Norovirus GII.Pe Genotype: Tracking a Foodborne Outbreak on a Cruise Ship Through Molecular Epidemiology,Brazil, 2014

Norovirus (NoV) is recognized as the most common cause of foodborne outbreaks. In 2014, an outbreak of acute gastroenteritis occurred on a cruise ship in Brazil, and NoV became the suspected etiology. Here we present the molecular identification of the NoV strains and the use of sequence analysis to determine modes of virus transmission. Food (cream cheese, tuna salad, grilled fish, orange mousse, and vegetables soup) and clinical samples were analyzed by ELISA, conventional RT-PCR, qRT-PCR, and sequencing. Genogroup GII NoV was identified by ELISA and conventional RT-PCR in fecal samples from 5 of 12 patients tested (41.7%), and in the orange mousse food sample by conventional RT-PCR and qRT-PCR. Two fecal GII NoV samples and the orange mousse GII NoV sample were successfully genotyped as GII.Pe (ORF 1), revealed 98.0–98.8% identities among them, and shared phylogenetically distinct cluster. Establishing the source of a NoV outbreak can be a challenging task. In this report, the molecular analysis of the partial RdRp NoV gene provided a powerful tool for genotyping (GII.Pe) and tracking of outbreak-related samples. In addition, the same fast and simple extraction methods applied to clinical samples could be successfully used for complex food matrices, and have the potential to be introduced in routine laboratories for screening foods for presence of NoV.     

Use of Preservative Agents and Antibiotics for Increased Poliovirus Survival on Positively Charged Filters

Environmental surveillance of poliovirus (PV) and other non-enveloped viruses can help identify silent circulation and is necessary to certify eradication. The bag-mediated filtration system is an efficient method to filter large volumes of environmental waters at field sites for monitoring the presence of viruses. As filters may require long transit times to off-site laboratories for processing, viral inactivation or overgrowth of bacteria and fungi can interfere with virus detection and quantification (Miki and Jacquet in Aquatic Microb Ecol 51(2):195–208, 2008). To evaluate virus survival over time on ViroCap^? filters, the filters were seeded with PV type 1 (PV1) and/or MS2 and then dosed with preservatives or antibiotics prior to storage and elution. These filters were stored at various temperatures and time periods, and then eluted for PV1 and MS2 recovery quantification. Filters dosed with the preservative combination of 2% sodium benzoate and 0.2% calcium propionate had increased virus survival over time when stored at 25 °C, compared to samples stored at 25 °C with no preservatives. While elution within 24 h of filtration is recommended, if storage or shipping is required then this preservative mixture can help preserve sample integrity. Addition of an antibiotic cocktail containing cephapirin, gentamicin, and Proclin^? 300 increased recovery after storage at 4 and 25 °C, when compared to storage with no antibiotics. The antibiotic cocktail can aid sample preservation if access to appropriate antibiotics storage is available and sample cold chain is unreliable. This study demonstrated that the use of preservatives or antibiotics is a simple, cost-effective method to improve virus detection from ViroCap cartridge filters over time.     

DNA Heat Treatment for Improving qPCR Analysis of Human Adenovirus in Wastewater

PCR inhibitory substances in complex sample matrices can cause false negatives or under-estimation of target concentration. This study assessed DNA heat treatment for reducing inhibition during qPCR analysis of human adenovirus (HAdV) in wastewater samples. Inhibition was reduced by heat treating DNA, where mean HAdV concentration was increased by 0.71 log₁₀ GC/L (and up to 3.04 log₁₀ GC/L in one case), and replicate variability and false negatives were reduced. DNA heat treatment should be further investigated for improving reliability of HAdV concentration estimates in water, which can support more accurate assessment of health risks associated with viral pathogen exposure.     

Assessment and Evaluation of an Integrated Hybrid Anaerobic–Aerobic Sewage Treatment System for the Removal of Enteric Viruses

