The study aimed to identify the long noncoding RNA (lncRNA) responsible for regulating muscle development in Tan sheep. RNA-seq analysis was conducted on longissimus dorsi samples from 1-day-old and 60-day-old Tan sheep to investigate the molecular processes involved in muscle development. A total of 5517 lncRNAs and 2885 mRNAs were found to be differentially expressed in the 60-day-old Tan sheep. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis revealed that these differentially expressed lncRNAs and mRNAs were linked to pathways crucial for muscle development, such as MAPK, cAMP, and calcium-mediated signaling pathways. Key genes like CDKN1A, MAPK14, TGFB1, MEF2C, MYOD1, and CD53 were identified as significant players in muscle development. The study validated the RNA-seq results through RT-qPCR, confirming the consistency of expression levels of differentially expressed lncRNAs and mRNAs. These findings indicate that lncRNA-mRNA networks produce a remarked effect on modulating muscle development in Tan sheep, such as lncRNAs (MSTRG.12808.1/MSTRG.22662.3/MSTRG.18310.1) and mRNAs (MSTRG.10027/MSTRG.10029/MSTRG.10258/MSTRG.11011/MSTRG.10354), laying the groundwork for future research in this area.

Maternal nutritional status during late gestation and lactation significantly influences the development and physiological metabolism of the offspring. Supplemental rumen-protected methionine (RPM) has the potential to enhance the development and growth of both ewes and their lambs. This study was performed to investigate the effects of maternal RPM supplementation on growth performance, blood metabolites, rumen morphology and fermentation parameters, as well as microbial composition of offspring lambs. Twenty Hu ewes were subjected to simultaneous estrus by inserting sponge containing progesterone into the vagina. At fifty-five days before the expected parturition date, the pregnant ewes (2-3 years of age, 53.7 +/- 2.77 kg of weight) were randomly assigned to two groups: 1) Ewes were fed a basal diet with no RPM supplementation (CON, n = 7), 2) Basal diet supplementation with 0.25 % DM of RPM (MET, n = 5). After lambing, the lambs were fed milk from their mothers and fed starter from d 14 to d 42 of life. The results indicated that MET ewes had higher colostrum fat content than CON ewes. Birth weight (BW), average daily gain (ADG), and weaning weight of lambs showed increase in response to RPM added in the maternal diets (P < 0.05). Similarly, concentrations of insulin-like growth factor-1 (IGF-1) was elevated with maternal RPM addition (P < 0.05). Moreover, GSH-PX showed an increasing trend, while AST showed a decreasing trend in MET lambs. The length, width and total epithelia thickness of rumen papillae were higher in the MET compared with the CON (P < 0.05). In term of proliferative genes expression, MET lambs had greater TJP1 and PCNA expression than in CON (P < 0.05). The ruminal total volatile fatty acid was not affected, whereas iso-butyrate and iso-valeric levels increased with maternal RPM addition (P < 0.05). Moreover, the 16S rRNA sequencing profile showed that relative abundance of Actinobacteria was decreased while Prevotella-1 was increased with RPM supplementation (P < 0.05). These results indicated that maternal RPM supplementation promoted growth performance of the lambs by improving colostrum composition of ewes, and serum parameters, rumen fermentation, and rumen micro- biota of lambs.

BACKGROUNDHeat stress (HS) has been shown to affect reproductive performance and muscle development negatively in animals. N-Acetylcysteine (NAC) plays a pivotal role in enhancing the antioxidant performance in animals as a recognized antioxidant. The present study assesses the potential of NAC to modulate the reproductive performance and antioxidant function in pregnant mice exposed to HS. The role of NAC in muscle development of offspring mice was also explored. RESULTSThe results showed that NAC supplementation from day 12 to day 18 of gestation increased the number of litters and enhanced the antioxidant function in pregnant mice under HS exposure. It improved the weight and body condition significantly in the offspring mice (P < 0.05). The alleviation of HS-induced muscle impairment with NAC was consistent with the alleviation of apoptosis, the enrichment of the proliferation and differentiation in the offspring mice muscle. N-Acetylcysteine also reversed HS-induced reduction in the cross-sectional area of the leg muscle and increased the proportion of myosin heavy chain IIx (MYHCIIx) in the muscle fiber. CONCLUSIONThe results of the present study support the use of NAC at a dose of 100 mg kg-1 body weight as supplement for protecting the offspring derived from pregnant mice exposed to HS from muscle impairment by accelerating proliferation and differentiation. (c) 2024 Society of Chemical Industry.

