It takes several hours for mammalian sperm to migrate from the ejaculation or insemination site to the fertilization site in the female reproductive tract in which glucose, amino acids, and fatty acids are regarded as the primary substrates for ATP generation. The present study was designed to investigate whether oleic acid and palmitic acid were beneficial to boar sperm in vitro; and if yes, to elucidate the mechanism that regulates sperm motility. Therefore, the levels of oleic acid and palmitic acid, motility, membrane integrity, acrosome integrity, and apoptosis of sperm were evaluated. Moreover, the enzymes involved in mitochondrial β-oxidation (CPT1: carnitine palmitoyltransferase 1; ACADVL: long-chain acyl-coenzyme A dehydrogenase) were detected with immunofluorescence and Western blotting. Consequently, the ATP content and the activities of CPT1, ACADVL, malate dehydrogenase (MDH), and succinate dehydrogenase (SDH) were also measured. We observed that CPT1 and ACADVL were expressed in boar sperm and localized in the midpiece. The levels of oleic acid and palmitic acid were decreased during storage at 17 °C. The addition of oleic acid and palmitic acid significantly increased sperm motility, progressive motility, straight-line velocity (VSL), membrane integrity, and acrosome integrity with a simultaneous decrease in sperm apoptosis after seven days during storage. When sperm were incubated with oleic acid and palmitic acid at 37 °C for 3 h, the activities of CPT1 and ACADVL, the ATP level, the mitochondrial membrane potential, the activities of MDH and SDH, as well as sperm motility patterns were significantly increased compared to the control (p < 0.05). Moreover, the addition of etomoxir to the diluted medium in the presence of either oleic acid or palmitic acid and the positive effects of oleic acid and palmitic acid were counteracted. Together, these data suggest that boar sperm might utilize oleic acid and palmitic acid as energy substrates for ATP production via β-oxidation. The addition of these acids could improve sperm quality.

Sperm are susceptible to excessive reactive oxygen species (ROS). Spermine and spermidine are secreted in large amounts by the prostate and potent natural free radical scavengers and protect cells against redox disorder. Thus, we used boar sperm as a model to study the polyamines uptake and elucidate whether polyamines protected sperm from ROS stress. Seven mature and fertile Duroc boars (aged 15–30 months) were used in this study. In Exp.1, spermine and spermidine (3.6 ± 0.3 mmol/L and 3.3 ± 0.2 mmol/L, respectively) were abundant in seminal plasma, and the content of polyamine decreased (P < 0.05) after preservation at 17 °C for 7 d or incubation at 37 °C for 6 h. In Exp.2, using labeling of spermine or spermidine by conjugation with fluorescein isothiocyanate (FITC) and Ultra-High-Performance Liquid Chromatography (UHPLC), we found that the accumulation of spermine or spermidine in sperm was inhibited by quinidine and DL-Tetrahydropalmatine (THP, organic cation transporters(OCT) inhibitors, P < 0.05), but not mildronate and L-carnitine (organic cation/carnitine transporter (OCTN) inhibitors, P > 0.05). In Exp.3, the addition of spermine or spermidine (0.5 mmol/L) in extender resulted in higher motility, plasma membrane and acrosome integrity and lower ROS level after preservation in vitro at 17 °C for 7 d (P < 0.05). In Exp.4, in the condition of oxidative stress (treatment with H₂O₂ at 37 °C for 2 h), addition of spermine (1 mmol/L) or spermidine (0.5 mmol/L) in extender increased activities of glutathione peroxidase, glutathione reductase, and glutathione S-transferase, and reduced glutathione/oxidized glutathione ratio (P < 0.05), and alleviate oxidative stress-induced lipid peroxidation (LPO), DNA damage, mitochondrial membrane potential (ΔΨ_m) decline, adenosine triphosphate (ATP) depletion, and intracellular calcium concentration ([Ca ²⁺]_i) overload (P < 0.05), thereby improving boar sperm motility, the integrity of plasma membrane and acrosome (P < 0.05) in vitro. These data suggest that spermine and spermidine alleviate oxidative stress via the antioxidant capacity, thereby improving the efficacy of boar semen preservation.

Inosine monophosphate (IMP) is a key factor affecting the fleshy flavor of meat; meanwhile, the free-range mode is an efficient strategy to improve muscular IMP content. To assess expression differences in IMP metabolism-related genes under different feeding patterns, Illumina Nextseq 500 sequencing was used to catalog the global gene expression profiles of muscle samples from Lueyang black-bone chicken under free-range and caging conditions. A total of 15510 unigenes were assembled, with 13423 (86.54%) and 6088 (39.25%) unigenes correctly annotated in the GO and KOG databases, respectively. Next, the "purine metabolism" pathway in the "nucleotide metabolism group" was assessed in depth. Through Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis, we retrieved 172 nucleotide- and 5 purine- metabolism related genes that were differentially expressed in muscle samples from free-range and caged chickens. At 60-day-old, AMPD1, NT5C1A and ENTPD8 showed higher levels in the free-range group, while only ENTPD8 was upregulated in 120-day-old chickens. In addition, GART, GARS and ADSL in free-range chickens showed higher levels compared with caged animals. Furthermore, IMPDH levels in free-range chicken were lower than those of caged chicken. Real-time quantitative polymerase chain reaction (qPCR) was used to validate the above findings. These results revealed a set of differentially expressed genes potentially related to IMP metabolism in chicken under different breeding modes, providing novel insights into controlling IMP levels in chicken meat.

