Single Sire Research Articles

PSIX-22 Maternal methane, carbon dioxide, and oxygen flux compared with progeny gas flux

Abstract Red Angus heifers [n = 19; body weight (BW) = 370 ± 8.1 kg) and their paired offspring (BW = 283 ± 4.2 kg; 18 heifers, 1 steer) were used to compare the respiration gas parameters between dams and their progeny over 2 calving cycles (8 pairs in the 1st cycle, 11 in the 2nd). The methane (CH4), carbon dioxide (CO2) emissions and oxygen (O2) consumption of the dams were measured at 14 mo of age over a 70-d period using an automated head-chamber system (AHCS) that measured the flux of these 3 gases. Dams grazed native mixed-grass prairie and were offered a daily supplement of alfalfa (Medicago sativa L.) pellets (1.0 kg) via the AHCS. Dams were artificially inseminated using sexed semen (female) from a single Black Angus sire. The gas fluxes of the progeny were measured, beginning at 11 mo of age, over a 70-d period. Like the dams, during their measurement period, the progeny grazed native mixed-grass prairie and were offered a daily supplement (1.0 kg) of alfalfa pellets via the AHCS. Pearson correlations were developed between the 70-d means of daily CH4 and CO2 emissions, O2 consumption, CH4:CO2 ratio, respiratory quotient (RQ), and heat of production estimates for the dams and their progeny by regressing maternal parameters on the parameters of the progeny, using beginning dam and progeny BW as covariates, and year as an indicator variable. A dam’s CH4 emission was very strongly correlated with that of their offspring (ρ = 0.86). The CO2 emission and O2 consumption of the dams were both also very strongly (ρ = 0.92 and ρ = 0.97, respectively) correlated, with that of their progeny. The heat production and CH4:CO2 ratio of the dams were very strongly (ρ = 0.96) and fairly (ρ = 0.50) correlated, respectively, with that of their progeny. Lastly, the RQ of the dam was also very strongly correlated (ρ = 0.87) with the RQ of the progeny. These results indicate that CH4 and CO2 emissions, O2 consumption, CH4:CO2 ratio, and heat of production of the dam are fairly to very strongly correlated to the gas flux parameters of their progeny. Hence, even with this small dataset, there is evidence that beef cattle traits related to greenhouse gas emissions are somewhat heritable and could potentially be selected in breeding animals to decrease greenhouse gas emission intensity.

PSIII-6 Multi-sire artificial insemination of beef heifers in the Nebraska Sandhills effect on pregnancy rate compared with single-sire insemination

Abstract The objective of this study was to compare pregnancy outcomes resulting from artificial insemination (AI) with multi-sire semen (MS) compared with single-sire semen (SS) in beef heifers. Another study observed pregnancy and paternity outcomes to sexed MS without a control treatment. In the first study, unrelated ABS Global Angus bulls (n = 3) were selected for AI to 441 (Yr 1) and 391 (Yr 2) Angus crossbred heifers. Heifers were synchronized with the melengestrol acetate – prostaglandin F2α timed-AI protocol. Conventional semen, from each bull, were labeled A, B, or C. Remaining semen was pooled in equal amounts to form conventional MS straws. Heifers were AI using alternating treatment of MS or SS: A, B, or C with one-half of inseminated heifers receiving either MS or SS. The GLIMMIX procedure in SAS 9.4 analyzed responses to treatments using the BIN distribution option and reported as LSMEANS. Contrasts and estimates analyzed responses between treatments. Ultrasound determined SS pregnancy rate averaged 60% (SS-A = 64%, SS-B = 58%, SS-C = 58%) and MS average 62% (P = 0.63). Paternity was determined in 57 MS calves and confirmed in 49 SS calves. In Yr 1, Bull A sired 7%, bull B sired 56%, and bull C sired 37%. Although bull A was much less successful than bull B or C, the values are not significantly different due to the small sample size (P = 0.11 and P = 0.24, respectively). Few conclusions can be made from the ongoing study, but it is hypothesized MS may improve consistency of pregnancy rate by equilibrating semen quality in herds subjected to timed AI. In the observational study, multiple 5-way cross bulls born from the ranch herd were selected for AI of 951 ± 41 heifers for 3 yr. Heifers were synchronized using the previously mentioned protocol with split-time AI. Unequal amounts of semen were collected, pooled, and sexed to favor heifer progeny at an expected ratio of 9:1. Semen from 2 to 4 random bulls were pooled making 3 to 4 MS groups each year. Heifers exhibiting estrus were administered sexed MS, but there was no control treatment. The observed ultrasound determined average pregnancy rate of heifers administered sexed MS was 65% (Yr 1), 75% (Yr 2), and 72% (Yr 3). Previous literature suggests average pregnancy rate to AI with sexed SS is 53%. Paternity testing in Yr 2 found semen contribution mirrored paternity ratios within some groups but the sire group with 2 bulls found one sired 15% and the other sired 85%. This study exemplified a viable method for AI that requires further research. Further research into semen quality of a single sire, and pooled semen could lead to optimization of fertilization among individuals within a herd.

429 Maternal rate of gain and one-carbon metabolites supplementation alters the gene expression profile of fetal brain during early gestation in beef heifers

Abstract 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 bta-mir-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.

