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To improve animal performance and modify growth by increasing lean tissue accretion, beef cattle production has relied on use of growth promoting technologies such as beta-adrenergic agonists. These synthetic catecholamines, combined with the variable inclusion of rumen degradable (RDP) and undegradable protein (RUP), improve feed efficiency and rate of gain in finishing beef cattle. However, research regarding the impact of beta-adrenergic agonists, protein level, and source on the ruminal microbiome is limited. The objective of this study was to determine the effect of different protein concentrations and beta-adrenergic agonist (ractopamine hydrochloride; RAC) on ruminal bacterial communities in finishing beef heifers. Heifers (n = 140) were ranked according to body weight and assigned to pens in a generalized complete block design with a 3 × 2 factorial arrangement of treatments of 6 different treatment combinations, containing 3 protein treatments (Control: 13.9% CP, 8.9% RDP, and 5.0% RUP; High RDP: 20.9% CP, 14.4% RDP, 6.5% RUP; or High RUP: 20.9% CP, 9.7% RDP, 11.2% RUP) and 2 RAC treatments (0 and 400 mg/day). Rumen samples were collected via orogastric tubing 7 days before harvest. DNA from rumen samples were sequenced to identify bacteria based on the V1-V3 hypervariable regions of the 16S rRNA gene. Reads from treatments were analyzed using the packages 'phyloseq' and 'dada2' within the R environment. Beta diversity was analyzed based on Bray-Curtis distances and was significantly different among protein and RAC treatments (P < 0.05). Alpha diversity metrics, such as Chao1 and Shannon diversity indices, were not significantly different (P > 0.05). Bacterial differences among treatments after analyses using PROC MIXED in SAS 9 were identified for the main effects of protein concentration (P < 0.05), rather than their interaction. These results suggest possible effects on microbial communities with different concentrations of protein but limited impact with RAC. However, both may potentially act synergistically to improve performance in finishing beef cattle. [ABSTRACT FROM AUTHOR]

New findings in developmental programming show an increased importance of methyl donor availability; however, effects of changing methyl donor concentration on the fetal transcriptome have yet to be identified. Differential gene expression (DEG) was used to determine the impact of maternal supplementation of methionine (MET), an obligate methyl donor, and guanidinoacetic acid (GAA), a methyl donor consumer, during the periconceptual period on bovine fetal development. MARC II heifers [n = 80; initial body weight (BW) = 346 ± 8.28 kg] receiving the same mixed ration and targeting the same gain (0.68 kg/d) were assigned to one of four treatments totaling 100 g/d of supplement with a ground corn carrier: MET (10 g/d), GAA (40 g/d), MET+GAA (10 g/d Met + 40 g/d GAA), and only ground corn carrier for control (CON). Supplementation began 63 d before breeding and concluded 63 d after breeding. Heifers pregnant with male offspring (CON, n = 10; MET, n = 8; GAA, n = 7; MET+GAA, n = 10) were slaughtered on d 63 of gestation. Transcript abundance was measured using RNA-Seq from extracted total RNA of fetal hindlimb samples (n = 35). The DEG analysis identified 227 upregulated and 121 downregulated genes from MET vs CON, 483 upregulated and 40 downregulated genes from GAA vs CON, and 672 upregulated and 34 downregulated genes from MET+GAA vs CON treated heifers (P ≤ 0.05). We identified 315 upregulated and 114 downregulated genes from MET vs GAA, 202 upregulated and 204 downregulated genes from GAA vs MET+GAA, and 35 upregulated and 133 downregulated genes from MET vs MET+GAA treated heifers (P ≤ 0.05). Over-representation analysis of DEGs highlighted immune response pathways that were found amongst the GAA vs CON, MET vs GAA, and GAA vs MET+GAA comparisons. Genes associated with cell inflammatory and immune response, including CD14, Toll-like receptor (TLR) 6, CD86, and TLR8, were upregulated in the GAA treatment. The TLR genes regulate expression of several pro-inflammatory cytokines, whereas CD genes are responsible for cell inflammatory response. Both are important for regulation of lipopolysaccharide (LPS)- binding protein, which plays a subsequent role in immune recognition when bound with LPS, resulting in genes from these families being recognized as markers for inflammatory response in beef cattle. Between MET vs CON and MET+GAA vs CON comparisons, 15 pathways including skeletal system development, alpha-amino acid catabolic process, and acylglycerol catabolic processes were shared. The FABP1 gene was upregulated in both aforementioned comparisons and has an integral role in transport and metabolism of fatty acids within cattle. Providing supplementation of methyl donors to the maternal environment results in over-representation of fetal muscle genes acting on immune response as well as fatty acid and amino acid metabolism. USDA is an equal opportunity provider and employer. [ABSTRACT FROM AUTHOR]

