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.     

生态高负荷土地快速渗滤系统处理猪场废水的效能及微生态

以经过厌氧反应器处理后的猪场废水为研究对象,探讨了新型生态高负荷土地快速渗滤系统对废水中COD、氨氮及总氮的去除情况;同时分析了该系统中的微生物群落结构以及过氧化氢酶、脲酶、硝酸盐还原酶的含量.结果表明,在水力负荷为11 cm·d~(-1)、进水COD为700 mg·L~(-1)左右的条件下,一级土地渗滤柱与二级土地渗滤柱对COD的去除率分别为78.8%与63.0%,总去除率达到了92.6%;而当水力负荷增大到22 cm·d~(-1)时,总的COD去除率仍在90.0%以上;该系统对于氨氮的去除更为显著,去除率可达99%左右.一级土地快速渗滤系统中上、中、下部的过氧化氢酶含量分别为1.899、0.990、0.323 m L·g~(-1),表明猪场废水中的有机物主要在该系统的上部与中部得以去除;二级土地快速渗滤系统上、中、下部的硝酸盐还原酶含量分别是3.453、3.831、1.971 m L·g~(-1),表明脱氮作用主要发生二级土地快速渗滤系统的上部与中部.该土地快速渗滤系统中微生物以Gram Negative与Gram Positive为主,特别是在二级土地快速渗滤系统中,AM Fungi与Actinomycetes占有一定比例,为猪场废水中难降解有机物的去除提供了保障.     

锌冶炼区耕地土壤和农作物重金属污染状况及风险评价

对贵州省某典型锌冶炼区耕地土壤和主要谷类农作物(稻米、玉米和小麦)进行取样调查,测定了土壤和谷类农作物中重金属Pb、Cd、Zn和Cu的含量,采用单因子污染指数和综合污染指数法评价了土壤和作物籽粒中重金属的污染状况,并分别采用潜在生态风险指数(RI)和危险商(HQ)法评价了土壤重金属污染的潜在生态风险程度和作物中重金属对成人和儿童的健康风险.结果表明:(1)冶炼区耕地土壤受到了重金属不同程度的污染,污染程度排序:玉米地水稻田小麦地,且CdCuZnPb,内梅罗综合污染指数分析结果表明,玉米地污染程度为重污染,水稻田和小麦地为轻污染,且均以Cd对综合指数的贡献最大;(2)土壤Cd存在极强生态风险,4种重金属潜在生态危害大小顺序为:CdPbCuZn;生态风险指数(RI)研究结果表明,研究区处于轻微、中等、强的和很强的生态风险程度的采样点比例分别为1.41%、21.1%、35.2%和42.3%.(3)冶炼区农作物稻米中重金属Pb、Cd、Zn、Cu的平均含量分别为0.145、0.017、16.97和2.704 mg·kg~(-1);玉米中重金属Pb、Cd、Zn、Cu的平均含量分别为0.094、0.055、26.81和4.464 mg·kg~(-1);小麦中重金属Pb、Cd、Zn、Cu的平均含量分别为0.048、0.085、35.37和5.426 mg·kg~(-1).(4)研究区稻米、玉米和小麦均存在重金属超标现象,重金属污染程度为小麦最重,稻米和玉米的污染程度相当,稻米和玉米污染等级均为安全,小麦处于警戒线.(5)各重金属元素每日摄入量均低于美国环保署的参考暴露剂量,安全性较好,但食用该区域谷类农作物引起复合重金属污染对成人和儿童均存在健康风险.(6)耕地土壤重金属含量与谷类农作物可食部分重金属含量间无明显相关性.     

正常污泥和膨胀污泥中EPS膜污染特性及其与膜表面作用能分析

胞外聚合物(Extracellular polymeric substance,EPS)是导致膜污染的重要物质,且不同形态污泥的EPS具有不同的性质,研究正常污泥和膨胀污泥的EPS膜污染特性具有重要意义.因此,实验考察了正常污泥和膨胀污泥的膜污染速率.结果表明,膨胀污泥膜污染速率远高于正常污泥,膨胀污泥EPS(Bulking sludge-EPS,BS-EPS)浓度((172.9±10.4) mg·g-1)明显高于正常污泥EPS(Normal sludge-EPS,NS-EPS)浓度((95.9±6 6) mg·g-1),且膨胀污泥的蛋白质/多糖(Protein/Carbohydrate,P/C)值(2.26)高于正常污泥的P/C值(1 97)此外,通过序批式吸附实验研究了正常污泥和膨胀污泥的EPS膜污染差异,并基于Derjaguine-Landaue-Verweye-Overbeek (XDLVO)理论计算了EPS与PVDF膜表面之间的作用能结果表明,EPS-膜系统的总界面能为负值,即EPS-膜系统之间会发生自吸附行为,而膜与BS-EPS的总界面能(-63.943mJ.m2)较膜与NS-EPS(-56.366 mJ·m2)具有较高的能量,因此,BS-EPS与膜的吸附力较强;NS-EPS和BS-EPS主要能量壁垒分别为8.89 kT和7.51 kT,表明NS-EPS与膜之间的排斥力更强,因而BS-EPS更容易造成膜污染