Metagenomics and metabolomics reveal that gut microbiome adapts to the diet transition in Hyla rabbits.

There is still a lack of longitudinal dynamic studies on the taxonomic features, functional reserves, and metabolites of the rabbit gut microbiome. An experiment was conducted to characterize the bacterial community of rabbits. By combining metagenomics and metabolomics, we have comprehensively analyzed the longitudinal dynamics of the rabbit gut microbiota and its effect on host adaptability. Our data reveal an overall increasing trend in microbial community and functional gene diversity and richness during the pre-harvest lifespan of rabbits. The introduction of solid feed is an important driving factor affecting rabbit gut microbiological compositions. Clostridium and Ruminococcus had significantly higher relative abundances in the solid feed stage. Further, the starch and fiber in solid feed promote the secretion of carbohydrate-degrading enzymes, which helps the host adapt to dietary changes. The rabbit gut microbiota can synthesize lysine, and the synthase is gradually enriched during the diet transformation. The gut microbiota of newborn rabbits has a higher abundance of lipid metabolism, which helps the host obtain more energy from breast milk lipids. The rabbit gut microbiota can also synthesize a variety of secondary bile acids after the introduction of solid feed. These findings provide a novel understanding of how the gut microbiota mediates adaptability to environment and diet in rabbits and provide multiple potential strategies for regulating intestinal health and promoting higher feed efficiency.

Vitamin A regulates dermal papilla cell proliferation and apoptosis under heat stress via IGF1 and Wnt10b signaling.

Heat stress (HS) negatively affects the development of hair follicles. The present study investigated the effect of vitamin A (VA) on the development of rabbit dermal papilla cells (DPCs) under HS and the underlying regulatory mechanisms. Addition of 0.4 mg/L VA to the culture medium significantly enhanced cell proliferation (P < 0.001) and inhibited the apoptosis of DPCs (P < 0.01). VA decreased the proportion of DPCs in G0/G1 stage of the cell cycle under HS along with the expression of caspase 3, heat shock protein 70 (HSP70), and microRNA 195 (miR-195) (P < 0.05). VA also activated the insulin-like growth factor 1 (IGF1) and Wnt10b/ß-catenin signaling pathways. The results of the dual luciferase reporter assay showed that IGF1 expression was modulated by miR-195-5p. Over-expression of miR-195-5p in DPCs with HS+VA treatment significantly reduced cell viability and IGF1 signaling (P < 0.01) and increased apoptosis (P < 0.01) compared with the HS+VA group. The positive effects of VA on proliferation and apoptosis of DPCs under HS were significantly attenu-ated by blocking Wnt10b and ß-catenin signaling with IWP-2 and XAV-939, respectively. These results demonstrate that VA can promote hair follicle development following HS via modulation of miR-195/IGF1 and Wnt10b/ß-catenin signaling pathways.

Combined Omics Analysis Further Unveils the Specific Role of Butyrate in Promoting Growth in Early-Weaning Animals.

Abnormal mutations in the microbial structure of early-weaning mammals are an important cause of enteritis. Based on the multiple known beneficial functions of butyrate, we hypothesized that butyrate would alleviate the imbalance of intestinal homeostasis induced by early weaning in animals. However, the mechanisms of action between butyrate and intestinal microbes are still poorly explored. In this study, we aimed to investigate whether butyrate exerts beneficial effects on the structure of the intestinal flora of weanling rabbits and their intestinal homeostasis, growth and development, and we attempted to elucidate the potential mechanisms of action through a combined omics analysis. We found that dietary butyrate upregulated the transcription of tight junction-related proteins in the epithelial barrier and improved the intestinal microbial structure by suppressing harmful bacteria and promoting beneficial ones. Intestinal and plasma metabolomes were also altered. The bile acid secretion, α-linolenic acid, apoptotic, and prostate cancer pathways responded to the positive dietary butyrate-induced metabolic changes in the weanling rabbits, resulting in the inhibition of inflammation, improved antioxidant capacity, increased rates of cell proliferation and survival, and decreased levels of apoptosis. Additionally, dietary butyrate suppressed the release of pro-inflammatory factors and enhanced positive appetite regulation, which increased the average daily gain of the rabbits. These results demonstrated that dietary butyrate can help maintain the integrity of the intestinal epithelial barrier, improve the structural composition of the intestinal microflora, enhance organismal metabolism, inhibit inflammation, reduce post-weaning anorexia, and promote growth and development in early-weaning rabbits. These positive effects of dietary butyrate were exerted via the modulation of the microbe-gut-brain axis.

Effects of the SLC38A2-mTOR Pathway Involved in Regulating the Different Compositions of Dietary Essential Amino Acids-Lysine and Methionine on Growth and Muscle Quality in Rabbits.