Simple Summary: The rumen makes up a large portion of the digestive tract of beef cattle and is responsible for the absorption of nutrients and microbial by-products. The rumen papillae interact with feed, microbial populations, and fermentation products important to cattle nutrition. Variation in the animal's ability to take up and utilize these nutrients affects feed efficiency. This study was performed to identify genes involved in feed efficiency that are expressed in two unrelated and physically distant populations of Angus and Hereford crossbred steers. A total of 83 genes were identified that may be useful indicators of feed efficiency in cattle. Differentially expressed genes were involved in a protein turnover pathway and a stomach lining turnover pathway. The use of meta-analysis for the two populations of cattle with different sire lines, management and handling techniques, and feed ingredients should allow the identification of genes that are involved in feed efficiency across cattle populations rather than those identified in a single population. In cattle, the rumen is an important site for the absorption of feed by-products released by bacterial fermentation, and variation in ruminal function plays a role in cattle feed efficiency. Studies evaluating gene expression in the rumen tissue have been performed prior to this. However, validating the expression of genes identified in additional cattle populations has been challenging. The purpose of this study was to perform a meta-analysis of the ruminal transcriptome of two unrelated populations of animals to identify genes that are involved in feed efficiency across populations. RNA-seq data from animals with high and low residual feed intake (RFI) from a United States population of cattle (eight high and eight low RFI) and a Canadian population of cattle (nine high and nine low RFI) were analyzed for differences in gene expression. A total of 83 differentially expressed genes were identified. Some of these genes have been previously identified in other feed efficiency studies. These genes included ATP6AP1, BAG6, RHOG, and YPEL3. Differentially expressed genes involved in the Notch signaling pathway and in protein turnover were also identified. This study, combining two unrelated populations of cattle in a meta-analysis, produced several candidate genes for feed efficiency that may be more robust indicators of feed efficiency than those identified from single populations of animals. [ABSTRACT FROM AUTHOR]

Abscess is the highest cause of liver condemnation and is estimated to cost the beef industry US$64 million annually. Fusobacterium necrophorum , commonly found in the bovine rumen, is the primary bacteria associated with liver abscess in cattle. Theoretically, damage to the rumen wall allows F. necrophorum to invade the bloodstream and colonize the liver. The objective of this study was to determine the changes in gene expression in the rumen epithelium and microbial populations adherent to the rumen epithelium and in the rumen contents of beef cattle with liver abscesses compared with those with no liver abscesses. Rumen epithelial tissue and rumen content were collected from 31 steers and heifers with liver abscesses and 30 animals with no liver abscesses. Ribonucleic acid (RNA) sequencing was performed on the rumen epithelium, and a total of 221 genes were identified as differentially expressed in the animals with liver abscesses compared with animals with no abscesses, after removal of genes that were identified as a result of interaction with sex. The nuclear factor kappa-light-chain enhancer of activated B cells signaling and interferon signaling pathways were significantly enriched in the differentially expressed gene (DEG) set. The majority of the genes in these pathways were downregulated in animals with liver abscesses. In addition, RNA translation and protein processing genes were also downregulated, suggesting that protein synthesis may be compromised in animals with liver abscesses. The rumen content bacterial communities were significantly different from the rumen wall epimural bacterial communities. Permutational multivariate analysis of variance (PERMANOVA) analysis did not identify global differences in the microbiome of the rumen contents but did identify differences in the epimural bacterial communities on the rumen wall of animals without and with liver abscesses. In addition, associations between DEG and specific bacterial amplicon sequence variants of epimural bacteria were observed. The DEG and bacterial profile on the rumen papillae identified in this study may serve as a method to monitor animals with existing liver abscesses or to predict those that are more likely to develop liver abscesses. [ABSTRACT FROM AUTHOR]

Mesenteric fat is a visceral fat depot that increases with cattle maturity and can be influenced by diet. There may be a relationship between the accumulation of mesenteric fat and feed efficiency in beef cattle. The purpose of this study was to identify genes that may be differentially expressed in steers with high and low BW gain and feed intake. RNA-Seq was used to evaluate the transcript abundance of genes in the mesenteric fat from a total of 78 steers collected over 5 different cohorts. A meta-analysis was used to identify genes involved with gain, feed intake or the interaction of both phenotypes. The interaction analysis identified 11 genes as differentially expressed. For the main effect of gain, a total of 87 differentially expressed genes (DEG) were identified (PADJ<0.05), and 24 were identified in the analysis for feed intake. Genes identified for gain were involved in functions and pathways including lipid metabolism, stress response/protein folding, cell proliferation/growth, axon guidance and inflammation. The genes for feed intake did not cluster into pathways, but some of the DEG for intake had functions related to inflammation, immunity, and/or signal transduction (JCHAIN, RIPK1, LY86, SPP1, LYZ, CD5, CD53, SRPX, and NF2). At PADJ<0.1, only 4 genes (OLFML3, LOC100300716, MRPL15, and PUS10) were identified as differentially expressed in two or more cohorts, highlighting the importance of evaluating the transcriptome of more than one group of animals and incorporating a meta-analysis. This meta-analysis has produced many mesenteric fat DEG that may be contributing to gain and feed intake in cattle. [ABSTRACT FROM AUTHOR]