N-acetylcysteine (NAC) is known for its beneficial effects on health due to its antioxidant and antiapoptotic properties. This study explored the protective effects of NAC against oxidative stress in heat-stressed (HS) skeletal muscle cells and its role in promoting muscle development. NAC reduced the heat shock response by decreasing the expression of heat shock protein 70 (HSP70) in HS-induced muscle cells during proliferation and differentiation. NAC also mitigated HS-induced oxidative stress via increasing the antioxidant enzyme levels and reducing oxidant enzyme levels. Treatment with NAC at 2 mM increased cell viability from 43.68% f 5.14%- 66.69% f 14.43% and decreased the apoptosis rate from 7.89% f 0.53%-5.17% f 0.11% in skeletal muscle cells. Additionally, NAC promoted the proliferation and differentiation of HS-induced skeletal muscle cells by upregulating the expression of PAX7, MYF5, MRF4 and MYHC. These findings suggest that NAC alleviates HSinduced oxidative damage in skeletal muscle cells and support muscle development.

MicroRNAs (miRNAs) are endogenous noncoding RNAs that produce a remarked effect on regulating posttranscriptional gene expression. Our previous study identified a decrease in the expression of oar-miR-411a-5p from umbilical plasma in intrauterine growth restriction (IUGR) Hu lambs subjected to maternal heat stress. In this study, we demonstrated that oar-miR-411a-5p could modulate skeletal muscle development. Overexpression of oar-miR-411a-5p significantly enhanced proliferation and differentiation in heat-stressed Hu sheep myoblasts while suppressing apoptosis. Conversely, inhibition of oar-miR-411a-5p resulted in opposing effects. Subsequently, RNAhybrid analysis revealed targeted sites between oar-miR-411a-5p and the 3 ' untranslated region (UTR) of SMAD2. This suggested that SMAD2 is a direct target gene of oar-miR-411a-5p, as its expression was negatively modulated by oar-miR-411a-5p, a finding corroborated by dual-luciferase assay and RT-qPCR. Furthermore, co-transfection of oar-miR-411a-5p and SMAD2 into Hu sheep myoblasts indicated that oar-miR-411a-5p modulated heat-stressed myoblast growth by targeting SMAD2. In conclusion, these findings elucidate the function of oar-miR-411a-5p in promoting the development of heat-stressed Hu sheep myoblasts, thereby enhancing our understanding of how miRNAs influence skeletal muscle growth in heat-stressed Hu sheep.

During the summer, pregnant ewes experience heat stress, leading to the occurrence of IUGR lambs. This study aims to explore the biomarkers of exosomal miRNAs derived from umbilical plasma in both IUGR and normal Hu lambs. We establish a heat-stressed Hu sheep model during mid-late gestation and selected IUGR and normal lambs for analysis. Exosomes from umbilical plasma were separated and small RNA sequencing is used to identify differentially expressed miRNAs. Next, we utilize MiRanda to predict the target genes of the differentially expressed miRNAs. To further understand the biological significance of these miRNAs, we conduct GO and KEGG pathway enrichment analysis for their target genes. The study’s findings indicate that oar-miR-411a-5p is significantly downregulated in exosomes derived from umbilical plasma of IUGR lambs, while oar-miR-200c is significantly upregulated in the HS-IUGR group ( P < 0.05). Furthermore, GO and KEGG enrichment analysis demonstrate that the target genes are involved in the Wnt, TGF-beta, and Rap1 signaling pathways. miRNAs found in exosomes have the potential to be utilized as biomarkers for both the diagnosis and treatment of IUGR fetuses.