In recent years, ensuring food security has been an important challenge for the world. It is important to make good use of China's domestic local feed resources to provide safe, stable, efficient, and high-quality rabbit meat products for China and the world. Lysine and methionine are the two most limiting essential amino acids in the rabbit diet. However, little is known about the rational composition of lysine and methionine in rabbit diets and the mechanisms that affect growth and development. Accordingly, in this study, we sought to address this knowledge gap by examining the effects of different compositions of lysine and methionine in rabbit diets. Subsequently, the growth status, nitrogen metabolism, blood biochemical indexes, muscle development, muscle quality, and the growth of satellite cells were evaluated in the animals. The results showed that diets containing 0.80% Lys and 0.40% Met improved average daily weight gain, feed conversion, nitrogen use efficiency, and muscle quality in the rabbits (p < 0.05). Additionally, it altered the amino acid transport potential in muscle by upregulating the expression of the SLC7A10 gene (p < 0.05). Meanwhile, the cell viability and the rate of division and migration of SCs in the 0.80% Lys/0.40 % Met composition group were increased (p < 0.05). SLC38A2 and P−mTOR protein expression was upregulated in the 0.80% lysine/0.40% methionine composition group (p < 0.05). In conclusion, 0.80% Lys/0.40% Met was the most suitable lysine and methionine composition in all tested diets. SLC38A2 acted as an amino acid sensor upstream of mTOR and was involved in the 0.80% Lys/0.40% Met regulation of muscle growth and development, thus implicating the mTOR signaling pathway in these processes.

Copper Depletion Strongly Enhances Ferroptosis via Mitochondrial Perturbation and Reduction in Antioxidative Mechanisms.

Copper serves as a co-factor for a host of metalloenzymes, particularly cytochrome c oxidase (COX). Although it is known that impaired COX function can lead to the excessive accumulation of reactive oxygen species (ROS), the mechanisms underlying how copper depletion leads to cell damage are poorly understood. Here, we have investigated the role of copper depletion during ferroptosis. The bathocuproinedisulfonic (BCS) treatment depolarized the mitochondrial membrane potential, increased the total cellular ROS levels, stimulated oxidative stress, and reduced the glutathione levels. Moreover, the depletion of copper limited the protein expression of glutathione peroxidase 4 (GPX4), which is the only enzyme that is known to prevent lipid peroxidation. Furthermore, we found that copper depletion decreased the sensitivity of the dermal papilla cells (DPCs) to erastin (an inducer of ferroptosis), and the ferroptosis inhibitor ferrostatin-1 (Fer-1) partially prevented BCS-mediated cell death. Overall, these findings establish a direct link between copper and ferroptosis; BCS-mediated copper depletion strongly enhances ferroptosis via mitochondrial perturbation and a reduction in antioxidative mechanisms.

mTORC1 Mediates the Processes of Lysine Regulating Satellite Cells Proliferation, Apoptosis, and Autophagy.

Lysine (Lys) is essential for skeletal muscle growth and protein synthesis in mammals. However, the regulatory network underlying Lys-regulated skeletal muscle development is unknown. To determine whether any cross-talk occurs among mammalian targets of rapamycin complex 1 (mTORC1) and Lys in the regulation of muscle satellite cells (SCs) proliferation, we applied the treatment rapamycin (a mTORC1 inhibitor) and MHY1485 (a mTORC1 activator) on Lys-added or -deficient SCs. The results show Lys deprivation significantly decreases SCs viability, protein synthesis, and cell cycling, increases autophagy and apoptosis, and inhibits the mTORC1 signaling pathway. Restoration of Lys content significantly attenuates this effect. mTORC1 signaling pathway activation during Lys deprivation or mTORC1 signaling pathway inhibition during Lys addition attenuates the effect of Lys deprivation or addition on SCs viability, protein synthesis, cell cycling, autophagy, and apoptosis. In conclusion, Lys could improve SCs proliferation, and inhibit SCs apoptosis and autophagy, via the mTORC1 signaling pathway.

Copper Modulates Mitochondrial Oxidative Phosphorylation to Enhance Dermal Papilla Cells Proliferation in Rex Rabbits.

Copper (Cu) is an important coenzyme factor in cell signaling, such as cytochrome c oxidase (Complex IV). Metabolism plays an important role in regulating the fate of mammalian cells. The aim of this study is to experimentally investigate the effect of copper on cell metabolism in the dermal papilla cells of the Rex rabbit. In this study, Cu promoted proliferation of dermal papilla cells (p = 0.0008) while also increasing levels of cellular CIII, CIV, Complex IV and ATP. Moreover, fifty metabolites that were significantly different between Cu and controls were identified as potential biomarkers of Cu stimulation. Copper-stimulated cells had altered levels of arachidonic acid derivatives, S-glutamic acid, and citric acid, which were primarily linked to two different pathways: arachidonic acid metabolism (p < 0.0001) and alanine, aspartate and glutamate metabolism (p = 0.0003). The addition of Cu can increase the proliferation of Rex rabbit dermal papilla cells. Increased levels of ubiquinol-cytochrome c reductase complex core protein 2 (CIII) and cytochrome c oxidase subunit 1 (CIV) were associated with the increased levels of cellular cytochrome c oxidase (Complex IV) and adenosine triphosphate (ATP). In a word, copper promotes cell proliferation by maintaining the function of the cellular mitochondrial electron transport chain (ETC) pathway.

