The aim of this study was to identify the c.495C > T polymorphism within exon 1 of the osteopontin gene (OPN), and to analyze its association with susceptibility to ketosis in Polish Holstein–Friesian (HF) cows. The study utilized blood samples from 977 HF cows, for the determination of β-hydroxybutyric acid (BHB) and for DNA isolation. The c.495C > T polymorphism of the bovine osteopontin gene was determined by PCR–RFLP. The CT genotype (0.50) was deemed the most common, while TT (0.08) was the rarest genotype. Cows with ketosis most often had the CC genotype, while cows with the TT genotype had the lowest incidence of ketosis. To confirm the relationship between the genotype and ketosis in cows, a weight of evidence (WoE) was generated. A very strong effect of the TT genotype on resistance to ketosis was demonstrated. The distribution of the ROC curve shows that the probability of resistance to ketosis is > 75% if cows have the TT genotype of the OPN gene (cutoff value is 0.758). Results suggest that TT genotype at the c.495C > T locus of the OPN gene might be effective way to detect the cows with risk of ketosis.

Epigenetic mechanisms of gene regulation are important for the proper differentiation of cells used for therapeutic and regenerative purposes. The primary goal of the present study was to investigate the impacts of 5-aza-2′ deoxycytidine (5-AZA-dc)- and/or trichostatin A (TSA)-mediated approaches applied to epigenomically modulate the ex vivo expanded equine chondrocytes maintained in monolayer culture on the status of chondrogenic cytodifferentiation at the transcriptome level. The results of next-generation sequencing of 3′ mRNA-seq libraries on stimulated and unstimulated chondrocytes of the third passage showed no significant influence of 5-AZA-dc treatment. Chondrocytes stimulated with TSA or with a combination of 5-AZA-dc+TSA revealed significant expressional decline, mainly for genes encoding histone and DNA methyltransferases, but also for other genes, many of which are enriched in canonical pathways that are important for chondrocyte biology. The TSA- or 5-AZA-dc+TSA-induced upregulation of expanded chondrocytes included genes that are involved in histone hyperacetylation and also genes relevant to rheumatoid arthritis and inflammation. Chondrocyte stimulation experiments including a TSA modifier also led to the unexpected expression incrementation of genes encoding HDAC3, SIRT2, and SIRT5 histone deacetylases and the MBD1 CpG-binding domain protein, pointing to another function of the TSA agent besides its epigenetic-like properties. Based on the transcriptomic data, TSA stimulation seems to be undesirable for chondrogenic differentiation of passaged cartilaginous cells in a monolayer culture. Nonetheless, obtained transcriptomic results of TSA-dependent epigenomic modification of the ex vivo expanded equine chondrocytes provide a new source of data important for the potential application of epigenetically altered cells for transplantation purposes in tissue engineering of the equine skeletal system.

The effect of the g.4290 C>G substitution in the FADS2 gene and g.285 C>T in the FABP4 gene on carcass quality, meat quality, and fatty acid profile of the pectoralis superficialis muscle of 238 male broiler chickens reared up to 45 days of age was analyzed. A significant influence of g.4290 C>G in the FADS2 gene on the pectoralis superficialis muscle fatty acid profile was demonstrated. Chickens with the GG genotype were characterized by the highest content of conjugated linoleic acid, amino acids, eicosapentaenoic acids, docosapentaenoic acid, docosahexaenoic acids. and the lowest value of the linoleic acid/alpha-linolenic acid ratio. The FABP4 polymorphism determined only the content of C18:1n-9, C18:2n-6 and docosahexaenoic acid. There was no effect of the studied genotypes on final body weight, carcass quality traits, or quality of broiler pectoral muscles.

Nanopore sequencing is a third-generation biopolymer sequencing technique that relies on monitoring the changes in an electrical current that occur as nucleic acids are passed through a protein nanopore. Increasing quality of reads generated by nanopore sequencing systems encourages their application in genome-wide polymorphism detection and genotyping. In this study, we employed nanopore sequencing to identify genome-wide polymorphisms in the horse genome. To reduce the size and complexity of genome fragments for sequencing in a simple and cost-efficient manner, we amplified random DNA fragments using a modified DOP-PCR and sequenced the resulting products using the MinION system. After initial filtering, this generated 28,426 polymorphisms, which were validated at a 3% error rate. Upon further filtering for polymorphism and reproducibility, we identified 9495 SNPs that reflected the horse population structure. To conclude, the use of nanopore sequencing, in conjunction with a genome enrichment step, is a promising tool that can be practical in a variety of applications, including genotyping, population genomics, association studies, linkage mapping, and potentially genomic selection.

Despite extensive research on the cryopreservation of stallion semen, the process still requires improvement. Several diluent additives have been developed to overcome cryopreservation issues, however, semen from valuable individuals often shows poor quality after thawing. Mesenchymal stem cells (MSCs) are regarded as factors that may play an important role in the regenerative process of sperm by secreting extracellular vesicles (EVs) that are actively involved in intercellular communication, tissue repair and cell membrane maintenance. The study aimed to determine the effect of the addition of EV-MSCs obtained from equine adipose tissue on various parameters of cryopreserved stallion semen. All cells met the minimum criteria established by the ISCT, being a characteristic surface profile of the presence of CD90 and lack of MHC II, differentiation into three mesodermal lineages, and adherence. EVs were collected from the expanded cells by ultracentrifugation, and their characteristics were confirmed using nanoparticle tracking analysis (NTA) and flow cytometry to detect CD63 and CD81 antigens. Semen was collected from 11 stallions of the Hucul breed, and each parameter of sperm quality with (25 µg/ml, 50 µg/ml) and without EV addition (motility, morphology and viability) was analyzed. The results showed statistically significant differences in sperm motility, progressive movement and viability following EV treatment. While the amplitude of lateral head displacement (ALH) and morphology parameters did not change, the addition of EVs does not affect these sperm features. The results provide an initial insight into the role of EVs in the maintenance and regeneration of horse sperm after cryopreservation-induced shock.