Estimation of Human Norovirus Infectivity from Environmental Water Samples by In Situ Capture RT-qPCR Method

Human noroviruses (HuNoVs) are highly infectious viruses for which water is an important medium of transmission. In this study, we explored a new in situ capture RT-qPCR (ISC-RT-qPCR) methodology to estimate the infectivity of HuNoV in environmental water samples. This assay was based on capturing encapsidated HuNoV by viral receptors, followed by in situ amplification of the captured viral genomes by RT-qPCR. We demonstrated that the ISC-RT-qPCR did not capture and enable signal amplification of heat-denatured Tulane Virus (TV) and HuNoVs. We further demonstrated that the sensitivity of ISC-RT-qPCR was equal or better than that of conventional RT-qPCR procedures for the detection of HuNoV GI and GII. We then utilized the ISC-RT-qPCR to detect HuNoV in environmental water samples for comparison against that from a conventional RT-qPCR procedure. TV was used as a process control virus. While complete inhibition of TV genomic signal was observed in 27% of samples tested by RT-qPCR, no inhibition of TV genomic signal was observed by ISC-RT-qPCR. From 72 samples tested positive for HuNoV GI signal by RT-qPCR, only 20 (27.8%) of these samples tested positive by ISC-RT-qPCR, suggesting that 72.2% of RT-qPCR-positive samples were unlikely to be infectious. From 16 samples tested positive for HuNoV GII signal by RT-qPCR, only one of these samples tested positive by ISC-RT-qPCR. Five samples that had initially tested negative for HuNoV GII signal by RT-qPCR, was tested as positive by ISC-RT-qPCR. Overall, ISC-RT-qPCR method provided an alternative assay to estimate infectivity of HuNoV in environmental samples.     

Comparative Virucidal Efficacy of Seven Disinfectants Against Murine Norovirus and Feline Calicivirus,Surrogates of Human Norovirus

Human noroviruses (HuNoV) are the leading cause of acute non-bacterial gastroenteritis in humans and can be transmitted either by person-to-person contact or by consumption of contaminated food. A knowledge of an efficient disinfection for both hands and food-contact surfaces is helpful for the food sector and provides precious information for public health. The aim of this study was to evaluate the effect of seven disinfectants belonging to different groups of biocides (alcohol, halogen, oxidizing agents, quaternary ammonium compounds, aldehyde and biguanide) on infectious viral titre and on genomic copy number. Due to the absence of a cell culture system for HuNoV, two HuNoV surrogates, such as murine norovirus and feline calicivirus, were used and the tests were performed in suspension, on gloves and on stainless steel discs. When, as criteria of efficacy, a log reduction >3 of the infectious viral titre on both surrogates and in the three tests is used, the most efficacious disinfectants in this study appear to be biocidal products B, C and D, representing the halogens, the oxidizing agents group and a mix of QAC, alcohol and aldehyde, respectively. In addition, these three disinfectants also elicited a significant effect on genomic copy number for both surrogate viruses and in all three tests. The results of this study demonstrate that a halogen compound, oxidizing agents and a mix of QAC, alcohol and aldehyde are advisable for HuNoV disinfection of either potentially contaminated surfaces or materials in contact with foodstuffs.     

Performance of Manufacturer Cleaning Recommendations Applied to 3D Food Ink Capsules for the Control of a Human Norovirus Surrogate

