Aldosterone (ALD) is a mineralocorticoid, an active component of the renin-angiotensin system. It has been reported that the concentration of ALD in large follicles is higher than that in small follicles. However, whether it has a beneficial effect on the maturation and development of oocytes from small follicles is currently unclear. Here, we showed that ALD supplementation during in vitro maturation promotes the maturation and development of oocytes from small follicles. Specifically, we found that 1 μg/mL ALD significantly enhanced oocyte quality by promoting cumulus expansion and polar body extrusion, increasing mitochondrial membrane potential and blastocyst rate, and reducing oxidative stress and early apoptosis. Notably, ALD supplementation enhanced the maturation and development of small follicle oocytes by inhibiting the Wnt/β-catenin pathway rather than the mineralocorticoid receptor (MR). Altogether, our findings demonstrate that ALD represents a viable strategy to enhance the quality of oocytes derived from small follicles, thereby providing a scientific basis for optimizing the in vitro maturation system for porcine oocytes.

Serine is crucial for maintaining proper immune system function in both humans and animals. This study aimed to investigate the effects of dietary serine supplementation on microbial interactions and intestinal immunity in weaned piglets. In Exp. 1, a total of 35 weaned piglets (age 21 d, initial body weight [BW] 7.01 ± 0.07 kg, n = 7) were allocated to five dietary treatments for 42 d, a basal diet with no serine and supplemented with 0.25%, 0.50%, 0.75%, or 1.00% serine. The results showed that supplementation with 1.00% serine in the diet increased final BW and significantly elevated serum concentrations of immunoglobulin (Ig) A, IgG, and IgM (P < 0.05), supporting its selection for subsequent experiments. In Exp. 2, a total of 21 weaned piglets (age 21 d, initial BW 7.09 ± 0.09 kg, n = 7) were divided into three groups and fed for 28 d: basal diet (Ctrl), basal diet with D-galactose (D-Gal), and basal diet with 1.00% serine and D-galactose (Ser). D-galactose treatment significantly reduced average daily feed intake and aggravated intestinal morphological damage (P < 0.05), whereas serine supplementation effectively alleviated these effects (P < 0.05). Serine also enhanced systemic humoral immunity by increasing serum IgG and IgM on d 14 (P < 0.001) and serum IgA on d 28 (P = 0.003), and improved intestinal barrier function as evidenced by higher villus height to crypt depth ratios in the jejunum (P = 0.041) and ileum (P = 0.033). Furthermore, serine supplementation significantly downregulated the ileal mRNA expression of autophagy markers (becline, microtubule associated protein 1 light chain 3 beta [Lc3b-1], enhancer of polycomb 2 [Ep2c], and sequestosome 1 [P62]) and reduced phosphorylated- nuclear factor-kappa B (NF-κB) p65 subunit protein levels in the ileum of D-galactose-treated piglets (P < 0.05). It also enhanced microbial diversity and increased Bacteroidetes abundance while lowering Firmicutes abundance and the Firmicutes/Bacteroidetes ratio (P < 0.05). In addition, Spearman correlation analysis showed that ileal secretory immunoglobulin G (SIgG) was positively correlated with Acidobacteria, Chloroflexi, Streptococcus, and Actinobacteria, but negatively correlated with Firmicutes and Lactobacillus (P < 0.05). Collectively, these findings indicated that serine improves growth performance, mitigates chronic oxidative stress, and regulates NF-κB signaling and gut microbiota composition in weaned piglets.