Introduction: Heat stress is harmful to the health of humans and animals, more and more common, as a consequence of global warming, while the mechanism that heat stress modulates skeletal development remains unknown. Hence, we conducted a model of heat stress in vitro.Methods: We used Hu sheep myoblasts as the research object, real-time quantitative PCR (RT-qPCR) and western blot (WB) were conducted to detect the expression of mRNA and protein in heat-stressed myoblasts. The would-healing assay was used to detect the migration of myoblasts. The mitochondria were observed by a transmission electron microscope.Results: mRNA and protein expression of HSP60 was significantly enriched in the heat-stressed myoblasts during proliferation and differentiation (p < 0.05). In our study, we indicated that heat stress enriched the intracellular ROS of the myoblasts (p < 0.001), leading to an increase in autophagy in the myoblasts to induce apoptosis. The results demonstrated that the protein expression of LC3B-1 and BCL-2 was significantly increased in myoblasts under heat stress during proliferation and differentiation (p < 0.05). Additionally, heat stress inhibited mitochondrial biogenesis and function and reduced the mitochondrial membrane potential and downregulated the expression of mtCo2, mtNd1 and DNM1L (p < 0.05) in myoblasts during proliferation and differentiation. Consequently, heat stress inhibited the proliferation and differentiation of the myoblasts, in accordance with the downregulation of the expression of PAX7, MYOD, MYF5, MYOG and MYHC (p < 0.05). Moreover, heat stress also inhibited the cell migration of the myoblasts.Discussion: This work demonstrates that heat stress inhibits proliferation and differentiation, and accelerates apoptosis by impairing mitochondrial function and promoting autophagy, which provides a mechanism to understand heat stress affects the development of the skeletal muscle.

IntroductionThis study aims to investigate the long-term effects of spirulina supplementation in a high-fat diet (HFD) on rumen morphology, rumen fermentation, and the composition of rumen microbiota in lambs. Spirulina is a blue-green microalgae that has been shown to have high nutritional value for livestock. MethodsFifty-four lambs were randomly divided into three groups: a normal chow diet (NCD) group, a high-fat diet (HFD) group, and a high-fat diet supplemented with 3% spirulina (HFD+S) group. Rumen morphology, rumen fermentation, and rumen microbiota were analyzed at the end of the study. ResultsSpirulina supplementation improved the concentration of volatile fatty acids and rumen papilla length. Additionally, there was a tendency for an increase in rumen weight and an upregulation of the genes Claudin-1, Claudin-4, and Occludin in the HFD+S group. Pyrosequencing of the 16S ribosomal RNA gene also showed that spirulina supplementation significantly changed the rumen microbiota composition in the HFD group, with a decrease in richness and diversity. Specifically, the relative abundance of Prevotella 9 and Megasphaera was significantly increased in the HFD group compared to the NCD group, while spirulina supplementation reversed these changes. DiscussionThis study suggests that 3% spirulina supplementation can improve rumen development and fermentation, and effectively relieve rumen microbe disorders in lambs caused by a high-fat diet. However, further research is needed to confirm the findings and to examine the long-term effects of spirulina supplementation in different types of livestock and under different dietary conditions.

