高通量测序分析血根碱对中华鳖肠道菌群结构的影响

High-throughput sequencing analysis of the effects of sanguinarine on Trionyx sinensis intestinal microbiota

中文关键词: 中华鳖  血根碱  高通量测序  肠道菌群

英文关键词: Trionyx sinensis  Sanguinarine  High-throughput sequencing  Intestinal microbiota

中文摘要:

      为研究血根碱对中华鳖(Trionyx sinensis)肠道菌群多样性及丰度的影响，选取480只初始体重为(31.75±7.20) g的中华鳖，设置添加0的对照组(RS1)和添加50 (RS2)、100 (RS3)和150 mg/kg (RS4)的3个实验组。实验结束后，提取中华鳖中肠组织的总DNA，进行16S rRNA V3-4区基因组测序。结果显示，实验组的总操作分类单元(Operational taxonomic units, OTUs)数量显著高于对照组，肠道菌群复杂度及丰富度较对照组有明显增加。肠道菌群结构组成分析发现，RS1与RS2结构较为接近，而RS1与RS3、RS4较远。在门分类水平上，变形菌门(Proteobacteria)和衣原体门(Chlamydiae)是优势菌群；在属分类水平上，螺杆菌属(Helicobacter)占绝对优势。通过比较发现，实验组中螺杆菌属丰度较对照组显著下降，而不动杆菌属(Acinetobacter)、乳杆菌属(Lactobacillus)和双歧杆菌属(Bifidobacterium)丰度均有明显上升。研究表明，在饲料中添加适量的血根碱(100~150 mg/kg)能有效提高中华鳖肠道菌群多样性及丰富度，并能改善肠道菌群结构。

英文摘要:

      To study the effect of sanguinarine on intestinal microbiota diversity in Trionyx sinensis, 480 individuals [initial weight (31.75±7.20) g] were randomly selected and divided into four groups. Three experimental diets RS2, RS3, and RS4 were formulated with 50, 100, and 150 mg/kg sanguinarine, respectively, and no sanguinarine was included in the control group. At the end of the experiment, the total DNA of the midgut tissue was extracted and the 16S rRNA V3-4 region was sequenced. The results showed that the total number of operational taxonomic units (OTU) in experimental groups was significantly higher than that in the control group, and the complexity and richness of intestinal flora was also significantly higher than that of the control group. Analysis of the structure and composition of the intestinal flora revealed that RS1 and RS2 were closer in structure, whereas RS1 was farther away from RS3 and RS4. Proteobacteria and Chlamydiae were the dominant phyla, and Helicobacter was the dominant genus. Helicobacter abundance in experimental groups was significantly lower than that in the control group, whereas the abundance of Acinetobacter, Lactobacillus, and Bifidobacterium were all significantly higher than that in the control. Therefore, the study demonstrated that adding an appropriate amount of sanguinarine (100~150 mg/kg) in the feed could effectively promote intestinal flora diversity and richness, and improve the community structure of T. sinensis intestinal flora.