Monpoeho S, Maul A, Bonnin C, Patria L, Ranarijaona S, Billaudel S, Ferré V
Laboratoire de Virologie, UPRES 1156, Centre Hospitalier Universitaire, Nantes, France.
Appl Environ Microbiol. 2004 Sep;70(9):5434-40. doi: 10.1128/AEM.70.9.5434-5440.2004.
Sludges derived from wastewater treatment are foul-smelling, biologically unstable substances. As well as containing numerous pathogenic microorganisms, they also consist of organic matter that can be used as agricultural fertilizer. Legislation nevertheless requires sludges to be virologically tested prior to spreading by the counting of infectious enterovirus particles. This method, based on culture of enterovirus on BGM cells, is lengthy and not very sensitive. The aim of this study was to propose an alternative method of genome quantification for all enteroviruses that is applicable to verifying the elimination of viruses in complex samples such as sludges. Our complete protocol was compared to the official method, consisting of enterovirus enumeration with the most probable number of cythopathic unit (MPNCU) assay through the study of four stabilization procedures: liming, composting, heat treatment, and mesophile anaerobic digestion. Enterovirus quantities at the start of the stabilization procedures were between 37 and 288 MPNCU/g on the one scale and between 4 and 5 log genome copies/g on the other. It was shown that all procedures except mesophile anaerobic digestion were highly effective in the elimination of enterovirus particles and genomes in wastewater sludges. Reduction of viruses by mesophile anaerobic digestion was by only 1 log (infectious particles and genomes). In conclusion, stabilization processes can indeed be checked by virological quality control of sludges with gene amplification. However, the infectivity of genomes needs to be confirmed with cell culture or a correlation model if the virological risk inherent in the agricultural use of such sludges is to be fully addressed.
来自污水处理的污泥是有恶臭、生物不稳定的物质。除了含有众多致病微生物外，它们还包含可用作农业肥料的有机物。然而，法规要求在污泥撒施前通过计数传染性肠道病毒颗粒进行病毒学检测。这种基于肠道病毒在BGM细胞上培养的方法耗时且灵敏度不高。本研究的目的是提出一种适用于验证复杂样品（如污泥）中病毒消除情况的所有肠道病毒基因组定量的替代方法。我们完整的方案与官方方法进行了比较，官方方法包括通过研究四种稳定化程序（石灰处理、堆肥、热处理和中温厌氧消化），用最可能数目的细胞病变单位（MPNCU）测定法对肠道病毒进行计数。稳定化程序开始时，肠道病毒数量在一个尺度上为37至288 MPNCU/g，在另一个尺度上为4至5 log基因组拷贝/g。结果表明，除中温厌氧消化外，所有程序在消除污水污泥中的肠道病毒颗粒和基因组方面都非常有效。中温厌氧消化对病毒的减少仅为1 log（感染性颗粒和基因组）。总之，稳定化过程确实可以通过对污泥进行基因扩增的病毒学质量控制来检查。然而，如果要充分解决此类污泥农业使用中固有的病毒学风险，则需要通过细胞培养或相关模型来确认基因组的感染性。
