Rodríguez Roberto A, Gundy Patricia M, Gerba Charles P
Department of Soil, Water, and Environmental Science, University of Arizona, P.O. Box 210038, Tucson, AZ 85721-0038, USA.
Appl Environ Microbiol. 2008 May;74(9):2583-7. doi: 10.1128/AEM.00626-07. Epub 2008 Mar 7.
The objective of this study was to compare PLC/PRF/5 and BGM cell lines for use in a total culturable viral assay (TCVA) of treated sewage effluents. Samples were collected before and after chlorination from an activated sludge wastewater treatment plant and from the effluent of a high-rate enhanced flocculation system, followed by UV light disinfection. Cell monolayers were observed for cytopathic effect (CPE) after two passages of 14 days each. Monolayers exhibiting viral CPE were tested for the presence of adenoviruses and enteroviruses by PCR or reverse transcription-PCR. Eight percent of the samples exhibited CPE on BGM cells, and 57% showed CPE on PLC/PRF/5 cells. Only enteroviruses were detected on the BGM cells, while 30% and 52% of the samples were positive for enteroviruses and adenoviruses, respectively, on the PLC/PRF/5 cells. Thirty percent of the samples were positive for both adenoviruses and enteroviruses in chlorinated activated sludge effluent. Thirty percent of the samples were positive for adenoviruses in the UV treatment effluent, but no enteroviruses were detected. In conclusion, the PLC/PRF/5 cells were more susceptible than BGM cells to viruses found in treated sewage. The use of BGM cells for TCVA may underestimate viral concentration in sewage effluent samples. The PLC/PRF/5 cells were more susceptible to adenoviruses, which is important in the evaluation of UV disinfection systems because adenoviruses are highly resistant to UV inactivation.
本研究的目的是比较PLC/PRF/5细胞系和BGM细胞系在处理后的污水总可培养病毒检测(TCVA)中的应用。从活性污泥废水处理厂以及高速强化絮凝系统的出水(经紫外线消毒)中,在氯化处理前后采集样本。每个细胞单层培养14天,传代两次后观察细胞病变效应(CPE)。对出现病毒CPE的细胞单层,通过聚合酶链反应(PCR)或逆转录聚合酶链反应检测腺病毒和肠道病毒的存在情况。8%的样本在BGM细胞上出现CPE,57%的样本在PLC/PRF/5细胞上出现CPE。在BGM细胞上仅检测到肠道病毒,而在PLC/PRF/5细胞上,分别有30%和52%的样本肠道病毒和腺病毒呈阳性。在氯化活性污泥出水中,30%的样本腺病毒和肠道病毒均呈阳性。在紫外线处理出水中,30%的样本腺病毒呈阳性,但未检测到肠道病毒。总之,PLC/PRF/5细胞比BGM细胞对处理后污水中的病毒更敏感。在TCVA中使用BGM细胞可能会低估污水样本中的病毒浓度。PLC/PRF/5细胞对腺病毒更敏感,这在评估紫外线消毒系统时很重要,因为腺病毒对紫外线灭活具有高度抗性。