生物医药企业废水致突变性和菌群构建的监测
Monitoring on Mutagenicity and Bacterial Community Structure of the Sewage from Biomedical Enterprises
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摘要:
目的 通过对来自生物医药集聚区废水细菌种群构成和诱变效能的检测,以期在生物医药研发企业排放废水的处置和管理中,应用有针对性的技术手段来改进和完善其检测与监测方法。 方法 生物医药企业排放污水中所含细菌总数和大肠菌群数量的检测,采用3M细菌总数测试片和快速大肠菌群测试片;致突变能力检测采用鼠伤寒沙门氏菌回复突变试验(Ames试验)进行;同时采用PCR克隆技术建立排放污水中细菌16S rRNA基因文库,通过对文库构成的分析,监测排放污水中细菌种群的构成。 结果 部分污水排放点细菌总数超过108 CFU/mL,最低为105 CFU/mL:其中大肠菌群在不同排放点基本一致,约为103~104个/mL。16S rRNA基因克隆文库序列经与GeneBank 16S rRNA序列比对,共鉴定出23个属的细菌,主要为红环菌科、丛毛单胞菌科和黄杆菌科内的菌属,并从其中一份废水检出人类致病菌军团菌。3份污水浓缩样能直接抑制测试菌的生长,2份Ames试验显示含有移码型直接突变物,另有2份Ames试验显示可能含有移码型直接突变物,碱基置换型突变物均阴性。 结论 对生物医药研发废水的管理策略的制订需要建立在长期、稳定、多方面监测的基础上,采用16S rRNA基因文库分析了解菌群构成和变迁,采用Ames试验检测菌群致突变能力的变化,从而由整体效应评估污水的污染状况,可以应用于生物医药研发企业污水监测。 Abstract:Objective Through detecting the structure of bacterial community and the effects of mutation to monitor and regulate the treatment of sewage from biomedical enterprises. Methods The study used 3M petrifilm aerobic count plates and 3M petrifilm rapid coliform count plates to count the number of bacteria and coliform groups. Sewage was concentrated and its mutagenic effects tested with the Ames test. Polymerase chain reaction (PCR) was conducted to build the 16S rRNA gene library, which was used to monitor the structure of bacterial community. Results Aerobic bacteria counting showed 3 sampling sites containing more than 108 CFU/mL bacteria. All sampling sites showed a consistent result on coliform count from 103-104/mL. Twenty three species were found in the samples by 16S rRNA gene clone method, mainly including Rhodocyclaceae, Comamonadaceae and Flavobacteriaceae, etc. Human pathogenic Legionella was detected in one sample. Two concentrated sewage samples showed frame shift mutation effect, and dose-effect relationship was shown in another two. Other samples were negative in Ames test. Conclusion The monitoring of sewage from biomedical enterprises should be established on a strategy of long-term, stable and multiple aspects. Ames test and 16S rRNA gene library analysis need to be involved in evaluating the comprehensive situation and mutagenic ability of sewage as a whole, and actually these two methods could be used in such monitoring.
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