428 Influence of melatonin supplementation in pregnant multiparous crossbred beef cows grazing endophyte-infected tall fescue on dam weight, milk yield, and pre-weaning growth of steer calves

Abstract The objective was to evaluate the influence of melatonin supplementation in pregnant beef cattle grazing toxic, endophyte-infected tall fescue (E+) on post-calving dam body weight (BW), milk production, and preweaning growth of steer calves. Multiparous cows were artificially inseminated with sex-sorted semen (Y chromosome-bearing sperm) from a single Red Angus sire and maintained on pastures free from E+ fescue prior to the start of the treatment period. Pregnant cows were assigned to one of three treatment groups with two replications per treatment: non-toxic, novel endophyte-infected (NE+) tall fescue without melatonin supplementation (NE+/NM; n = 15), E+ fescue with melatonin supplementation (E+/M; n = 16), and E+ fescue without melatonin supplementation (E+/NM; n = 18). The treatment period was over 70 d between May and July (d 147 ± 1 to 217 ± 1 of gestation). A commercially available feed was mixed with melatonin (100 µg/kg BW) or without melatonin and hand-fed to each cow daily. At the conclusion of the treatment period (56 ± 18 d before parturition), cattle were moved to a single group and maintained on pastures free from E+ fescue for the remainder of the study. Dams and steer calves were weighed at d 0, 28, 56, 84, 112, 140, and 168 relative to calving (d 0 = calving). Four-hour milk yields on d 28 ± 3 and d 56 ± 3 were used to estimate daily milk volume and weight. Dam and calf BW were analyzed using the MIXED procedure of SAS specific for repeated measures with treatment, time, and the interaction as fixed effects and replicate as random. Calf birth weight, dam milk volume, and dam milk weight were analyzed using the MIXED procedure of SAS with treatment as a fixed effect and replicate as random. On d 28, estimated daily milk volume and weight were decreased (P < 0.05) in E+/NM dams relative to NE+/NM dams. On d 56, estimated daily milk volume and weight were decreased (P < 0.03) in E+/NM and E+/M dams relative to NE+/NM dams. There was an effect of time for dam and calf weights (P < 0.01). There was a treatment by time interaction for calf body weight, with calves born to E+/NM dams having decreased body weight relative to calves born to NE+/NM and E+/M dams from d 112 to d 168 (P = 0.02). There were no other effects of treatment, time, or the interaction (P > 0.05). These data suggest melatonin supplemented to pregnant crossbred multiparous beef cows grazing endophyte-infected tall fescue during mid-late gestation may increase preweaning growth of steer calves.

227 Moderate body weight loss reduces total antioxidant capacity in pregnant beef heifers and one-carbon metabolite supplementation does not mitigate this response

Abstract In beef heifers, body weight (BW) gain and physiological stress are important considerations for reproductive outcomes and whole herd production efficiencies. The objective of this study was to determine if dietary one-carbon metabolite (OCM) supplementation in moderate BW loss pregnant heifers alters circulating total antioxidant capacity, superoxide dismutase, and glutathione peroxidase activity. In a replicated 2 × 2 factorial, Angus-cross heifers (n = 81) were stratified by BW to gain 0.45 kg/d (CON) or lose 0.23 kg/d (RES) and receive corn carrier with OCM supplements (+OCM) or without (−OCM). The OCM supplements consisted of vitamin B12 (20 mg) and folate (320 mg) injections weekly and dietary rumen-protected methionine (7.4 g/d) and choline (44.4 g/d) from breeding (d 0) to d 63 of gestation. The four treatment groups were: CON−OCM (n = 20), CON+OCM (n = 21), RES−OCM (n = 21), and RES+OCM (n = 19). Heifers underwent estrus synchronization and were bred via artificial insemination using female sexed semen to a single sire. Pregnancy was diagnosed and fetal sex was verified on d 35 of gestation. Blood was collected and assayed for total antioxidant capacity, superoxide dismutase, and glutathione peroxidase activity on d −2 (baseline), 35, and 63 of gestation for all three replicates. Data were analyzed using the MIXED procedure of SAS with repeated measures to determine the effect of gain, supplement, day, and their two-way and three-way interactions. The gain × supplement interaction remained in all models, but other interactions were removed if P > 0.10. Replicate and heifer BW were included in the model as covariates and covariance structure was determined by lowest AIC and BIC. Statistical significance was considered at P ≤ 0.05 and tendencies at 0.05 < P ≤ 0.10. There was no BW gain × supplement interaction, or main effect of OCM supplements in total antioxidants. There was a gain × day effect (P ≤ 0.01), in which total antioxidants were increased on d 35 and further increased on d 63 in CON compared with RES treatments. There was no gain × supplement interaction, or main effect of gain or OCM supplements in superoxide dismutase and glutathione peroxidase. However, both superoxide dismutase and glutathione peroxidase were decreased (P ≤ 0.01) on d 63 compared with d 0 and 35. These data suggest that moderate BW loss reduces total antioxidant capacity in pregnant beef heifers, which could affect redox homeostasis. However, OCM supplements did not affect total antioxidant capacity, superoxide dismutase, or glutathione peroxidase. These findings are somewhat surprising considering oxidative stress has been well characterized in nutrient-restricted pregnancies, and given the interconnection between energy metabolism and one-carbon metabolism. Future research is warranted to fully elucidate the therapeutic effects of one-carbon metabolites.

