Mitochondria hold redox homeostasis and energy metabolism as a crucial factor during oocyte maturation, while the exposure of estrogenic mycotoxin zearalenone causes developmental incapacity in porcine oocyte. This study aimed to reveal a potential resistance of phytoalexin resveratrol against zearalenone during porcine oocyte maturation and whether its mechanism was related with PTEN-induced kinase 1 (PINK1)/Parkin-mediated mitophagy. Porcine oocytes were exposed to 20 μM zearalenone with or without 2 μM resveratrol during in vitro maturation. As for the results, zearalenone impaired ultrastructure of mitochondria, causing mitochondrial depolarization, oxidative stress, apoptosis and embryonic developmental incapacity, in which mitophagy was induced in response to mitochondrial dysfunction. Phytoalexin resveratrol enhanced mitophagy through PINK1/Parkin in zearalenone-exposed oocytes, manifesting as enhanced mitophagy flux, upregulated PINK1, Parkin, microtubule-associated protein light-chain 3 beta-II (LC3B-II) and downregulated substrates mitofusin 2 (MFN2), voltage-dependent anion channels 1 (VDAC1) and p62 expressions. Resveratrol redressed zearalenone-induced mitochondrial depolarization, oxidative stress and apoptosis, and accelerated mitochondrial DNA copy during maturation, which improved embryonic development. This study offered an antitoxin solution during porcine oocyte maturation and revealed the involvement of PINK1/Parkin-mediated mitophagy, in which resveratrol mitigated zearalenone-induced embryonic developmental incapacity.

Mycotoxins such as zearalenone (ZEN), deoxynivalenol (DON) and T-2 toxin (T-2) are the most poisonous biological toxins in food pollution. Mycotoxin contaminations are a global health issue. The aim of the current study was to use porcine Leydig cells as a model to explore the toxic effects and underlying mechanisms of ZEN, DON and T-2. The 50% inhibitory concentration (IC₅₀) of ZEN was 49.71 μM, and the IC₅₀ values of DON and T-2 were 2.49 μM and 97.18 nM, respectively. Based on the values of IC₅₀, ZEN, DON and T-2 exposure resulted in increased cell apoptosis, as well as disrupted mitochondria membrane potential and cell cycle distribution. The results also showed that ZEN and DON significantly reduced testosterone and progesterone secretion in Leydig cells, but T-2 only reduced testosterone secretion. Furthermore, the expression of steroidogenic acute regulatory (StAR) protein and 3β-hydroxysteroid dehydrogenase (3β-HSD) were significantly decreased by ZEN, DON and T-2; whereas the protein expression of cholesterol side-chain cleavage enzyme (CYP11A1) was only significantly decreased by ZEN. Altogether, these data suggest that the ZEN, DON and T-2 toxins resulted in reproductive toxicity involving the inhibition of steroidogenesis and cell proliferation, which contributes to the cellular apoptosis induced by mitochondrial injury in porcine Leydig cells.

Chinese indigenous pig breeds have unique genetic characteristics and a rich diversity; however, effective breed identification methods have not yet been well established. In this study, a genotype file of 62,822 single-nucleotide polymorphisms (SNPs), which were obtained from 1059 individuals of 18 Chinese indigenous pig breeds and 5 cosmopolitan breeds, were used to screen the discriminating SNPs for pig breed identification. After linkage disequilibrium (LD) pruning filtering, this study excluded 396 SNPs on non-constant chromosomes and retained 20.92~−27.84% of SNPs for each of the 18 autosomes, leaving a total of 14,823 SNPs. The principal component analysis (PCA) showed the largest differences between cosmopolitan and Chinese pig breeds (PC1 = 10.452%), while relatively small differences were found among the 18 indigenous pig breeds from the Yangtze River Delta region of China. Next, a random forest (RF) algorithm was used to filter these SNPs and obtain the optimal number of decision trees (ntree = 1000) using corresponding out-of-bag (OOB) error rates. By comparing two different SNP ranking methods in the RF analysis, the mean decreasing accuracy (MDA) and mean decreasing Gini index (MDG), the effects of panels with different numbers of SNPs on the assignment accuracy, and the statistics of SNP distribution on each chromosome in the panels, a panel of 1000 of the most breed-discriminative tagged SNPs were finally selected based on the MDA screening method. A high accuracy (>99.3%) was obtained by the breed prediction of 318 samples in the RF test set; thus, a machine learning classification method was established for the multi-breed identification of Chinese indigenous pigs based on a low-density panel of SNPs.

