Mastitis is a common inflammation of mammary glands that has a significantly impact on dairy production and animal health, causing considerable economic burdens worldwide. Elevated reactive oxygen species (ROS) followed by oxidative stress, apoptosis, inflammatory changes and suppressed immunity are considered the key biomarkers observed during mastitis. The Nrf2/KEAP1 signaling pathway plays a critical role in regulating antioxidant responses and cellular defense mechanisms. When activated by bioactive compound treatment, Nrf2 translocates to the nucleus and induces the expression of its target genes to exert antioxidant responses. This reduces pathogen-induced oxidative stress and inflammation by inhibiting NF-kB signaling in the mammary glands, one of the prominent pro-inflammatory signaling pathway. Here, we summarize recent studies to highlight the therapeutic potential of Nrf2/KEAP1 pathway in the prevention and treatment of mastitis. Collectively this review article aims to explore the potential of bioactive compounds in mitigating mastitis by targeting the Nrf2/KEAP1 signaling pathway.

It has been well-established that the number of vertebrae is associated with body size and meat productivity. In current study we utilized a digital radiography (DR) technology to detect the number of thoracolumbar vertebrae in live donkeys. For this purpose, we introduced for the first time a groundbreaking device designed by our team for assessing thoracolumbar vertebrae number traits in equids, employing a sample of 1,000 donkeys sourced from five distinct donkey farms. This assessment incorporates a range of crucial body metrics, including body height, length, and various other measurements. Subsequently, our study determined the number of thoracolumbar vertebrae in 112 donkeys, utilizing the DR system. These findings were further validated through post-mortem evaluations conducted by slaughtering the donkeys. Our findings demonstrated a remarkable resemblance between the thoracolumbar vertebrae numbers visualized through the DR system in live donkeys and those obtained via slaughter verification. In conclusion, this research underscores the accuracy and effectiveness of the DR system for the detection of thoracolumbar vertebrae in live donkeys, which might be helpful for assessing the body size and meat productivity. We also recommended the utilization of DR system for counting thoracolumbar vertebrae in other animals in live state and could be a useful addition to livestock business industry for the prediction of body size and meat productivity efficiency.

IntroductionOxidative stress, resulting from environmental changes, significantly affects female fertility. Developing a mouse model to study oxidative stress lays the groundwork for research into human reproductive health and livestock fertility.Materials and methodsIn this study, we established and evaluated an oxidative stress model by administering hydrogen peroxide (H2O2) to mice. ICR mice of similar age (7–8 weeks old) and average body weight (31.58 ± 1.12 g) were randomly assigned to four groups (A, B, C, and D). Group A served as the control and was injected with a saline solution, while groups B, C, and D received saline solutions containing 0.75%, 1.50%, and 3.0% H2O2, respectively, over one week. We measured the body weights of all mice before and after the experimental period.Results and discussionOur findings showed that the average body weight of mice in groups A and B increased, while groups C and D experienced weight loss. Group C showed a significantly lower average weight gain compared to groups A and B, and group D exhibited an even more pronounced reduction in weight gain. Although group D had a high mortality rate, there was no significant difference in mortality rates among groups B, C, and D. Serum malondialdehyde (MDA) content increased with higher concentrations of H2O2, with a significant difference noted between groups C and A. Catalase (CAT) activity in group B was significantly higher than in group A, while superoxide dismutase (SOD) activity in group C was notably elevated compared to groups A and B. Conversely, glutathione peroxidase (GSH-Px) activity in group C was significantly lower than in both group A and group B. Hematoxylin and eosin (HE) staining revealed changes in ovarian morphology and follicle dynamics. The percentage of atretic follicles in group C was significantly higher than in the control group, and group D had a significantly lower total number of healthy follicles compared to the untreated group. Increased H2O2 content resulted in a reduction of ovary size and an irregular appearance in group D.ConclusionBased on our findings, treatment with 1.50% H2O2 effectively established an oxidative stress model in mice within 1 week. This model serves as a valuable reference for future clinical studies on oxidative stress and reproductive disorders in female animals and humans.

