminal-associated communities (0.64 ≥ R 2 ≥ 0.12, p ≤ 0.003). The ruminal fluid microbial community was the only one that differed by treatment ( p < 0.01). However, differences ( p < 0.05) by treatment were detected in microbial richness (vagina); diversity (ruminal tissue, fluid, and eye); composition at the phylum and genus level (ruminal tissue, fluid, and vagina); and in total bacterial abundance (eye and vagina). From serum cytokines evaluated, concentration of chemokine IP-10 was greater ( p = 0.02) in VTM calves compared to control calves. Overall, our results suggest that upon birth, the whole-body of newborn calves are colonized by relatively rich, diverse, and site-specific bacterial communities. Noticeable differences were observed in ruminal, vaginal, and ocular microbiota of newborn calves in response to prenatal VTM supplementation. These findings can derive future hypotheses regarding the initial microbial colonization of different body sites, and on maternal micronutrient consumption as a factor that may influence early life microbial colonization., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Luecke, Holman, Schmidt, Gzyl, Hurlbert, Menezes, Bochantin, Kirsch, Baumgaertner, Sedivec, Swanson, Dahlen and Amat.)

A growing number of studies have investigated the feasibility of utilizing hemp by-products as livestock feedstuffs; however, their impact on livestock microbiomes remains unexplored. Here, we evaluated the effects of feeding hempseed cake on the gastrointestinal, respiratory, and reproductive microbiota in beef heifers. Angus-crossbred heifers (19-months old, initial body weight = 494 ± 10 kg [SE]) were fed a corn-based finishing diet containing 20% hempseed cake as a substitute for 20% corn dried distillers' grains with solubles (DM basis; Control; n = 16/group) for 111 days until slaughter. Ruminal fluid and deep nasopharyngeal swabs (days 0, 7, 42, 70 and 98), and vaginal and uterine swabs (at slaughter) were collected, and the microbiota assessed using 16S rRNA gene sequencing. Diet affected the community structure of the ruminal (d 7-98; 0.06 ≤ R 2  ≤ 0.12; P < 0.05), nasopharyngeal (d 98; R 2  = 0.18; P < 0.001), and vaginal (R 2  = 0.06; P < 0.01) microbiota. Heifers fed hempseed cake had increased microbial diversity in the rumen, reduced microbial richness in the vagina, and greater microbial diversity and richness in the uterus. In addition to the distinct microbial communities in the rumen, nasopharynx, vagina and uterus, we identified 28 core taxa that were shared (≥ 60% of all samples) across these sampling locations. Feeding hempseed cake appeared to alter the bovine gut, respiratory and reproductive microbiota. Our results suggest that future research aiming to evaluate the use of hemp by-products in livestock diet should consider their impact on animal microbiome and microbiome mediated animal health and reproductive efficiency. Our findings also highlight the need for research evaluating the impact of hemp-associated food and personal care products on the human microbiome., (© 2023. The Author(s).)

Maternal mineral nutrition during the critical phases of fetal development may leave lifetime impacts on the productivity of an individual. Most research within the developmental origins of the health and disease (DOHaD) field is focused on the role of macronutrients in the genome function and programming of the developing fetus. On the other hand, there is a paucity of knowledge about the role of micronutrients and, specifically, minerals in regulating the epigenome of livestock species, especially cattle. Therefore, this review will address the effects of the maternal dietary mineral supply on the fetal developmental programming from the embryonic to the postnatal phases in cattle. To this end, we will draw a parallel between findings from our cattle model research with data from model animals, cell lines, and other livestock species. The coordinated role and function of different mineral elements in feto-maternal genomic regulation underlies the establishment of pregnancy and organogenesis and, ultimately, affects the development and functioning of metabolically important tissues, such as the fetal liver, skeletal muscle, and, importantly, the placenta. Through this review, we will delineate the key regulatory pathways involved in fetal programming based on the dietary maternal mineral supply and its crosstalk with epigenomic regulation in cattle.