Vitamin D and vitamin D receptor (VD/VDR) plays a vital role in the development of spermatozoa, which is largely determined by the testosterone level in serum. Testosterone biosynthesis is closely related to lipid metabolism in gonadal adipose around testes. VDR could regulate lipid metabolism in adipocytes as well. However, it still remains unknown how VDR regulates lipid metabolism to impact testosterone biosynthesis in testis. Hereby, various parameters of male fertility were compared between wildtype (WT) and Vdr knockout (Vdr-KO) male mouse. For Vdr-KO mice, the size of testis and gonadal adipose was smaller than that of WT, and the sperm quality and testosterone level were lower than WT. Subsequently, testis proteome data between Vdr-KO and WT mice indicated that dysregulation of lipid metabolism was closely associated with decreased testosterone biosynthesis in Vdr-deficient mouse. And further evaluation of VDR functions in Leydig cells verified that VDR impacted lipid metabolism and regulated the expression of a range of genes involved in testosterone biosynthesis. Knockdown VDR could significantly decrease testosterone synthesis and secretion in Leydig cells. Meanwhile, expression of genes involved in androgen synthesis was decreased but genes related to lipolysis were up-regulated. Collectively, the present study unveiled the relationship between lipid metabolism and testosterone biosynthesis mediated by VDR in mouse testis and its effect on male fertility. These findings will greatly enhance our current understanding of VDR intermediate in lipid metabolism and androgen synthesis.

This study aimed to understand the genetic and metabolic traits of a Lactiplantibacillus plantarum JS21 strain and its probiotic abilities through laboratory tests and computer analysis. L. plantarum JS21 was isolated from a traditional fermented food known as “Jiangshui” in Hanzhong city. In this research, the complete genetic makeup of JS21 was determined using Illumina and PacBio technologies. The JS21 genome consisted of a 3.423 Mb circular chromosome and five plasmids. It was found to contain 3023 protein-coding genes, 16 tRNA genes, 64 rRNA operons, 40 non-coding RNA genes, 264 pseudogenes, and six CRISPR array regions. The GC content of the genome was 44.53%. Additionally, the genome harbored three complete prophages. The evolutionary relationship and the genome collinearity of JS21 were compared with other L. plantarum strains. The resistance genes identified in JS21 were inherent. Enzyme genes involved in the Embden–Meyerhof–Parnas (EMP) and phosphoketolase (PK) pathways were detected, indicating potential for facultative heterofermentative pathways. JS21 possessed bacteriocins plnE/plnF genes and genes for polyketide and terpenoid assembly, possibly contributing to its antibacterial properties against Escherichia coli (ATCC 25922), Escherichia coli (K88), Staphylococcus aureus (CMCC 26003), and Listeria monocytogenes (CICC 21635). Furthermore, JS21 carried genes for Na⁺/H⁺ antiporters, F₀F₁ ATPase, and other stress resistance genes, which may account for its ability to withstand simulated conditions of the human gastrointestinal tract in vitro. The high hydrophobicity of its cell surface suggested the potential for intestinal colonization. Overall, L. plantarum JS21 exhibited probiotic traits as evidenced by laboratory experiments and computational analysis, suggesting its suitability as a dietary supplement.

In order to protect the genetic resource of native horse breeds, the genetic diversity of mitochondrial DNA (mtDNA) D-loop of three native horse breeds in western China were investigated. Forty-three 600 bp mtDNA D-loop sequences were analyzed by PCR and sequencing techniques, 33 unique haplotypes with 70 polymorphic sites were detected in these horses, which account for 11.67% of 600 bp sequence analyzed, showing the abundant genetic diversity of the three native horse breeds in western China. The Neighbour-Joining (NJ) phylogenetic tree based on 247 bp of 43 D-loop sequences demonstrated the presence of seven major lineages (A to G), indicating that the three native horse breeds in western China originated from multiple maternal origins. Consistent with the front, the NJ phylogenetic tree based on 600 bp of mtDNA D-loop sequences of 43 Chinese western native horses and 81 sequences of six horse breeds from GenBank indicated that the three horse breeds had distributed into the seven major lineages (A to G). The structure of the phylogenic tree is often blurred because the variation in a short segment of the mitochondrial genome is often accompanied by high levels of recurrent mutations. Consequently, longer D-loop sequences are helpful in achieving a higher level of molecular resolution in horses.