AIMS:This trial was performed to investigate the effects of combined feeding of Candida utilis CICC 31170, Bacillus coagulans R11, and Lactobacillus acidophilus NCFM and a multi-enzyme complex on the growth performance, immune parameters, feed digestibility, and rumen microbiota of weaned goats.METHODS AND RESULTS:Thirty weaned goats were randomly divided into CON, PRB, and COB groups and fed different diets. End weight and ADG increased significantly in the PRB and COB groups (P < 0.05), and ADFI increased significantly in COB (P < 0.05). On day 80, there was a significant increase in IL-10 content in PRB and COB compared to the CON (P < 0.05). Highly significant increases in rumen papilla width, epithelial cell thickness, stratum spinosum+basale thickness, and stratum corneum thickness were found in PRB and COB (P < 0.05). COB group significantly increased the gene expression of HMGCL and MCT1 in rumen epithelium (P < 0.001). The COB group had the tendency to increase the feed digestibility of dry matter and crude fat compared with the CON group (P < 0.10). The abundance of Prevotellaceae_unclassified was significantly higher in PRB (P < 0.05), and the abundance of Fibrobacteres was significantly higher in COB in comparison to those in CON (P < 0.05).CONCLUSIONS:The results indicate that the dietary potential probiotics and enzymes complex could modulate the growth performance, immunity, feed digestibility, and rumen microbiota in weaned goats.

It has been reported that N6-methyladenosine (m6A) methyltransferase-like 3 (METTL3) plays an important role in zygote genome activation during embryonic development, but the effects of METTL3 under oxidative stress in the early development of goat embryos remain largely unknown. In this study, zygotes were monitored at 72 and 168 h after fertilization, and they developed to the 8-cell stage and blastocyst stage under hypoxic conditions and normoxic conditions. Single-cell transcriptome sequencing was performed at the 8-cell stage and the blastocyst stage in the goat embryos, the differentially expressed METTL3 was screened from the sequencing results. We found that microinjection of small interfering RNA (siRNA) against METTL3 caused developmental arrest, both 8-cell rates (37.45 ± 2.21% vs. 47.09 ± 1.38%; P < 0.01) and blastocyst rates of Si-METTL3 (12.17% ± 2.84 vs. 20.83 ± 3.61%; P < 0.01) in Si-METTL3 group were significantly decreased compared with that of control under hypoxic conditions, significant changes were found in the m6A-related genes and the expression levels of critical transcription factors, such as, NANOG, GATA3, CDX2 and SOX17, were decreased. This study revealed the key role of METTL3 in the regulation of embryonic development under oxidative stress, and laid the foundation for further study of the crucial mechanism of oxidative stress during the early embryonic development of goats.

Aerobic composting is currently the effective recycling way to use sheep manure. This research aims at exploring the effects of the excipients and microbial agents on the maturity of sheep manure aerobic composting. We used sheep manure, rice straw and fungus bran as raw materials, added exogenous inoculants,measured the physical and chemical indicators, biological indicators and heavy metal indicators in each treatment. The average temperature was Z4>Z3>Z2>Z1, the highest temperature was 68.1℃, 69.1℃, 69.3℃and 71.1℃, respectively. pH increased first and then decreased. At the end of composting, pH was 8.53, 8.25,8.43 and 8.35, respectively. C/N decreased gradually and was 15, 16, 15 and 14 respectively at the end of composting. The moisture content gradually decreased, and the moisture content of Z2 was the lowest among the four treatments at the end of composting, furthermore the loss rate was the largest which was 66.82%. The total potassium content gradually increased, and the total potassium content of Z4 increased by 196.58%. GI gradually increased, reaching 110% on day 50, Z4 reached it first. The content of NH4+-N decreased with fluctuation. The heavy metal content did not exceed the national standards. The effect of composting sheep manure with rice straw was better than that with fungus bran, and the combination of mixed agents and fungus bran treatment entered the thermophilic stage in advance, reached the maturity rapidly in the sheep manure composting.