Herein, we present a dataset based on the RNA-Seq analysis of liver tissue from bovine female fetuses at day 83 of gestation. The findings were reported in the main article, "Periconceptual maternal nutrition affects fetal liver programming of energy- and lipid-related genes" [1]. These data were generated to investigate the effects of periconceptual maternal vitamin and mineral supplementation and rates of body weight gain on the transcript abundance of genes associated with fetal hepatic metabolism and function. To this end, crossbred Angus beef heifers ( n  = 35) were randomly assigned to 1 of 4 treatments in a 2 × 2 factorial design. The main effects tested were 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 - 0.28 kg/d) or moderate (MG - 0.79 kg/d) - from breeding to day 83). The fetal liver was collected on day 83 ± 0.27 of gestation. After total RNA isolation and quality control, strand-specific RNA libraries were prepared and sequenced on the Illumina® NovaSeq 6000 platform to generate paired-end 150-bp reads. After read mapping and counting, differential expression analysis was performed with edgeR. We identified 591 unique differentially expressed genes across all six vitamin-gain contrasts (FDR ≤ 0.1). To our knowledge, this is the first dataset investigating the fetal liver transcriptome in response to periconceptual maternal vitamin and mineral supplementation and/or the rate of weight gain. The data described in this article provides genes and molecular pathways differentially programming liver development and function., Competing Interests: Authors J. C. Forcherio and R. Scott are employees of Purina Animal Nutrition LLC (Land O'Lakes, Inc., Arden Hills, MN, USA), which sponsored the sample analysis for this experiment. Purina Animal Nutrition LLC manufactured the Purina® Wind & Rain® Storm® AllSeason 7.5 Complete mineral, the VTM and NoVTM pellets, and the protein/energy supplement used in this study. The funders had no role in the collection, analyses, or interpretation of data, in the writing of the manuscript, or in the decision to publish the results that were done entirely independently by North Dakota State University and Auburn University personnel. All the other authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2023 The Author(s).)

In this study, we evaluated the seminal and fecal microbiota in yearling beef bulls fed a common diet to achieve moderate (1.13 kg/day) or high (1.80 kg/day) rates of weight gain. Semen samples were collected on days 0 and 112 of dietary intervention ( n  = 19/group) as well as postbreeding ( n  = 6/group) using electroejaculation, and the microbiota was assessed using 16S rRNA gene sequencing, quantitative PCR (qPCR), and culturing. The fecal microbiota was also evaluated, and its similarity with seminal microbiota was assessed. A subset of seminal bacterial isolates ( n  = 33) was screened for resistance against 28 antibiotics. A complex and dynamic microbiota was detected in bovine semen, and the community structure was affected by sampling time ( R 2  = 0.16, P <  0.001). Microbial richness increased significantly from day 0 to day 112, and diversity increased after breeding ( P >  0.05). Seminal microbiota remained unaffected by the differential rates of gain, and its overall composition was distinct from fecal microbiota, with only 6% of the taxa shared between them. A total of 364 isolates from 49 different genera were recovered under aerobic and anaerobic culturing. Among these seminal isolates were pathogenic species and those resistant to several antibiotics. Overall, our results suggest that bovine semen harbors a rich and complex microbiota which changes over time and during the breeding season but appears to be resilient to differential gains achieved via a common diet. Seminal microbiota is distinct from the fecal microbiota and harbors potentially pathogenic and antibiotic-resistant bacterial species. IMPORTANCE Increasing evidence from human and other animal species supports the existence of a commensal microbiota in semen and that this seminal microbiota may influence not only sperm quality and fertility but also female reproduction. Seminal microbiota in bulls and its evolution and factors shaping this community, however, remain largely underexplored. In this study, we characterized the seminal microbiota of yearling beef bulls and its response to the bull age, different weight gains, and mating activity. We compared bacterial composition between seminal and fecal microbiota and evaluated the diversity of culturable seminal bacteria and their antimicrobial resistance. Our results obtained from sequencing, culturing, and antibiotic susceptibility testing provide novel information on the taxonomic composition, evolution, and factors shaping the seminal microbiota of yearling beef bulls. This information will serve as an important basis for further understanding of the seminal microbiome and its involvement in reproductive health and fertility in cattle.

