Jinshun Zhan

Through an integrated multi-omics analysis of rumen microbial communities, muscle transcriptomes, metabolic profiles, and liver metabolic profiles, this study systematically compared high- and low-performing lambs to elucidate their divergent effects on meat quality attributes and growth development. A total of 100 male lambs with similar birth weight (3.07 ± 0.06 kg) were selected within 72 h. All test lambs were synchronized weaning at 45 days of age and uniformly fed the same diet (total mixed ration) in the same pen until 180 days of age, with ad libitum access to food and water throughout this period. Subsequently, the eight lambs with the highest (HADG) and lowest (LADG) average daily gains were slaughtered for performance evaluation and multi-omics analysis. This study found that HADG lambs increased body weight, muscle fiber diameter, eye muscle area, improved amino acid (histidine, arginine, valine, isoleucine, essential amino acid/total amino acid, and essential amino acid/nonessential amino acid), and fatty acid (linoleic acid, behenic acid, and arachidonic acid) composition enhanced rumen enzymes (pepsase, lipase, xylanase, amylase, and carboxymethyl cellulose) and promoted efficient fermentation (p < 0.05). Analysis of microbial populations indicated a notable increase in Prevotella levels within the rumen of HADG lambs. Furthermore, the rumen markers Schwartzia and Streptococcus exhibited significant correlations with differential meat quality traits. Analysis of the muscle transcriptome indicated a significant correlation between the turquoise module and host phenotypes, particularly body weight. Additionally, muscle metabolism is primarily concentrated within the black module; however, it exhibits a significant correlation with the host body phenotype in the yellow module (p < 0.05). Moreover, liver metabolites, rumen microbes, host phenotype, and muscle transcripts were significantly correlated (p < 0.05). In conclusion, the interactions among rumen microbes, muscle, and liver in lambs promote rumen fermentation, which in turn regulate muscle transcriptional activity and modify metabolic profiles in both the liver and muscle. Moreover, PCK1, SPP1, FGF7, NR4A1, DUSP5, GADD45B, etc., can be candidate genes for muscle growth and development. This finding provides a theoretical basis for further exploiting the production potential of Hu lambs.

This study aimed to evaluate the effect of early weaning (EW) on the growth performance, gastrointestinal development, serum parameters, and metabolomics of Hu sheep lambs. Twenty-four male Hu lambs were initially ewe-reared. A total of 12 lambs were weaned at 30 d of age (D30) as the EW group, and the remaining 12 lambs were weaned at 45 d of age (D45) as the control (CON) group. Serum samples were collected from six lambs per treatment on D30, D33, D36, and D45, and the lambs were slaughtered on D45 to collect the rumen and small intestine. The results showed that, compared with the CON group, the average daily gain (ADG), final body weight (p < 0.001), as well as average daily feed intake (ADFI) of lambs in the EW group significantly decreased in the first (p = 0.004) and second (p = 0.013) 5 days of treatment. Additionally, EW increased the ruminal weight and papillae length but reduced the duodenal villus height on D45 (p < 0.05). As for the serum parameters, the concentrations of glucose on D33, D36, and D45 (p < 0.001), and the IL-6 content on D45 (p = 0.018) were observed to be lower, while the levels of immunoglobulin A (IgA) (p = 0.027), IgG (p = 0.035), and IgM (p = 0.002) on the four ages were all higher in the EW group than those in CON group. Additionally, both treatment and age interactively affected the levels of GLU (p = 0.001), TP (p = 0.041), and IL-6 (p = 0.016). Additionally, the serum metabolomics analysis on D45 showed that the contents of 5-HT and arachidonic acid were increased, while L-phenylalanine, L-tyrosine, and L-glutamic acid were reduced in the EW group (p < 0.05). These differential metabolites were enriched in the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways, including inflammatory mediator regulation, protein digestion and absorption, and phenylalanine and tyrosine biosynthesis. The current results identify that EW at D30 decreased the growth performance (ADG and ADFI) of Hu lambs within two weeks post-weaning, which might be associated with impaired duodenal morphology and glucose metabolism. The serum metabolomics analysis revealed that EW altered the concentrations of 5-HT, phenylalanine, tyrosine, and arachidonic acid, which could serve as potential regulatory targets for modulating the health of EW Hu lambs.