PSIII-12 Influence of early gestational heat stress on biomarkers of mammary gland development in replacement gilts genomically selected for thermotolerance or thermosensitivity

Abstract Heat stress (HS) alters physiological and metabolic processes in lactating sows leading to decreased milk production, which is a major factor limiting growth and survivability of piglets. While genomic selection for thermotolerance may be a viable solution to alleviate the negative effects of HS on pig welfare, it may be linked to a reduction in milk production and subsequently, litter growth performance. Therefore, the study objective was to evaluate the effects of genomic selection for thermotolerance and its interaction with early gestational HS on biomarkers of mammary gland development in replacement gilts. We hypothesized that HS exposure as well as genomic selection for thermotolerance would negatively affect mammary epithelial proliferation rate. A total of 36 Landrace (33%) × Large White (67%) crossbred gilts divergently selected for thermotolerance (TOL; n = 18) or thermosensitivity (SEN; n = 18) were balanced by body weight, bred to a single Duroc sire, and then exposed to either thermoneutral (TN; constant 17 to 22°C) or cyclical HS (26 to 36°C) conditions until d 65 of gestation. From d 66 of gestation until farrowing, all pregnant gilts were exposed to TN conditions. Of the 36 total gilts bred, only 28 became pregnant yielding 15 HS gilts (n = 8 SEN and 7 TOL) and 13 TN gilts (n = 7 SEN and 6 TOL). On d 105 of gestation, a mammary biopsy was taken from all gilts, mammary tissue was placed into 10% buffered formalin, and KI67 immunohistochemical staining was performed to identify proliferating mammary epithelial cells (MEC). Proliferating and non-proliferating MEC populations were counted using ImageJ tool. Data were analyzed using PROC GLM in SAS 9.4 with individual gilt as the experimental unit. Overall, early gestational HS decreased (P < 0.01; 16.53 ± 2.12%) the percent of proliferating MEC relative to those kept under TN conditions (26.42 ± 2.39%). However, no effect of genomic line divergence was detected with any comparison (P > 0.05). In conclusion, early gestation HS, but not genomic selection for thermotolerance, had a negative impact on biomarkers of mammary gland development in replacement gilts. These data may suggest that early gestation HS could have a reductive effect on milk production capacity, which may negatively affect litter growth.

PSIV-A-7 Effects of late gestational nutrient restriction of beef heifers on small intestinal crypt depth in neonatal calves

Abstract We hypothesized that late gestational nutrient restriction would impair small intestinal development in neonatal calves, including crypt cell depth and proliferation. Fall-calving crossbred beef heifers [body weight (BW) = 466 ± 30 (SD) kg; BCS: 5.4 ± 0.5] bred to a single sire were individually-fed 100% (control; n = 10) or 70% (nutrient restricted; n = 12) of NASEM metabolizable energy and metabolizable protein requirements for maintenance, pregnancy, and growth from d 160 of gestation to calving. Calves were removed from their dams immediately post-calving and fed artificial colostrum and milk replacer relative to body weight until euthanasia at 48.7 ± 0.4 h of age. The gastrointestinal tract was removed, and proximal jejunum samples were dissected from each calf and immersion-fixed in 10% neutral buffered formalin. Tissues were embedded in paraffin blocks and cut into 5-µm sections that were mounted on glass slides. Slides were incubated with Ki-67 (marker of nuclear proliferation) and counterstained with Hematoxylin using an autostainer. Images were taken at 5× magnification using an upright microscope. QuPath software was used to measure crypt depth, starting at the base of the crypt region and ending at the furthest cluster of Ki-67 positive cells, perpendicular to the base of the mucosa. For each calf, 10 crypt depth measurements were taken from 4 images, resulting in 40 crypt depth measurements that were averaged. Nutritional plane was a fixed effect in the model, and maternal sire, date of study initiation, and calf sex (when P < 0.25) were considered covariates. We previously reported that calves born to nutrient restricted dams weighed less (P ≤ 0.05) at birth and 48 h than calves born to control dams. Small intestinal mass tended to be less (P = 0.11) in offspring of nutrient restricted dams but was not affected (P = 0.58) when expressed relative to body weight. Calves born to nutrient restricted dams also tended to have less (P = 0.11) small intestinal mass relative to brain weight. Maternal nutrient restriction did not affect (P = 0.95) jejunal crypt depth. Additionally, crypt depth relative to body weight was not affected (P = 0.20) by nutrient restriction. Although crypt depth was not affected, further analysis of proliferation and morphology will help to understand the relationship between maternal nutrient restriction and small intestine development in neonatal calves.

Influence of Maternal Nutrition and One-Carbon Metabolites Supplementation during Early Pregnancy on Bovine Fetal Small Intestine Vascularity and Cell Proliferation.

To investigate the effects of nutrient restriction and one-carbon metabolite (OCM) supplementation (folate, vitamin B12, 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.