Our objectives were to develop a Mobile Cow Command Center (MCCC) capable of precision monitoring of grazing heifers to 1) examine the relationship between supplement intake on concentrations of liver mineral and blood metabolites and 2) examine activity, reproductive, and health behavior. Yearling crossbred Angus heifers ( N = 60; initial BW = 400.4 ± 6.2 kg) were fitted with radio frequency identification ear tags that allowed access to electronic feeders (SmartFeed system; C-Lock Inc., Rapid City, SD), and with activity monitoring tags (CowManager B.V., the Netherlands) that monitored reproductive, feeding, and health-associated behaviors. Heifers were assigned randomly to one of three treatments for a 57-day monitoring period: 1) no supplement ( CON ; N = 20), 2) free choice mineral ( MIN ; Purina Wind and Rain Storm [Land O'Lakes, Inc.], N = 20), or 3) free choice energy and mineral supplement ( NRG ; Purina Accuration Range Supplement 33 with added MIN [Land O'Lakes, Inc.], N = 20). Consecutive day body weights, blood, and liver biopsies were collected at pasture turnout and final day of monitoring. By design, mineral intake was greatest in MIN heifers (49 ± 37 g/d) and energy supplement intake was greatest in NRG heifers (1,257 ± 37 g/d). Final BW and ADG were similar among treatments ( P > 0.42). Concentrations of glucose on day 57 were greater ( P = 0.01) in NRG compared with CON and MIN heifers. Liver concentrations of Se and Fe on day 57 were greater ( P < 0.05) in NRG heifers than CON, with MIN being intermediate. Activity tags reported NRG heifers spent less time eating ( P < 0.0001) and more time ( P < 0.0001) being "highly active" than MIN with CON heifers being intermediate. Data retrieved from activity tags identified 16 of 28 pregnant heifers exhibiting some type of estrus-associated behavior even after confirmation of established pregnancy. The activity monitoring system triggered a total of 146 health alerts from 34 of the 60 heifers monitored, but only 3 heifers of the heifers initiating an electronic health alert needed clinical treatment. However, animal care staff identified nine additional heifers that required treatment for which no electronic health alert was generated. The electronic feeders successfully controlled intake of individual heifers managed in groups pastures; however, the activity monitoring system misrepresented estrus and health events., (© The Author(s) 2023. Published by Oxford University Press on behalf of the American Society of Animal Science.)

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.

