不同饱血状态微小牛蜱中肠和唾液菌群结构的分析

doi:10.11843/j.issn.0366-6964.2017.03.017

摘要/Abstract

摘要：

旨在探明微小牛蜱随吸血时间的延长，中肠和唾液微生物菌群结构的差异，以及优势菌属在中肠和唾液中增殖和迁移的特点。在无菌条件下采集半饱血、饱血微小牛蜱唾液和中肠内容物；提取细菌总DNA，PCR扩增其16S rDNA V3-V4区，Illumina MiSeq双端测序；对序列进行拼接和过滤、OTUs聚类、物种注释及多样性分析。结果显示，共得到134 317条序列，聚类后获得1 304个OTUs，半饱血雌成蜱唾液（SP）、饱血雌成蜱唾液（SF）、半饱血雌成蜱中肠内容物（MP）、饱血雌成蜱中肠内容物（MF）分别获得336、332、269、289个OTUs，其中152个OTUs为4个样本共有；4个样品在门水平上，以变形菌门和厚壁菌门为优势菌门，变形菌门在中肠中的含量大于唾液，而厚壁菌门反之；在属水平上，4个样本以柯克斯体属、不动杆菌属、甲基杆菌属、短波单胞菌属、立克次体属、埃希菌属为优势菌属，其中柯克斯体属、不动杆菌属在4个样本中含量均较大，柯克斯体属在半饱血状态下，含量高于饱血状态；不动杆菌属在唾液中的含量高于中肠。结果表明，微小牛蜱中肠和唾液中有复杂的微生物菌群结构；细菌在不同组织中的分布不同，其含量随吸血时间的延长而发生变化。

Abstract:

The aim of this study was to investigate the difference of bacterial flora and diversity of the saliva and midgut contents of Rhipicephalus microplus with the extension of feeding time, and the proliferation and migration characteristics of dominant genera in saliva and midgut. The saliva (S) and midgut contents (M) collected from partially (P) or fully (F) engorged adult female ticks under sterile environment, the total DNA of bacteria were extracted and PCR amplification. Using Illumina MiSeq sequencing platform, two-terminal sequencing of V3-V4 areas of 16S rDNA was carried. Then OTUs (operational taxonomic units) picking, taxonomy assignment, and diversity analysis were carried on the effective Tags. The results showed that 134 317 sequences were obtained, the total of OTUs were 1 304, OTUs obtained from four samples (SP, SF, MP and MF) were 336, 332, 269, 289, respectively. 152 OTUs had high similarity in 4 samples. Proteobacteria and Firmicutes were the most predominant phyla in all samples. Proteobacteria had higher abundance in midgut than saliva. On the contrary, Firmicutes was highly abundant in saliva. Coxiella, Acinetobacter, Methylobacterium, Brevundimonas, Rickettsia and Escherichia were the major genera. Coxiella and Acinetobacter were highly abundant in all samples. The abundance of Coxiella was higher in partially engorged than fully engorged ticks. Acinetobacter had higher abundance in midgut than saliva. These findings suggested that the saliva and midgut of R. microplus had complicated microbial community structure. The distribution of bacteria were different in different tissues, and the abundance of bacteria changed with the feeding time.

中图分类号:

S852.746

段德勇，程天印. 不同饱血状态微小牛蜱中肠和唾液菌群结构的分析[J]. 畜牧兽医学报, 2017, 48(3): 530-537.

DUAN De-yong，CHENG Tian-yin. Bacterial Flora Analysis of Saliva and Midgut Contents from Partially or Fully Engorged Female Adult Rhipicephalus microplus[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2017, 48(3): 530-537.

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