厌氧水环境中噬菌体MS2的存活和团聚特性及理化因素的影响

为探究病毒在厌氧水环境中的存活特性,以噬菌体MS2为模式病毒,采用双层平板法进行噬菌体MS2的定量检测,研究温度、pH值、悬浮颗粒物、乙酸等理化条件对噬菌体MS2的影响,分析其衰减动力学特征,并通过测定Zeta电位和噬菌斑直径考察噬菌体颗粒在不同条件下的聚集状况.结果表明,在厌氧水环境中,噬菌体MS2的衰减符合一阶指数衰减模型.在众多研究因素中,温度是影响噬菌体MS2存活的最主要因素.噬菌体MS2在4,17,25和35℃时的T₉₀分别为20.36,6.14,5.15和0.46d.35℃条件下12h后噬菌体失活率高达2.44lg,而4℃条件下7d后噬菌体失活率仅有0.78lg.增加乙酸浓度能明显提高噬菌体MS2的衰减速率.低pH值和悬浮颗粒物条件会促进噬菌体的团聚,使噬菌体颗粒Zeta电位降低,水力学直径增大,但悬浮颗粒物浓度过高会影响颗粒间的静电作用.噬菌体的团聚也增加了噬菌斑的直径,pH=6和20mg/L的悬浮颗粒物条件下,直径1.0mm以上的大噬菌斑数量占比分别达到了45.61%和57.74%.明确厌氧水环境中各种理化因素对噬菌体MS2的影响,可为水环境病毒控制提供科学依据.     

影响病毒在物体表面和空气中生存的因素分析

新型冠状病毒肺炎（COVID-19）疫情严重危害人类生命安全和影响国家经济发展.截至目前,关于环境条件（湿度与温度）、空气污染及人为活动对2019-nCoV传播的文献鲜见报道.由于新型冠状病毒（2019-nCoV）采样困难,难以开展直接试验研究,因此通过对国内外其他类似病毒文献调研,探究影响病毒传播和生存的影响因素,以期为疫情防控和控制2019-nCoV传播提供指导.结果表明:①冠状病毒在低温低湿的环境中能够保持较长的存活时间与传播力,尤其在干燥物体表面停留时间更长,如冠状病毒能在温度为22~25℃、相对湿度（RH）为40%~45%的光滑物体表面存活5 d以上;②同时,冠状病毒气溶胶在低温低湿的条件下也具有高存活率,提示在密闭环境中2019-nCoV气溶胶可能在温度为20~25℃、相对湿度为30%~50%的条件下具有较高的存活率;③颗粒物浓度越高,2019-nCoV传播的风险可能随之增加;④感染者的行为活动也对2019-nCoV的传播起着不可忽视的作用,应避免在人群中打喷嚏、咳嗽,并且用手、手肘或其他物品加以防护,降低病毒传播的可能性.综上,提出几点疫情防控的相关建议,如改善室内微环境和开展2019-nCoV的检测、灭活、影响因素、传播规律等相关研究工作.      

水环境中病毒污染及监测问题探讨

水环境中广泛存在着病毒污染,而现行的水环境标准对于病毒缺乏相关的规定,许多病毒引起的疾病介水传播,这就可能造成致病微生物对人体健康的潜在威胁.在此,阐述水环境中病毒的危害,探讨可行的水体中病毒的指示物、监测手段,以及对病毒的灭活等相关问题.     

藻细胞和高岭土的存在对病毒MS2存活的影响

选用病毒MS2作为水中肠道病毒的指示病毒,高岭土和铜绿微囊藻分别作为无机颗粒物和有机颗粒物,研究颗粒物浓度、pH值、不同价态离子浓度、天然有机物(NOM)等水质条件下,无机(高岭土)、有机(铜绿微囊藻)颗粒物存在对病毒MS2存活的影响.结果表明,无机颗粒物高岭土对病毒MS2的存活无明显影响,但当水体钙硬度(钙离子产生的硬度)较大时,病毒MS2的表观存活量增加1个对数;铜绿微囊藻的存在会导致病毒MS2的存活量降低1个对数左右,但当溶液的pH值大于4.0或铜绿微囊藻的浓度小于1.0×106cells·L-1时,藻类对病毒的生存无明显影响;当水体钙硬度较大时,藻反而会增加病毒MS2的存活对数.因此在高浊水、高藻水中,水的钙硬度增加会使水体中病毒生存能力变强,进而增加饮用水的安全风险.     