Adequate maternal nutrition is key for proper fetal development and epigenetic programming. One-carbon metabolites (OCM), including vitamin B12, folate, choline, and methionine, play a role in epigenetic mechanisms associated with developmental programming. This study investigated the presence of B12 and folate in maternal serum, allantoic fluid (ALF), and amniotic fluid (AMF), as well as how those concentrations in all three fluids correlate to the concentrations of methionine-folate cycle intermediates in heifers receiving either a control (CON) or restricted (RES) diet for the first 50 d of gestation and fetal hepatic gene expression for methionine-folate cycle enzymes. Angus cross heifers (n = 43) were estrus synchronized, bred via artificial insemination with semen from a single sire, and randomly assigned to one of two nutrition treatments (CON = 20, RES = 23). Heifers were ovariohysterectomized on either day 16 (n = 14), 34 (n = 15), or 50 of gestation (n = 14), where samples of maternal serum (n = 42), ALF (n = 29), and AMF (n = 11) were collected and analyzed for concentrations of folate and B12. Concentrations of B12 and folate in ALF were greater (P < 0.05) in RES compared to CON. For ALF, folate concentrations were also greater (P < 0.01) on day 34 compared to day 50. There was a significant (P = 0.04) nutrition × fluid interaction for B12 concentrations where concentrations were greatest in restricted ALF, intermediate in control ALF, and lowest in CON and RES serum and AMF. Folate concentrations were greatest (P < 0.01) in ALF, intermediate in serum, and lowest in AMF. Additionally, positive correlations (P < 0.05) were found between ALF and AMF folate concentrations and AMF concentrations of methionine, serine, and glycine. Negative correlations (P < 0.05) between AMF folate and serum homocysteine were also observed. Both positive and negative correlations (P < 0.05) depending on the fluid evaluated were found between B12 and methionine, serine, and glycine concentrations. There was a downregulation (P = 0.05) of dihydrofolate reductase and upregulation (P = 0.03) of arginine methyltransferase 7 gene expression in RES fetal liver samples compared with CON fetal liver on day 50. Combined, these data show restricted maternal nutrition results in increased B12 and folate concentrations present in fetal fluids, and increased expression of genes for enzymes within one-carbon metabolism., (© The Author(s) 2023. Published by Oxford University Press on behalf of the American Society of Animal Science. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Developmental programming is the concept that 'stressors' during development (i.e. pregnancy, the perinatal period and infancy) can cause long-term changes in gene expression, leading to altered organ structure and function. Such long-term changes are associated with an increased risk of a host of chronic pathologies, or non-communicable diseases including abnormal growth and body composition, behavioural or cognitive dysfunction, metabolic abnormalities, and cardiovascular, gastro-intestinal, immune, musculoskeletal and reproductive dysfunction. Maternal nutrition during the periconceptual period, pregnancy and postnatally can have profound influences on the developmental program. Animal models, including domestic livestock species, have been important for defining the mechanisms and consequences of developmental programming. One of the important observations is that maternal nutritional status and other maternal stressors (e.g. environmental temperature, high altitude, maternal age and breed, multiple fetuses, etc.) early in pregnancy and even periconceptually can affect not only embryonic/fetal development but also placental development. Indeed, altered placental function may underlie the effects of many maternal stressors on fetal growth and development. We suggest that future directions should focus on the consequences of developmental programming during the offspring's life course and for subsequent generations. Other important future directions include evaluating interventions, such as strategic dietary supplementation, and also determining how we can take advantage of the positive, adaptive aspects of developmental programming.

Reproductive failure is a major economical drain on cow-calf operations across the globe. It can occur in both males and females and stem from prenatal and postnatal influences. Therefore, the cattle industry has been making efforts to improve fertility and the pregnancy rate in cattle herds as an attempt to maintain sustainability and profitability of cattle production. Despite the advancements made in genetic selection, nutrition, and the implementation of various reproductive technologies, fertility rates have not significantly improved in the past 50 years. This signifies a missing factor or factors in current reproductive management practices that influence successful fertilization and pregnancy. Emerging lines of evidence derived from human and other animals including cattle suggest that the microbial continuum along the male and female reproductive tracts are associated with male and female fertility-that is, fertilization, implantation, and pregnancy success-highlighting the potential for harnessing the male and female reproductive microbiome to improve fertility in cattle. The objective of this narrative review is to provide an overview of the recent studies on the bovine seminal and vagino-uterine microbiome and discuss individual and interactive roles of these microbial communities in defining cattle fertility., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Luecke, Webb, Dahlen, Reynolds and Amat.)