The capability of a cost-effective and a small size decentralized pilot wastewater treatment plant (WWTP) to remove enteric viruses such as rotavirus, norovirus genogroup I (GGI), norovirus genogroup II (GGII), Hepatitis E virus (HEV), and adenovirus was studied. This pilot plant is an integrated hybrid anaerobic/aerobic setup which consisted of anaerobic sludge blanket (UASB), biological aerated filter (BAF), and inclined plate settler (IPS). Both the UASB and BAF are packed with a non-woven polyester fabric (NWPF). Results indicated that the overall log₁₀ reductions of enteric viruses’ genome copies through the whole system were 3.1 ± 1, 3.3 ± 0.5, and 2.6 ± 0.9 log₁₀ for rotavirus, norovirus GGI, and adenovirus, respectively. Reduction efficiency for both norovirus GGII and HEV after the different treatment steps could not be calculated because there were no significant numbers of positive samples for both viruses. The overall reduction of rotavirus infectious units through the whole system was 2.2 ± 0.8 log₁₀ reduction which is very close to the overall log₁₀ reduction of adenovirus infectious units through the whole system which was 2.1 ± 0.8 log₁₀ reduction. There was no considerable difference in the removal efficiency for different rotavirus G and P types. Adenovirus 41 was the only type detected in the all positive samples. Although the pilot WWTP investigated is cost effective, has a small footprint, does not need a long distance network pipes, and easy to operate, its efficiency to remove enteric viruses is comparable with the conventional centralized WWTPs.     

苏州市PM2.5中水溶性离子的季节变化及来源分析

2015年在苏州市城区采集大气细颗粒物PM_(2.5)样品共87套,用重量法分析了PM_(2.5)的质量浓度,离子色谱法分析了颗粒物中F-、Cl-、NO_3~-、SO_4~(2-)、Na~+、NH_4~+、K~+、Mg~(2+)和Ca~(2+),共9种水溶性无机离子.观测期间,苏州市PM_(2.5)的年均质量浓度为(74.26±38.01)μg·m-3,其季节特征为冬季春季秋季夏季;9种水溶性离子的总质量浓度为(43.95±23.60)μg·m~(-3),各离子的浓度高低顺序为NO_3~-SO_4~(2-)NH_4~+Na~+Cl~-K~+Ca~(2+)F-Mg~(2+);SNA(SO_4~(2-)、NO_3~-和NH_4~+三者的简称)是最主要的水溶性离子;SO_4~(2-)、NO_3~-和NH_4~+三者之间具有显著的相关性,它们在PM_(2.5)中主要是以NH_4NO_3和(NH_4)_2SO_4的结合方式存在.苏州市PM_(2.5)中水溶性离子的主要来源包括工业源、燃烧源、二次过程和建筑土壤尘等.     

南京冬季市区和郊区气溶胶中PAHs浓度的昼夜特征及粒径分布

为了研究南京市区与郊区气溶胶中多环芳烃(PAHs)污染状况和分布特征,利用气-质联用仪(GC-MS)分析了2010年1月1～10日日间和夜间分别在南京大学和南京信息工程大学采集的气溶胶样品,得到南京市区与郊区17种PAHs浓度,总浓度分别为41.36～220.35 ng.m-3和45.10～200.86 ng.m-3,其中约66%～67%分布于细粒子(Dp≤2.1μm)中.研究发现,南京市区和郊区气溶胶中PAH总浓度均处于较高的水平;但两者昼夜变化趋势不同,即市区PAH总浓度日间高于夜间,郊区PAH总浓度日间低于夜间.主导风向的改变和高压天气系统对PAH浓度变化影响较大;在市区其影响主要表现在细粒子部分,而郊区主要表现在粗粒子部分.市区和郊区不同环数的PAHs粒径分布不同;2～3环PAHs,郊区含量高于市区;而4～6环PAHs,市区含量高于郊区.高环数(4～6环)PAHs在粗模态出现较大浓度峰可能是由于南京地区粗模态气溶胶中碳含量较高.市区和郊区相似的特征比值说明两者的PAHs具有相同污染来源,主要为生物质及煤的燃烧和汽车尾气,表明南京市区PAHs受到郊区工业源排放影响较大.     

粉煤灰综合利用过程中汞的二次释放规律研究

我国粉煤灰年产生量4亿t左右,近年来利用率稳定在65%～68%,主要利用方式包括建材生产、道路施工、建筑工程和农业利用.建材生产包括高温工序,可能存在粉煤灰中汞的二次释放.本研究设计实验模拟了水泥生产、蒸养砖生产的主要环境因素,利用程序升温脱附的方法研究粉煤灰利用过程中的汞迁移转化规律,并对全国范围内粉煤灰利用过程汞的二次释放量做出估算.研究发现,粉煤灰中的汞以HgCl2(Hg2Cl2)、HgS和HgO的形式存在;水泥生产过程中,粉煤灰中98%以上的汞会释放;蒸养砖生产过程中,粉煤灰中汞的平均释放率为28%,释放率主要受到粉煤灰中的HgCl2(Hg2Cl2)比例的影响.我国粉煤灰利用过程中的汞二次排放量由2002年的4.07 t.a-1增至2008年的9.18 t.a-1,其中水泥行业的贡献率占到96.6%.