OBJECTIVE:This study was conducted to reveal the role of nuclear poly(A) binding protein 1 (PABPN1) in the proliferation of preadipocytes, and to reveal the relationship between PABPN1 and cAMP response element (CRE)-binding protein (CREB) in the regulation of preadipocyte proliferation. METHODS:Vectors overexpressing and siRNAs against PABPN1/CREB were transiently transfected into both porcine preadipocytes and mouse 3T3-L1 cells. Preadipocyte proliferation was measured with cell counting kit-8, 5-ethynyl-2'-deoxyuridine, real-time quantitative polymerase chain reaction, Western blotting, and flow cytometry analyses. Additionally, the transcriptional regulation of CREB on PABPN1 were analyzed with dual-luciferase reporter gene and electrophoretic mobility shift assay. RESULTS:Overexpression of PABPN1 inhibits, and knockdown of PABPN1 promotes, the proliferation of both porcine preadipocytes and 3T3-L1 cell lines. PABPN1 overexpression increased, while knockdown decreased, the cell population in the G0/G1 phase. These indicates that PABPN1 repressed preadipocyte proliferation by inhibiting cell cycle progress. Additionally, it was revealed that CREB regulated the expression of PABPN1 through binding to the promoter and that CREB inhibited preadipocyte proliferation by repressed cell cycle progress. Furthermore, we showed that PABPN1 functions as a downstream gene of CREB to regulate the proliferation of preadipocytes. CONCLUSION:PABPN1 inhibits preadipocyte proliferation by suppressing the cell cycle. We also found that CREB could promote PABPN1 expression by binding to a motif in the promoter. Further analysis confirmed that PABPN1 functions as a downstream gene of CREB to regulate the proliferation of preadipocytes. These results suggest that the CREB/PABPN1 axis plays a role in the regulation of preadipocyte proliferation, which will contribute to further revealing the mechanism of fat accumulation.

Astaxanthin, a naturally occurring exogenous antioxidant, possesses a unique molecular structure defined by conjugated double bonds and polar end groups, giving it unparalleled antioxidant capacity - exceeding β-carotene and α-tocopherol by orders of magnitude. Notably, astaxanthin surpasses traditional antioxidants not only in radical scavenging potency, but also in multiple metabolic modulating effects. Accumulating evidence confirms that dietary astaxanthin accumulates in intestinal tissues and the systemic circulation, where it orchestrates cellular signaling pathways to restore homeostasis under pathophysiological stress. Despite the well-established broad-spectrum bioactivities of astaxanthin, the multi-target regulatory mechanisms underlying its metabolic modulation remain incompletely elucidated. This review systematically unravels the structure-activity relationships that govern astaxanthin's biological potency, focusing on the mechanisms by which astaxanthin regulates oxidative stress, inflammation, autophagy, apoptosis, ferroptosis and gut microbiota-host interactions via metabolically activated molecular signaling during critical windows of health development in humans and animals. A deeper understanding of astaxanthin's mechanisms may pave the way for improved astaxanthin-based interventions in the future, ultimately advancing health management and prevention strategies.

The effects of furfural and 5-methyl-2-furfural produced by the Maillard reaction on PhIP formation were investigated in chemical models and roasted pork patties. In the chemical models, the results indicated that increasing levels of furfural (r = −0.7338, R2 = 0.9557) and 5-methyl-2-furfural (r = −0.7959, R2 = 0.9864) significantly reduced PhIP formation, displaying a strong linear correlation. The effects of furfural and 5-methyl-2-furfural on the precursors of phenylalanine (Phe) and phenylacetaldehyde showed a significant reduction in the Phe level, while the level of phenylacetaldehyde was not increased. In addition, neither furfural nor 5-methyl-2-furfural could significantly reduce creatinine or PhIP. Further mechanism studies showed that furfural (5-methyl-2-furfural) directly captured Phe to form the corresponding Schiff base compounds a (2-((furan-2-ylmethylene) amino)-3-phenylpropanoic acid) and b (2-(((5-methylfuran-2-yl)methylene)amino)-3-phenylpropanoic acid). This process reduced the production of phenylacetaldehyde, thereby inhibiting the PhIP formation pathway. More importantly, these two compounds were detected in roasted pork patties to which glucose was added. The above pathway was finally confirmed in roasted pork patties. These results revealed that furfural and 5-methyl-2-furfural, formed during the Maillard reaction, play a significant role in inhibiting the formation of PhIP by reacting with Phe.