本试验旨在研究枯草芽孢杆菌与地衣芽孢杆菌单独饲喂与联合饲喂对断奶山羊生长性能、腹泻率、血清生化指标以及粪便菌群的影响.选取60只3月龄海门山羊,随机分为4组,每组3个重复,每个重复5只羊.对照组(C组)饲喂基础饲粮;枯草芽孢杆菌组(BS组)饲喂基础饲粮+400 mg/kg的枯草芽孢杆菌(有效活菌数为1×1011 CFU/g);地衣芽孢杆菌组(BL组)饲喂基础饲粮+400 mg/kg的地衣芽孢杆菌(有效活菌数为1×1011 CFU/g);联合饲喂组(BS_BL组)饲喂基础饲粮+400 mg/kg的枯草芽孢杆菌+400 mg/kg的地衣芽孢杆菌(有效活菌数为1×1011 CFU/g).预试期10 d,正试期30 d.结果表明:1)试验第10天,BS_BL组断奶山羊的体重极显著高于C组、BS组与BL组(P<0.01);试验第30天,C组、BS组、BL组与BS_BL组的断奶山羊的体重无显著差异(P>0.05);试验第1～10天,BS_BL组的平均日增重(ADG)极显著高于C组、BS组与BL组(P<0.01);试验第21～30天,BL组与BS_BL组的ADG极显著高于C组与BS组(P<0.01);试验第1～30天,试验组与C组的ADG无显著差异(P>0.05);试验第21～30天,BS_BL组的平均日采食量(ADFI)极显著高于C组、BS组与BL组(P<0.01);试验第1～30天,BS_BL组的ADFI显著高于C组、BS组与BL组(P<0.05);试验第1～30天,试验组与C组的料重比(F/G)差异不显著(P>0.05).2)试验第1～10天、第11～20天,BS组、BL组与BS_BL组的腹泻率显著低于C组(P<0.05);第1～30天,BS_BL组的腹泻率显著低于C组(P<0.05).3)试验第30天,BS组的血清葡萄糖(GLU)含量、碱性磷酸酶(ALP)和乳酸脱氢酶(LDH)活性显著低于C组(P<0.05),血清谷草转氨酶(AST)活性和白蛋白(ALB)含量极显著低于C组(P<0.01);BL组血清尿素氮(UN)含量显著高于C组(P<0.05),BS_BL组血清AST活性和GLU含量显著低于C组(P<0.05).4)粪便进行16S rDNA测序发现,Chao1和ob-served-species指数显示BS_BL组所含物种数目较多,Shannon和Simpson指数显示BS_BL组多样性高.C组细菌的优势菌门为变形菌门、拟杆菌门、放线菌门;BS组细菌的优势菌门为放线菌门、变形菌门、拟杆菌门;BL组细菌的优势菌门为拟杆菌门、变形菌门、浮霉菌门;BS_BL组细菌的优势菌门为拟杆菌门、变形菌门、厚壁菌门.C组细菌的优势菌属为堆囊菌属、土地杆菌属、球孢菌属;BS组细菌的优势菌属为野野村菌属、嗜红杆菌属、堆囊菌属;BL组细菌的优势菌属为副足杆菌属、野野村菌属、扁平菌属;BS_BL组细菌的优势菌属为Marinilabiliaceae_unclassi-fied、假单胞菌属、Clostridiales_vadinBB60_group_unclassified.综上所述,枯草芽孢杆菌与地衣芽孢杆菌联合饲喂能显著提高断奶山羊的生长性能,降低腹泻率,改善血清生化指标,改变粪便菌群的多样性以及丰富度.

Carbon is the crucial source of energy during aerobic composting. There are few studies that explore carbon preservation by inoculation with microbial agents during goat manure composting. Hence, this study inoculated three proportions of microbial agents to investigate the preservation of carbon during goat manure composting. The microbial inoculums were composed of Bacillus subtilis, Bacillus licheniformis, Trichoderma viride, Aspergillus niger, and yeast, and the proportions were B1 treatment (1:1:1:1:2), B2 treatment (2:2:1:1:2), and B3 treatment (3:3:1:1:2). The results showed that the contents of total organic carbon were enriched by 12.21%, 4.87%, and 1.90% in B1 treatment, B2 treatment, and B3 treatment, respectively. The total organic carbon contents of B1 treatment, B2 treatment, and B3 treatment were 402.00 ± 2.65, 366.33 ± 1.53, and 378.33 ± 2.08 g/kg, respectively. B1 treatment significantly increased the content of total organic carbon compared with the other two treatments (p < 0.05). Moreover, the ratio of 1:1:1:1:2 significantly reduced the moisture content, pH value, EC value, hemicellulose, and lignin contents (p < 0.05), and significantly increased the GI value and the content of humic acid carbon (p < 0.05). Consequently, the preservation of carbon might be a result not only of the enrichment of the humic acid carbon and the decomposition of hemicellulose and lignin, but also the increased OTU amount and Lactobacillus abundance. This result provided a ratio of microbial agents to preserve the carbon during goat manure aerobic composting.