DNA methylation represents a predominant epigenetic modification with broad implications in various biological functions. Its role is particularly significant in the process of collagen deposition, a fundamental aspect of dermal development in donkeys. Despite its critical involvement, the mechanistic insights into how DNA methylation influences collagen deposition in donkey skin remain limited. In this study, we employed whole genome bisulfite sequencing (WGBS) and RNA sequencing (RNA-seq) to investigate the epigenetic landscape and gene expression profiles in the dorsal skin tissues of Dezhou donkeys across three developmental stages: embryonic (YD), juvenile (2-year-old, MD), and mature (8-year-old, OD). Our analysis identified numerous differentially methylated genes that play pivotal roles in skin collagen deposition and overall skin maturation, including but not limited to COL1A1, COL1A2, COL3A1, COL4A1, COL4A2, GLUL, SFRP2, FOSL1, SERPINE1, MMP1, MMP2, MMP9, and MMP13. Notably, we observed an inverse relationship between gene expression and DNA methylation proximal to transcription start sites (TSSs), whereas a direct correlation was detected in regions close to transcription termination sites (TTSs). Detailed bisulfite sequencing analyses of the COL1A1 promoter region revealed a low methylation status during the embryonic stage, correlating with elevated transcriptional activity and gene expression levels. Collectively, our findings elucidate key genetic markers associated with collagen deposition in the skin of Dezhou donkeys, underscoring the significant regulatory role of DNA methylation. This research work contributes to the foundational knowledge necessary for the genetic improvement and selective breeding of Dezhou donkeys, aiming to enhance skin quality attributes.

The primary focus of donkey hide gelatin processing lies in the dermal layer of donkey hide due to its abundant collagen content. However, the molecular mechanism involved in collagen organization and skin development in donkey skin tissue across various developmental stages remains incomplete. The current study aims to investigate the transcriptomic screening of lncRNAs and mRNA associated with skin development and collagen organization across different ages in Dezhou donkeys’ skin. In the pursuit of this objective, we used nine skin tissue samples obtained from Dezhou donkeys at various ages including 8-month fetal stage, followed by 2 and 8 years. RNA-seq analysis was performed for the transcriptomic profiling of differentially expressed genes (DEGs) and lncRNAs associated with skin development in different age groups. Our investigation revealed the presence of 6,582, 6,455, and 405 differentially expressed genes and 654, 789, and 29 differentially expressed LncRNAs within the skin tissues of Dezhou donkeys when comparing young donkeys (YD) vs. middle-aged donkeys (MD), YD vs. old donkeys (OD), and MD vs. OD, respectively. Furthermore, we identified Collagen Type I Alpha 1 Chain (COL1A1), Collagen Type III Alpha 1 Chain (COL3A1), and Collagen Type VI Alpha 5 Chain (COL6A5) as key genes involved in collagen synthesis, with COL1A1 being subject to cis-regulation by several differentially expressed LncRNAs, including ENSEAST00005041187, ENSEAST00005038497, and MSTRG.17248.1, among others. Interestingly, collagen organizational and skin development linked pathways including Protein digestion and absorption, metabolic pathways, Phosphatidylinositol 3-Kinase-Protein Kinase B signaling pathway (PI3K-Akt signaling pathway), Extracellular Matrix-Receptor Interaction (ECM-receptor interaction), and Relaxin signaling were also reported across different age groups in Dezhou donkey skin. These findings enhance our comprehension of the molecular mechanisms underlying Dezhou donkey skin development and collagen biosynthesis and organization, thus furnishing a solid theoretical foundation for future research endeavors in this domain.