Aspartate (Asp) metabolism-mediated antioxidant functions have important implications for neonatal growth and intestinal health; however, the antioxidant mechanisms through which Asp regulates the gut microbiota and influences RIP activation remain elusive. This study reports that chronic oxidative stress disrupts gut microbiota and metabolite balance and that such imbalance is intricately tied to the perturbation of Asp metabolism. Under normal conditions, in vivo and in vitro studies reveal that exogenous Asp improves intestinal health by regulating epithelial cell proliferation, nutrient uptake, and apoptosis. During oxidative stress, Asp reduces Megasphaera abundance while increasing Ruminococcaceae. This reversal effect depends on the enhanced production of the antioxidant eicosapentaenoic acid mediated through Asp metabolism and microbiota. Mechanistically, the application of exogenous Asp orchestrates the antioxidant responses in enterocytes via the modulation of the RIP3-MLKL and RIP1-Nrf2-NF-κB pathways to eliminate excessive reactive oxygen species and maintain mitochondrial functionality and cellular survival. These results demonstrate that Asp signaling alleviates oxidative stress by dynamically modulating the gut microbiota and RIP-dependent mitochondrial function, providing a potential therapeutic strategy for oxidative stress disease treatment.

Wild boars exhibit genetic and phenotypic diversity shaped by migrations and local adaptations. Their expansion across Eurasia, especially in Central Asia, remains underexplored. Here, we present newly sequenced whole-genome data of 47 wild boars from Eastern Asia, Central Asia, and Europe, combined with 49 existing genomes, creating a comprehensive dataset of 96 individuals. Our analyses show that Asian wild boars and Southeast Asian Suids split ∼3.6 million years ago (mya), with Central Asian and Southern Chinese ancestors diverging ∼1.8 mya. The split between Central Asian and European-Near East ancestors occurred ∼0.9 mya, followed by a European-Near East divergence ∼0.6 mya. We identify signatures of local adaptation in Central Asian populations, including two positively selected variants in LPIN1, associated with lipid metabolism, and a missense mutation in ALPK2, linked to meat traits. These findings provide insights into wild boar dispersal and adaptation and shed light on domestic pig breeding.

Glutathione (GSH) is a potent antioxidant regulating oxidative stress, but whether exogenous GSH supplementation mitigates stress injury through host-microbiome and liver interactions remains unclear. This study aimed to determine the regulatory mechanism of GSH using in vivo (28-day-old weaned piglets) and in vitro (alpha mouse liver 12 (AML12) cells) stress injury models. Thirty-five healthy weaned piglets (mean body weight (9.52±0.20) kg) were fed diets supplemented with 0.01

After exposure to cold stress, animals enhance the production of beige adipocytes and expedite thermogenesis, leading to improved metabolic health. Although brown adipose tissue in rodents is primarily induced by β3-adrenergic receptor (ADRB3) stimulation, the activation of major β-adrenergic receptors (ADRBs) in pigs has been a topic of debate. To address this, we developed overexpression vectors for ADRB1, ADRB2, and ADRB3 and silenced the expression of these receptors to observe their effects on the adipogenic differentiation stages of porcine preadipocytes. Our investigation revealed that cold stress triggers the transformation of subcutaneous white adipose tissue to beige adipose tissue in pigs by modulating adrenergic receptor levels. Meanwhile, we found that ADRB3 promotes the transformation of white adipocytes into beige adipocytes. Notably, ADRB3 enhances the expression of beige adipose tissue marker genes, consequently influencing cellular respiration and metabolism by regulating lipolysis and mitochondrial expression. Therefore, ADRB3 may serve as a pivotal gene in animal husbandry and contribute to the improvement of cold intolerance in piglets.

We have hypothesized that the TAP2 gene is associated with lipid metabolism. Here, 10 Min-pig tissues were collected to detect the expression of TAP2 in different tissues. We obtained dorsal subcutaneous structural vascular fraction (SVF) cells from the Min-pig’s back adipose tissue and induced SVF cells into mature adipocytes. By overexpression and interference, the effect of TAP2 on fat deposition in Min-pig SVF cells was studied. Recombinant human insulin, dexamethasone, indomethacin, 3-isobutyl-1-methylxanthine, triiodothyronine, and rosiglitazone could successfully induce SVF cells into mature adipocytes, and the induction efficiency was above 50