PSII-12 Influence of Dietary Methionine and Guanidinoacetic Acid on Estrous Cycles and Early Pregnancy in Beef Heifers

Abstract Interest is growing in methods to apply precision nutritional supplements strategically during the peri-conceptual period to enhance progeny performance of beef females. Studies where supplemental methionine (MET) was provided to bovine females in the peri-conceptual period demonstrated epigenetic modifications in muscle of the progeny. Guanidinoacetic acid (GAA) has been fed directly to livestock to increase creatine synthesis, but has not been evaluated for use in fetal programming. We fed MET and GAA to cross-bred heifers during the peri-conceptual period as a 2 x 2 factorial design to influence carcass characteristics of their steer calves. It was, therefore, important to determine if the interactions of dietary MET and GAA in the peri-conceptual period would alter reproductive efficiency. The null hypothesis was that these supplements would not alter reproductive efficiency of beef heifers when fed in the peri-conceptual period. Heifers were divided into four breeding groups for management purposes. On day -63, heifers (n = 20/diet, age = 384.1 ± 1.1 d) were assigned HeatTime collars and individually fed MET (10 g/d), GAA (40 g/d), the combination (MET + GAA), or ground corn carrier (CON) for 126 days. On day -9, heifers received 100 mg of gonadotropin releasing hormone and a CIDR. On day -2, the CIDR was removed and luteolysis was initiated with 25 mg of prostaglandin F2a. All heifers were inseminated with male sexed semen from a single sire and administered a second 100 mg injection of gonadotropin releasing hormone on day 0. On day 63, a blood sample was collected from all pregnant heifers (CON n = 10; MET n = 8; GAA n = 7; MET + GAA n =10) to evaluate circulating progesterone concentrations. Reproductive tracts were collected to determine CL weight, gravid uterine weight, and placentome number. Data were analyzed using the MIXED procedure of SAS with group, MET, GAA, and the interaction of MET and GAA as fixed effects. Days to first estrus were greater and number of estrus events prior to synchronization were fewer in CON heifers compared with supplemented heifers (P < 0.05). Luteal weight tended to be greater in heifers supplemented with MET compared with CON and GAA heifers (P = 0.09), but peak intensity of estrus, concentration of progesterone at day 63 of pregnancy, gravid uterine weight, and placentome number did not differ due to diet (P > 0.10). Supplementation with MET or GAA may improve onset of puberty but does not negatively alter reproductive performance in cross-bred beef heifers. These supplements can be used for fetal programming of muscle development in steer progeny without negatively impacting reproductive performance in the breeding herd. USDA is an equal opportunity provider and employer. Funded in part by USDA-NIFA-AFRI # 2022-67015-36196.

243 Translating Maternal Vitamin and Mineral Supplementation Effect on Early Developing Fetus Through Hepatic Long Non-Coding Rnas Profile in Beef Heifers

Abstract Maternal nutrition is critical during pregnancy as the fetus relies on the nutrients from the dam. Fetal organogenesis starts during the first trimester. Thus, an appropriate supply of nutrients, including vitamins and minerals, is indispensable. In the field of fetal developmental programming, there is still a lack of knowledge based on appropriate livestock models to explain the role of regulatory factors in translating the effect of maternal minerals and vitamins on fetal development and metabolic function. Therefore, this study was designed to investigate the effect of maternal vitamin and mineral supplementation (VTM -d 71 periconceptual to d 83 of pregnancy) on fetal liver expression of long non-coding RNAs (lncRNAs) and their potential regulatory metabolic pathways. Angus crossbreed beef heifers (n = 16) were supplemented (VTM, n = 8) or not (NoVTM, n = 8) with 113 g· heifer-1· day-1 of mineral premix (Purina Wind & Rain Storm All-Season 7.5 Complete) and all heifers were bred with female sexed semen from a single sire. Transcriptomic profiles were determined with RNA-Seq in fetal livers collected on d 83 ± 0.27 of gestation. After data quality control and read mapping, lncRNAs were predicted using the FEELnc pipeline. Using the DESeq2 software, we identified 50 differentially expressed novel lncRNAs (expression of 32 lncRNAs was upregulated and 18 were downregulated; FDR < 0.05) located either downstream, upstream, or in exonic, or intronic regions of 51 genes (including PFKP, SLC30A10, KCNH8, ZNF451, MINPP1, BCL9, PAH, ITPR2, AHCY, and ABCG8), potentially regulating their expression. These genes could affect pathways (FDR < 0.1) such as PI3K signaling, mineral storage and transport, and amino acids biosynthetic pathways (i.e., cysteine and methionine biosynthesis). Collectively, these results suggest that periconceptual maternal vitamin and mineral supplementation affects the expression of lncRNAs and potentially regulates the expression of genes involved in fetal liver development and function.

PSXIV-19 Gas Flux By Calves from Dams Identified as Either High Or Low Methane Emitters

Abstract Eight (BW = 398 ± 8 kg) Red Angus heifers were used to examine the similarity between respiration gas parameters of dams and their offspring. The CH4, CO2 emissions, and O2 consumption of heifers were measured over a 70-day period using an automated head-chamber system that measured CO2, CH4, and O2 fluxes. Heifers grazed native mixed-grass prairie and were given a daily supplement of 1.025 kg of alfalfa (Medicago sativa L.) pellets via the automated head-chamber system. Heifers were then artificially inseminated using sexed semen from a single Black Angus sire. The emissions of the offspring (BW = 284 ± 7 kg; 7 heifers, 1 steer) were measured at 11 months of age over a 70-day period. Like the dams, during their measurement period, the offspring grazed native mixed-grass prairie and were given a daily supplement of 1.025 kg of alfalfa pellets via the automated head-chamber system. Means for CH4, CO2 emissions, O2 consumption, CH4:CO2 ratio, respiratory quotient, and heat production were compared between dams and their calf by regressing maternal parameters on the parameters of the calves and using beginning dam and offspring BW as covariates. Methane emissions of the dams and their offspring had an excellent Pearson’s correlation (ρ = 0.79). Carbon dioxide emissions and O2 consumption had an excellent Pearson’s correlation among dams and their offspring (ρ = 0.88 and 0.91, respectively). Heat production as well as the CH4:CO2 ratio also had an excellent and good Pearson’s correlation (ρ = 0.91 and 0.67, respectively). The respiratory quotient correlation was good (ρ = 0.60). These results indicated that CH4 and CO2 emissions, O2 consumption, the CH4:CO2 ratio and HP production from offspring have a good Pearson’s correlation with the dam’s emissions. Hence, even with this small dataset, there is evidence that beef cattle can be selected for lower greenhouse gas emissions.