In vertebrates and invertebrates, selenium (Se) is an essential micronutrient, and Se deficiency or excess is associated with gonadal insufficiency and gamete dysfunction in both males and females, leading to implantation failure, altered embryonic development and, ultimately, infertility. During pregnancy, Se excess or deficiency is associated with miscarriage, pre-eclampsia (hypertension of pregnancy), gestational diabetes, fetal growth restriction and preterm birth. None of this is surprising, as Se is present in high concentrations in the ovary and testes, and work in animal models has shown that addition of Se to culture media improves embryo development and survival in vitro in association with reduced reactive oxygen species and less DNA damage. Selenium also affects uterine function and conceptus growth and gene expression, again in association with its antioxidant properties. Similarly, Se improves testicular function including sperm count, morphology and motility, and fertility. In animal models, supplementation of Se in the maternal diet during early pregnancy improves fetal substrate supply and alters fetal somatic and organ growth. Supplementation of Se throughout pregnancy in cows and sheep that are receiving an inadequate or excess dietary intake affected maternal whole-body and organ growth and vascular development, and also affected expression of angiogenic factors in maternal and fetal organs. Supplemental Se throughout pregnancy also affected placental growth, which may partly explain its effects on fetal growth and development, and also affected mammary gland development, colostrum yield and composition as well as postnatal development of the offspring. In conclusion, Se supplementation in nutritionally compromised pregnancies can potentially improve fertility and pregnancy outcomes, and thereby improve postnatal growth and development. Future research efforts should examine in more detail and more species the potential benefits of Se supplementation to reproductive processes in mammals., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Dahlen, Reynolds and Caton.)

The objective of this study was to evaluate the effects of feeding heifers a vitamin and mineral supplement and targeting divergent rates of weight gain during early gestation on the fetal liver amino acid, carbohydrate, and energy profile at d 83 of gestation. Seventy-two crossbred Angus heifers were randomly assigned in a 2 × 2 factorial arrangement to one of four treatments comprising the main effects of vitamin and mineral supplementation (VTM or NOVTM) and feeding to achieve different rates of weight gain (low gain [LG] 0.28 kg/day vs. moderate gain [MG] 0.79 kg/day). Thirty-five gestating heifers with female fetuses were ovariohysterectomized on d 83 of gestation and fetal liver was collected and analyzed by reverse phase UPLC-tandem mass spectrometry with positive and negative ion mode electrospray ionization, as well as by hydrophilic interaction liquid chromatography UPLC-MS/MS with negative ion mode ESI for compounds of known identity. The Glycine, Serine, and Threonine metabolism pathway and the Leucine, Isoleucine, and Valine metabolism pathway had a greater total metabolite abundance in the liver of the NOVTM-LG group and least in the VTM-LG group (p < 0.01). Finally, both the TCA Cycle and Oxidative Phosphorylation pathways within the Energy Metabolism superpathway were differentially affected by the main effect of VTM, where the TCA cycle metabolites were greater (p = 0.04) in the NOVTM fetal livers and the Oxidative Phosphorylation biochemicals were greater (p = 0.02) in the fetal livers of the VTM supplemented heifers. These data demonstrate that the majority of metabolites that are affected by rate of weight gain or vitamin/mineral supplementation are decreased in heifers on a greater rate of weight gain or vitamin/mineral supplementation.

Fetal programming is established early in life, likely through epigenetic mechanisms that control gene expression. Micronutrients can act as epigenetic modifiers (EM) by modulating the genome through mechanisms that include DNA methylation and post-translational modification of chromatin. Among the EM, methionine, choline, folate, and vitamin B 12 have been suggested as key players of DNA methylation. However, the effects of supplementing these four EM, involved in the methionine folate cycle on DNA methylation, are still under investigation. This manuscript provides the genome-wide DNA methylation dataset (GSE180362) of bovine embryonic fibroblast cells exposed to different supplementation levels of glucose and methionine, choline, folate, and vitamin B 12 (collectively named as Epigenetic Modifiers - EM). The DNA methylation was measured using MSP-I digestion and Reduced Representation Bisulfite Sequencing. Bioinformatics analyses included data quality control, read mapping, methylation calling, and differential methylation analyses. Supplementary file S1 and data analysis codes are within this article. To our knowledge, this is the first dataset investigating the effects of four EM in bovine embryonic fibroblast DNA methylation profiles. Furthermore, this data and its findings provide information on putative candidate genes responsive to DNA methylation due to EM supplementation., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships which have or could be perceived to have influenced the work reported in this article., (© 2022 The Author(s). Published by Elsevier Inc.)

