With the comprehensive prohibition of antibiotics in the feed industry in China, the incidence of diarrhea in rabbits increased, such as loss of appetite, vomiting, and excretion of atheromatous feces. In order to explore the pathological and the molecular mechanisms of the diarrhea in the rabbitry fed with antibiotic-free diet, we used microbial metagenomics, transcriptome, and non-targeted metabolomics sequencing. The results showed that the Firmicutes level was significantly decreased (p < 0.001) and the Proteobacteria level was significantly increased (p < 0.05). The functional enrichment of cecum revealed that most differentially expressed genes (DEGs) were expressed in immune, inflammatory, and metabolic processes. The enrichment of the cecal fecal metabolites focused on the bile secretion, antifolate resistance, and tryptophan metabolism pathways, which are mainly associated with inflammation. The results of correlation analysis showed that Fournierella was positively correlated with myricetin, ursolic acid, and furtherly might cause bile secretion and tryptophan metabolism disorder, aggravate intestinal inflammation, change intestinal permeability, and reduce host immunity, leading to diarrhea in rabbits. This study provides a theoretical basis for illustrating the reason for diarrhea and developing new feeds for the health of rabbits.
Imbalances in the gut microbiota composition may lead to several reproductive disorders and diseases during pregnancy. This study investigates the fecal microbiome composition between primiparous and multiparous cows during non-pregnancy and pregnancy to analyze the host-microbial balance at different stages. The fecal samples obtained from six cows before their first pregnancy (BG), six cows during their first pregnancy (FT), six open cows with more than three lactations (DCNP), and six pregnant cows with more than three lactations (DCP) were subjected to 16S rRNA sequencing, and a differential analysis of the fecal microbiota composition was performed. The three most abundant phyla in fecal microbiota were Firmicutes (48.68%), Bacteroidetes (34.45%), and Euryarchaeota (15.42%). There are 11 genera with more than 1.0% abundance at the genus level. Both alpha diversity and beta diversity showed significant differences among the four groups (p < 0.05). Further, primiparous women were associated with a profound alteration of the fecal microbiota. The most representative taxa included Rikenellaceae_RC9_gut_group, Prevotellaceae_UCG_003, Christensenellaceae_R_7_group, Ruminococcaceae UCG-005, Ruminococcaceae UCG-013, Ruminococcaceae UCG-014, Methanobrevibacter, and [Eubacterium] coprostanoligenes group, which were associated with energy metabolism and inflammation. The findings indicate that host-microbial interactions promote adaptation to pregnancy and will benefit the development of probiotics or fecal transplantation for treating dysbiosis and preventing disease development during pregnancy.
MicroRNAs (miRNAs) are a class of small non-coding RNAs with the capability of modulating gene expression at the post-transcriptional level either by inhibiting messenger RNA (mRNA) translation or by promoting mRNA degradation. A large number of studies have shown that miR-383-5p plays a key role in the development of cancer cells, but the mechanism of miR-383-5p regulating adipogenesis is still poorly understood. In our study, we found that the expression of miR-383-5p in perirenal adipose tissue of rabbits aged 35 and 70 days was higher, which was significantly different from other tissues (p < 0.05). In addition, cell counting kit 8 (CCK-8), flow cytometry and 5-ethynyl-2’-deoxyuridine (EDU) experiments were used to identify the effect of miR-383-5p on the proliferation of rabbit preadipocytes. The results showed that miR-383-5p promoted the proliferation of rabbit preadipocytes. Western Blotting (WB), qRT-PCR and Oil Red O staining experiments were used to identify the effect of miR-383-5p on the differentiation of rabbit preadipocytes. After overexpression of miR-383-5p, the expression of fat differentiation markers PPARγ, FABP4, and CEBP/α were significantly higher than that of the control group, while the inhibitory group showed the opposite trend. The results of oil red o staining showed that the number of lipid droplets in the overexpression group was significantly higher than that in the control group, while the number of lipid droplets in the inhibition group was opposite. WB experiment also showed the same trend. These results showed that miR-383-5p promoted the differentiation of rabbit preadipocytes. Moreover, 1642 (721 up-regulated and 921 down-regulated) known differentially expressed (DE) genes were screened from miR-383-5p mimic groups and miR-383-5p negative control (NC) groups by transcriptome sequencing technology. Gene Ontology Resource (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enriched the pathways related to lipid metabolism. Protein catabolism, organic nitrogen compound biosynthesis, and lipid biosynthesis were significantly enriched. Finally, using bioinformatics websites to predict the discovery of RAD51AP1 as a potential target for miR-383-5p, their target relationship was demonstrated after its dual luciferase reporting experiment. Overall, our data indicate that miR-383-5p plays a positive regulatory effect on the proliferation and differentiation of rabbit preadipocytes.
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