Lactic acid bacteria are widely regarded as safe alternatives to antibiotics in livestock and poultry farming and have probiotic potential. Ligilactobacillus agilis (L. agilis) is a prominent component of pigeon crop microbiota; however, its function is unknown. In this study, a strain of L. agilis 1003 from pigeon cecum was identified by combining whole genome sequencing and phenotypic analysis, and its safety and probiotic properties were studied. Whole-genome sequencing revealed that the L. agilis 1003 genome length is 2.58 Mb, its average percent guanine-cytosine is 40.43%, and it encodes 1,757 protein-coding genes. Annotation of clusters of orthologous groups classified predicted proteins from the assembled genome as having cellular, metabolic, and information-related functions. A gene cluster associated with the synthesis of a broad-spectrum antimicrobial compound confirmed by antibacterial spectrum testing was identified using genome mining tools. Based on hemolysis test results, the strain was determined to be safe. This strain exhibited a high survival rate in the presence of bile salts and acidic conditions and a significant self-aggregation propensity and hydrophobicity. In vivo animal experiments showed that L. agilis 1003 exhibits probiotic and antibacterial effects and that the substances exerting antibacterial effects are organic acids. Metabolomics analysis revealed that L. agilis 1003 supernatant contained seven organic acids, including butyric acid. L. agilis 1003 showed good safety and probiotic potential in genomics, physiological biochemistry, and animal experiments, and could be considered a suitable candidate for promoting livestock and poultry health.

Probiotics are a kind of living microorganisms, and taking probiotics in moderation is beneficial to the health of the host. Probiotics have the functions of antitoxin, anti-pathogenic bacteria, and diarrhea reduction, which can improve the balance of intestinal flora, promote the digestion and absorption of nutrients, and improve host immunity. Adding probiotics to livestock and poultry diets can supplement intestinal microbial population, improve animal production performance, and restore the immune system function of the host. In this paper, the screening criteria, action mechanism, and application progress of probiotics in intestinal health of livestock and poultry were summarized, and the effects of probiotics on intestinal structure, immune function, and production performance were discussed, which provided a reference for the application of probiotics in livestock and poultry production.

Many efforts have been made to reveal the mechanisms underlying skeletal muscle development because of its importance in animals. However, knowledge on chromatin accessibility, a prerequisite for gene expression, remains limited. Here, dynamic changes in chromatin accessibility were analyzed in the skeletal muscles of Min pigs at the ages of 30, 90, and 210 d using an assay for transposase-accessible chromatin with high-throughput sequencing (ATAC-seq). A total of 16,301 differentially accessible regions (DARs) associated with 7455 genes were identified among three developmental stages. Seven out of eight DARs selected for a functional analysis were found to regulate reporter gene expression significantly (p < 0.05), indicating that DARs are active in gene expression. A total of 2219 differentially expressed genes (DEGs) were identified with RNA sequencing (RNA-seq). Through integrated analyses of ATAC-seq and RNA-seq data, 54 DEG_DAR_genes and 61 transcription factors (TFs) were characterized as critical for muscle development. Among them, Kruppel-like factor 5 (KLF5), targeted to 36 DEG_DAR_genes, was the most important TF. The effects of KLF5 on DEG_DAR_gene expression were then analyzed with molecular biology techniques. KLF5 was found to regulate SLPI (secretory leukocyte proteinase inhibitor) expression by directly binding to the promoter; KLF5 was also involved in APOA1 (apolipoprotein A-I) expression through affecting the regulatory role of DAR located in the intron. These results indicate that the TFs identified were functional. Altogether, the chromatin accessibility region, TFs, and genes important for muscle development in Min pigs were identified. The results provide novel data for further revealing the mechanisms underlying the epigenetic regulation of muscle development.