Epigenetic modifiers (EM; methionine, choline, folate, and vitamin B 12 ) are important for early embryonic development due to their roles as methyl donors or cofactors in methylation reactions. Additionally, they are essential for the synthesis of nucleotides, polyamines, redox equivalents, and energy metabolites. Despite their importance, investigation into the supplementation of EM in ruminants has been limited to one or two epigenetic modifiers. Like all biochemical pathways, one-carbon metabolism needs to be stoichiometrically balanced. Thus, we investigated the effects of supplementing four EM encompassing the methionine-folate cycle on bovine embryonic fibroblast growth, mitochondrial function, and DNA methylation. We hypothesized that EM supplemented to embryonic fibroblasts cultured in divergent glucose media would increase mitochondrial respiration and cell growth rate and alter DNA methylation as reflected by changes in the gene expression of enzymes involved in methylation reactions, thereby improving the growth parameters beyond Control treated cells. Bovine embryonic fibroblast cells were cultured in Eagle's minimum essential medium with 1 g/L glucose (Low) or 4.5 g/L glucose (High). The control medium contained no additional OCM, whereas the treated media contained supplemented EM at 2.5, 5, and 10 times (×2.5, ×5, and ×10, respectively) the control media, except for methionine (limited to ×2). Therefore, the experimental design was a 2 (levels of glucose) × 4 (levels of EM) factorial arrangement of treatments. Cells were passaged three times in their respective treatment media before analysis for growth rate, cell proliferation, mitochondrial respiration, transcript abundance of methionine-folate cycle enzymes, and DNA methylation by reduced-representation bisulfite sequencing. Total cell growth was greatest in High ×10 and mitochondrial maximal respiration, and reserve capacity was greatest ( p < 0.01) for High ×2.5 and ×10 compared with all other treatments. In Low cells, the total growth rate, mitochondrial maximal respiration, and reserve capacity increased quadratically to 2.5 and ×5 and decreased to control levels at ×10. The biological processes identified due to differential methylation included the positive regulation of GTPase activity, molecular function, protein modification processes, phosphorylation, and metabolic processes. These data are interpreted to imply that EM increased the growth rate and mitochondrial function beyond Control treated cells in both Low and High cells, which may be due to changes in the methylation of genes involved with growth and energy metabolism., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Crouse, Caton, Claycombe-Larson, Diniz, Lindholm-Perry, Reynolds, Dahlen, Borowicz and Ward.)

Recent developments call for further research on the timing and mechanisms involved in the initial colonization of the fetal/infant gut by the maternal microbiome and its role in Developmental Origins of Health and Disease (DOHaD). Although progress has been made using primarily preterm infants, ethical and legal constraints hinder research progress in embryo/fetal-related research and understanding the developmental and mechanistic roles of the maternal microbiome in fetal microbial imprinting and its long-term role in early-life microbiome development. Rodent models have proven very good for studying the role of the maternal microbiome in fetal programming. However, some inherent limitations in these animal models make it challenging to study perinatal microbial colonization from a biomedical standpoint. In this review, we discuss the potential use of bovine animals as a biomedical model to study the maternal microbiome, in utero microbial colonization of the fetal gut, and their impact on offspring development and DOHaD., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Amat, Dahlen, Swanson, Ward, Reynolds and Caton.)