The gut–liver axis and its interactions are essential for host physiology. Thus, we examined the jejunal microbiota, fermentation parameters, digestive enzymes, morphology, and liver metabolic profiles in different growth development lambs to investigate the liver–gut axis’s role in their development. One hundred male Hu lambs of similar birth weight and age were raised under the same conditions until they reached 180 days of age. Subsequently, the eight lambs with the highest (HADG) and lowest (LADG) average daily weight gains were slaughtered for index assessment. The study indicates that the body weight, carcass weight, propanoic acid, butyric acid, propanoic acid ratio, butyric acid ratio, and digestive enzymes (beta-glucosidase, microcrystalline cellulase, xylanase, and carboxymethyl cellulase) were significantly higher in HDAG lambs than in LADG lambs (p < 0.05). Additionally, there were no significant differences in the jejunal microbiota’s structure and function among lambs at different growth development stages (p > 0.05). Overall, our analysis revealed that HADG lambs compared to LADG lambs exhibited an up-regulation of metabolites (such as spermine, cholic acid, succinic acid, betaine, etc.) that were positively correlated with the butyric acid ratio, propanoic acid ratio, propanoic acid, xylanase, microcrystalline cellulase, beta-glucosidase, amylase, carboxymethyl cellulase, carcass weight, and body weight, while these metabolites were negatively correlated with the kidney, acetic acid, acetic acid/ propanoic acid, and acetic acid ratio. Furthermore, there was a significant correlation between liver metabolism and jejunal microbiota. This study revealed significant differences in hepatic metabolites and jejunal fermentation among lambs at different growth stages, which may inform targeted regulation strategies to enhance lamb productivity.

This study investigated the impact of tea polyphenols on serum indices, rumen microorganisms, rumen metabolism, and liver metabolism in Hu sheep. Sixty healthy lambs, aged three months and with similar average weights, were chosen and randomly assigned to control (CON), TP400, TP800, and TP1200 groups, each consisting of fifteen lambs. The control group received a basal diet, while the experimental groups were provided with basal diet supplemented with 400 mg/kg, 800 mg/kg, and 1200 mg/kg of tea polyphenols, respectively. Compared with the CON group, the addition of tea polyphenols to the diet significantly increased serum IgA, GSH−Px, and TSOD. In addition, tea polyphenols were able to increase rumen pH but had no significant effect on the rumen NH₃−N, VFA molar content, and the microbial top 10 phylum and genus levels. Moreover, Firmicutes predominated in the network map of the top 80 abundant microorganisms at the genus level, identifying 13 biomarkers at the genus level. In addition, strong correlations were observed between liver and rumen metabolites, particularly between rumen succinic acid and liver alanyl−serine and methylmalonic acid. Furthermore, tea polyphenol additions changed the enrichment of liver and rumen metabolites in the top five KEGG metabolic pathways, but 400−1200 mg/kg additions had no negative impact on the liver and rumen. In summary, TP significantly influences rumen and liver metabolites in Hu sheep, enhancing lamb immunity and antioxidant capacity, with 400 mg/kg being the most effective dosage.

The objective of this experiment was to explore the effects of three-way hybridization on rumen microbes and metabolites in sheep using rumen metagenomics and metabolomics. Healthy Hu and CAH (Charolais × Australian White × Hu) male lambs of similar birth weight and age were selected for short-term fattening after intensive weaning to collect rumen fluid for sequencing. Rumen metagenomics diversity showed that Hu and CAH sheep were significantly segregated at the species, KEGG-enzyme, and CAZy-family levels. Moreover, the CAH significantly increased the ACE and Chao1 indices. Further, correlation analysis of the abundance of the top 80 revealed that the microorganisms were interrelated at the species, KEGG-enzyme, and CAZy-family levels. Overall, the microbiome significantly affected metabolites of the top five pathways, with the strongest correlation found with succinic acid. Meanwhile, species-level microbial markers significantly affected rumen differential metabolites. In addition, rumen microbial markers in Hu sheep were overall positively correlated with down-regulated metabolites and negatively correlated with up-regulated metabolites. In contrast, rumen microbial markers in CAH lambs were overall negatively correlated with down-regulated metabolites and positively correlated with up-regulated metabolites. These results suggest that three-way crossbreeding significantly affects rumen microbial community and metabolite composition, and that significant interactions exist between rumen microbes and metabolites.

This experiment was conducted to investigate the effect of three–way hybrid sheep and Hu sheep on serum indicators, rumen fermentation, rumen enzyme activity, and microorganisms in sheep. Healthy and similar birth weights from three groups (Hu, n = 11; Charolais × Australian White × Hu, CAH, n = 11; Charolais × Dorper × Hu, CDH, n = 11) were selected to be fed by the ewes until 45 days of age. Subsequently, they were weaned intensively and underwent short–term fattening for 3 months along with selected male lambs fed intensively. During this period, they were fed and watered ad libitum. Blood and rumen fluid were collected and analyzed for serum indicators and rumen fluid microorganisms, enzyme activity, and VFA, respectively, at the end of the fattening period. Compared with Hu lamb, the offspring of the three–way hybrid lamb showed significant improvements in body weight, serum lactate dehydrogenase, and creatinine content. However, there was no significant effect on serum immunity and antioxidant indices. In addition, the rumen fluid volatile fatty acid (VFA) molar concentration and microcrystalline cellulose and lipase content were significantly lower in the three–way hybrid lamb compared to Hu lamb, but β–glucosidase, amylase, pepsin, and VFA molar ratio were not significantly affected. Subsequently, 16S rRNA sequencing diversity analysis revealed that three–way hybrid lamb significantly increased rumen microbial ACE and Chao1 indices compared to Hu lamb. Meanwhile, the abundance of Verrucomicrobiota and Synergistota significantly increased at the phylum level. Correlation analysis showed that Prevotella had the highest proportion, while Rikenellaceae_RC9_gut_group correlated most closely with others genus. The microbial communities isovaleric acid molar concentration and proportion were strongly correlated. In addition, there were significant differences in correlations between microbial communities and isobutyric acid, butyric acid and valeric acid content, and their molar proportion, but they were not significantly correlated with digestive enzymes. From the functional enrichment analysis, it was found that hybrid progeny were mainly enriched in the pyruvate metabolism, microbial metabolism in diverse environments, carbon metabolism, and quorum sensing pathways. In contrast, the Hu sheep were primarily enriched in the cysteine and methionine, amino sugar and nucleotide sugar, and biosynthesis of secondary metabolite pathways. These results suggest that hybridization can play a role in regulating organismal metabolism and improve animal production performance by influencing the structure and characteristics of microbial communities.

