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Extremes in body condition reduce fertility and overall productivity in beef cattle herds, due in part to altered systemic metabolic conditions that influence the intrafollicular and uterine environment. Follicular fluid and serum metabolome profiles are influenced by body composition in women and dairy cattle; however, such information is lacking in beef cattle. We hypothesized that body condition score (BCS)-related alterations in the metabolome of preovulatory follicular fluid and serum may influence oocyte maturation while impacting the oviductal or uterine environment. Therefore, we performed a study with the objective to determine the relationship between BCS and the metabolome of follicular fluid and serum in lactating beef cattle. We synchronized the development of a preovulatory follicle in 130 cows of varying BCS. We collected blood and performed transvaginal follicle aspirations to collect follicular fluid from the preovulatory follicle ~18 h after gonadotropin-releasing hormone administration to stimulate the preovulatory gonadotropin surge. We then selected follicular fluid and serum samples from cows with BCS 4 (Thin; n = 14), BCS 6 (Moderate; n = 18), or BCS8 (Obese; n = 14) for ultra-high performance liquid chromatography-high resolution mass spectrometry. We identified differences in the follicular fluid or serum of thin, moderate, and obese animals based on multiple linear regression. MetaboAnalyst 5.0 was used for enrichment analysis of significant metabolites. We identified 38 metabolites in follicular fluid and 49 metabolites in serum. There were no significant differences in follicular fluid metabolite content among BCS classifications. There were 5, 22, and 1 serum metabolites differentially abundant between thin-obese, moderate-thin, and moderate-obese classifications, respectively (false discovery rate [FDR]0.10). These metabolites were enriched in multiple processes including "arginine biosynthesis," "arginine/proline metabolism," and "D-glutamine/D-glutamate metabolism" (FDR0.04). Pathways enriched with serum metabolites associated with BCS indicate potentially increased reactive oxygen species (ROS) in serum of thin cows. ROS crossing the blood follicular barrier may negatively impact the oocyte during oocyte maturation and contribute to the reduced pregnancy rates observed in thin beef cows.Extremes in body condition affect fertility and pregnancy outcomes in beef cows. Much research has been done in women and dairy cows to evaluate body condition’s effect on oocyte and embryo quality, pregnancy rates, and pregnancy outcomes. However, little work of this type has been done in beef cows and most studies do not focus on the preovulatory time period. The preovulatory time period is an essential time for the oocyte, as final stages of prematuration and the completion of oocyte maturation take place in the peri-ovulatory follicle. The follicular fluid provides the microenvironment for oocyte maturation and exchanges substances with maternal circulation at the blood follicular barrier. Alterations in maternal circulation due to extremes in body condition may pass into the follicular fluid and affect the oocyte during the preovulatory time period. Such conditions may contribute to the reduced fertility seen in beef cows with extreme body condition.

Protein supplementation to replacement heifers is utilized to reach targeted body weights, which may affect the animal’s gut and reproductive microbiomes and potentially impact breeding outcomes. The objective of the study was to evaluate the effects of protein supplementation on the ruminal and uterine bacterial communities of developing heifers. Thirty-nine pre-pubertal commercial Angus heifers were blocked by body weight (BW) and randomly assigned to one of three supplementation groups: 10% crude protein (CP), 20% CP, or 40% CP. Supplements were provided four times weekly for 140d. Blood and BW were taken every 14d to monitor development. Every 56d, uterine flushes and rumen content were collected for bacterial identification. Uterine flushes were collected via 20 mL sterile saline flushed through a Foley catheter, and rumen content by esophageal tubing. Bacterial DNA was extracted and amplified targeting the V1-V3 region of the 16S rRNA gene. Microbiome analyses were performed in R 4.1 with dada2 and phyloseq packages. Statistical analyses were performed in R and SAS 9.4. The GLIMMIX procedure was used including fixed effects of protein, month, and pubertal status. Random effects included BW, interaction of BW and protein, and heifer within the interaction, with repeated measures of month. Alpha diversity differed by month in the rumen, and by month and pubertal status in the uterus (P < 0.05). Bray Curtis analyses indicated clustering by month and pubertal status (P < 0.001) in the uterus, and by month in the rumen (P < 0.001). Multiple uterine and ruminal bacteria abundances differed over time and by protein supplementation and pubertal status (P < 0.05). The impact of pubertal status and month of development may indicate maturation of the uterine bacterial communities through development. Protein supplementation affecting the uterine bacterial communities may provide opportunity to manipulate the uterine microbiome, potentially affecting future reproductive success.

