广西蚕沙细菌组成多样性解析和VBNC菌群的发掘

【目的】解析广西蚕沙中细菌群落组成与多样性,为蚕沙中菌种资源发掘和蚕沙的综合利用提供科学依据。【方法】通过高通量测序技术研究细菌群落组成特征,同时利用常规稀释涂布平板法和基于复苏促进因子(Rpf)的MPN培养系统解析并筛选蚕沙中可培养和活的非可培养(VBNC)状态的优势菌群,并经16SrRNA基因测序对筛选得到的菌株作初步分类鉴定。【结果】高通量测序表明,广西蚕沙样品中细菌归属于10个门、18个纲、27个目、57个科、96个属,其中4个属的丰度达1%以上,优势菌群为变形菌门(Proteobacteria)肠杆菌属(Enterobacter);通过稀释涂布平板法共获得14个属的33株可培养细菌,其中4个属(Citrobacter、Weissella、Chitinophaga、Pseudoclavibacter)在高通量测序中未被检测到;而在MPN培养系统中,基于复苏促进因子的处理组细菌总数最大检出丰度提高了100倍,并从中共检出21株对Rpf敏感的VBNC菌株,其中6个属(Paenibacillus、Caulobacter、Roseomonas、Pantoea、Erwinia、Acine...

Bacterial diversity and VBNC bacteria in silkworm excrement from Guangxi

[Objective] To analyze the composition and diversity of bacterial in silkworm excrement from Guangxi, and to explore microbial resource of silkworm excrement. [Methods] Through high-throughput sequencing technology, we analyzed the composition of bacterial communities in silkworm excrement. Meanwhile, we isolated culturable bacteria and those in viable but non-culturable (VBNC) state in silkworm excrement using spread-plate method and resuscitation promoting factor (Rpf)-based MPN method, and then classified and identified the isolated strains through 16S rRNA gene sequence analysis. [Results] The bacteria detected from silkworm excrement of Guangxi belonged to 10 phyla, 18 classes, 27 orders, 57 families and 96 genera, in which the bacteria abundance of 4 genera was greater than 1% and the dominant microflora was Enterobacter (Proteobacteria). We obtained 33 culturable bacteria from 14 genera through the spread-plate method, in which 4 genera (Citrobacter, Weissella, Chitinophaga, Pseudoclavibacter) were not detected by high-throughput sequencing. The detection abundance of bacteria increased by up to 100 times by Rpf-based MPN method. Also, we found 21 Rpf-sensitive and resuscitation-cultivable VBNC strains from the Rpf-treatment group, in which 6 genera (Paenibacillus, Caulobacter, Roseomonas, Pantoea, Erwinia, Acinetobacter) and 2 genera (Paenibacillus and Caulobacter) were not found through spread-plate method and high-throughput sequencing, respectively. [Conclusion] Our results provide a new perspective and a new approach for exploring and utilizing the microbial resources in silkworm excrement.