Objective: Rutin, also called vitamin P, is a flavonoids from plants. Previous studies have indicated that rutin can alleviate the injury of tissues and cells by inhibiting oxidative stress and ameliorating inflammation. There is no report on the protective effects of rutin on goat rumen epithelial cells (GRECs) at present. Hence, we investigated whether rutin can alleviate lipopolysaccharide (LPS)-induced damage in GRECs.Methods: GRECs were cultured in basal medium or basal medium containing 1 μg/mL LPS, or 1 μg/mL LPS and 20 μg/mL rutin. Six replicates were performed for each group. After 3-h culture, the GRECs were harvested to detect the relevant parameters.Results: Rutin significantly enhanced the cell activity (p<0.05) and transepithelial electrical resistance (TEER) (p<0.01) and significantly reduced the apoptosis rate (p<0.05) of LPSinduced GRECs. Rutin significantly increased superoxide dismutase, glutathione peroxidase, and catalase activity (p<0.01) and significantly decreased lactate dehydrogenase activity and reactive oxygen species and malondialdehyde (MDA) levels in LPS-induced GRECs (p<0.01). The mRNA and protein levels of interleukin 6 (IL-6), IL-1β, and C-X-C motif chemokine ligand 8 (CXCL8) and the mRNA level of tumor necrosis factor-α (TNF-α) and chemokine C-C motif ligand 5 (CCL5) were significantly increased in LPS-induced GRECs (p<0.05 or p<0.01), while rutin supplementation significantly decreased the mRNA and protein levels of IL-6, TNF-α, and CXCL8 in LPS-induced GRECs (p<0.05 or p<0.01). The mRNA level of toll-like receptor 2 (TLR2), and the mRNA and protein levels of TLR4 and nuclear factor κB (NF-κB) was significantly improved in LPS-induced GRECs (p<0.05 or p<0.01), whereas rutin supplementation could significantly reduce the mRNA and protein levels of TLR4 (p<0.05 or p<0.01). In addition, rutin had a tendency of decreasing the protein levels of CXCL6, NF-κB, and inhibitor of nuclear factor kappa-B alpha (0.05< p<0.10). Rutin could significantly decreased interferon regulatory factor 3 mRNA expression in LPS-induced GRECs (p<0.05), whereas interferon induced protein with tetratricopeptide repeats 3 (IFIT3) and toll-interacting protein (TOLLIP) mRNA expression was not significantly different between the groups. LPS reduced the tight junction protein zonula occludin 1 (ZO-1) level in GRECs whereas rutin enhanced it. Rutin significantly improved tight junction protein Claudin-1 mRNA expression in LPS-induced GRECs (p<0.01), but could not affect tight junction protein Occludin mRNA expression.Conclusion: Rutin alleviated LPS-induced barrier damage in GRECs by improving oxidation resistance and anti-inflammatory activity, which may be related to TLR/NF-κB signaling pathway inhibition.

Previous studies found that rutin can improve production performance of sheep and dairy cows. However, it is not clear whether rutin has similar effects in goats. Hence, the aim of this experiment was to study the effects of rutin supplementation on growth performance, slaughter performance, serum parameters, and meat quality of Nubian goats. A total of 36 healthy Nubian ewes were randomly divided into three groups. Goats were fed the basal diet supplemented with 0 (R0), 25 (R25), and 50 (R50) mg rutin per kg of diet. The growth performance and slaughter performance of goats had no significant difference among three groups. The meat pH_45min and moisture were significantly higher in the R25 group than the R50 group (p < 0.05), but the color value b* and contents of C14:0, C16:0, C18:0, C18:1n9c, C20:1, saturated fatty acid (SFA), and monounsaturated fatty acid (MSFA) in meat had an opposite outcome. The dressing percentage had an increasing tendency in the R25 group compared with the R0 group (0.05 < p < 0.10), but the shear force, water loss rate and crude protein of meat had opposite results. In conclusion, rutin could not affect the growth performance and slaughter performance of goats; low levels could possibly improve meat quality.