PSIV-14 Effects of Maternal Nutrient Restriction During Late Gestation on Expelled Cotyledonary Mrna Expression in Primiparous Beef Females

Abstract To determine the effects of late gestational nutrient restriction on expelled cotyledonary mRNA expression, fall-calving Hereford × Simmental-Angus heifers [single-sired; BW: 451 ± 28 (SD) kg; BCS: 5.4 ± 0.7] bred to a single sire were allocated by fetal sex and expected calving date to receive either 100% (control; CON; n = 12) or 70% (nutrient restricted; NR; n = 11) of NASEM metabolizable energy and metabolizable protein requirements for maintenance, pregnancy, and growth. Beginning on d 160 of gestation, heifers were individually fed chopped sorghum sudan hay (1.74 Mcal ME/kg, 6.69% CP, 72.0% NDF; DM basis) and based on individual intakes, supplemented to meet targeted nutritional planes. Calving was observed for all females and placentas were collected at natural expulsion. Immediately after collection (average: 4.5 h post-calving; range: 2.3 to 12.1 h post-calving), 2 representative cotyledons on the ipsilateral side were cleaned with phosphate buffered saline, and cotyledonary tissue was flash frozen. Total RNA was extracted, and RT-qPCR was performed for mRNA expression of glucose transporters (SLC2A1, SLC2A3, SLC2A4), a fructose transporter (SLC2A5), amino acid transporters (SLC1A1, SLC3A2, SLC7A1, SLC7A5, SLC7A8, SLC38A2), angiogenic factors/receptors (VEGFA, NOS3, FLT1, KDR, GUCY1B3), and pregnancy-associated glycoprotein 2 (PAG2). Relative mRNA expression was analyzed with nutritional plane, treatment initiation date, time to cotyledon sampling, and calf sex (when P < 0.25) as fixed effects. We have previously reported that total uterine blood flow during late gestation was not affected (P ≥ 0.20) by nutritional plane or the nutritional plane × day of gestation interaction. Post-calving, NR dams were 64 kg BW less (P < 0.001) and 2.0 BCS less (P < 0.001), but calf birth weight was not affected (P = 0.72) by nutritional plane. Contralateral placental dry weight tended to be less (P = 0.07) in NR dams, but total placental weight was not affected (P = 0.86) by late gestational nutritional plane. Cotyledonary relative mRNA expression of SLC2A3 and SLC38A2 was greater (P ≤ 0.05) and relative expression of SLC2A1, SLC2A4, and NOS3 tended to be greater (P ≤ 0.07) in NR dams compared with CON. Late gestational nutritional plane did not affect (P ≥ 0.13) cotyledonary relative mRNA expression of FLT1, GUCY1B3, KDR, PAG2, SLC1A1, SLC2A5, SLC3A2, SLC7A1, SLC7A5, SLC7A8, and VEGFA. In summary, first parity beef females who experienced late gestational nutrient restriction maintained uterine blood flow and placental mass and had 4 nutrient transporters and 1 angiogenic factor upregulated in cotyledons, all which likely contributed to their conservation of fetal growth.

242 Methionine and Guanidinoacetic Acid Supplementation During the Periconceptual Period of Gestation Shifts Methionine Metabolism of Fetal Bull Calves at D 63 of Gestation

Abstract Recent research has demonstrated that feeding rumen protected methionine (MET) products increases circulating methionine while feeding guanidinoacetic acid (GAA) may result in a methyl deficiency. Therefore, we hypothesized that maternal MET or GAA supplementation from 63 days before breeding until d 63 of gestation would increase fetal hepatic methyl donors or result in a fetal hepatic methyl donor deficiency, respectively. Eighty MARC II heifers (n = 20 per treatment; initial BW = 344 ± 8 kg; day -63) were weighed and assigned to one of four breeding groups and four treatments stratified by age and starting weight. Heifers were fed a total mixed ration to gain 0.68 kg/d and received 100 g of one of four treatments top dressed: control supplement of ground corn (CON), MET (10 g/d) in a ground corn carrier, GAA (40 g/d) in a ground corn carrier, or MET + GAA (10 g/d MET + 40 g/d GAA) in a ground corn carrier. At breeding (d 0), all heifers were bred to a single sire using male sexed semen. At d 63 of gestation, 35 heifers (CON n = 10, MET n = 8, GAA n = 7, MET + GAA, n = 10) confirmed pregnant with bull calves were humanely slaughtered and fetal liver tissues were collected and sent to Metabolon Inc. for global metabolomics analysis. Data were analyzed as a 2 x 2 factorial with 2 levels of GAA and 2 levels of MET with breeding group included in the model. For this abstract, only metabolites from the methionine cycle and adjacent pathways will be reported and only metabolites with significant P-values will be discussed (P ≤ 0.05). There were no main effects or interactions for the abundance of methionine, GAA, creatine, SAM, SAH or homocysteine. Glycine was decreased and sarcosine increased in fetuses from MET supplemented dams. Serine was affected by a MET x GAA interaction being decreased in livers of CON, MET, and MET + GAA fetuses compared with GAA. Cystathione, S-methylcysteine, s-methylcysteine sulfoxide, and hypotaurine were greater in fetuses of MET supplemented heifers and pyridoxal phosphate was reduced in fetal liver of MET supplemented heifers. The abundance of urea was greater in GAA supplemented fetuses. These results indicate that fetuses from dams fed methionine may be able to maintain balance in methionine supply by shunting methionine through adjacent pathways including the transsulfuration pathway. We also interpret these data to imply that excess methionine is not utilized by the fetus and is metabolized to products that can be excreted. Lastly, GAA did not alter the fetal methyl supply. USDA is an equal opportunity provider and employer. Funded by (USDA-NIFA-AFRI # 2022-67015-36196).