BACKGROUND:At present, donkey meat in the market shows an imbalance between supply and demand, and there is an urgent need to cultivate a meat-type Dezhou donkey breed. On the one hand, it can improve the imbalance in the market, and on the other hand, it can promote the rapid development of the donkey industry. This study aimed to reveal significant genetic variation in the NK1 homeobox 2 gene (NKX1-2) of Dezhou donkeys and investigate the association between genotype and body size in Dezhou donkeys. RESULTS:In this study, a SNP (g.54704925 A > G) was identified at the exon4 by high-depth resequencing of the Dezhou donkey NKX1-2 gene. The AA genotype is the dominant genotype. The g.54704925 A > G site was significantly associated with body length, thoracic girth, and hide weight (P < 0.05), while it was highly significantly associated with body height and carcass weight (P < 0.01) in Dezhou donkeys. CONCLUSION:Overall, the results of this study showed that the NKX1-2 gene could be a candidate gene for breeding meat-type Dezhou donkeys, and the g.54704925 A > G locus could be used as a marker locus for selection and breeding.

Background Previous studies have shown that the protein kinase cGMP-dependent 2 (PRKG2) gene is associated with dwarfism in humans, dogo Argentines, and Angus cattle, as well as with height and osteoblastogenesis in humans. Therefore, the PRKG2 gene was used as the target gene to explore whether this gene is associated with several thoracolumbar vertebrae and carcass traits in Dezhou donkeys.Results In this study, fifteen SNPs were identified by targeted sequencing, all of which were located in introns of the PRKG2 gene. Association analysis illustrated that the g.162153251 G > A, g.162156524 C > T, g.162158453 C > T and, g.162163775 T > G were significantly different from carcass weight. g.162166224 G > A, g.162166654 T > A, g.162167165 C > A, g.162167314 A > C and, g.162172653 G > C were significantly associated with the number of thoracic vertebrae. g.162140112 A > G was significantly associated with the number and the length of lumbar vertebrae, and g.162163775 T > G was significantly associated with the total number of thoracolumbar vertebrae.Conclusion Overall, the results of this study suggest that PRKG2 gene polymorphism can be used as a molecular marker to breed high-quality Dezhou donkeys.

The main component of donkey meat is skeletal muscle, and different muscle fibers have been found to be associated with meat quality. However, RNA-seq technology has yet to be used to profile the transcriptomic changes of different muscles of the donkey. In this study, the characterizations of different muscles on the gene expression profiles of Dezhou donkey were obtained, the aim was to identify the important genes in donkey muscles, and aid in improving donkey meat quality via RNA-seq. In the donkey gluteus (DG) and donkey longissimus dorsi (DL) group, GO enrichment indicated that DEGs were mainly involved in the biological regulation and metabolic process, and KEGG analysis shows that a total of 427 DEGs were mapped to 216 KEGG pathways and 23 KEGG pathways were significantly enriched such as the ribosome, glycolysis/gluconeogenesis, glucagon signaling pathway and biosynthesis of amino acids pathways. Meanwhile, 504 DEGs were mapped to 223 KEGG pathways, in which 17 were significantly enriched including cardiac muscle contraction and oxytocin signaling pathway in donkey hamstring muscles (DH) and DL group. In addition, the tenderness in donkey meat might involve muscle fiber type and glucose metabolism, which might profit from the DEGs including MYH1, MYH7, TNNC1, TNNI3, TPM3, ALDOA, ENO3, and PGK1. The genes found in this study will provide some ideas for further understanding the molecular mechanism of donkey meat quality.

随着现代农业的发展,驴的传统役用功能大大降低,逐渐向产皮、产肉、产奶以及休闲娱乐方向转变,驴产品的消费需求也不断扩大.然而,在自然本交条件下,由于驴的妊娠期长、发情持续时间长、受孕率低和产驹率低,驴的繁殖效率表现不高,驴存栏量不断下降.近年来,随着驴的规模化养殖逐渐兴起,驴精液的冷冻保存、人工授精和胚胎移植等高效繁殖技术也逐步应用于驴的生产实践.深入开展驴的繁殖生理特征及繁殖生物学技术的应用研究,对于提高驴的繁殖效率、实现驴的快速扩繁以及遗传改良具有重要意义.本文重点综述了相关驴繁殖生理特征与驴繁殖生物学技术应用的研究进展,为有效提高驴的繁殖效率及研究应用提供参考.