N6-methyladenosine (m6A) modification plays a critical role in mammalian development. However, the role of m6A in the skeletal muscle development remains largely unknown. Here, we report a global m6A modification pattern of goat skeletal muscle at two key development stages and identified that the m6A modification regulated the expression of the growth arrest and DNA damage-inducible 45B (GADD45B) gene, which is involved in myogenic differentiation. We showed that GADD45B expression increased during myoblast differentiation, whereas the downregulation of GADD45B inhibits myogenic differentiation and mitochondrial biogenesis. Moreover, the expression of GADD45B regulates the expression of myogenic regulatory factors and peroxisome proliferator-activated receptor gamma coactivator 1 alpha by activating the p38 mitogen-activated protein kinase (MAPK) pathway. Conversely, the inactivation of p38 MAPK abolished the GADD45B-mediated myogenic differentiation. Furthermore, we found that the knockdown of fat mass and obesity-associated protein (FTO) increases GADD45B m6A modification and decreases the stability of GADD45B mRNA, which impairs myogenic differentiation. Our results indicate that the FTO-mediated m6A modification in GADD45B mRNA drives skeletal muscle differentiation by activating the p38 MAPK pathway, which provides a molecular mechanism for the regulation of myogenesis via RNA methylation.

Background: The aim of this study was to explore the effects of growth performance, slaughter performance, serum biochemical, immune and antioxidant indexes and rumen microflora fed with a basal diet (CON group), added B. subtilis and B. licheniformis (PRO group), supplemented with B. subtilis, B. licheniformis and enzyme preparations (COM group) on fattening goats.Methods: 39 male goats were randomly divided into 4 groups with 13 individuals in each group for feed period of 80 d. Goats were fed as follows: CON diet, PRO diet with B. subtilis and B. licheniformis, and COM diet with B. subtilis, B. licheniformis and compound enzymes.Results: ADFI of COM group significantly increased compared with CON group and PRO group (P < 0.01), as well as COM group and PRO group dramatically promoted ADG versus with CON group (P < 0.05). As a consequence, the body weight of fattening goats in the COM group was predominantly higher than that in the CON group (P < 0.01). In addition, the PRO group and COM group enhanced the TNF-α (P < 0.05) and IL-10 content (P < 0.01) in the serum. No differences were observed in serum biochemical and antioxidant indexes of three groups (P > 0.05). Likewise, the GR values of PRO group and COM group were noteworthy improved in comparison with CON group (P < 0.01). The VFA contents in rumen fluid were insignificantly different (P > 0.05). COM group also enriched the relative abundance of Proteobacteria compared with CON group and PRO group (P < 0.05). Nevertheless, the relative abundance of Actinobacteria decreased of PRO group and COM group in rumen fluid microorganisms (P < 0.05). Apparently, COM group significantly enriched nitrogen metabolism, glycolysis and TCA cycle (P < 0.05), whereas nucleotides biosynthesis was notably reduced (P < 0.05).Conclusion: The combined feed of probiotics and enzymes had more profound effects than probiotics feed. Consequently, supplementation with B. subtilis and B. licheniformis and enzymes in the basal diet of fattening goats, which could improve growth performance, slaughter performance, immunity and accommodate rumen microbiota.