BackgroundIndependent studies have shown that several single nucleotide polymorphisms (SNP) in the human FTO (fat mass and obesity associated) gene are associated with obesity. SNP have also been identified in the pig FTO gene, among which some are associated with selected fat-deposition traits in F2 crosses and commercial populations. In this study, using both commercial pig populations and an experimental Meishan × Pietrain F2 population, we have investigated the association between one FTO SNP and several growth and carcass traits. Association analyses were performed with the FTO polymorphism either alone or in combination with polymorphisms in flanking loci.MethodsSNP (FM244720:g.400C>G) in exon 3 of porcine FTO was genotyped by PCR-RFLP and tested for associations with some growth, carcass and fat-related traits. Proportions of genetic variance of four pig chromosome 6 genes (FTO, RYR1, LIPE and TGFB1) on selected traits were evaluated using single- and multi-locus models.ResultsLinkage analysis placed FTO on the p arm of pig chromosome 6, approximately 22 cM from RYR1. In the commercial populations, allele C of the FTO SNP was significantly associated with back fat depth and allele G with muscling traits. In the Meishan × Pietrain F2 pigs, heterozygotes with allele C from the Pietrain sows and allele G from the Meishan boar were more significantly associated with fat-related traits compared to homozygotes with allele G from the Pietrain and allele G from the Meishan breed. In single- and multi-locus models, genes RYR1, TGFB1 and FTO showed high associations. The contribution in genetic variance from the polymorphism in the FTO gene was highest for back fat depth, meat area on the musculus longissimus lumborum et thoracis tissues and metabolite glucose-6-phosphate dehydrogenase.ConclusionsOur results show that in pig, FTO influences back fat depth in the commercial populations, while in the Meishan × Pietrain F2 pigs with a CG genotype, heterosis occurs for several fat-related traits.

Abstract:The current knowledge of factors regulating voluntary feed intake in pigs is quite limited. The objective of this study was to test the influence of the missense mutation p.Asp298Asn (AF087937:c.746G>A) of the MC4R gene on selected production traits in pig crosses. These crosses are commonly used on commercial farms in the Czech Republic. The allele frequencies of c.746G>A were as follows: G allele -0.59 and A allele -0.41. We detected statistically significant differences in the content of intramuscular fat in the musculus longissimus lumborum et thoracis, and a similar trend was observed in shoulder and neck. A allele correlated with higher values of fatness and G allele with a higher percentage of lean meat. However, we did not find any significant influence on either feed intake or growth rate in this study. For another mutation, p.Arg236His (NM_214173.1:c.707G>A), frequencies of alleles were disproportional (A allele -0.02 and G allele -0.98), only two genotypes were observed (AG and GG) and linkage disequilibrium was not detected. Therefore, we assume that the effect of this polymorphism on growth rate and fatness in the Czech population of pigs is negligible.

The emergence and spread of resistant tuberculosis (TB) pose a threat to public health, so it is necessary to diagnose the drug-resistant forms in a clinically short time frame and closely monitor their transmission. In this study, we carried out a first whole genome sequencing (WGS)-based analysis of multidrug resistant (MDR) M. tuberculosis strains to explore the phylogenetic lineages diversity, drug resistance mechanisms, and ongoing transmission chains within the country. In total, 65 isolates phenotypically resistant to at least rifampicin and isoniazid collected in the Czech Republic in 2005–2020 were enrolled for further analysis. The agreement of the results obtained by WGS with phenotypic drug susceptibility testing (pDST) in the determination of resistance to isoniazid, rifampicin, pyrazinamide, streptomycin, second-line injectables and fluoroquinolones was more than 80%. Phylogenetic analysis of WGS data revealed that the majority of MDR M. tuberculosis isolates were the Beijing lineage 2.2.1 (n = 46/65; 70.8%), while the remaining strains belonged to Euro-American lineage. Cluster analysis with a predefined cut-off distance of less than 12 single nucleotide polymorphisms between isolates showed 19 isolates in 6 clusters (clustering rate 29.2%), located mainly in the region of the capital city of Prague. This study highlights the utility of WGS as a high-resolution approach in the diagnosis, characterization of resistance patterns, and molecular-epidemiological analysis of resistant TB in the country.