With the widespread availability of 3D food printing systems for purchase, users can customize their food in new ways. Manufacturer recommendations for cleaning these machines remain untested with regard to the prevention of foodborne pathogen transmission. This study aimed to determine if manufacturer cleaning recommendations for food ink capsules utilized in 3D food printers are adequate to control human norovirus (HuNoV). A HuNoV surrogate, Tulane virus (TuV; ~?6 log₁₀ PFU/mL), was inoculated onto the interior surface of stainless steel food ink capsules. Capsules were either unsoiled or soiled with one of the following: butter, protein powder solution, powdered sugar solution, or a mixture containing all three food components. The capsules were allowed to dry and then one of three hygienic protocols was applied: manual washing (MW), a dishwasher speed cycle (DSC), or a dishwasher heavy cycle (DHC). The interaction effect between DSC and pure butter was a significant predictor of log reduction (P?=?0.0067), with the pure butter and DSC combination achieving an estimated mean log reduction of 4.83 (95% CI 4.13, 5.59). The DSC was the least effective method of cleaning when compared with MW and the DHC. The 3-way interaction effects between wash type, soil, and capsule position were a significant predictor of log reduction (P?=?0.00341). Capsules with butter in the DSC achieved an estimated mean log reduction of 2.81 (95% CI 2.80, 2.83) for the front-most position versus 6.35 (95% CI 6.33, 6.37) for the back-most position. Soil matrix, cleaning protocol, and capsule position all significantly impact capsule cleanability and potential food safety risk. The DHC is recommended for all capsules, and the corners should be avoided when placing capsules into the dishwasher. The current study seeks to provide recommendations for users of additive manufacturing and 3D food printing including consumers, restaurants, industry, and regulatory industries.

     

Prevalence of Human Noroviruses in Frozen Marketed Shellfish,Red Fruits and Fresh Vegetables

Noroviruses (NoVs), currently recognised as the most common human food-borne pathogens, are ubiquitous in the environment and can be transmitted to humans through multiple foodstuffs. In this study, we evaluated the prevalence of human NoV genogroups I (GI) and II (GII) in 493 food samples including soft red fruits (n = 200), salad vegetables (n = 210) and bivalve mollusc shellfish (n = 83), using the Bovine Enterovirus type 1 as process extraction control for the first time. Viral extractions were performed by elution concentration and genome detection by TaqMan Real-Time RT-PCR (RT-qPCR). Experimental contamination using hepatitis A virus (HAV) was used to determine the limit of detection (LOD) of the extraction methods. Positive detections were obtained from 2 g of digestive tissues of oysters or mussels kept for 16 h in seawater containing 2.0–2.7 log₁₀ plaque-forming units (PFU)/L of HAV. For lettuces and raspberries, the LOD was, respectively, estimated at 2.2 and 2.9 log₁₀ PFU per 25 g. Of the molluscs tested, 8.4 and 14.4 % were, respectively, positive for the presence of GI NoV and GII NoV RNA. Prevalence in GI NoVs varied from 11.9 % for the salad vegetables samples to 15.5 % for the red soft fruits. Only 0.5 % of the salad and red soft fruits samples were positive for GII NoVs. These results highlight the high occurrence of human NoVs in foodstuffs that can be eaten raw or after a moderate technological processing or treatment. The determination of the risk of infection associated with an RT-qPCR positive sample remains an important challenge for the future.     

Virucidal Efficacy of a Hydrogen Peroxide Nebulization Against Murine Norovirus and Feline Calicivirus,Two Surrogates of Human Norovirus

Human noroviruses (HuNoV) are amongst the leading causes of acute non-bacterial gastroenteritis in humans and can be transmitted via person-to-person contact, via contact with contaminated surfaces or by consumption of contaminated food. Contaminated surfaces in healthcare settings contribute to the transmission of viruses. No-touch automated room disinfection systems might prevent such a spread of contamination and thus their virucidal effect needs to be evaluated. The aim of this study was to assess the efficacy of a nebulization system spraying hydrogen peroxide on two main surrogates of HuNoV, namely murine norovirus (MNV) and feline calicivirus (FCV). The viruses were dried on cover glasses and on stainless steel discs and exposed to nebulization. The number of infectious viral particles and genomic copies before and after the nebulization was compared. The efficacy in reducing infectivity of both surrogates was demonstrated. For the infectious viral titre of MNV and FCV, a log₁₀ reduction factor ≥4.84 and 4.85 was observed after nebulization, respectively, for tests on cover glasses and ≥3.90 and 5.30, respectively, for tests on stainless steel discs. Only low reductions in genomic copy numbers were observed for both surrogates. The nebulization of hydrogen peroxide showed a clear virucidal effect on both HuNoV surrogates, MNV and FCV, on two different carriers and the use of nebulization should be promoted in complementarity with conventional disinfection methods in healthcare settings and food processing facilities to reduce viral load and spread of contamination.     