A recent study reported the existence of a diverse microbiota in 5-to-7-month-old calf fetuses, suggesting that colonization of the bovine gut with so-called "pioneer" microbiota may begin during mid-gestation. In the present study, we investigated 1) the presence of microbiota in bovine fetuses at early gestation (12 weeks), and 2) whether the fetal microbiota is influenced by the maternal rate of gain or dietary supplementation with vitamins and minerals (VTM) during early gestation. Amniotic and allantoic fluids, and intestinal and placental (cotyledon) tissue samples obtained from fetuses ( n = 33) on day 83 of gestation were processed for the assessment of fetal microbiota using 16S rRNA gene sequencing. The sequencing results revealed that a diverse and complex microbial community was present in each of these fetal compartments evaluated. Allantoic and amniotic fluids, and fetal intestinal and placenta microbiota each had distinctly different (0.047 ≥ R 2 ≥ 0.019, P ≤ 0.031) microbial community structures. Allantoic fluid had a greater ( P < 0.05) microbial richness (number of OTUs) (Mean 122) compared to amniotic fluid (84), intestine (63), and placenta (66). Microbial diversity (Shannon index) was similar for the intestinal and placental samples, and both were less diverse compared with fetal fluid microbiota ( P < 0.05). Thirty-nine different archaeal and bacterial phyla were detected across all fetal samples, with Proteobacteria (55%), Firmicutes (16.2%), Acidobacteriota (13.6%), and Bacteroidota (5%) predominating. Among the 20 most relatively abundant bacterial genera, Acidovorax , Acinetobacter , Brucella , Corynebacterium , Enterococcus , Exiguobacterium , and Stenotrophomonas differed by fetal sample type ( P < 0.05). A total of 55 taxa were shared among the four different microbial communities. qPCR of bacteria in the intestine and placenta samples as well as scanning electron microscopy imaging of fetal fluids provided additional evidence for the presence of a microbiota in these samples. Minor effects of maternal rate of gain and VTM supplementation, and their interactions on microbial richness and composition were detected. Overall, the results of this study indicate that colonization with pioneer microbiota may occur during early gestation in bovine fetuses, and that the maternal nutritional regime during gestation may influence the early fetal microbiota., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Amat, Holman, Schmidt, McCarthy, Dorsam, Ward, Borowicz, Reynolds, Caton, Sedivec and Dahlen.)

Maternal nutritional status affects conceptus development and, therefore, embryonic survival, growth, and development. These effects are apparent very early in pregnancy, which is when most embryonic losses occur. Maternal nutritional status has been shown to affect conceptus growth and gene expression throughout the periconceptual period of pregnancy (the period immediately before and after conception). Thus, the periconceptual period may be an important "window" during which the structure and function of the fetus and the placenta are "programmed" by stressors such as maternal malnutrition, which can have long-term consequences for the health and well-being of the offspring, a concept often referred to as Developmental Origins of Health and Disease (DOHaD) or simply developmental programming. In this review, we focus on recent studies, using primarily animal models, to examine the effects of various maternal "stressors," but especially maternal malnutrition and Assisted Reproductive Techniques (ART, including in vitro fertilization, cloning, and embryo transfer), during the periconceptual period of pregnancy on conceptus survival, growth, and development. We also examine the underlying mechanisms that have been uncovered in these recent studies, such as effects on the development of both the placenta and fetal organs. We conclude with our view of future research directions in this critical area of investigation., (© 2022. Springer Nature Switzerland AG.)

Assisted reproductive techniques (ART) and parental nutritional status have profound effects on embryonic/fetal and placental development, which are probably mediated via "programming" of gene expression, as reflected by changes in their epigenetic landscape. Such epigenetic changes may underlie programming of growth, development, and function of fetal organs later in pregnancy and the offspring postnatally, and potentially lead to long-term changes in organ structure and function in the offspring as adults. This latter concept has been termed developmental origins of health and disease (DOHaD), or simply developmental programming, which has emerged as a major health issue in animals and humans because it is associated with an increased risk of non-communicable diseases in the offspring, including metabolic, behavioral, and reproductive dysfunction. In this review, we will briefly introduce the concept of developmental programming and its relationship to epigenetics. We will then discuss evidence that ART and periconceptual maternal and paternal nutrition may lead to epigenetic alterations very early in pregnancy, and how each pregnancy experiences developmental programming based on signals received by and from the dam. Lastly, we will discuss current research on strategies designed to overcome or minimize the negative consequences or, conversely, to maximize the positive aspects of developmental programming.

