基于生物反应器制药废水中活的但非可培养状态细菌的分离

目的]探究制药废水中活的但非可培养（viable but non-culturable，VBNC）状态细菌的组成与系统关系。方法]利用土质滤材构建生物反应器，基于复苏促进因子（resuscitation promoting factor，Rpf）对 VBNC细菌的复苏刺激生长作用，采用 MPN （most probable number）法，稀释平板法分离其中复苏可培养化的 VBNC 菌株并解析其16S rRNA基因系统关系。结果]在 MPN培养系中，Rpf对部分细菌有生长刺激作用。制药废水中存在对 Rpf敏感、复苏可培养化的 VBNC优势菌群；制药废水中对 Rpf敏感的 VBNC优势细菌主要有革兰氏阳性放线菌Microbacterium、Gordonia 和 Leucobacter 属近缘菌，革兰氏阴性菌 Candidimonas、Xanthobacter和 Aminobacter属近缘菌，其中菌株 ZYM1、ZYM3、ZYZR4和 ZYXR1可能属于新种。结论]研究结果揭示了制药废水中存在 VBNC状态细菌，为研究其形成机理、复苏活性化机制以及制药废水的深度处理等提供重要的思路与方法。

Viable but Non-culturable Bacteria in Bioreactor-based Pharmaceutical Wastewater

We aimed to investigate the composition and phylogenetic rela-tionships of the viable but non-culturable （VBNC） state bacteria in pharmaceutical wastewater. Method] Soil filter was used for constructing bioreactor. Based on the resuscitation- and growth-promoting function of Resuscitation Promoting Factor （Rpf） for VBNC bacteria, VBNC bacteria were isolated by most probable number （MPN） method and dilution-plating method and 16S rRNA gene phylogenetic analysis was carried out. Result] In MPN culture system, Rpf could promote the resuscitation and growth of some bacteria. There were VBNC advantage floras that sensitive to Rpf in pharmaceutical wastewater. The culturable VBNC bacteria in pharmaceutical wastewater consisted of high-GC gram-positive actinomycetes including genera Mi-crobacterium, Gordonia and Leucobacter, and gram-negative bacteria including gen-era Candidimonas, Xanthobacter and Aminobacter. Four strains （ZYM1, ZYM3, ZYZR4, ZYXR1） could be potential novel species. Conclusion] This research re-vealed there were VBNC bacteria in pharmaceutical wastewater. These results could provide important ideas and methods for further studies on VBNC bacteria in the pharmaceutical wastewater, especial y the formation mechanism and recovery mech-anism of VBNC bacteria and the advanced degradation process improvement of pharmaceutical wastewater.