Application of a Swab Sampling Method for the Detection of Norovirus and Rotavirus on Artificially Contaminated Food and Environmental Surfaces

Noroviruses and rotaviruses are the leading causes of non-bacterial gastroenteritis in humans worldwide. Virus-contaminated food and surfaces represent an important risk to public health. However, established detection methods for the viruses in food products are laborious and time-consuming. Here, we describe a detailed swabbing protocol combined with real-time RT-PCR for norovirus and rotavirus detection on artificially contaminated food and environmental surfaces. Recovery rates between 2 and 78% for norovirus and between 8 and 42% for rotavirus were determined for contaminated food surfaces of apple, pepper, cooked ham and salami. From contaminated environmental surfaces (stainless steel, ceramic plate, polyethylene, wood), recovery rates between 26 and 52% (norovirus) and between 10 and 58% (rotavirus) were determined. The results demonstrate the suitability of the swab sample method for virus detection on food and environmental surfaces. Compared to other methods, it is easy to perform and significantly time-saving, predestining it for routine testing.     

两种水源性诺如病毒富集方法的比较

评估阳离子膜吸附-洗脱法和阴离子膜吸附-洗脱法在富集水源性诺如病毒（NV）过程中的病毒回收效果、消耗时间及使用成本,可以获得更适用于水源性诺如病毒实际监测工作的较优富集方法.以无锡市疾病预防控制中心保存的诺如病毒阳性标准质粒为模板,采用荧光定量RT-PCR（反转录酶-聚合酶链反应）方法绘制病毒定量标准曲线,并应用于病毒的绝对定量.将含诺如病毒的粪便悬液先稀释成3个不同浓度,并加入灭菌水中,分别经过阴离子膜（硝酸纤维素滤膜）或阳离子膜（Nanoceram膜）吸附洗脱,最终相同体积洗脱液提取核酸后用荧光定量RT-PCR方法进行绝对定量,并进一步计算最终病毒回收率.结果表明:①样本病毒浓度为8×101~8×1010 copies/μL时,病毒拷贝数的lg对数值与C_t（荧光定量PCR中达到设定阈值所经过的循环数）呈线性关系.②随着样本病毒浓度的增加,阴离子膜法和阳离子膜法的病毒回收率都相应增加.③当检测样本中病毒浓度分别为8×101、4×102 copies/μL时,阴离子膜法回收率分别为22.2%±4.1%和36.8%±6.2%,阳离子膜法回收率分别为17.4%±1.5%和28.8%±6.1%,说明阴离子膜吸附-洗脱法对病毒的富集回收率显著高于阳离子膜吸附-洗脱法（P < 0.05）.研究显示,综合富集效果、耗费时间和成本等因素,阴离子膜吸附-洗脱法比阳离子膜吸附-洗脱法更适用于日常水源性诺如病毒的富集工作.      

Survival of Norovirus Surrogate on Various Food-Contact Surfaces

Norovirus (NoV) is an environmental threat to humans, which spreads easily from one infected person to another, causing foodborne and waterborne diseases. Therefore, precautions against NoV infection are important in the preparation of food. The aim of this study was to investigate the survival of murine norovirus (MNV), as a NoV surrogate, on six different food-contact surfaces: ceramic, wood, rubber, glass, stainless steel, and plastic. We inoculated 10⁵ PFU of MNV onto the six different surface coupons that were then kept at room temperature for 28 days. On the food-contact surfaces, the greatest reduction in MNV was 2.28 log₁₀ PFU/coupon, observed on stainless steel, while the lowest MNV reduction was 1.29 log₁₀ PFU/coupon, observed on wood. The rank order of MNV reduction, from highest to lowest, was stainless steel, plastic, rubber, glass, ceramic, and wood. The values of d _R (time required to reduce the virus by 90 %) on survival plots of MNV determined by a modified Weibull model were 277.60 h (R ² = 0.99) on ceramic, 492.59 h (R ² = 0.98) on wood, 173.56 h on rubber (R ² = 0.98), 97.18 h (R ² = 0.94) on glass, 91.76 h (R ² = 0.97) on stainless steel, and 137.74 h (R ² = 0.97) on plastic. The infectivity of MNV on all food-contact surfaces remained after 28 days. These results show that MNV persists in an infective state on various food-contact surfaces for long periods. This study may provide valuable information for the control of NoV on various food-contact surfaces, in order to prevent foodborne disease.     