农业农村部2021—2035全国畜禽遗传改良计划指出:畜禽种业是畜牧业发展的根基,是畜牧业核心竞争力的重要体现.在"三品一标"行动方案中,品种培优居于首位.山东省是驴产业发展强省,全国90％的阿胶产自山东,具有阿胶、驴肉、驴奶、化妆品和保健品等完整的产业链条.德州驴是我国五大大型驴品种之一,随着政府支持力度的加强,德州驴的养殖逐渐向规模化、标准化和产业化方向发展,并且发挥了支持医养健康,特色文化振兴的良好作用.驴是单胎动物,生长周期和妊娠期长,而我国地方驴的品种数量和产业规模均位居世界首位,但国内外仍没有一个驴的培育品种出现.因此,加大驴的种质资源保护力度,尽快启动遗传改良工作具有重要意义.

The number of thoracolumbar vertebrae is a quantitative trait positively correlated with the economic traits of livestock. More thoracolumbar vertebrae individuals could genetically be used to improve the livestock population, as more thoracolumbar vertebrae means a longer carcass, which could bring more meat production. Nuclear receptor subfamily 6 group A member 1 (NR6A1) is considered a strong candidate gene for effecting the number of vertebrae in livestock. The purposes of this study are as follows: (a) Analyzing the effect of TLN variation on body size and carcass traits of Dezhou donkey; (b) Studying the distribution of seven single nucleotide variants (SNVs) in NR6A1 gene of Dezhou donkey; (c) Exploring the relationship between latent SNVs and TLN, the body size and carcass traits. We examined the thoracic and lumbar vertebrae number and seven SNVs in NR6A1 gene of 455 Dezhou donkeys, and analyzed the relationships between them. Five types of thoracolumbar combinations (T17L5 (individual with 17 thoracic and five lumbar vertebrae) 2.4%, T18L5 75.8%, T19L5 1.1%, T17L6 11.9%, and T18L6 8.8%) of Dezhou donkeys were detected in this study. For one thoracolumbar vertebra added, the body length of Dezhou donkey increases by 3 cm and the carcass weight increases by 6 kg. Seven SNVs (g.18093100G > T, g.18094587G > T, g.18106043G > T, g.18108764G > T, g.18110615T > G, g.18112000C > T and g.18114954T > G) of the NR6A1 gene were found to have a significant association with the TLN, body size and carcass traits of Dezhou donkey (p < 0.05), respectively. For instance, g.18114954C > T is significantly associated with lumber vertebrae number, the total number of thoracolumbar, and carcass weight, and individuals with TT genotype had significantly larger value than CC genotype (p < 0.05). Using these 7SNVs, 16 different haplotypes were estimated. Compared to Hap3Hap3, individuals homozygous for Hap2Hap2 showed significantly longer length in one thoracic spine (STL), the total thoracic vertebrae and one thoracolumbar spine. Our study will not only extend the understanding of genetic variation in the NR6A1 gene of Dezhou donkey, but also provide useful information for marker assisted selection in donkey breeding program.

Donkey meat has been recognized as a nutritive food for human consumption in China because of its protein, vitamins, and mineral contents. However, the genetic mechanisms governing muscle growth and meat quality in donkeys are poorly understood, particularly in terms of proteomics. Our study was the first to use data-independent acquisition (DIA)-based quantitative proteomics to investigate differentially abundant proteins (DAPs) in donkey muscles. A total of 111 and 127 proteins in the semitendinosus (ST)/longissimus thoracis (LT) and gluteus maximus (GM)/LT muscles were differentially expressed. The most significant potential pathways in meat quality were the GnRH and MAPK pathways between ST and LT and the fat digestion and absorption and regulation of actin cytoskeleton pathways between GM and LT. Several DAPs were closely related to muscle growth and meat quality. These findings could contribute to an improved understanding of the key proteins and processes involved in muscle growth and meat quality in donkey.