Research has shown that pigs from different regions exhibit varying responses to cold stimuli. Typically, cold stimuli induce browning of white adipose tissue mediated by adrenaline, promoting non-shivering thermogenesis. However, the molecular mechanisms underlying differential response of pig breeds to norepinephrine are unclear. The aim of this study was to investigate the differences and molecular mechanisms of the effects of norepinephrine (NE) treatment on adipocytes of Min pigs (a cold-resistant pig breed) and Duroc-Landrace-Yorkshire (DLY) pigs. Real time-qPCR, western blot, and immunofluorescence were performed following NE treatment on cell cultures of adipocytes originating from Min pigs (n = 3) and DLY pigs (n = 3) to assess the expressions of adipogenesis markers, beige fat markers, and mitochondrial biogenesis markers. The results showed that NE did not affect browning of adipocytes in DLY pigs, whereas promoted browning of adipocytes in Min pigs. Further, the expression of ADRB1 (Adrenoceptor Beta 1, ADRB1) was higher in subcutaneous adipose tissue and adipocytes of Min pigs than those of DLY pigs. Overexpression of ADRB1 in DLY pig adipocytes enhanced sensitivity to NE, exhibiting decreased adipogenesis markers, upregulated beige fat markers, and increased mitochondrial biogenesis. Conversely, adipocytes treated with ADRB1 antagonist in Min pigs resulted in decreased cellular sensitivity to NE. Further studies revealed differential CpG island methylation in ADRB1 promoter region, with lower methylation levels in Min pigs compared to DLY pigs. In conclusion, differential methylation of the ADRB1 promoter region leads to different ADRB1 expression, resulting in varying responsiveness to NE in adipocytes of two pig breeds. Our results provide new insights for further analysis of the differential cold responsiveness in pig breeds from different regions.

The wide geographical distribution of Eurasian wild boar ( Sus scrofa ) offers a natural experiment to study the thermoregulation. Here, we conducted whole-genome resequencing and chromatin profiling experiments on the local populations from cold regions (northern and northeastern Asia) and warm regions (southeastern Asia and southern China). Using genome-wide scans of four methods, we detected candidate genes underlying cold-adaptation with significant enrichment of pathways related to thermogenesis, fat cell development, and adipose tissue regulation. We also found two enhancer variants under positive selection, an intronic variant of IGF1R (rs341219502) and an exonic variant of BRD4 (rs327139795), which showed the highest differentiation between cold and warm region populations of wild boar and domestic pigs. Moreover, these rare variants were absent in outgroup species and warm-region wild boar but nearly fixed in cold-region populations, suggesting their de novo origins in cold-region populations. The experiments of CUT&Tag chromatin profiling showed that rs341219502 of IGF1R is associated with the gain of three novel transcription factors involving regulatory changes in enhancer function, while rs327139795 of BRD4 could result in the loss of a phosphorylation site due to amino acid alteration. We also found three genes ( SLCO1C1, PDE3A , and TTC28 ) with selection signals in both wild boar and native human populations from Siberia, which suggests convergent molecular adaptation in mammals. Our study shows the adaptive evolution of genomic molecules underlying the remarkable environmental flexibility of wild boar. ### Competing Interest Statement The authors have declared no competing interest.

Cold stress in low-temperature environments can trigger changes in gene expression,but epigenomics regulation of temperature stability in vital tissues,including the fat and diencephalon,is still unclear.Here,we explore the cold-induced changes in epigenomic features in the diencephalon and fat tissues of two cold-resistant Chinese pig breeds,Min and Enshi black(ES)pigs,utilizing H3K27ac CUT&Tag,RNA-seq,and selective signature analysis.Our results show significant alterations in H3K27ac modifications in the diencephalon of Min pigs and the fat of ES pigs after cold exposure.Dramatic changes in H3K27ac modifications in the diencephalon of Min pig are primarily associated with genes involved in energy metabolism and hormone regulation,whereas those in the fat of ES pig are primarily associated with immunity-related genes.Moreover,transcription factors PRDM1 and HSF1,which show evidence of se-lection,are enriched in genomic regions presenting cold-responsive alterations in H3K27ac modification in the Min pig diencephalon and ES pig fat,respectively.Our results indicate the diversity of epigenomic response mechanisms to cold exposure between Min and ES pigs,providing unique epigenetic resources for studies of low-temperature adaptation in large mammals.

