Kalmyk cattle are an important meat breed in Kazakhstan, valued for their strong physique, genetic stability, and adaptability. In this study, we investigated the mRNA expression of ELOVL fatty acid elongase 6 (ELOVL6) and CREB-regulated transcription coactivator 2 (CRTC2) across multiple tissues, and further examined their genetic variations and associations with growth and carcass traits in 200 Kalmyk cattle. Expression analysis showed that CRTC2 was most highly expressed in the heart and liver, whereas ELOVL6 was predominantly expressed in the spleen and large intestine, highlighting their tissue-specific expression patterns. In addition, we identified a polymorphic SNP (g.16511290A > G) in the 3′UTR of ELOVL6, with three genotypes (AA, AG, GG) and the G allele being dominant (0.520). Polymorphism information content (PIC) analysis indicated high genetic diversity at this locus. Importantly, this SNP was significantly associated with live weight and body oblique length (p < 0.05), and individuals carrying heterozygous AG genotype showed higher body weight and length. Collectively, these findings suggest that g.16511290A > G within ELOVL6 may serve as useful molecular markers for body measurements and meat traits, providing valuable resources for marker-assisted selection in beef cattle breeding programs.

This study investigates the role of Perilipin1 (PLIN1) in milk fat synthesis in bovine mammary epithelial cells (BMECs) and its regulatory mechanisms, aiming to provide a foundation for improving milk fat content through molecular breeding. BMECs were used as a model to analyze the effects of PLIN1 overexpression (OE-PLIN1) and interference (si-PLIN1) on milk fat synthesis and lipid-related gene expression using RT-qPCR, Western blot, and Oil Red O staining. Results show that OE-PLIN1 significantly enhances triglyceride (TAG) accumulation in BMECs (P < 0.01), upregulates lipid synthesis-related genes (such as PPARγ, C/EBPα, C/EBPβ, FABP4, FASN) (P < 0.05), and downregulates the mRNA expression of lipid breakdown-related genes (HSL, ATGL) (P < 0.05). Conversely, si-PLIN1 significantly reduces TAG accumulation (P < 0.05) and lowers the expression of lipid synthesis and breakdown genes (P < 0.05). Additionally, OE-PLIN1 combined with SREBP1 siRNA interference (si-SREBP1) did not have a significant impact on the mRNA and protein levels of SREBP1, but it significantly altered SREBP1’s phosphorylation, indicating that SREBP1 interference inhibits PLIN1’s effect on milk fat synthesis. This study suggests that PLIN1 promotes milk fat synthesis in BMECs via regulating SREBP1 activity, offering a new strategy for enhancing milk fat content in dairy cattle.

In pigs, the effect of sex on production and reproductive traits has been largely reported, however, whether sex exerts its influence through regulating mitochondrial function is still unclear. In this study, we constructed 15 male cells and 15 female fibroblasts derived from 35-day and 50-day fetuses, newborn piglets and 1-year-old pigs to identify the sex effect on mitochondrial functions. Results indicated significant differences on cellular and molecular characteristics between male and female cells, including energy metabolic trait, mitochondrial DNA (mtDNA) replication and transcription, and mRNA expressions of mitochondrial biogenesis genes and mitoprotease genes. Referring to sex, males exhibited significantly higher oxygen consumption rate productions, levels of reactive oxygen species (ROS) and mtDNA copy numbers than those with females in muscle and ear fibroblasts. And the expressions of mtDNA, mitochondrial biogenesis genes (POLG, PPARGC1A, TFAM and TWNK) and XPNPEP3 were higher in males than females in ear fibroblasts derived from 1-year-old adult pigs (EFA cells). While, the cell proliferation and expressions of genes related to ROS metabolism were not influenced by sex. The results highlight the effect of sex on mitochondrial function and gene expression, and provide important data for a comprehensive understanding of the mechanisms underlying sex regulation of energy metabolism-related traits in pigs.