There are many applications of soybean lecithin (SL) and cholesterol-loaded cyclodextrin (CLC) in sperm freezing processes. To the best of our knowledge, there have been few cases of the combined use of SL and CLC in freezing rooster semen. We investigated the effects of CLC, SL, and their combination on rooster sperm cryodamage. Three experiments were conducted: experiment 1, SL (0.5%, 1%, 1.5%, 2.0%); experiment 2, CLC (1.25 mg, 2.5 mg, 5 mg, 10 mg, 15 mg); and experiment 3, CLC + SL (2.5 mg + 0.25%, 2.5 mg + 0.5%, 2.5 mg + 1%, 2.5 mg + 1.5%). Semen samples were cryopreserved using a programmed cryostat, followed by the determination of post-thaw sperm quality, antioxidant indices, and hatching. The results showed that the combination of 2.5 mg CLC + 0.5% SL had the most significant synergistic effect on cryodamage, and the viability (56.69%), motility (54.35%), mitochondrial activity (54.23%), plasma membrane integrity (53.52%), acrosome integrity (54.71%), and antioxidant activity (MDA concentration: 5.65 nmol/mL; SOD activity: 152.73 U/mL) were significantly greater than those of the other combinations (p < 0.05). Nevertheless, the combined CLC and SL addition group did not substantially increase the fertilization and hatching rates of frozen semen compared with the addition of 2.5 mg CLC. In conclusion, the addition of 2.5 mg CLC and 2.5 mg CLC + 0.5% SL enhanced the quality and fertility of frozen rooster sperm.

This study developed a 60K single-nucleotide polymorphism (SNP) liquid-phase chip (‘Shenxin I’) based on genotyping by target sequencing (GBTS) technology. The chip was used to genotype 1451 individuals spanning five conserved local pig breeds: Fengjing (FJ), Meishan (MMS), Pudong White (PD), Shaowutou (SW), and Shanghai White (SHW), and newborn purebred Meishan offspring (MMS_New). The study assessed the genetic diversity, population structure, and genomic breed composition (GBC) of these breeds. The GBC analysis provided insights into genomic introgression within admixed individuals. Results indicate that PD and FJ breeds exhibited the higher fraction of the genome covered by runs of homozygosity (F_ROH) and slower linkage disequilibrium (LD) decay rate, suggesting higher genetic inbreeding level in these two breeds. The study conducted genetic distance of identity-by-state (IBS) analysis and molecular pedigree construction for the MMS population. Subsequently, purebred mating was implemented by selecting boars from different pedigrees to mate with sows and excluding individuals with high inbreeding coefficients from the breeding program. This resulted in a significant reduction in the overall inbreeding level of the born progeny and an increase in genetic diversity compared to the original population. Consequently, the study concludes that utilizing liquid-phase chip technology for genotyping, constructing molecular pedigrees, and optimizing mating combinations in small populations of locally conserved pig breeds contributes to enhancing the conservation effectiveness.

This study combined liquid-phase chip technology with a genome-wide association study (GWAS) to identify candidate functional genes for litter size in Meishan pigs. Genotyping of 198 Meishan sows using the “Shenxin No. 1” 60K porcine SNP chip, followed by quality control with PLINK v1.90, yielded 31,430 autosomal SNPs. Principal component analysis revealed no significant population stratification. Using a mixed linear model in GEMMA (v 0.98.1), we identified 9 significant SNP loci and 52 candidate genes on chromosomes 5 and 14, including AEBP2, P2RX2, ANKLE2, PGAM5, and EP400, which are implicated in embryonic development, ovarian function, and cell proliferation. Isolated porcine ovarian granulosa cells were validated by FSHR immunofluorescence, showing 99% purity. Overexpression of AEBP2 significantly upregulated its protein expression (Western blot) and promoted granulosa cell proliferation (CCK-8). Collectively, these results pinpoint potential candidate genomic regions and functional genes associated with litter size and suggest that AEBP2 may enhance reproductive performance by promoting granulosa cell proliferation and follicular development. This provides new targets for elucidating the genetic mechanisms underlying the high fecundity of Meishan pigs and for advancing molecular breeding efforts.