燃煤污染源排放颗粒物采样方法实验室比对研究

为研究适用于我国燃煤污染源排放颗粒物的分级采样标准测试方法,采用民用小煤炉排放装置结合烟气采集系统,对比研究了直接采样法(双级虚拟撞击PM₁₀/PM_2.5采样器、旋风采样器、总烟尘采样器)和稀释采样法(低压荷电撞击器ELPI配备稀释系统)对烟气中不同粒径颗粒物测试结果的稳定性、相关性、仪器可操作性等特点,并分析了煤质对颗粒物分级测试结果的影响. 结果表明:①从决定系数和残差平方和角度分析,稀释采样法测试结果自身拟合性相对直接采样法差,实测数据点相对分散,95%置信带较宽. 直接采样法中,基于本文确定的清洗和收集方式,旋风采样器测试得到的PM₁₀和PM_2.5浓度拟合度高达0.999;双级虚拟撞击PM₁₀/PM_2.5采样器测试结果稳定性也较高,其测试得到的PM₁₀和PM_2.5浓度拟合度也为0.999. ②不同颗粒物采样器对烟气中PM_2.5、PM₁₀、TSP的浓度测试结果均表明,稀释采样法与直接采样法测试结果相关性较低;而各直接采样法之间呈高度相关,Pearson相关系数在0.993~0.999之间. ③稀释采样法分级测试结果显示,当颗粒物排放浓度较低时,各级滤膜称量误差叠加可导致PM₁₀、PM_2.5浓度测试结果误差较大. 因此,针对常温且颗粒物浓度较低的烟气,建议采用直接采样法;针对高温烟气,稀释采样法可捕集稀释降温过程中形成的可凝结颗粒物. ④各采样方法测试结果表明,相对于燃用无烟煤(如蜂窝煤),燃用烟煤产生的烟气中颗粒物浓度相对较高,且PM_2.5占比较高. 研究显示:稀释采样法能模拟燃煤污染源高温烟气排入大气环境中可凝结颗粒物的形成过程,测试结果更接近真实排放情况;对于常温颗粒物浓度较低的烟气,更适宜采用直接采样法.      

扬尘估算的采样方法对比

AP-42方法是目前较为权威的一种扬尘总量估算方法,泥沙承载量作为该方法中重要参数之一,采样方法对其结果有非常大的影响.在详细介绍了'AP-42提供的采样方法'与'TRAKER方法'两种采样方法的具体实施步骤基础上,对两种方法进行对比,发现'AP-42提供的采样方法'操作规范,但工作量大,易受外部影响;而'TRAKER方法'更为先进,对于科学研究具有较大的利用价值,但采样结果转换公式是经验公式,存在一定误差.     

Molecular Study of the Persistence of Infectious Human Norovirus on Food-Contact Surfaces

The aim of this study was to investigate the effect of relative humidity (RH) and temperature on norovirus (NoV) persistence as infectious particles on food-contact surfaces such as stainless steel and polyvinyl chloride (PVC). For this purpose, a new method combining enzymatic digestion with molecular beacon-based NASBA targeting the ORF1-ORF2 domain was developed to discriminate between infectious and noninfectious NoV. Stainless steel and PVC disks were contaminated with known amounts of human NoV and kept for 56 days at 7 and 20°C at high (86% ± 4%) and low (30% ± 10%) RH. NoV retained its putative infectivity for 56 days on PVC and for 49 days on stainless steel at 7°C and for 7 and 28 days, respectively, at low and high RH at 20°C on both tested surfaces. These results confirm that NoV persists in an infective state on inert surfaces for long periods of time and consequently may cause illness. The new molecular approach to detecting infectious NoV on inert surfaces may provide valuable information for evaluating environmental surface decontamination strategies.     