72 Effects of Differing Planes of Nutrition and one-Carbon Metabolite Supplementation of Beef Heifers on B12, Folate, and Folate Intermediate Concentrations in the dam and Fetus

Abstract Vitamin B12 and folate are involved in the metabolic process known as one-carbon metabolism, which among other things, has a role in DNA, RNA, and protein methylation. This methylation connects one-carbon metabolism to epigenetics, which influences fetal development. Thus, an adequate supply of one-carbon metabolites (OCM) is important during fetal development. We hypothesized that differing planes of nutrition, along with supplementation of OCM, would impact concentrations of B12, folate, and folate intermediates in maternal liver, maternal serum, fetal liver, allantoic fluid (ALF), and amniotic fluid (AMF). Specifically, OCM supplementation would increase circulating and hepatic B12 and folate concentrations compared with non-supplemented heifers. Heifers (n = 72) were individually fed on one of two planes of nutrition: control (CON; 0.45 kg/d gain) or restricted (RES; -0.23 kg/d gain). To complete the 2 × 2 factorial arrangement, heifers were also given either OCM supplementation (10 g/d rumen-protected methionine + 60 g/d rumen-protected choline in the diet plus 20 mg B12 + 320 mg folic acid; weekly intramuscular injections) or no supplementation (grain carrier + saline injection). Thus, the final treatments were CON-OCM, CON+OCM, RES-OCM, and RES+OCM. Heifers were bred to a single sire via artificial insemination with sexed semen. Heifers pregnant with female calves (n = 31) were selected for slaughter on d 63. Samples of maternal liver, ALF, AMF, and fetal liver were collected at slaughter. Maternal serum was collected at breeding (d 0) and at slaughter. For B12 and folate concentration analysis, maternal serum was sent to IDEXX BioAnalytics (N. Grafton, MA). For analysis of folate intermediate concentrations in maternal and fetal liver, ALF, and AMF, samples were analyzed using LC-MS/MS. There were treatment × day interactions (P ≤ 0.05) for B12 and folate in maternal serum; on d 63, R+OCM was greater compared with C-OCM and R-OCM and all treatments at d 0 (Table 1). For folate intermediate concentrations, no plane of nutrition × supplement interactions (P ≥ 0.06) were found in fetal liver, ALF, or AMF; however, 5-methyl THF in ALF tended (P = 0.06) to be greater in R+OCM compared with all other treatments (Table 2). No interactions were found for concentrations in maternal liver when corrected for total liver weight; however, there was a supplement effect (P ≤ 0.05) where the concentrations of folic acid, dihydrofolate, tetrahydrofolate, 5-methyltetrahydrofolate, 5,10-methylenetetrahydrofolate and 5,10-methenyltetrahydrofolate were increased in +OCM compared with -OCM, and a plane of nutrition effect (P ≤ 0.05) with increased 5,10-methenyltetrahydrofolate and 5,10-methylenetetrahydrofolate concentrations in RES. We conclude that supplementing OCM increases concentrations of B12, folate and its intermediates in maternal liver and serum. USDA is an equal opportunity provider and employer.

71 Effects of Late Gestational Nutrient Restriction on Calf pre-Weaning Growth, Backgrounding and Feedlot Performance, and Carcass Composition

