Background: Pyometra, involves pus accumulation that can lead to death if detected late, making early diagnosis and management crucial, and it is more common in older dogs. The Axiom™ Canine HD Array was used in the study to determine single nucleotide polymorphisms (SNPs) associated with pyometra, with the aim of providing foundational data for the future development of SNP markers. Methods: Samples were collected from 95 dogs of 26 breeds in South Korea, and SNP genotypes were analyzed for the final two SNPs (AX-168186923, AX-168208364). To verify the sequences and genotypes of the selected SNPs, we conducted a sequencing analysis of 96 dogs from 11 breeds previously reported to have been affected by the disease. To confirm the linkage disequilibrium (LD) between the selected SNPs and the influence of non-synonymous SNP (nsSNP), candidate gene function and, protein structure change prediction analyses were performed. Results: Sequence and genotype analyses of the selected SNPs revealed differences in genotype frequencies among the varieties. A strong LD block was formed between the SNPs, confirming that both SNPs were genetically associated. Candidate gene ABCC4 analysis revealed that ABCC4 transports prostaglandins. An analysis of the three-dimensional protein structure affected by the nsSNP (AX-168186923) showed that the amino acid changed from Methionine to Valine, but the overall protein structure remained unchanged. Conclusions: The SNPs selected in this study can be used to identify genetic factors, associated with pyometra and vulnerable breeds. This is expected to reduce companion animal related costs and the psychological burden on owners.

The vitrification of embryos is essential for animal reproduction and significantly contributes to assisted reproductive technologies, enabling fast cryopreservation without ice crystal formation. Mitochondria, vital organelles in cellular metabolism, are responsible for critical functions like ATP synthesis, calcium regulation, and apoptotic signaling. Preserving mitochondrial integrity is essential for ensuring embryonic strength. Studies demonstrate that vitrification, a widely used cryopreservation method, can markedly impair mitochondrial function in mammalian embryos. This study examines the efficacy of novel/modified antifreeze peptide as a biocompatible agent when used in an appropriate concentration with base vitrification media. Blastocysts vitrified in base media as well as supplemented with the peptide exhibited significantly enhanced post-thaw survival rates, attaining re-expansion and hatching rates of 96.89 ± 4.2% and 88.31 ± 1.3%, respectively, in contrast to 79.38 ± 3.7% and 52.57.9 ± 0.8% observed in the control group. Furthermore, peptide-treated BLs demonstrated elevated expression of PGC1α, BCL2, and Sirt-1, which are the key genes related to mitochondrial membrane potential and anti-apoptotic factors. The mitochondrial function was maintained, and the levels of reactive oxygen species (ROS) and the expression of genes such as Cyto-c, caspases 3, and caspase 9 were markedly diminished in the embryos vitrified with peptide. These findings highlight the ability of this modified peptide to preserve mitochondrial integrity and reduce oxidative stress, hence enhancing the survival of blastocysts post-vitrification.

This study examined the effects of pre-soaking solvents and repeated steaming-drying (SD) cycles on the antioxidant activity and active compound content of Rehmanniae Radix Preparata(RRP), the processed root of Rehmannia glutinosa Libosch. SD treatments were conducted for 1 to 9 cycles using four different pre-soaking solvents: Takju (a traditional rice wine), Spirits, Honey, and Sugar solution. The results showed no significant differences in DPPH and ABTS radical scavenging activities or in total polyphenol and flavonoid contents among the pre-soaking treatments, although samples pre-soaked in honey and Takju exhibited slightly higher levels. Polyphenol and flavonoid contents increased progressively with the number of SD cycles, reaching levels 2–3 times higher after nine cycles. Catalpol content remained relatively constant regardless of treatment, whereas aucubin content increased in all groups, with the highest accumulation observed in the Takju treatment. Similarly, 5-hydroxymethylfurfural (5-HMF) content increased with the number of SD cycles, with the highest levels found in the honey group, followed by Takju, Sugar, and Spirits. In conclusion, while the type of pre-soaking solvent had minimal influence on antioxidant activities and catalpol content, both aucubin and 5-HMF contents increased with additional SD cycles, with Takju proving particularly effective in enhancing their accumulation.

Fetal Bovine Serum (FBS) plays a crucial role in animal cell culture; however, the increasing number of bovine fetuses used and sacrificed solely for FBS collection has raised ethical concerns globally. The welfare of fetuses during FBS blood collection has become a key focus of debate among animal welfare and ethics organizations worldwide. Previous studies indicate that heat-inactivated coelomic fluid (HI-CF) from the earthworm Perionyx excavatus may serve as a viable FBS alternative in adherent cell cultures. This study evaluates the potential of HI-CF as an FBS substitute during the in vitro maturation (IVM) stage of bovine embryo culture, with a focus on improving developmental rate through antioxidation effects. In this study, 2% HI-CF was incorporated into IVM media, assessing its impact on cell growth, differentiation, and the expression of genes related to antioxidation. The group of 2% of HI-CF exhibited a trend toward increased cleavage and blastocyst development rates compared to the control group. Although antioxidant genes such as NRF2 and GSR showed no statistically significant differences between the control and treatment groups, a trend toward increased expression was observed. Conversely, GPX1 displayed a trend of decreased expression. Notably, IGF1 and NQO1 were significant upregulated (p < 0.05) in the 2% HI-CF group. Additionally, oocytes stained with H2DCFDA showed a significantly reduced ROS levels (p < 0.05) in the 2% HI-CF group compared with controls. These findings suggest that HI-CF's antioxidative effects support enhanced cell growth and blastocyst development rate, surpassing those observed with FBS. Consequently, HI-CF shows promise as an effective alternative to FBS in vitro maturation of bovine oocytes.