不同粉尘采样方法结果比较及相互转换

介绍了个体接触浓度采样法和环境浓度采样法,总粉尘浓度采样法和呼吸性粉尘浓度采样法,最高容许浓度(MAC)采样法与时间加权平均容许浓度(PC-TWA)采样法的结果比较与相互转换,对已有的研究结果做了概述.     

The Optimization of Methods for the Collection of Aerosolized Murine Norovirus

Globally, norovirus is the most common gastroenteritis causing pathogen. Annually, norovirus causes 685 million cases of acute gastroenteritis and 200,000     

固定源排放颗粒物采样方法的研究进展

固定源是大气可吸入颗粒物的主要来源之一,而目前国内对固定源排放颗粒物的监测和研究还相对比较薄弱,缺乏必要的技术手段和设备.文章总结了国内外固定源标准采样方法,分析了现有采样技术和采样设备的不足之处.而模拟烟气排放到大气真实环境中发生的物理化学变化的稀释采样正在成为研究固定污染源颗粒物排放特征的重要手段.文章阐述了固定源...     

Adhesion of Norovirus to Surfaces: Contribution of Thermodynamic and Molecular Properties Using Virus-Like Particles

Food and Environmental Virology - The aim of the study was to assess human norovirus and feline calicivirus (FCV) surface free energy, hydrophobicity, and ability to interact with fresh foods and...     

不同复苏方法对阴树脂复苏效果的影响研究

研究了直接酸洗复苏法、树脂转为氯型后酸洗复苏法、氯化钠转型后用次氯酸钠氧化复苏法、次氯酸钠氧化后用盐碱混液复苏法、双氧水氧化后用盐碱混液复苏法等五种不同的复苏方法对铁和有机物污染树脂复苏效果的影响。结果表明：复苏方法不同时，亚铁离子污染树脂的复苏效果和铁离子污染树脂的复苏效果是不相同的，同时可看出，双氧水氧化后盐碱混液复苏法复苏铁污染树脂的效果最好；五种不同的复苏方法对腐植酸污染的树脂都有较好的复苏效果，且复苏效果相差不大。     

The Influence of Temperature on Norovirus Inactivation by Monochloramine in Potable Waters: Testing with Murine Norovirus as a Surrogate for Human Norovirus

Human noroviral infections are generally more common during winters in temperate regions. This study used a murine norovirus (MNV) as a human norovirus surrogate to test the effect of water temperature (4 and 25°C) on virus survival and its susceptibility to the levels of monochloramine (~1.89 ppm) to terminally disinfect municipally treated potable waters. The titre of MNV remained essentially unchanged for at least 24 h in raw river water at both temperatures. The virus became undetectable in <2 h in monochloramine-containing samples held at 25°C, but its titre remained virtually unaltered at 4°C (P < 0.05) under the same conditions. These findings strongly suggest that water temperature can influence the norovirucidal activity of monochloramine and its possible impact on the seasonality of outbreaks of noroviral infections.     

TSP(IP)采样方法研究及比较

对现行的颗粒物采样的几种方法进行了对比研究。用几种方法同地点进行同步采样，样品微观观测结果表明；几种采样方法所采集的样品中，最大颗粒物的几何尺寸有明显的不同；样品分析及统计结果表明：几种采样方法所得监测结果之间存在着非常好的相关性。通过计算可相互通用，具有可比性；同时给出几种方法相互间的计算公式，可信度区间及可信度。     

道路交通扬尘采样方法研究进展

道路交通扬尘是我国北方城市环境空气中PM10和PM2.5污染的重要来源之一,要改善环境空气质量,进一步降低颗粒物浓度,必须采取有效措施控制道路交通扬尘,而控制扬尘的第一步是确定采样方法。目前我国还没有标准化的道路交通扬尘采样方法,本文归纳了当前国内外应用的主要方法,重点介绍了降尘法、积尘负荷法和快速检测法三种采样方法,并对三种方法进行了比较,分析了各自具有的优势和存在的问题。最后指出,快速检测法是最有潜力的采样方法,未来还需进一步的研究,争取早日制订出适合我国国情的标准化的道路交通扬尘采样方法。     

沉积物及水界面硝化与反硝化作用研究

河道是迁移转化来自陆源N素迁移的重要途径.准确测定河道中硝化、反硝化作用对流域N的收支平衡核算有重要意义.本文分析了河道中沉积物-水界面硝化、反硝化作用的原理并分析讨论了N2通量法、15N同位素法、抑制剂法的优缺点.建立科学、合理的实验方法将进一步完善流域N循环研究机制,为建立科学的流域管理措施提供技术支持.     