The milk microbiome in swine has been recently demonstrated to develop and alter the growing gut microbiome in offspring. Milk is the primary nutritional source for piglets and the established microbial communities from initial diet remain stable over time, thus being critical for long-term effects on the animal. Since little is known about the ontogeny of the milk microbiome, the objective of this study was to determine differences in the sow milk microbiome among multiple parities and days in lactation. A total of 32 sows were grouped by parity, 1–4, with 8 sows per parity. Sows were on the same diet and had ad libitum access to water. At sampling, teats were cleaned and stripped. Milk samples were manually collected on one side from a fore, mid, and hind teat, then combined for one whole milk sample per animal. Milk was sampled on days 2 and 16 to determine differences between days in lactation. Samples were sequenced using an Illumina MiSeq for V1-V3 regions of the bacterial 16S rRNA gene and downstream analyses were conducted in the R environment using ‘phyloseq’ and ‘DADA2’ packages. Statistical analyzes utilized a completely randomized design and a mixed model analysis of variance, with fixed effects of parity and days in lactation and examining a parity by day interaction. The genera Corynebacterium were greater (P < 0.05) abundance in parity 4. There was an increase (P < 0.05) in Staphylococcus at day 2 compared to day 16. Core microbial communities, Lactobacillus and Romboutsia increased (P < 0.05) by day 16. Identifying microbial communities present in the milk microbiome in sows can allow producers to understand how these microbes may influence the gut microbiome in growing piglets to augment gain. USDA is an equal opportunity provider and employer.

Fescue toxicosis causes substantial financial losses to the beef industry due to its negative effects on reproduction, growth and feed efficiency. Recent research has demonstrated that the negative effects of fescue toxicosis may be mitigated by supplementation of red clover isoflavones. To determine impacts on the ruminal environment, the objective of this study was to evaluate the effect of red clover isoflavones and endophyte-infected tall fescue on ruminal metabolites. Thirty-six Angus steers weighing 250±20 kg were randomly assigned to treatments in a 2×2 factorial arrangement, consisting of endophyte-free or endophyte-infected tall fescue seed with or without red clover isoflavones. For the 21d trial, steers were provided a basal diet supplemented with fescue seed targeting a minimum of 0.011 mg × kg of BW−1 × d−1 of total alkaloids. A total of 943 mg isoflavones were administered daily via bolus. Following the trial, 50mL of rumen fluid was collected via orogastric tubing. Untargeted metabolomic analysis was performed using an Exactive Plus Orbitrap MS, and 10µL sample was introduced using electrospray ionization into the Dionex UltiMate 3000 UPLC system. Peaks were identified in MAVEN, then normalized and analyzed in MetaboAnalyst 4.0, and 106 known metabolites were identified. Differences in seed type × isoflavone were analyzed by one-way ANOVA and main effects were analyzed by t-test. A total of 11 metabolites differed based on seed type × isoflavone treatment group (P ≤ 0.05), 11 differed by seed type (P ≤ 0.5), and seven differed by isoflavone treatment (P ≤ 0.05), namely amino acids or intermediates of amino acid metabolism. Methane (P = 0.03) and sulfur (P = 0.03) metabolism pathways were both impacted by isoflavone treatment, whereas 20 metabolic pathways were impacted as a result of differences in seed type (P ≤ 0.05). The rumen metabolome appears to be more affected by seed type, but a longer trial will likely result in more profound impacts.

Beef cattle is the most consumed red meat in the United States, and the United States is the largest producer and consumer of beef cattle globally. Feed is one of the largest input costs for the beef cattle industry, accounting for 40–60% of the total input costs. Identifying methods for improving feed efficiency in beef cattle herds could result in decreased cost to both producers and consumers, as well as increased animal protein available for global consumption. Identification of feed efficiency biomarkers could aid in the selection of more feed-efficient animals. To identify potential biomarkers of feed efficiency, rumen fluid was collected from low- (n = 14) and high-RFI (n = 15) Angus steers based on RFI ± 0.5 SD around the mean following a 70-d feed efficiency trial to determine rumen metabolites associated with variations in feed efficiency. Steers were 7 mo of age, weighing 264±2.7 kg. Rumen fluid was collected via gastric tubing, filtered through a 0.22 µM syringe filter, extracted for metabolomic analysis using 0.1% formic acid in acetonitrile:water:methanol (2:2:1), and injected into the Dionex UltiMate 3000 UHPLC system with an Exactive Plus Orbitrap MS. Peaks for known metabolites were identified using MAVEN and analyzed using MetaboAnalyst 4.0 and SAS. Significance was determined using an α ≤ 0.05. Based on fold change (high/low RFI), eight metabolites were greater in low-RFI steers compared to high-RFI steers, including 3,4-dihydroxyphenylacetate, 4-pyridoxate, citraconate, hypoxanthine, succinate/methylmalonate, thymine, uracil, and xylose (P ≤ 0.05), while 12 metabolites differed by ≥ 2-fold. These metabolites were predominantly involved in amino acid and lipid metabolism. Rumen fluid metabolomes differed in steers of varying feed efficiencies. These metabolites may be used as biomarkers of feed efficiency and may provide insight as to factors contributing to differences in feed efficiency that may be exploited to improve feed efficiency in beef cattle herds.