Chen Xiaobing, Di Panpan, Wang Hongming, Li Bailin, Pan Yingjie, Yan Shuling, Wang Yongjie
Laboratory of Quality and Safety Risk Assessment for Aquatic Products on Storage & Preservation, Ministry of Agriculture, Shanghai, China; College of Food Science and Technology, Shanghai Ocean University, Shanghai, China.
College of Food Science and Technology, Shanghai Ocean University, Shanghai, China.
PLoS One. 2015 Apr 13;10(4):e0123990. doi: 10.1371/journal.pone.0123990. eCollection 2015.
Chinese mitten crab (CMC, Eriocheir sinensis) is an economically valuable species in South-East Asia that has been widely farmed in China. Characterization of the intestinal bacterial diversity of CMC will provide insights into the aquaculturing of CMCs. Based on the analysis of cloned 16S rRNA genes from culture-independent CMC gut bacteria, 124 out of 128 different clones reveal >95% nucleotide similarity to the species belonging to the four phyla of Tenericutes, Bacteroidetes, Firmicutes and Proteobacteria; one clone shows 91% sequence similarity to the member of TM7 (a candidate phylum without cultured representatives). Fluorescent in situ hybridization also reveals the abundance of Bacteroidetes in crab intestine. Electron micrographs show that spherical and filamentous bacteria are closely associated with the microvillus brush border of the midgut epithelium and are often inserted into the space between the microvilli using a stalk-like cell appendage. In contrast, the predominant rod-shaped bacteria in the hindgut are tightly attached to the epithelium surface by an unusual pili-like structure. Both 16S rRNA gene denaturing gel gradient electrophoresis and metagenome library indicate that the CMC Mollicutes group 2 appears to be present in both the midgut and hindgut with no significant difference in abundance. The CMC Mollicutes group 1, however, was found mostly in the midgut of CMCs. The CMC gut Mollicutes phylotypes appear to be most closely related to Mollicutes symbionts detected in the gut of isopods (Crustacea: Isopoda). Overall, the results suggest that CMCs harbor diverse, novel and specific gut bacteria, which are likely to live in close relationships with the CMC host.
中华绒螯蟹(CMC,Eriocheir sinensis)是一种在东南亚具有经济价值的物种,在中国已被广泛养殖。对中华绒螯蟹肠道细菌多样性的表征将为中华绒螯蟹的水产养殖提供见解。基于对来自未培养的中华绒螯蟹肠道细菌的克隆16S rRNA基因的分析,128个不同克隆中的124个与柔膜菌门、拟杆菌门、厚壁菌门和变形菌门四个门的物种显示出>95%的核苷酸相似性;一个克隆与TM7(一个没有培养代表的候选门)的成员显示出91%的序列相似性。荧光原位杂交也揭示了蟹肠道中拟杆菌门的丰度。电子显微镜照片显示,球形和丝状细菌与中肠上皮的微绒毛刷状缘紧密相关,并且经常使用茎状细胞附属物插入微绒毛之间的空间。相比之下,后肠中占主导地位的杆状细菌通过一种不寻常的菌毛状结构紧密附着在上皮表面。16S rRNA基因变性凝胶梯度电泳和宏基因组文库均表明,中华绒螯蟹柔膜菌2组似乎同时存在于中肠和后肠中,丰度没有显著差异。然而,中华绒螯蟹柔膜菌1组主要在中华绒螯蟹的中肠中发现。中华绒螯蟹肠道柔膜菌的系统发育型似乎与在等足类动物(甲壳纲:等足目)肠道中检测到的柔膜菌共生体关系最为密切。总体而言,结果表明中华绒螯蟹含有多样、新颖和特定的肠道细菌,它们可能与中华绒螯蟹宿主存在密切关系。