Abstract Fall-calving Hereford-SimAngus heifers [single-sired; BW: 451 ± 28 (SD) kg; BCS: 5.4 ± 0.7] bred to a single sire were individually fed 100% (control; CON; n = 12) or 70% (nutrient restricted; NR; n = 13) of NASEM metabolizable energy and metabolizable protein requirements for maintenance, pregnancy, and growth from day 160 of gestation to calving. Post-calving, all females were fed chopped tall-fescue hay supplemented to meet lactational nutrient requirements in Calan gates to constrain diets of calves to milk only until day 147, then pen-fed in drylots until weaning at day 242 ± 3 (SD). Post-weaning, calves were fed a tall fescue hay-based backgrounding diet in the same large drylots for 53 ± 12 d, then moved to partially covered pens with GrowSafe feeding systems in 2 groups and fed a corn-based feedlot diet (formulated to be 18.3% CP, 1.21 Mcal NEg/kg, DM basis; 89% DM) for 255 ± 3 d (similar days on feed rather than carcass endpoint). Cattle were harvested in a commercial plant, and camera carcass grading was performed. Data were analyzed with maternal treatment, calving date, and calf sex (when P < 0.25) as fixed effects; feedlot group was included for feedlot and carcass measures. We previously reported that birth weight was not affected (P = 0.72), but NR decreased (P < 0.04) milk production by 15%. Calves born to NR dams had reduced ADG from birth to day 147 (P = 0.02), day 147 to weaning (P = 0.08), and birth to weaning (P = 0.003), resulting in NR calves having 13% lighter (P = 0.004) weaning weight. Backgrounding ADG was not affected (P = 0.80) by maternal treatment; thus, feedlot entry BW was 11% less (P = 0.009) in NR calves. On day 28, 56, 84, and 112 on feed, NR calves weighed less (P ≤ 0.09) than control. Final BW was not affected (P = 0.91) by maternal treatment. Feedlot ADG was greater for NR calves from day 168 to slaughter (P = 0.02) and the overall feeding period (P = 0.03), but not before day 168 on feed (P ≥ 0.22). Calves born to NR dams had less (P = 0.02) dressing percentage, but improved (P = 0.02) yield grade and backfat thickness. Hot carcass weight, ribeye area, marbling score, and KPH were not affected (P ≥ 0.26) by maternal treatment. Percent prime, yield grade ≤ 2, and yield grade ≥ 4 were not affected (P ≥ 0.29) by maternal treatment. Overall, late gestational nutrient restriction decreased growth pre-weaning but increased late finishing growth, without affecting backgrounding or early feedlot growth. These cattle had leaner carcasses after similar days on feed, suggesting that late gestational nutrient restriction altered growth patterns or body composition.

PSIV-7 Multi-Sire Artificial Insemination of Beef Heifers in the Nebraska Sandhills May Increase Pregnancy Rate Compared with Single-Sire Insemination

Abstract The objective of these studies was to observe pregnancy outcomes resulting from artificial insemination (AI) with multi-sire semen (MS) compared with single-sire semen (SS) in beef heifers. One study compared MS AI to SS AI and the other study observed the pregnancy rate to AI using sexed MS semen. In the first study, three bulls were selected for AI of 441 heifers in year one. Heifers were synchronized with the melengestrol acetate – PGF2α – GnRH protocol using estrus detection and timed AI. Seventy-five single-sire straws of semen, from each bull, were labeled A, B, or C. Remaining semen was pooled together to form multi-sire semen straws. Heifers were artificially inseminated using alternating treatment of MS or SS; A, B, or C. Ultrasound determined pregnancy rate of heifers by treatment averaged 63% ± 5.5% (SS-A), 64% ± 5.5% (SS-B), 65% ± 5.5% (SS-C), and 69% ± 3.3% (MS). Heifers administered MS averaged 4.5% ± 4.5% greater than SS (P = 0.32) and 5.9% ± 6.4% greater than the least performing sire (P = 0.36). The observed differences between MS and SS pregnancy rate are less than previous research suggests. It is hypothesized the best fertilization period for a sire and dam may differ between individuals and herds leading to variability in pregnancy rates. In the second study, multiple unproven bulls were selected for AI of 868 ± 37 heifers over two years. Heifers were synchronized using the previously mentioned protocol with split-time AI. Semen was collected and sexed to favor heifer progeny at an expected ratio of 9:1. The sexing process has been shown to decrease pregnancy rate due to sperm damage and decreased quantity. Semen from 3 random bulls were pooled together making 4 pools or 3 pools of multi-sire semen straws in year 1 and 2 respectively. Multi-sire sexed semen was only administered to heifers who exhibited estrus. The observed ultrasound determined pregnancy rate of heifers administered multi-sire sexed semen was 65% in year 1 and 75% in year 2, which is 12% and 22% greater than the average pregnancy rate reported in previous studies using single-sire sexed semen. Observed differences lends credence to the possibility of improving pregnancy rate by utilizing multiple sires for AI when semen quality is lower. Ultimately these two studies may suggest that multiple sires may outperform pregnancy rate of single sires to AI. This difference may be more pronounced when semen quality is lower. Further research into semen quality of single sires and pooled semen could lead to optimization of fertilization among individuals within a herd.

70 Nutrient Restriction During Late Gestation Alters Maternal Circulating Amino Acid Concentrations in Primiparous Beef Females

