在这项研究中,我们调查了丁酸梭菌(A组)的作用,枯草芽孢杆菌(B组),和免疫增强剂藻类β-1,3葡聚糖(C组)对Reeves\'turtleMauremysreevesii肠道菌群的影响以及丁酸梭菌对Reevesii脾免疫组织转录组的影响。Reeve\'海龟被分配到四组,每个包含来自18个样品的三个重复。初始体重为106.35±0.03g的幼龟饲喂不含益生菌的基本饮食(D组),或含有丁酸梭菌TF20201120,枯草芽孢杆菌的基本饮食,或藻类β-1,3葡聚糖补充剂,分别。在实验周期的60、90和120d喂养海龟后,16SrRNA基因的高通量测序显示,在饲喂60天时,四组之间的α多样性没有显着差异(P>0.05),在90天,A组的α多样性差异显著(P<0.05),香农指数上升26.62%,辛普森指数下降83.33%;在120d,A组的α多样性(香农指数)呈递减趋势,B,C,在门一级,大量的拟杆菌,变形杆菌,A组Fusobacterium随着饲喂时间的增加而显著增加(P<0.05),在属一级,与其他三组相比,A组的Ruminocycaceae和Anaerotracus的丰度显着增加(P<0.05)。转录组分析表明384个基因在瑞维氏杆菌脾脏中差异表达,195个基因上调,189个基因下调,和丁酸梭菌TF201120调节脾造血细胞谱系信号通路(P<0.05)。通过qPCR证实了几种鉴定的免疫相关基因的调节。这些结果表明丁酸梭菌,B.枯草杆菌,免疫增强剂藻类β-1,3葡聚糖可以改善瑞维氏杆菌的肠道菌群,其中丁酸梭菌TF20201120是最有效的,并且显着增强了M.reevesii的免疫力。
In this study, we investigated the effects of Clostridium butyricum (group A), Bacillus subtilis (group B), and the immune enhancer algal β-1,3 glucan (group C) on the intestinal flora of Reeves\' turtle Mauremys reevesii and the effects of C. butyricum on the transcriptome of M. reevesii splenic immune tissues. Reeve\' turtles were assigned to four groups, each containing three replicates from 18 samples. Juvenile turtles with an initial weight of 106.35 ± 0.03 g were fed a basic diet containing no probiotics (group D), or a basic diet containing C. butyricum TF20201120, B.subtilis, or algal β-1,3 glucan supplement, respectively. After the turtles had been fed for 60, 90, and 120 d of the experimental period, high-throughput sequencing of the 16S rRNA gene revealed no significant difference in alpha diversity among the four groups at 60 days of feeding (P > 0.05), and at 90 days, the alpha diversity in group A was significantly different (P < 0.05), with an increase of 26.62% in the Shannon index and a decrease of 83.33% in the Simpson index; at 120 d, the alpha diversity (Shannon index) showed a decreasing trend in order for groups A, B, and C, At the phylum level, the abundance of Bacteroidetes, Proteobacteria, and Fusobacteria in group A increased significantly with increasing feeding time (P < 0.05), At the genus level, the abundance of Ruminococcaceae and Anaerotruncus in group A increased significantly compared with that in the other three groups (P < 0.05). Transcriptome analysis showed that 384 genes were differentially expressed in the spleen of M. reevesii, 195 genes were upregulated and 189 genes were downregulated, and C. butyricum TF201120 regulated the hematopoietic cell lineage signaling pathway in the spleen of M. reevesii (P < 0.05). The regulation of several identified immune-related genes was confirmed by qPCR. These results showed that C. butyricum, B. subtilis, and the immune enhancer algal β-1,3 glucan can improve the intestinal flora of M. reevesii, with C. butyricum TF20201120 being the most effective and significantly enhancing the immunity of M. reevesii.