The aim of this study was to identify for the first time single nucleotide polymorphisms (SNPs) associated with Haemonchus contortus resistance in Florida Native sheep, using a targeted sequencing approach. One hundred and fifty-three lambs were evaluated in this study. At the start of the trial, phenotypic records for fecal egg count (FEC), FAMACHA score, body condition score (BCS), and weight were recorded and deworming of sheep with levamisole (18 mg/kg of body weight) was performed. Ten days post-deworming (baseline) and 28 d post-baseline, a full hematogram of each sheep was obtained and FEC, FAMACHA score, BCS, and weight were assessed. Average daily gain was calculated at the end of the trial. Out of 153 animals, 100 sheep were selected for genotyping using a targeted sequencing approach. Targeted sequencing panel included 100 candidate genes for immune response against H. contortus. SNPs were discarded if call rate <95% and minor allele frequency ≤0.05. A mixed model was used to analyze the response variables and included the identity by state matrix to control for population structure. A contemporary group (age, group, and sex) was included as fixed effect. Bonferroni correction was used to control for multiple testing. Eighteen SNPs on chromosomes 1, 2, 3, 4, 6, 7, 11, 15, 18, 20, 24, and 26 were significant for different traits. Our results suggest that loci related to Th17, Treg, and Th2 responses play an important role in the expression of resistant phenotypes. Several genes including ITGA4, MUC15, TLR3, PCDH7, CFI, CXCL10, TNF, CCL26, STAT3, GPX2, IL2RB, and STAT6 were identified as potential markers for resistance to natural H. contortus exposure. This is the first study that evaluates potential genetic markers for H. contortus resistance in Florida Native sheep.

Background RNA sequencing (RNA-seq) has allowed for transcriptional profiling of biological systems through identification of differentially expressed (DE) genes and pathways. Results A total of 80 steers were selected from the multibreed Angus-Brahman herd of the University of Florida. Sensory panel tenderness, juiciness and connective tissue as well as marbling, WBSF and cooking loss were assessed in longissimus dorsi muscle. Nuclear RNA was extracted from muscle and an RNA-seq library for each sample was constructed, multiplexed, and sequenced based on protocols by Illumina HiSeq 3000 PE100 platform to generate 2 × 101 bp paired-end reads. On average, 34.9 million high-quality paired reads were uniquely mapped to the Btau_4.6.1 reference genome and a total of 8,799 genes were analyzed. Including all 80 animals, gene and exon expression analysis was carried out using a meat quality index as a continuous response variable. The expression of 208 genes and 3,280 exons from 1,565 genes was associated with the meat quality index (p-value ≤ 0.05). Out of the 80 samples sequenced, 40 animals with extreme low and high WBSF, tenderness and marbling values were selected for a differential expression (DE) analysis for gene and isoforms. A total of 676 (adjusted p-value ≤ 0.05), 70 (adjusted p-value ≤ 0.1) and 198 (adjusted p-value ≤ 0.1) genes were DE for WBSF, tenderness and marbling, respectively. A total of 106 isoforms from 98 genes for WBSF, 13 isoforms from 13 genes for tenderness and 43 isoforms from 42 genes for marbling (FDR ≤ 0.1) were DE. Conclusion A number of cytoskeletal and transmembrane anchoring related genes and pathways were identified in the expression, DE and gene enrichment analyses, and these proteins can have a direct effect on meat quality. Cytoskeletal proteins and transmembrane anchoring molecules can influence meat quality by allowing cytoskeletal filament interaction with myocyte and organelle membranes, contributing to cytoskeletal structure, microtubule network stability, and cellular architecture maintenance during the postmortem.

Background: Transcription has a substantial genetic control and genetic dissection of gene expression could help us understand the genetic architecture of complex phenotypes such as meat quality in cattle. The objectives of the present research were: 1) to perform eQTL and sQTL mapping analyses for meat quality traits in longissimus dorsi muscle; 2) to uncover genes whose expression is influenced by local or distant genetic variation; 3) to identify expression and splicing hot spots; and 4) to uncover genomic regions affecting the expression of multiple genes. Results: Eighty steers were selected for phenotyping, genotyping and RNA-seq evaluation. A panel of traits related to meat quality was recorded in longissimus dorsi muscle. Information on 112,042 SNPs and expression data on 8,588 autosomal genes and 87,770 exons from 8,467 genes were included in an expression and splicing quantitative trait loci (QTL) mapping (eQTL and sQTL, respectively). A gene, exon and isoform differential expression analysis previously carried out in this population identified 1,352 genes, referred to as DEG, as explaining part of the variability associated with meat quality traits. The eQTL and sQTL mapping was performed using a linear regression model in the R package Matrix eQTL. Genotype and year of birth were included as fixed effects, and population structure was accounted for by including as a covariate the first PC from a PCA analysis on genotypic data. The identified QTLs were classified as cis or trans using 1 Mb as the maximum distance between the associated SNP and the gene being analyzed. A total of 8,377 eQTLs were identified, including 75.6% trans, 10.4% cis, 12.5% DEG trans and 1.5% DEG cis; while 11,929 sQTLs were uncovered: 66.1% trans, 16.9% DEG trans, 14% cis and 3% DEG cis. Twenty-seven expression master regulators and 13 splicing master regulators were identified and were classified as membrane-associated or cytoskeletal proteins, transcription factors or DNA methylases. These genes could control the expression of other genes through cell signaling or by a direct transcriptional activation/repression mechanism. Conclusion: In the present analysis, we show that eQTL and sQTL mapping makes possible positional identification of gene and isoform expression regulators.

Background: Transcription has a substantial genetic control and genetic dissection of gene expression could help us understand the genetic architecture of complex phenotypes such as meat quality in cattle.Results: A total of 80 steers were selected for phenotyping, genotyping and RNA-seq evaluation. A panel of traits related to meat quality were recorded. Information on 112,042 SNPs and expression data on 8,588 autosomal genes and 87,770 exons from 8,467 genes were included in an expression and splicing quantitative trait loci (QTL) mapping (eQTL and sQTL, respectively). Expression of 1,352 genes was previously identified as associated with meat quality traits using a gene, exon and isoform differential expression (DE) analysis. The R package Matrix eQTL was used to perform the QTL mapping using linear regression. The identified QTLs were classified as cis or trans using 1 Mb as maximum distance between the associated SNP and the gene. Polymorphisms associated with expression of at least 20 genes, and splicing of at least 20 exons were considered QTL hot spots. A total of 8,377 eQTLs were identified, including 75.6% trans, 10.4% cis, 12.5% DE trans and 1.5% DE cis; 11,929 sQTLs were uncovered: 66.1% trans, 16.9% DE trans, 14% cis and 3% DE cis. Twenty seven expression master regulators and 13 splicing master regulators were identified and were classified as membrane associated or cytoskeletal proteins, transcription factors or DNA methylases These genes could control expression of other genes through cell signaling or by a direct transcriptional activation/repression mechanism. The ZNF804A, ALAD, OR13F1 and ENSBTAG00000000336 genes were identified as both expression and splicing master regulators.Conclusion: In the present analysis, we show that eQTL and sQTL mapping makes possible positional identification of gene and isoform expression regulators. Additionally, this mapping provides new insight into the regulatory network architecture in longissimus dorsi muscle in an Angus-Brahman multibreed population.