In donkeys, the gestation period is a dynamic and precisely coordinated process involving systemic and local alterations. Both the gut microbiota and its link with blood metabolites are thought to play significant roles in maintaining maternal health and supporting fetal development during the gestation period. This study was conducted to evaluate gut microbiota changes and the correlation with plasma metabolites in Dezhou donkeys during the gestation period. The donkeys were divided into the four following groups according to their pregnancy stages: the non-pregnant (NP), early stage of pregnancy (P1), middle stage of pregnancy (P2), and late stage of pregnancy (P3) groups. A total of 24 (n = 6 per group) samples of donkey feces and plasma were collected. The results showed that the diversity (Shannon index) of fecal bacteria significantly increased throughout the gestation period. The phyla Spirochaetota and Fibrobacterota varied significantly according to the stages of pregnancy (p < 0.05). At the genus level, the abundance of Treponema in pregnant donkeys was greater than that in non-pregnant donkeys (p < 0.05), and the genus Streptococcus reached its maximum abundance in the P2 period (p < 0.05). The abundance of Ruminococcaceae_NK4A214_group and norank_f_norank_o_WCHB1-41 linearly increased with the progression of pregnancy (p < 0.05). In addition, the host plasma metabolome was altered significantly during the gestation period. Testolic acid, estradiol-17beta 3-sulfate, equol 7’-o-glucuronide, equol 4’-o-glucuronide, estrone, estrone 3-glucuronide, and estradiol were the most significant differential enriched metabolites, and they increased gradually as gestation progressed. The altered metabolites were mainly enriched in pathways matched to bile secretion, ABC transporters, amino acid metabolism, protein digestion and absorption, mineral absorption, fatty acid degradation, glycerophospholipid metabolism, and steroid hormone biosynthesis. We also found a significant correlation between the shifts in donkey fecal bacteria and changes in the host metabolism. In summary, this study provided systematic data on the fecal bacterial changes and host plasma metabolism of donkeys throughout pregnancy. The results indicated that host–bacteria interactions during the gestation period influence the host metabolism. These interactions benefit the pregnant donkeys by providing a sufficient supply of nutrients and energy for fetal growth and maternal health.

Heat cooking of meat gives it a specific taste and flavor which are favored by many consumers. While the characteristic taste components of chicken, duck, pig, and seafood have been studied, there is a lack of information about the molecular components that give donkey meat its unique taste. Here, the characterization profiles of raw donkey meat (RDM) and cooked donkey meat (CDM) meat by metabonomics and lipidomics. The results showed that a total of 186 metabolites belonging to 8 subclasses were identified in CDM and RDM, including carbohydrates (27.42%), amino acids (17.20%), lipids (13.44%), and nucleotides (9.14%). In total, 37 differential metabolites were identified between CDM and RDM. Among these, maltotriose, L-glutamate, and L-proline might predominantly contribute to the unique umami and sweet taste of donkey meat. Comprehensive biomarker screening detected 9 potential metabolite markers for the discrimination among RDM and CDM, including L-glutamate, gamma-aminobutyric acid, and butane-1, 2, 3, 4-tetrol. Moreover, a total of 992 and 1,022 lipids belonging to 12 subclasses were identified in RDM and CDM, respectively, mainly including triglycerides (TGs) and glycerophospholipids (GPs). Of these lipids, 116 were significantly different between CDM and RDM. The abundances of 61 TGs rich in saturated and monounsaturated fatty acids were retained in CDM, whereas the abundances of 37 GPs rich in polyunsaturated fatty acids were reduced, suggesting that TGs and GPs might be the predominant lipids for binding and generating aroma compounds, respectively. A total of 13 lipids were determined as potential markers for the discrimination among RDM and CDM, including PC(O-16:2/2:0), LPE(22:5/0:0), and PC(P-16:0/2:0). In conclusion, this study provided useful information about the metabolic and lipid profiles of donkey meat which may explain its unique taste and flavor, which could serve as a basis for the development and quality control of donkey meat and its products.