In the present study, we evaluated whether the nasopharyngeal, ruminal, and vaginal microbiota would diverge (1) in virgin yearling beef heifers (9 months old) due to the maternal restricted gain during the first trimester of gestation; and (2) in pregnant beef heifers in response to the vitamin and mineral (VTM) supplementation during the first 6 months of pregnancy. As a secondary objective, using the microbiota data obtained from these two cohorts of beef heifers managed at the same location and sampled at the same time, we performed a holistic assessment of the microbial ecology residing within the respiratory, gastrointestinal, and reproductive tract of cattle. Our 16S rRNA gene sequencing results revealed that both α and β-diversity of the nasopharyngeal, ruminal and vaginal microbiota did not differ between virgin heifers raised from dams exposed to either a low gain (targeted average daily gain of 0.28 kg/d, n = 22) or a moderate gain treatment (0.79 kg/d, n = 23) during the first 84 days of gestation. Only in the vaginal microbiota were there relatively abundant genera that were affected by maternal rate of gain during early gestation. Whilst there was no significant difference in community structure and diversity in any of the three microbiota between pregnant heifers received no VTM ( n = 15) and VTM supplemented ( n = 17) diets, the VTM supplementation resulted in subtle compositional alterations in the nasopharyngeal and ruminal microbiota. Although the nasopharyngeal, ruminal, and vaginal microbiota were clearly distinct, a total of 41 OTUs, including methanogenic archaea, were identified as core taxa shared across the respiratory, gastrointestinal, and reproductive tracts of both virgin and pregnant heifers.

Crossbred Angus cow-calf pairs ( n = 28 pairs) at the Central Grasslands Research Extension Center (Streeter, ND) were used to evaluate an electronic feeder to monitor individual mineral intake and feeding behavior and their relationship with growth performance and liver mineral concentrations. Cows and calves were fitted with radio frequency identification ear tags that allowed access to an electronic feeder (SmartFeed system; C-Lock Inc., Rapid City, SD) and were provided ad libitum minerals (Purina Wind and Rain Storm, Land O'Lakes, Inc., Arden Hills, MN). Mineral intake, number of visits, and duration at the feeder were recorded over a 95-d monitoring period while pairs were grazing native range. Liver biopsies were collected from a subset of cows on the final day of monitoring and analyzed for mineral concentrations. Data were analyzed with the GLM procedure in SAS for mineral intake and feeding behavior with age class (cows vs. calves), intake category (high vs. low), and the interaction between class and category in the model. Correlations were calculated among cow feeding behavior and calf intake and growth performance with the CORR procedure, and a comparison of liver mineral concentrations among cows of high (>90 g/d; average 125.4 g/d) and low (<90 g/d; average 33.5 g/d) mineral intake was performed using the GLM procedure. High-intake calves (>50 g/d; average 72.2 g/d) consumed greater ( P < 0.001) amounts of minerals than low-intake calves (<50 g/d; average 22.2 g/d) intake calves. Cows and calves attended the mineral feeder a similar ( P = 0.71) proportion of the days during the experiment (overall mean of 20%, or once every 5 d). On days calves visited the feeder, they consumed less ( P < 0.01) minerals than cows (222 ± 27 vs. 356 ± 26 g/d, respectively). Over the grazing period, calves gained 1.17 ± 0.02 kg/d, whereas cows lost 0.35 ± 0.02 kg/d. Calf mineral intake was correlated with cow duration at the mineral feeder ( r = 0.403, P = 0.05). Cows with high mineral intake had greater ( P < 0.01) concentrations of Se (2.92 vs. 2.41 ug/g), Cu (247 vs. 116 ug/g), and Co (0.51 vs. 0.27 ug/g) compared with low mineral intake cows, but liver concentrations of Fe, Zn, Mo, and Mn did not differ ( P ≥ 0.22). We were able to successfully monitor individual mineral intake and feeding behavior with the electronic feeder evaluated, and the divergence in mineral intake observed with the feeder was corroborated by concentrations of minerals in the liver., (© The Author(s) 2021. Published by Oxford University Press on behalf of the American Society of Animal Science.)