Genetic and other molecular markers are powerful tools with the potential to improve livestock production traits, like feed efficiency. Feed efficiency is a complex biological trait mediated by many genes. Molecular based research, including transcriptome studies to evaluate gene expression profiles to determine the genes and pathways that contribute to feed efficiency in beef cattle have been widely performed over the last decade. Despite its widespread use, challenges remain in transcriptomic data analysis, in particular non-reproducibility of results between studies. These discrepancies exist for a number of reasons including small numbers of replicates, technical differences (e.g. sample preparation, sequencing platform), and biological differences (e.g., environmental, management, and genetic effects). Recent developments in technology and analytic tools provide an opportunity to integrate and synthesize the results of existing and future data. The potential to utilize these tools to have greater impact on feed efficiency will also be discussed. USDA is an equal opportunity provider and employer. [ABSTRACT FROM AUTHOR]

To improve animal performance and modify growth by increasing lean tissue accretion, beef cattle production has relied on use of growth promoting technologies such as beta-adrenergic agonists. These synthetic catecholamines, combined with the variable inclusion of rumen degradable (RDP) and undegradable protein (RUP), may improve feed efficiency and rate of gain in finishing beef cattle. However, the impact of beta-adrenergic agonists and protein level and source on the ruminal microbiome is limited. The objective of this study was to determine the effect of different protein concentrations and beta-adrenergic agonist (ractopamine hydrochloride; RAC) on ruminal bacterial communities in finishing beef heifers. Heifers (n = 140) were ranked according to body weight and assigned to pens in a randomized complete design to 6 different treatments, containing 3 protein treatments (Control: 13.9% CP, 8.8% RDP, and 5.0% RUP; High RDP: 20.9% CP, 13.4% RDP, 6.1% RUP; or High RUP: 20.9% CP, 9.1% RDP, 10.4% RUP) and 2 RAC treatments (0 and 400 mg/day). Rumen samples were collected from heifers by oral lavage 7 days before harvest. The DNA from the samples were sequenced to identify bacteria based on the V1-V3 hypervariable regions of the 16S rRNA gene using the Illumina MiSeq. Sequences and data from the treatments were analyzed using the R environment and PROC MIXED in SAS 9.4 (SAS Inst.; Cary, NC). Beta diversity was analyzed using PERMANOVA based on PCoA Bray-Curtis distances and were significant among the treatments (P < 0.05). Alpha diversity metrics such as Chao1 and Shannon diversity indices were not significantly different (P > 0.05). Differences among treatments at variable taxonomic levels after analyses through DESeq2 were significantly different for the main effects of protein and RAC, as well as their interaction (P < 0.05). These results suggest possible effects on the microbial communities, potentially acting synergistically to improve performance. [ABSTRACT FROM AUTHOR]

We have previously identified components of the immune system contributing to feed intake and gain in both the rumen and small intestine of beef steers. In this study, we examined the spleen, a major lymphatic organ near the digestive tract, to determine whether it was also influencing individual feed efficiency status through immune responses. Animals (n = 16) that were divergent for gain and intake were selected for tissue sampling. The spleen transcriptomes were evaluated by microarray. A total of 1216 genes were identified as differentially expressed. Genes were over-represented in Kyoto encyclopedia of genes and genomes (KEGG) pathways including biological regulation, protein folding, cell communication, immune systems process, response to stress, and RNA metabolic process. Several stress response or heat shock genes including HSPH1, HSPA1A, HSPA4, DNAJB4, DNAJA4, etc., were identified as a stress response functional gene cluster in the low gain-low intake animals. These genes were up-regulated amongst the low gain-low intake animals compared to all other groups. Canonical pathways associated with the differentially expressed genes included the coagulation system, extrinsic prothrombin activation, protein ubiquitination, unfolded protein response, and aldosterone signaling in epithelial cells. An analysis of expressed copy number variable (CNV) genes in the spleen produced some of the same genes and gene families that were differentially expressed. Our data suggests the splenic contribution to some of the underlying variation among gain and intake within this group of animals may be a result of immune function and stress response. In addition, some of the differences in immune response functions may be related to gene copy number. [ABSTRACT FROM AUTHOR]