长江口近岸水体悬浮颗粒物多环芳烃分布与来源辨析

对长江口近岸水体悬浮颗粒物中的多环芳烃(PAHs)进行了定量分析.结果表明,悬浮颗粒物PAHs总量为2 278.79～14 293.98 ng/g,排污口附近浓度最高,远离排污口浓度降低;就其组成特征而言,以4～6环PAHs为主,2～3环PAHs相对较少.聚类分析表明.除了城市排污外,河口水动力条件也对近岸PAHs分布特征产生一定影响.此外,悬浮颗粒物浓度、有机碳、炭黑含量也是控制近岸PAHs分布的重要影响因素.主成分分析和PAHs特征参数分析发现,近岸水环境中PAHs的主要来源为矿物燃料的不完全燃烧,此外还有少量石油输入.生态风险评价结果显示,大部分PAH化合物均超过ER-L值和ISQV-L值,表明长江口近岸水体悬浮颗粒物中的PAHs已具有不利的生物影响效应.     

病毒在无生命物体表面存活时间及其影响因素分析

2020年3月11日,世界卫生组织宣布新型冠状病毒肺炎（COVID-19,简称“新冠肺炎”）疫情全球大流行.病毒性传染病在人群中的发生需具备3个相互关联的条件,即传染源、传播途径和易感人群,其中传播途径是一个关键环节.从环境传播的角度,综述了包括SARS（重症急性呼吸综合征）病毒、MERS（中东呼吸综合征）病毒等病毒在无生命物体表面的存活时间及影响因素.结果表明:包括冠状病毒在内的一些具有传染性的病毒能在多种无生命物体表面存活一定时间,进而可能通过物体表面进行环境传播,造成潜在的健康风险;物体材质、温度、湿度以及病毒载量等因素是影响病毒在无生命物体表面存活的主要因素.在上述分析的基础上,对医疗废物和生活垃圾的收集、运输、处理等过程提出了针对性的防控措施,以期对新型冠状病毒（2019-nCoV）以及未来可能出现的其他病毒的环境传播防控对策提供参考.      

东湖通道工程施工废水的理化特性与处置对策研究

为认识东湖通道施工废水的基本特性,以不受施工干扰的湖心水样为对照,对东湖通道施工过程中产生的生产、生活废水进行了分析.结果发现,通道施工废水的主要特征是悬浮颗粒物、营养物质、还原性物质等物质的含量高,水体呈碱性;其中,坑道废水悬浮颗粒物多、浊度高;桩机打桩废水悬浮颗粒物多、浊度高、营养物质含量高;搅拌施工废水氧化还原电位高、悬浮颗粒物含量高;材料堆放废水高锰酸盐指数高,生活污水浊度高、营养物质和水中油的含量高,粪大肠杆菌群超标.坑道废水、桩机打桩废水、搅拌施工废水和生活污水均为劣Ⅴ类,材料堆放废水符合Ⅳ类水标准.针对东湖通道施工废水的理化特性,鉴于国家5A级景区对水质的较高要求,建议在施工区构建临时沉淀池、小型氧化塘等设施开展原位污废水处置,确保达标排放,避免影响水生态环境.     

模拟扰动条件下太湖水体悬浮物的结构特性

设计了1个水体扰动模拟实验装置, 通过该装置中的光散射分析仪和图象采集、分析系统,能在线记录底泥受到不同强度扰动时水体中悬浮颗粒物的变化过程.利用该装置,研究了太湖梅梁湾3个采样点的底泥在受到不同强度扰动后,水体浊度变化规律和悬浮颗粒物结构特性.结果表明,底泥含水率及粒度分布对底泥受扰动后再悬浮的潜力有较大影响,含水率越高,再悬浮的潜力越大.随着扰动强度增大,水体的浊度相应增加,水体中悬浮颗粒物中值粒径也增大,分别从6.63, 5.79, 5.51 μm上升到12.49, 11.38, 13.33 μm.悬浮颗粒物图象分析表明,颗粒物在扰动中发生聚集、絮凝过程.颗粒物图象的形态学分析表明,悬浮颗粒物形状呈现分形现象,颗粒物的长短比在1.27～1.52之间,分形维数D₂则在1.72～1.94之间.     

太湖水体散射特性及其与悬浮物浓度关系模型

水体散射特性与水环境参数如悬浮颗粒及其浓度密切相关.利用Wetlabs公司研制的水体固有光学特性测量系统,于2006-10～2006-11对太湖水体的散射系数和后向散射系数进行了测量.在对水体散射特性分析的基础上,建立了太湖水体颗粒物后向散射系数的光谱模型.此外,利用后向散射率计算水体折射系数,根据折射系数的变化范围,将水中颗粒物的主导因子分为3种类型：①浮游植物主导;②无机颗粒物主导;③两者共同主导.在对散射系数与无机悬浮物、有机悬浮物、总悬浮物浓度相关性分析的基础上,针对颗粒物主导因子的不同,分别建立了散射系数与无机悬浮物浓度的乘幂关系模型.     