Locating groundwater monitoring wells near cutoff walls

This paper presents a modeling study of two five-well monitoring networks designed for a rectangular landfill oriented oblique to regional groundwater flow. Respectively, a ‘linear’ and ‘non-linear’ network ignored and considered the hydraulic head field induced by a cutoff wall at the landfill's up-gradient boundary. All wells were located the same distance (measured parallel to groundwater flow) from the down-gradient boundary of the landfill, and there was an equal spacing (measured perpendicular to groundwater flow) between wells in each network. A simulation model was used to compute the detection efficiency of each monitoring network. In the non-linear network, wells clustered near the down-gradient corner of the landfill registered a 100% detection efficiency. This strategy was effective because the cutoff wall induced convergent groundwater flow beneath the landfill. Inter-well spacing was too high, and consequently detection efficiency was lower in the linear network. Two outer wells in that network detected no contaminant plumes. This study suggests that distorted hydraulic head fields induced by partial cutoff walls should be considered when designing detection monitoring networks at landfills.     

Variables influencing growth and morphology of colonies of cells from human amniotic fluid

Growth of cells from amniotic fluid was studied with respect to cell concentration in the inoculum, blood contamination of the fluid, fluid colour, fluid clarity, gestational age of the pregnancy, and growth factors. Dependent variables measured were colony formation, colony size, and colony morphology after 7, 11, and 14 days of culture. The following conclusions were established from these studies: small sample volumes are the most efficient for producing colonies; cells from very bloody or dark brown fluids have a slower rate of growth; growth of cells from cloudy (noncontaminated) fluids is better than growth of cells from clear fluids; the proportion of colonies that are epithelioid varies with gestational age; the stimulating effect of 100 ng/ml fibroblast growth factor on cells from amniotic fluid was confirmed.     

大气汞监测技术进展

为了切实履行《关于汞的水俣公约》,落实美丽中国建设战略,我国亟待加强对大气环境及各涉汞行业的监管.采用文献调研、分类梳理的方法,系统地分析和总结了近年来国内外常用的大气汞监测技术现状与问题,并探讨了未来技术发展方向.结果表明:为了满足大气环境中低浓度多形态汞的监测需求,越来越多功能的大气汞监测技术被研发,当前基于AFS（原子荧光光谱）和AAS（原子吸收光谱）等方法的监测技术对Hg0（元素汞）可实现ng/m3的检出限,对Hg2+（氧化态汞）和Hgp（颗粒态汞）有着低至pg/m3的检出限.自动化采样监测技术是未来汞监测的发展趋势,当前技术对大气中Hg0有着2.5~5.0 min水平的时间分辨率,对大气中Hgp可达到1~2 h的时间分辨率,同时可实现远程传输数据.人工采样监测和被动采样监测技术虽然时间分辨率较差,但在Hg2+和Hgp的监测方面有着一定优势.研究显示,完善大气汞监测管理体系,除了需要研发更强大的Hg2+和Hgp测定技术用于理解相关迁移转化过程,还亟待在相应的标定方法上实现技术革新,并应用这些技术组建地区和全球大气汞监测网络.      

济南市大气颗粒物短期暴露对老年人群血清淀粉样蛋白P组分影响的定群研究

为了解济南市不同粒径大气颗粒物暴露对老年人群血清淀粉样蛋白P组分（SAP）的急性影响,评估大气颗粒物暴露对人群神经系统的健康风险,以山东省济南市甸柳社区为研究地点,采用定群研究设计,对社区76名健康老年人进行5次重复测量,结合社区附近（< 2 km）环保监测超级站的PM_2.5和PM₁₀浓度数据,在控制年龄、性别等协变量情况下,利用线性混合效应模型分别分析PM_2.5和PM₁₀暴露对SAP的效应影响.结果表明:大气颗粒物短期暴露与老年人群SAP的升高呈正相关;PM_2.5累积滞后0~6 h,每升高1个四分位数间距（IQR）,SAP水平升高18.73%（95% CI为9.20%~29.08%）（FDR_H-P < 0.05）（其中,CI为可信区间,FDR_H-P为经多重校正后的P值）;PM₁₀累积滞后0~6 h,每升高1个IQR,SAP水平亦升高,但变化不显著（FDR_H-P>0.05）.研究显示,大气颗粒物暴露可以引起人群反应神经退行性病变的SAP指标升高,提示大气颗粒物对人群神经系统具有潜在威胁;PM_2.5对SAP的效应高于PM₁₀,提示小粒径颗粒物具有较高的健康危害,应加强对小粒径颗粒物的健康影响和干预防护研究.      

