Graczyk Thaddeus K, Kacprzak Malgorzata, Neczaj Ewa, Tamang Leena, Graczyk Halshka, Lucy Frances E, Girouard Autumn S
Department of Environmental Health Sciences, Division of Environmental Health Engineering, Johns Hopkins Bloomberg School of Public Health, Baltimore, MD 21205, USA.
Parasitol Res. 2007 Aug;101(3):569-75. doi: 10.1007/s00436-007-0515-x. Epub 2007 Mar 16.
Solid waste landfill leachate and sewage sludge samples were quantitatively tested for viable Enterocytozoon bieneusi, Encephalitozoon intestinalis, Encephalitozoon hellem, and Encephalitozoon cuniculi spores by the multiplexed fluorescence in situ hybridization (FISH) assay. The landfill leachate samples tested positive for E. bieneusi and the sludge samples for E. bieneusi and E. intestinalis. The effects of four sanitization treatments on the inactivation of these pathogens were assessed. Depending on the variations utilized in the ultrasound disintegration, sonication reduced the load of human-virulent microsporidian spores to nondetectable levels in 19 out of 27 samples (70.4%). Quicklime stabilization was 100% effective, whereas microwave energy disintegration was 100% ineffective against the spores of E. bieneusi and E. intestinalis. Top-soil stabilization treatment gradually reduced the load of both pathogens, consistent with the serial dilution of sewage sludge with the soil substrate. This study demonstrated that sewage sludge and landfill leachate contained high numbers of viable, human-virulent microsporidian spores, and that sonication and quicklime stabilization were the most effective treatments for the sanitization of sewage sludge and solid waste landfill leachates. Multiplexed FISH assay is a reliable quantitative molecular fluorescence microscopy method for the simultaneous identification of E. bieneusi, E. intestinalis, E. hellem, and E. cuniculi spores in environmental samples.
通过多重荧光原位杂交（FISH）测定法，对固体废物填埋场渗滤液和污水污泥样本中的活微小隐孢子虫、肠脑炎微孢子虫、海伦脑炎微孢子虫和兔脑炎微孢子虫孢子进行了定量检测。填埋场渗滤液样本中微小隐孢子虫检测呈阳性，污泥样本中微小隐孢子虫和肠脑炎微孢子虫检测呈阳性。评估了四种消毒处理对这些病原体灭活的效果。根据超声破碎中使用的变量，在27个样本中的19个（70.4%）中，超声处理将对人类有致病性的微孢子虫孢子负荷降低到无法检测的水平。生石灰稳定化处理的效果为100%，而微波能量破碎处理对微小隐孢子虫和肠脑炎微孢子虫的孢子无效。表土稳定化处理逐渐降低了两种病原体的负荷，这与污水污泥与土壤基质的系列稀释一致。本研究表明，污水污泥和填埋场渗滤液含有大量有活力的、对人类有致病性的微孢子虫孢子，并且超声处理和生石灰稳定化处理是污水污泥和固体废物填埋场渗滤液消毒最有效的处理方法。多重FISH测定法是一种可靠的定量分子荧光显微镜方法，用于同时鉴定环境样本中的微小隐孢子虫、肠脑炎微孢子虫、海伦脑炎微孢子虫和兔脑炎微孢子虫孢子。