The Qinghai–Tibet Plateau (QTP) wild boar is an excellent model for investigating high-altitude adaptation. In this study, we analyzed genome-wide data from 93 wild boars compiled from various studies worldwide, including the QTP, southern and northern regions of China, Europe, Northeast Asia, and Southeast Asia, to explore their phylogenetic patterns and high-altitude adaptation based on genome-wide selection signal analysis and run of homozygosity (ROH) estimation. The findings demonstrate the alignment between the phylogenetic associations among wild boars and their geographical location. An ADMIXTURE analysis indicated a relatively close genetic relationship between QTP and southern Chinese wild boars. Analyses of the fixation index and cross-population extended haplotype homozygosity between populations revealed 295 candidate genes (CDGs) associated with high-altitude adaptation, such as TSC2, TELO2, SLC5A1, and SLC5A4. These CDGs were significantly overrepresented in pathways such as the mammalian target of rapamycin signaling and Fanconi anemia pathways. In addition, 39 ROH islands and numerous selective CDGs (e.g., SLC5A1, SLC5A4, and VCP), which are implicated in glucose metabolism and mitochondrial function, were discovered in QTP wild boars. This study not only assessed the phylogenetic history of QTP wild boars but also advanced our comprehension of the genetic mechanisms underlying the adaptation of wild boars to high altitudes.

The genetic basis of complex traits and phenotypic differentiation remains unclear in pigs. Using nine genomes-seven of which were newly generated, high-quality de novo assembled genomes-and 1081 resequencing genomes, we built a pan-genome and identified 134.24 Mb nonredundant nonreference sequences, 1099 novel protein-coding genes, 187,927 structural variations (SVs) and 30,143,962 single-nucleotide polymorphisms (SNPs). Analysis of selective domestication revealed BRCA1 associated with enhanced adipocyte growth and fat deposition, and ABCA3 linked to an alleviated immune response and reduced lung injury. Integrating 162 transcriptomes and 162 methylomes of skeletal muscle across 27 developmental stages revealed the regulatory mechanism of phenotypic differentiation between Eastern and Western breeds. Artificial selection reshaped local DNA methylation status and imparted regulatory effects on the progression patterns of heterochronic genes such as GHSR and BDH1, particularly during embryonic development. Altogether, our work provides valuable resources for understanding molecular mechanisms behind phenotypic variations and enhancing the genetic improvement programs in pigs.

MicroRNAs play essential roles in biological processes by regulating gene expression at the post-transcriptional level. Our previous studies suggested the role of miR-26a in porcine fat accumulation. Here, through gain- and loss-of-function analyses, we first showed that miR-26a increased the proliferation of porcine preadipocytes by promoting cell division and that miR-26a inhibited the preadipocyte differentiation. Next, acyl-CoA dehydrogenase, medium chain (ACADM) was revealed to promote the proliferation and differentiation of preadipocytes for the first time. Then, it was revealed that miR-26a regulates adipogenesis by directly binding to the 3′ untranslated region of ACADM and the long-chain acyl-Co A synthetase 1 (ACSL1) gene, a previously known regulator of adipogenesis. Finally, RNA-sequencing, performed on preadipocytes overexpressing miR-26a, identified 337 differentially expressed genes in the early stage of adipogenesis; among them, nine genes were characterized as potential targets of miR-26a. The 337 genes were mainly involved in Gene Ontology terms related to cell division, indicating that cell cycle progression was also a major event regulated by miR-26a during adipogenesis. We provide novel data for understanding the molecular mechanisms underlying adipogenesis, which will contribute to controlling fat accumulation in animals.

Min pigs exhibit remarkable cold tolerance, where vitamin B1 synthesis by gut microbiota is crucial for the host's energy metabolism. However, the role of this synthesis in cold adaptation of Min pigs are not yet fully understood. This study utilized 16S rRNA amplicon and metagenomic sequencing to examine seasonal variations in the gut microbiota of Min pigs. Results indicated a significant rise in microbial diversity in winter, with the Bacteroidetes group being the most notably increased. The vitamin B1 biosynthetic pathway was significantly enriched during winter, with six significantly upregulated genes (ThiC, ThiD, ThiE, ThiG, ThiH, and ThiL) showing strong evidence of purifying selection. Among the six vitamin B1 synthesis genes significantly upregulated during winter, the increase was mainly due to a marked elevation in several sequences from specific microbial species. Binding energy analysis revealed that, except for ThiL, the average substrate binding energy of the top 10 sequences with the largest seasonal differences was significantly lower than those of the 10 sequences with the smallest differences. Furthermore, most of these sequences were uniquely prevalent in Min pigs and were not found in the homologous sequences of Duroc pigs. Bacteroidetes and Bacteroidales were identified as the primary contributors to these gene sequences. This research provides valuable insights for developing innovative cold-resistant feed and probiotics.