Due to its similarity with human milk, donkey milk is gaining interest as a nutritional milk source. The aim of current study was to compare the difference in the milk composition and metabolites of two Dezhou donkey strains (SanFen and WuTou). The results showed that fat, total solids, and solids-not fat of SanFen milk were higher than WuTou milk. Then we further identified the metabolites in the donkey milk by untargeted metabolomics using LC–MS, and showed that 49 significantly different metabolites between two milk samples were detected. 36 metabolites in the SanFen were significantly higher (p < 0.05), while 13 metabolites were significantly lower than the WuTou (p < 0.05). The results indicated that some different metabolites, such as l-glutamate, and l-glutamine, glucose, and choline, might be ascribed to the genetic differences of the two Dezhou donkey strains. The significantly different metabolites (SDM) were mainly enriched in arginine biosynthesis, d-glutamine and d-glutamate metabolism, alanine, aspartate and glutamate metabolism, and starch and sucrose metabolism. This is the first comparative study performed in SanFen and WuTou and indicates that stains differentially affect the milk composition and metabolites. Furthermore, the SDM and metabolic pathways at least partly explore the regulation mechanisms related to different milk quality and lactation traits between two donkey strains.

The maternal intestinal microbial community changes dramatically during pregnancy and plays an important role in animal growth, metabolism, immunity and reproduction. However, our understanding of microbiota compositional dynamics during the whole pregnancy period in donkey is incomplete. This study was carried out to evaluate gut microbiota alterations as well as the correlation with serum biochemical indices, comparing pregnant donkeys to non-pregnant donkeys. A total of 18 pregnant (including EP, early-stage pregnancy; MP, middle-stage pregnancy and LP, late-stage pregnancy) and six non-pregnant (C as a control) donkey blood samples and rectum contents were collected. The results showed that pregnant donkeys had higher microbial richness than non-pregnant donkeys and that the lowest microbial diversity occurred at the EP period. Moreover, the relative abundances of the families Clostridiaceae and Streptococcaceae were significantly higher in the EP group (p < 0.05) than that in the C and MP groups, while the relative abundances of the families Lachnospiraceae and Rikenellaceae were significantly lower in the EP group (p < 0.05) than that in the C group. The predicted microbial gene functions related to the inflammatory response and apoptosis, such as Staphylococcus aureus infection, the RIG-1-like receptor signaling pathway and apoptosis, were mainly enriched in EP. Furthermore, pregnant donkeys had higher glucose levels than non-pregnant donkeys, especially at EP period. EP donkeys had lower triglyceride, total protein and albumin levels but higher malondialdehyde, interleukin 1β, interleukin 6 and tumor necrosis factor-α levels than those in the C and MP groups. Additionally, there were strong correlations between inflammatory cytokine levels and the relative abundances of genera belonging to the Clostridiaceae and Streptococcaceae families. This is the first comparative study performed in donkeys that indicates that pregnancy status (especially in the early pregnancy period) alters the gut microbiota composition, which was correlated with serum biochemical parameters. These results could provide useful information for improving the reproductive management in Dezhou donkeys.

Dezhou donkey is an excellent local hide, meat and milk breed in Shandong Province. To accelerate the speed of breeding, reproduction and conversation, correlation and multiple regression analysis between body weight (BW) and body size of Dezhou donkey rearing under intensive farms was made by SAS 9.4 software (Statistical Analysis for Windows). A total of 162 donkeys of both gender of age 2 ~ 10 years old were used to investigate the relationships between BW and body dimensional traits (cm) including height of withers, body length; thoracic depth, thoracic girth (TG), thoracic width (TW), circumference of cannon bone (CB), height of rump, rump length (RL) and rump width (RW). The results showed that BW and body measurements have positive and great correlations with R2 value ranged from 0.58 to 0.88 (P <0.01). The R2 values from the single-parameter equations showed that the TG was highly related to BW (0.72, P < 0.01). The stepwise regression equations were applied to obtain the best prediction equations, and the results indicated that the prediction accuracy for BW was improved with the inclusion of more body measurement variables. The "best fit" models were: BW (kg) = 1.88 × TG + 1.27 × BL + 2.55 × TW + 4.61 × CB + 2.18 × RW + 1.78 × RL - 422.8 (R2 = 0.906, P < .01). The predicted BW from the present equations showed the nearest value to the real BW (R2 > 0.94, P < .01). In addition, the equations derived to predict the BW of donkeys in Britain, Morocco were less satisfactory for use with the present Dezhou donkey breed because they overestimated or underestimated the BW due to the different donkey breed.