Ketosis is a common metabolic disease in high-yield dairy cows. Key genes affecting ketosis need to be further explored by new methods. The gene expression profiling and clinical data of GSE92398, GSE104079, and GSE4304 were obtained from the gene expression omnibus (GEO) database. Core modules and genes associated with RFI (residual feed intake) and ADF (alternate day fasting) were identified by weighted gene co-expression network analysis (WGCNA). Subsequently, the key genes related to ketosis and RFI were determined by protein-protein interaction (PPI) networks, ROC curves, functional enrichment, and differential expression analysis, respectively. The results showed that the genes of ACACA, ELOVL6 and XPO7 could be used as regulators of ketosis induced by low feed intake in dairy cows. At the same time, three genes (HRFI, STAT3 and IFNAR1) were retained as additional RFI biomarkers that could be considered. We identified three key factors as candidate genes and biomarkers of ketosis and RFI, respectively. These factors may provide a theoretical basis for targeted therapy of ketosis in dairy cows.

The study investigates the relationship between biofilm formation and antibiotic resistance in Escherichia coli (E. coli) isolated from calves. Using biochemical and molecular methods, we identified the isolates and assessed their biofilm-forming ability through an improved crystal violet staining method. The minimum inhibitory concentrations (MICs) of 18 antibiotics against the isolates were determined using the broth microdilution method. The impact of cefoxitin on biofilm formation was analyzed using laser scanning confocal microscopy (LSCM). Additionally, qRT-PCR was employed to evaluate the expression levels of biofilm-related genes (luxS, motA, fliA, pfs, and csgD) in response to varying cefoxitin concentrations. Results indicated a significant correlation between antimicrobial resistance (AMR) and biofilm formation ability. Cefoxitin effectively reduced biofilm formation of multidrug-resistant E. coli isolates at 1/2 and 1 MIC, with enhanced inhibition at higher concentrations. The QS-related genes luxS, pfs, motA, and fliA were downregulated, leading to decreased csgD expression. At 1/2 MIC, csgD expression was significantly reduced. In conclusion, cefoxitin inhibits biofilm formation in multidrug-resistant E. coli by down-regulating key genes, offering a potential strategy to mitigate resistance and control infections in calves caused by biofilm-positive E. coli isolates.

Variation in litter size (LS) in sheep is linked to genetic factors, including the Zona pellucida-3 (ZP3) gene, which plays a role in ovine reproductive processes. This study examined the association between ZP3 gene variations and LS in Kari sheep. Two groups of 160 Kari ewes were analysed: one consistently producing singletons and another producing twins, with occasional triplets. Additionally, Madakhlasht sheep, which sometimes produce twins, and Balkhi sheep, which produce only singletons, were used as references. The entire ZP3 gene was amplified using PCR and sequenced at 30× with Next Generation Sequencing. Bioinformatics analysis identified 70 variants across the three breeds, located in upstream regions, introns, and exons. Notably, two point mutations and a six-nucleotide insertion were found upstream of the initiation codon in twin-producing Kari ewes, potentially affecting ZP3 expression and LS. Two missense mutations (I101L in exon 1 and R408H in exon 8) were heterozygous in twin-producing Kari ewes but homozygous in other groups, correlating with LS. Protein modelling suggested that the I101L mutation alters the binding site, potentially impacting protein function. These findings indicate that ZP3 gene variations influence reproductive efficiency and LS in sheep, with specific variants serving as potential markers for selective breeding to enhance LS.

Glycogen synthase 2 (GYS2) encodes liver glycogen synthase, a rate limiting enzyme in glycogen metabolism. Our preliminary work suggested that GYS2 was a candidate gene affecting milk production traits by analyzing the liver proteome of dairy cows. Herein, this research identified single nucleotide polymorphisms (SNPs) of GYS2, analyzed their genetic effects on traits of dairy cattle, and speculated the pathogenic mechanism through functional prediction of key mutation sites. Seven SNPs were found by resequencing and the association analysis showed that these SNPs were significantly associated with 305-day milk yield, fat yield, protein yield or fat percentage (p-value ≤ 0.0488). Six SNPs among them formed two haplotype blocks and they were associated with 305-day milk yield, fat yield, protein yield or fat percentage (p-value ≤ 0.0349). Furthermore, 5:g.88602007G > A and 5:g.88602026G > A were predicted to change the transcription factor binding sites (TFBSs), which might regulate the expression of GYS2. The missense mutation site, 5:g.88602535G > T, changed the secondary structure of mRNA and the secondary and tertiary structure of protein. In summary, the GYS2 was proved to have genetic effect on milk production traits, and its valuable seven SNPs, could provide more useful genetic information for molecular breeding of dairy cows.