Towards real-time detection of wastewater in surface waters using fluorescence spectroscopy

The presence of municipal wastewater at the intake of a major drinking water treatment facility located on Lake Ontario was examined using fluorescence data collected during a period of continuous monitoring. In addition, controlled mixing of lake water and wastewater sampled from a local treatment facility were conducted using a bench-scale fluorescence system to quantify observed changes in natural organic matter. Multivariate linear regression was applied to components derived from parallel factors analysis. The resulting mean absolute error for predicted wastewater level was 0.22% (V/V, wastewater/lake water), indicating that wastewater detection at below 1.0% (V/V) was possible. Analyses of sucralose, a wastewater indicator, were conducted for treated wastewater as well as surface water collected at two intake locations on Lake Ontario. Results suggested minimal wastewater contribution at the drinking water intake. A wastewater detection model using a moving baseline was developed and applied to continuous fluorescence data collected at one of the drinking water intakes, which agreed well with sucralose results. Furthermore, the simulated addition of 1.0% (V/V) of wastewater/wastewater was detectable in 89% of samples analyzed, demonstrating the utility of fluorescence-based wastewater monitoring.     

基于物联网的智能环境污染检测与控制系统研究

为了实行对环境污染的远程智能检测,提出基于物联网技术的智能环境污染检测与控制系统设计方法,采用无线传感组网技术进行智能环境污染检测系统物联网设计,实现对环境污染信息采样,采集的环境污染数据主要有水污染数据、土壤污染数据和大气污染数据等。将数据输入的集成控制单元中,通过上位机进行远程传输控制,采用TMS320VC5409A智能处理DSP芯片作为集成控制芯片进行控制系统硬件设计,环境污染检测与控制系统硬件模块分为物联网传感模块、AD模块、上位机通信模块和数据输出接口模块等。在嵌入式Linux内核中实现环境污染检测程序加载,实现污染检测和优化控制。测试结果表明,采用该方法进行环境污染检测与控制的智能性较好,准确度较高,实时性较好,总体性能较优。     

环境监测优化布点黑箱理论模型的研究

提出黑箱理论模型优化环境监测布点，根据Ｍｏｏｒｅ Ｐｅｎｒｏｓｅ广义逆理论提出二次多项式方程的最小范数算式。应用于大气监测优化布点的实例结果表明，该方法用于监测优化布点是可靠的     

电网视频远程监控系统在环境污染治理中的应用研究

为提高环境污染治理水平,设计了电网视频远程监控系统,将改进系统应用到环境污染治理中,分析其对环境污染治理效果的影响。依据电网视频远程监控系统的整体规划,对组管理服务器、采集服务器、客户端等系统硬件部分进行改进,优化软件部分的应用层、会话层及数据处理层各功能模块,完成电网视频远程监控系统的设计。通过系统清晰度检测,分析电网视频远程监控系统在环境污染治理中的应用效果。实验结果表明,该系统实时性好、清晰度高、监控耗能低、监控准确度高,能够为环境污染治理提供极大的帮助。     

Nucleic Acid Amplification-Based Methods for Detection of Enteric Viruses: Definition of Controls and Interpretation of Results

The most effective methods for virus detection in food and environmental samples are those based on nucleic acid amplification. Complex methods must be applied by the analyst in order to control for false negative results of virus detection assays in those samples that may be contaminated by virus concentrations above the detection level. The verification of analytical results is a necessity and this depends on using an appropriate suite of controls to monitor the efficacy of the critical steps in the method and allow correct result interpretations to be made. We describe the suite of controls necessary for analysing food and environmental samples for the presence of enteric viruses by nucleic acid amplification-based methods. To exclude false negative and positive interpretations of results, the inclusion of this suite of controls will be essential when considering incorporating monitoring of viruses in food or environmental safety management plans.