再生水补给型水体中病原微生物的来源、归宿及其风险评估

我国北方地区,以污水处理厂出水等非常规水源补给为主的河/湖逐渐成为城市群下游河流的主导类型;以污水处理厂尾水为补给水源的湿地公园近年来亦大量涌现.在节约城市用水、改善生态环境的同时,污水所携带的病原体对环境安全和人类健康带来潜在的威胁.为评估水环境病原微生物污染的健康风险,深入了解水体中病毒的类型以及病毒污染与环境因素之间的关系,认识病原微生物在水环境系统中的迁移轨迹、人暴露风险点及暴露途径具有重要的意义.针对典型再生水补给型水体存在的公共卫生安全问题,借鉴同时探索污水流行病学方法,并结合微生物定量风险评估（QMRA）分析,通过“污染-风险评价-风险预警”的流程,可以实现病原微生物污染全过程风险管理,为城市水环境的管理和良性运行提供理论支撑.     

One-year Surveillance of Human Enteric Viruses in Raw and Treated Wastewaters,Downstream River Waters,and Drinking Waters

Human enteric viruses are a major cause of waterborne diseases, and can be transmitted by contaminated water of all kinds, including drinking and recreational water. The objectives of the present study were to assess the occurrence of enteric viruses (enterovirus, norovirus, adenovirus, hepatitis A and E virus) in raw and treated wastewaters, in rivers receiving wastewater discharges, and in drinking waters. Wastewater treatment plants’ (WWTP) pathogen removal efficiencies by adenovirus quantitative real-time PCR and the presence of infectious enterovirus, by cell culture assays, in treated wastewaters and in surface waters were also evaluated. A total of 90 water samples were collected: raw and treated wastewaters (treated effluents and ultrafiltered water reused for industrial purposes), water from two rivers receiving treated discharges, and drinking water. Nested PCR assays were used for the identification of viral DNA/RNA, followed by direct amplicon sequencing. All raw sewage samples (21/21), 61.9 % of treated wastewater samples (13/21), and 25 % of ultrafiltered water samples (3/12) were contaminated with at least one viral family. Multiple virus families and genera were frequently detected. Mean positive PCRs per sample decreased significantly from raw to treated sewage and to ultrafiltered waters. Moreover, quantitative adenovirus data showed a reduction in excess of 99 % in viral genome copies following wastewater treatment. In surface waters, 78.6 % (22/28) of samples tested positive for one or more viruses by molecular methods, but enterovirus-specific infectivity assays did not reveal infectious particles in these samples. All drinking water samples tested negative for all viruses, demonstrating the effectiveness of treatment in removing viral pathogens from drinking water. Integrated strategies to manage water from all sources are crucial to ensure water quality.     

Pre-harvest Viral Contamination of Crops Originating from Fecal Matter

Pre-harvest contamination of fresh produce and fruits is a possible route for viral transmission which should not be ignored. The contamination originates from viruses shed in human or animal fecal materials which eventually reach crops through many steps and hurdles, including spread of viruses into the agricultural environment through leakage of septic tanks/pipes or runoff from animal lagoons, virus survival during biosolids and manure treatment, virus survival and transport in soil and subsequent contamination of irrigation water, and virus transmission to crops through irrigation water, etc. Initially, large quantities of virus particles may be released from infected humans or animals, and then in the environment viruses are gradually inactivated under various natural conditions (e.g., temperature, water activity, microbial activities, etc.) and only a tiny portion of viruses may reach crops and cause the contamination. However, the fact that the infectious dose of some foodborne viruses is as low as 10–100 particles makes the pre-harvest contamination still a threat to human health. In the USA, the Environmental Protection Agency (USEPA) and United States Department of Agriculture (USDA) regulations and guidances on proper treatment and usage of manure and biosolids for agricultural purpose may largely reduce the release of viruses to the environment; however, pre-harvest viral contamination due to fecal matter is not completely avoidable. This review focuses on the current knowledge of pre-harvest viral contamination and describes the complex transmission process regarding the presence and survival of virus in soil and fecal material, the effect of different biosolids/manure treatment on virus inactivation, the transmission of virus from soil to water, the contamination of crops by irrigation water and survival of virus on crops.     