Background Feed intake and gain are economically important traits in beef production. The rumen wall interacts with feed, microbial populations, and fermentation products important to cattle nutrition. As such, it is likely to be a critical component in the beef steer's ability to utilize feedstuffs efficiently. To identify genes associated with steer feed intake and body weight gain traits, and to gain an understanding of molecules and pathways involved in feed intake and utilization, RNA sequencing (RNA-Seq) was performed on rumen papillae from 16 steers with variation in gain and feed intake. Four steers were chosen from each of the four Cartesian quadrants for gain × feed intake and used to generate individual RNA-Seq libraries. Results Normalized read counts from all of the mapped reads from each of the four groups of animals were individually compared to the other three groups. In addition, differentially expressed genes (DEGs) between animals with high and low gain, as well as high and low intake were also evaluated. A total of 931 genes were differentially expressed in the analyses of the individual groups. Eighty-nine genes were differentially expressed between high and low gain animals; and sixty-nine were differentially expressed in high versus low intake animals. Several of the genes identified in this study have been previously associated with feed efficiency. Among those are KLK10 , IRX3 , COL1A1 , CRELD2 , HDAC10 , IFITM3 , and VIM . Conclusions Many of the genes identified in this study are involved with immune function, inflammation, apoptosis, cell growth/proliferation, nutrient transport, and metabolic pathways and may be important predictors of feed intake and gain in beef cattle. [ABSTRACT FROM AUTHOR]

Different dietary and feed additive strategies have been developed to reduce the liver abscess in feedlot cattle, but liver abscesses are still a major problem in beef production. We have limited knowledge about how rumen microbial communities interact with host epithelial gene expression in healthy and liver-abscessed animals. The objective of this study was to investigate the associations between the rumen content associated and rumen epimural microbiome and epithelial gene expression in liver-abscessed and healthy animals. To this end, we collected the ruminal contents and tissue samples from healthy (N=30; score=0, steers n=19 and heifers n=11) and liver-abscessed (N=30; score=A+, steers n=21 and heifers n=9) feedlot cattle at harvest. The bacterial community compositions in the ruminal contents and papillae were evaluated via 16S rDNA sequencing of the V4 region using the Illumina MiSeq platform. Additionally, total RNA was extracted from rumen epithelial tissues and sequenced using the Illumina NextSeq platform. The permutational analysis (PERMANOVA) on Bray Curtis distances matrices showed the microbial community in the ruminal contents was significantly different (P< 0.001) from the bacterial community observed in rumen papillae. The ruminal contents contained a higher abundance of Bacteroidetes and Proteobacteria while papillae contained higher abundance of Firmicutes. The epimural microbiota was different (P< 0.01) between healthy and liver abscessed animals while ruminal contents microbiome was not different between the two groups. The DeSeq2 algorithm identified differentially expressed genes (221) related to MAPK, NF-kappa B signaling pathway, immune and inflammatory response in liver-abscessed animals. Additionally, a wide range of epimural bacterial taxa were correlated (-0.52 to 0.67) with differentially expressed genes. These data demonstrate the interaction between epimural microbiota and the host and its effect on liver abscesses, and indicate the need to study the epimural microbiome for its impact on liver abscesses in feedlot cattle. USDA is an equal opportunity provider and employer. [ABSTRACT FROM AUTHOR]

Decreasing costs are making low coverage sequencing with imputation to a comprehensive reference panel an attractive alternative to obtain functional variant genotypes that can increase the accuracy of genomic prediction. To assess the potential of low-pass sequencing, genomic sequence of 77 steers sequenced to >10X coverage was downsampled to 1X and imputed to a reference of 946 cattle representing multiple Bos taurus and Bos indicus-influenced breeds. Genotypes for nearly 60 million variants detected in the reference were imputed from the downsampled sequence. The imputed genotypes strongly agreed with the SNP array genotypes ( r ¯ = 0.99 ) and the genotypes called from the transcript sequence ( r ¯ = 0.97 ). Effects of BovineSNP50 and GGP-F250 variants on birth weight, postweaning gain, and marbling were solved without the steers' phenotypes and genotypes, then applied to their genotypes, to predict the molecular breeding values (MBV). The steers' MBV were similar when using imputed and array genotypes. Replacing array variants with functional sequence variants might allow more robust MBV. Imputation from low coverage sequence offers a viable, low-cost approach to obtain functional variant genotypes that could improve genomic prediction. [ABSTRACT FROM AUTHOR]