Oocytes spontaneously resume meiosis following their liberation from follicles, preventing full competence acquisition. Biphasic IVM (CAPA-IVM) maintains oocytes in meiotic arrest to improve developmental competence, and it specially affects poorly developed oocytes. We assessed the effect of CAPA-IVM on oocytes from small (<3mm) and large (>3mm) follicles of prepubertal goats. Oocytes were cultured for 6h in pre-IVM with C-type natriuretic peptide (CNP) and estradiol as meiotic inhibitors, and germinal vesicle (GV) rate and chromatin configuration were assessed. Then, oocytes were cultured in conventional IVM (c-IVM) or CAPA-IVM (pre-IVM + c-IVM) and EGF receptor (EGFR) protein expression, intra-oocyte ROS and blastocyst development were assessed. GV rate was higher in CNP groups than control (69% vs 28%, and 67% vs 31%, small and large follicles, respectively), but GV chromatin configuration was similar. In large follicles, EGFR expression was higher in oocytes and cumulus cells after CAPA-IVM, and ROS levels were lower. In small follicles these differences were not observed. c-IVM and CAPA-IVM produced similar blastocyst rates in small (3.7% vs 2.6%, respectively) and large follicles (8.3% vs 2.5%). Overall, CAPA-IVM enhanced EGFR expression for EGF peptide signalling and antioxidant capacity in oocytes from large follicles but oocytes from small follicles were too immature to benefit from it.

The history of turkey (Meleagris gallopavo) domestication can be traced back to the period between 700 and 200 BC in Mexico. This process involved multiple contributors and resulted in the development of modern local turkey breeds. This research investigates the complete mitochondrial diversity across a diverse range of local turkeys. Seventy-three turkeys were sampled from various populations, including autochthonous Italian breeds, an American breed (Narragansett), as well as wild turkeys from the USA and Mexico. The mitochondrial DNA (mtDNA) was employed as a powerful tool for biodiversity and breed phylogeny investigation. An analysis of the entire mtDNA was conducted to identify breed-specific unique traits, mitochondrial-specific characteristics, and the phylogenetic relationship among turkey populations. A total of 44 polymorphic sites were identified. Brianzolo and Narragansett birds were characterized as genetically homogeneous populations. Thirty-two different haplotypes were identified when our samples were compared with mtDNA D-loop of 96 online available turkeys from various geographical countries. H1 and H2, differing for one mutation, were the most abundant, comprising 132 of the 185 sequences. H1 included samples coming from every region, while H2 was predominantly characterized by Italian samples. USA and Mexican samples appear to be more variable in their mtDNA than the other populations.

The PPARGC1A gene plays a fundamental role in regulating cellular energy metabolism, including adaptive thermogenesis, mitochondrial biogenesis, adipogenesis, gluconeogenesis, and glucose/fatty acid metabolism. In a previous study, our group investigated seven SNPs in Mediterranean buffalo associated with milk production traits, and the current study builds on this research by exploring the regulatory influences of the PPARGC1A gene in buffalo mammary epithelial cells (BuMECs). Our findings revealed that knockdown of PPARGC1A gene expression significantly affected the growth of BuMECs, including proliferation, cell cycle, and apoptosis. Additionally, we observed downregulated triglyceride secretion after PPARGC1A knockdown. Furthermore, the critical genes related to milk production, including the STATS, BAD, P53, SREBF1, and XDH genes were upregulated after RNAi, while the FABP3 gene, was downregulated. Moreover, Silencing the PPARGC1A gene led to a significant downregulation of β-casein synthesis in BuMECs. Our study provides evidence of the importance of the PPARGC1A gene in regulating cell growth, lipid, and protein metabolism in the buffalo mammary gland. In light of our previous research, the current study underscores the potential of this gene for improving milk production efficiency and overall dairy productivity in buffalo populations.