Abstract We hypothesized that maternal circulating AA concentrations would be reduced by late gestational nutrient restriction, indicating less available nutrients for transport by the uteroplacenta to the growing fetus. Fall-calving Hereford-SimAngus heifers [single-sired; BW: 451 ± 28 (SD) kg; BCS: 5.4 ± 0.7] bred to a single sire were allocated by fetal sex and expected calving date to receive either 100% (control; CON; n = 12) or 70% (NR; n = 13) of NASEM metabolizable energy and metabolizable protein requirements for maintenance, pregnancy, and growth from day 160 of gestation to calving. Heifers were individually fed chopped sorghum sudan hay (1.74 Mcal ME, 6.66% CP, 72.0% NDF; DM basis) and based on individual intakes, supplemented to meet targeted nutritional planes. Jugular blood samples were obtained on days 159 (Pre-treatment), 202, and 244 of gestation, and 1 h post-calving to determine serum concentrations of the 20 common AA (except cysteine), 1-methyl histidine, and 3-methyl histidine. Gestational and post-calving AA were analyzed separately with nutritional plane, treatment initiation date, and calf sex (when P < 0.25) as fixed effects. Gestational AA included day and nutritional plane x day using repeated measures. We have previously reported that dam BCS was less (P ≤ 0.04) by day 202 of gestation and dam BW was less (P ≤ 0.01) by day 223 of gestation for NR dams. Post-calving, NR dams were 64 kg BW and 2.0 BCS lless (P < 0.001), but calf birth weight was not affected (P = 0.72) by nutritional plane. Histidine, lysine, and methionine were greater (P ≤ 0.03) in NR dams on day 202 of gestation. Tryptophan and valine were less (P ≤ 0.05) while methionine and total AA tended to be less (P ≤ 0.08) on day 244 of gestation in NR dams. Aspartic acid, glutamic acid, and tyrosine were less (P ≤ 0.03), but 3-methyl histidine was greater (P ≤ 0.01) on days 202 and 244 of gestation in NR dams. 1-methyl histidine was greater (P ≤ 0.05) at all 3 gestational timepoints but leucine was less (P = 0.03) in NR dams during gestation. Post-calving, glutamic acid, leucine, and valine were less (P ≤ 0.04), and methionine, threonine, tryptophan, tyrosine, and total AA tended to be less (P ≤ 0.10) in NR dams. Histidine, 1-methyl histidine, 3-methyl histidine, and serine were greater (P ≤ 0.03) in NR dams post-calving. In summary, late gestational nutrient restriction reduced total maternal circulating AA and altered the maternal serum AA profile, but calf birth weight was not affected. Individual AA that were greater or unaffected in NR dams were likely indicative of body protein catabolism occurring to meet nutrient requirements.

11 Impact of a Cache Valley Fever Outbreak on Lambing Performance in a Landrace Hair Sheep Flock: A Case Study

Abstract Cache Valley Fever (CVF) is a mosquito-born, viral disease endemic to North America. While generally not associated with clinical symptoms in ewes, it affects fetal development during the first trimester of pregnancy resulting in malformations and pregnancy loss. A suspected outbreak of CVF was recorded in a flock of Barbados Blackbelly and St. Croix hair sheep managed under an 8-mo, accelerated mating system with ewes bred in March, July and November. The outbreak was associated with a July breeding, in a summer with unusually high rainfall and standing water in pastures. There had been no previous occurrence of CVF or other abortion storms in the flock (20+ years). Lambing outcome associated with the suspected outbreak of CVF was compared with that of the three prior July matings (CON). Ewes (n = 32-35) were managed in a pasture-based system under rotational grazing. Ewes were mated in two single sire groups of 16 to 18 ewes to like-breed sires each breeding season. Pregnancy was determined by transrectal ultrasound at end and 21 d after the 25-day breeding season. Ewes lambed on pasture, and lambs were tagged, and birth weights recorded within 24 hours. Blood samples were collected from all ewes following the CVF lambing and submitted for CVF antibody testing. All ewes showed CVF antibody titers. Pregnancy rate for the CVF lambing was not different from CON (85.7 vs. 89.8%; P = 0.512), whereas lambing rate was less in CVF for both ewes exposed (74.3 vs. 88.8%; P = 0.039) and ewes lambing (86.7 vs. 98.9%; P = 0.004). Incidence of stillborn and neonatal death of lambs was greater in CVF than CON (37.0 vs.5.6%; P = 0.002), as was the number of malformed lambs (13.0 vs. 0%; P < 0.001). Malformed lambs were associated with breech presentations. Litter size was not different in the CVF lambing (1.77 vs 1.86; P = 0.471), but lamb birth weight tended to be less (2.80 vs. 3.04 kg; P = 0.055) while ewe postpartum weight was not different (45.3 vs. 47.0 kg; P = 0.384). The outbreak of CVF reduced the lambing rate and increased the incidence of neonatal death and malformations. The outbreak occurred during a summer with unusually high rainfall and may be associated with changing weather patterns linked to climate change. Delaying summer breeding may be considered a mitigation strategy if feasible.

Concentrations of vitamin B12 and folate in maternal serum and fetal fluids, metabolite interrelationships, and hepatic transcript abundance of key folate and methionine cycle genes: the impacts of maternal nutrition during the first 50 d of gestation.

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.

Incorporating A.I. [Artificial Intelligence] into Boar Semen A.I. [Artificial Insemination

Analysis of the U.S swine herd shows variation in pregnancy rate is more attributable to male- factor subfertility than the dam. To date, a limited degree of correlations has been observed between conventional semen analysis parameters and actual fertility after standard quality cutoffs are met. Thus, a clear ability to predict male-factor fertility is lacking. Recent technological advances in flow cytometry, namely image-based flow cytometry, allows for high-throughput, single-cell phenotyping. This allows for multi-million sperm bioimage data sets to be easily attained. These datasets can then be analyzed utilizing machine & deep learning analytic methods and correlated with single sire field fertility data to open the black box of boar fertility prediction. In this presentation, we will discuss recent advancements our lab has made utilizing machine & deep learning, artificial intelligence into boar sperm quality analysis. This includes bioimage, deep learning analysis of boar sperm morphology as well as our latest results related to boar semen fertility prediction. Our findings establish a new era for boar sperm quality analysis and pave the way for accurate fertility prediction in future precision agriculture applications. Work performed here was supported by USDA’s National Institute of Food and Agriculture grant number 2019- 67012-29714 (KK) and 2022-67015-36298 (KK).