Eukaryotic translation elongation factor 1 alpha (EEF1A) plays a key role in protein synthesis. In higher vertebrates EEF1A occurs in two isoforms, EEF1A1 and EEF1A2, encoded by distinct genes. The purpose of this study was to compare the two porcine genes as for the genomic sequence, gene organization and mRNA expression in different tissues, as well as to search for polymorphism and chromosomal assignment. Standard methods of DNA and mRNA analysis were used. We determined the complete genomic sequence of the porcine EEF1A1 and EEF1A2 genes. The two genes differ in the lengths of transcription units (3102 and 8588 bp, respectively), but have similar genomic organization and their coding sequences are highly similar (78% identity of coding sequences and 92.4% identity of amino acid sequences). Several polymorphisms in the two genes were detected. EEF1A1 and EEF1A2 were mapped to SSC1p11.1 and SSC17q23.3, respectively. mRNA of EEF1A1 was expressed in all studied tissues (the highest expression was in 44-day fetal muscle and low expression in adult liver and brain), while EEF1A2 was expressed only in skeletal-muscle, tongue, heart, diaphragm and brain tissues. EEF1A2 was not expressed in fetal muscle tissue (44 days). In this paper results are provided on genomic sequences, genomic organization, polymorphism, chromosomal assignment and spatial and temporal expressions of the porcine EEF1A1 and EEF1A2 genes. Novel polymorphisms were described in both genes. Porcine EEF1A2 was studied for the first time.

Highlights The Euro-American lineage 4.7 was the most represented among the XDR-TB/MDR-TB resistant strains. Seven of 12 isolates (58%) belonged to the same recent transmission chain. Strains originally from Eastern European countries were diagnosed in patients without any epidemiological links with these countries. Incidence of resistant XDR-TB/MDR-TB in Slovakia, while low, still should be taken to monitor.

Interaction between gene expression and protein-coding genetic variation is increasingly being appreciated as an important source of adaptive phenotypic variation. In this study, we used reverse transcription-qPCR to test for gene expression variation in two β-globin paralogs (HBB-T1 and HBB-T2) of the Eurasian bank vole (Myodes glareolus), which both display the same structural polymorphism Ser52Cys responsible for variation in Cys-based antioxidant capacity of red blood cells (RBCs). We first demonstrated that HBB-T1 is the major expressed adult HBB gene in the bank vole accounting for ∼85% of total hemoglobin. We then measured the relative expression of the two homozygous genotypes in each gene and found that when present in HBB-T1, the oxidative-stress resistant Cys52 allele is significantly associated with higher expression ratio HBB-T1:HBB-T2. The results further indicated that the Cys52 allele present in HBB-T1 was associated with higher normalized expression of that gene compared to the Ser52 allele, although this difference was statistically significant only when using one reference gene but not the other. We argue that, altogether, our results indicate the presence of a cis-acting regulatory genetic variation modulating the expression of the two alleles in HBB-T1. Previous studies indicated that the resistant RBC phenotype is likely beneficial under conditions conducive to oxidative stress. The duplicate HBB genes of the bank vole thus may represent a novel example of gene-regulatory genetic variation interacting with a well-defined protein-coding variation to control an adaptive trait.

Cone-rod dystrophy is a progressive inherited retinal degenerative disorder that occurs in humans and dogs. The deletion in the nephronophthisis 4 (NPHP4) gene was established as a causative mutation in standard wire-haired Dachshunds. We analyzed all varieties of Dachshunds from the Czech Republic and five other dog breeds and found that the deletion in the NPHP4 (in heterozygous state) is present not only in standard-, but also in miniature wire-haired Dachshunds, but not in other varieties of Dachshunds or in other breeds.

ABSTRACT:Genes that are expressed in skeletal muscles may play a role in prenatal muscle development and postnatal muscle growth and can be considered candidates for economically important traits. Four porcine genes that were differentially expressed in skeletal muscles of Czech Large White and Wild Boar (SORT1, EMP3, IL18, and BTG2) were selected to search for polymorphism, linkage assignment, and association analysis with carcass traits. Through comparative sequencing of portions of the genes numerous polymorphisms were revealed (SORT1 -21, EMP3 -6, IL18 -41, BTG2 -9). Linkage analysis in a Meishan × Pietrain F 2 pedigree showed the positions of the genes relative to other genes and markers on the respective chromosomes -SORT1 on SSC4, EMP3 on SSC6, IL18 and BTG2 on SSC9. Preliminary association analysis in pig commercial crosses with selected SNPs showed associations with several carcass traits at nominal P value of < 0.05, which may indicate their involvement in muscle growth and fat deposition. The tested polymorphisms may not be causal for the associations, but they may be in linkage disequilibrium with causative mutations.