垃圾填埋场防渗层渗漏检测方法的比较

对目前国外几种垃圾填埋场防渗层渗漏检测方法(地下水检测法,扩散管法,电容传感法,追踪剂法,电化学感应电缆法,电学法)进行了详细的叙述和比较,并指出电学法是较好的检测方法.      

不同恢复方式对硝化颗粒污泥活性的影响

利用SBR反应器培养的成熟好氧硝化颗粒污泥,进行了硝化颗粒污泥临界活性以及不同氨氮浓度及曝气时间对储存的好氧硝化颗粒污泥活性恢复的影响研究.结果表明,储存不同时间的硝化菌活性(SOUR,O2/VSS)差别较大,储存前硝化颗粒污泥硝化菌SOUR为13.15 mg·(g·h)-1,储存20 d的硝化菌SOUR下降了1.26 mg·(g·h)-1,恢复运行了5个周期,氨氮去除率已经达到95%以上,恢复后活性为13.87 mg·(g·h)-1.但储存30 d的硝化菌SOUR降了11.63 mg·(g·h)-1,恢复运行51个周期后,氨氮去除率才达到92.64%,恢复后活性为14.92 mg·(g·h)-1,同时这种储存方法恢复时间较长,因此提出硝化颗粒污泥的临界活性为当硝化菌SOUR开始下降时,进行活性恢复.在临界活性的基础上,采用当硝化菌SOUR下降到临界活性时实施恢复,之后进入下一个储存周期,这种储存方式即为动态储存.当进水氨氮浓度分别为20、30、40 mg·L-1时,进行颗粒污泥活性恢复,进水氨氮浓度为40 mg·L-1恢复后硝化菌活性最高,经过3次动态储存后,其活性保持良好.当曝气时间分别为1、2、3 h时,进行颗粒污泥活性恢复,曝气时间为1 h时恢复后硝化菌活性最高,在动态储存过程中其活性一直保持较高水平.     

油田含油污水处理方法对比研究

石油因应用广泛,其开采量不断加大,而石油在开采过程中会产生大量的含油废水,油田采出水其水量大、水质成分复杂,若不有效处理,将对周围造成不利影响。通过论述油田含油污水的来源、分类、常用处理方法,并从三级处理、四个油形、COD去除效率三方面进行分析,最终得出富含浮油污水优选重力分离法进行处理;富含分散油污水优选浮选法、过滤法进行处理;富含乳化油污水优选膜分离法进行处理;富含溶解油污水优选活性污泥法进行处理。     

几种土壤重金属污染评价方法的对比

土壤重金属污染评价是土壤环境管理的基础性工作,采用合理的污染评价方法,才能识别土壤重金属污染状况,满足土壤环境管理和决策需要。选择单因子指数法、地质累积指数法、次生原生比值法和富集因子法四种方法进行对比分析,并进行实例计算和结果对比评价。结果表明:单因子指数法简单灵活,评价结果能直观反映土壤的污染情况,是比较实用的评价方法,但其将污染等级与元素实际含量简单地看成线性关系,导致评价结果较其它方法偏高。地质累积指数法是在单因子指数法基础上改进而来,计算公式较为简洁,污染评价结果较单因子指数法低。次生原生比值法突出元素形态的影响,未考虑元素总量和背景值,导致评价结果不能有效地与其它评价方法进行比较,在部分数据上还会出现污染浓度高却得到污染等级低的结论。富集因子法不仅考虑了元素的背景值,而且以地球化学过程中人为污染少、化学性质稳定、分析结果精确度高的Fe、Al、Si等元素作为参比元素,评价结果与地质累积指数法具有高度一致性,能够很好地体现土壤污染状况,但参比元素的实测值不易获取。研究结果可为土壤重金属污染评价方法的选取与改进提供基础数据。