Vitamin A alleviates heat stress-induced damage to hair follicle development in Rex rabbits.

BACKGROUND: Rex rabbits are important fur rabbits. Heat stress severely reduces the fur quality of Rex rabbits. The aim of this study was to experimentally investigate the effect of dietary vitamin A (VA) addition on hair follicle development and related signal pathways in Rex rabbits under heat stress. RESULTS: In the experiment, 90 Rex rabbits were randomly divided into three groups: control group (20-25 °C, fed basic diet), heat stress group (30-34 °C, fed basic diet), and heat stress + VA group (20-25 °C, fed 12 000 IU/kg VA in addition to the basic diet). VA could significantly increase the hair follicle density (P < 0.01), hair length (P < 0.05), and the ratio of secondary to primary hair follicles (P < 0.05). In addition, VA could significantly inhibit the expression of BMP2, BMP4, FGF5, TGF-ß1, and miR-214 in heat-stressed Rex rabbits and significantly increase the expression of noggin, IGF1, IGF1R, Wnt10b, CTNNB1, SHH, and miR-203 and the levels of Wnt10b and p-ß-catenin; however, there was no significant effect of VA on the expression of EGF and miR-205. CONCLUSION: The dietary addition of VA can increase the hair follicle density and fur quality of heat-stressed Rex rabbits. Wnt10/ß-catenin, insulin-like growth factor 1 (IGF1), fibroblast growth factor 5 (FGF5), noggin-BMP, and sonic hedgehog (SHH) signaling were associated with VA regulation under heat stress. It is possible that miR-205 and miR-194 contribute to the regulation of Wnt10/ß-catenin and bone morphogenetic protein (BMP) signaling. © 2021 Society of Chemical Industry.

Dietary lysine supplementation improves growth performance and skeletal muscle development in rabbits fed a low protein diet.

The purpose of this study was to investigate the effects on growth of Lysine (Lys) supplementation in a low protein diet. We also investigated the gene or protein expression related to skeletal muscle development and intestinal amino acid transporters, and determined the major signalling associated with Lys-regulating skeletal muscle development. 1000 healthy, weights averaging 938.6 ± 6.54 g weaned rabbits were randomly divided into five groups (five replicates in each group and 40 rabbits in each replicate). These groups consisted of the normal protein group (NP group, consuming a diet containing 16.27% protein), the low protein group (LP group, 14.15%-14.19% protein) and the LP group with an addition of 0.15%, 0.3% or 0.45% Lys. The trial included 7 d of pre-feeding and 28 d of exposure to the treatment. Compared with NP diet and LP diet, LP+0.3% Lys group improved growth performance (p < 0.05), full-bore weight and half-bore weight of rabbits (p < 0.05). The LP+0.3% Lys group also resulted in a decrease in the excretion of faecal nitrogen and urinary nitrogen (FN; UN; p < 0.05), and an increase in nitrogen utilisation rate (NUR; p < 0.05). LP diet increased the mRNA expression of MSTN and WWP1, and decreased the mRNA expression of IGF1 (p < 0.05). LP diet decreased the protein expression of P-P70S6K1, P-4EBP1 and P-S6 (p < 0.05). LP+0.3% Lys group attenuated the effects of LP diet on the expression of MSTN, WWP1, IGF1, P-P70S6K1, P-4EBP1 and P-S6 (p < 0.05). LP+0.3% Lys group resulted in an increase in mRNA expression of MyoD and protein expression of P-mTOR relative to the NP and LP groups (p < 0.05). In summary, the addition of Lys to a LP diet provides a theoretical basis for the popularisation and application of Lys in rabbit production.

Effect of sodium butyrate on slaughter performance, serum indexes and intestinal barrier of rabbits.

The purpose of this study was to investigate the effect of sodium butyrate on slaughter performance, serum indexes and the intestinal barrier in rabbits. Six hundred healthy weaned rabbits were randomly divided into three groups (5 replicates per group, 40 rabbits per replicate): control (fed a basal diet), sodium butyrate (fed a basal diet containing 0.5% sodium butyrate) and antibiotic (fed a basal diet containing 0.004% antibiotic). The trial lasted 35 days, including 7 days of pretesting and 28 days of formal testing. The results showed that dietary sodium butyrate supplementation increased the full-bore weight, the half-bore weight and the half-bore rate of rabbits. Meanwhile, the content of aspartate aminotransferase (AST) in serum was increased in rabbits fed the sodium butyrate diet. According to the intestinal barrier, after adding sodium butyrate to feed, the tight junction function of the rabbit intestine is enhanced, and the intestinal microbial composition is also improved. To sum up, after sodium butyrate was added to feed instead of antibiotics, slaughter performance was significantly enhanced, serum indexes were improved, and intestinal barrier function was also enhanced. Therefore, sodium butyrate can be added to feed as an additive and can replace antibiotics.