Detection of Human Enteric Viruses in Freshwater from European Countries

The transmission of water-borne pathogens typically occurs by a faecal–oral route, through inhalation of aerosols, or by direct or indirect contact with contaminated water. Previous molecular-based studies have identified viral particles of zoonotic and human nature in surface waters. Contaminated water can lead to human health issues, and the development of rapid methods for the detection of pathogenic microorganisms is a valuable tool for the prevention of their spread. The aims of this work were to determine the presence and identity of representative human pathogenic enteric viruses in water samples from six European countries by quantitative polymerase chain reaction (q-PCR) and to develop two quantitative PCR methods for Adenovirus 41 and Mammalian Orthoreoviruses. A 2-year survey showed that Norovirus, Mammalian Orthoreovirus and Adenoviruses were the most frequently identified enteric viruses in the sampled surface waters. Although it was not possible to establish viability and infectivity of the viruses considered, the detectable presence of pathogenic viruses may represent a potential risk for human health. The methodology developed may aid in rapid detection of these pathogens for monitoring quality of surface waters.     

二氧化氯对不同微生物的灭活特性及其对群落结构特征的影响

二氧化氯是一种性能优良、应用广泛的消毒剂,可通过破坏细胞或病毒的组成结构、阻碍细胞代谢等方式实现微生物灭活。在自配水条件下,以二氧化氯投加量×消毒时间计算,二氧化氯剂量在15（mg·min）/L时,可实现对常见病毒（包括肠病毒71型、大肠杆菌噬菌体MS2等）3 log以上灭活率,在60（mg·min）/L时,可实现对常见细菌（包括大肠杆菌、金黄色葡萄球菌等）1.5 log以上灭活率,但灭活隐孢子虫卵则需要更高的剂量（如1.9 log灭活率可能需约600（mg·min）/L剂量）;在实际污水厂进水中,30（mg·min）/L二氧化氯剂量只能分别实现0.8 log和0.5 log的大肠杆菌和总大肠菌群灭活率。二氧化氯消毒效果随温度升高显著提升,对于不同微生物,pH的变化对二氧化氯消毒效果的影响可能存在不同,而水中的有机物通常会因消耗二氧化氯而降低消毒效果,但在自然水体中也存在由于天然有机物可能的影响导致消毒效果优于自配水的情况。关于二氧化氯消毒后细菌群落结构的变化研究不多,仅有少量研究涉及市政污水、再生水、饮用水等。二氧化氯消毒一定时间后,悬浮态和生物膜上的微生物均可能出现再生长现象,但再生长过程中这些残生细菌的群落结构变化及其生长分泌特性仍有待研究。     

Occurrence of Human Enteric Viruses in Water Sources and Shellfish: A Focus on Africa

Enteric viruses are a diverse group of human pathogens which are primarily transmitted by the faecal–oral route and are a major cause of non-bacterial diarrhoeal disease in both developed and developing countries. Because they are shed in high numbers by infected individuals and can persist for a long time in the environment, they pose a serious threat to human health globally. Enteric viruses end up in the environment mainly through discharge or leakage of raw or inadequately treated sewage into water sources such as springs, rivers, dams, or marine estuaries. Human exposure then follows when contaminated water is used for drinking, cooking, or recreation and, importantly, when filter-feeding bivalve shellfish are consumed. The human health hazard posed by enteric viruses is particularly serious in Africa where rapid urbanisation in a relatively short period of time has led to the expansion of informal settlements with poor sanitation and failing or non-existent wastewater treatment infrastructure, and where rural communities with limited or no access to municipal water are dependent on nearby open water sources for their subsistence. The role of sewage-contaminated water and bivalve shellfish as vehicles for transmission of enteric viruses is well documented but, to our knowledge, has not been comprehensively reviewed in the African context. Here we provide an overview of enteric viruses and then review the growing body of research where these viruses have been detected in association with sewage-contaminated water or food in several African countries. These studies highlight the need for more research into the prevalence, molecular epidemiology and circulation of these viruses in Africa, as well as for development and application of innovative wastewater treatment approaches to reduce environmental pollution and its impact on human health on the continent.

     

UVC Inactivation of dsDNA and ssRNA Viruses in Water: UV Fluences and a qPCR-Based Approach to Evaluate Decay on Viral Infectivity

Disinfection by low-pressure monochromatic ultraviolet (UVC) radiation (253.7 nm) became an important technique to sanitize drinking water and also wastewater in tertiary treatments. In order to prevent the transmission of waterborne viral diseases, the analysis of the disinfection kinetics and the quantification of infectious viral pathogens and indicators are highly relevant and need to be addressed. The families Adenoviridae and Polyomaviridae comprise human and animal pathogenic viruses that have been also proposed as indicators of fecal contamination in water and as Microbial Source Tracking tools. While it has been previously suggested that dsDNA viruses may be highly resistant to UVC radiation compared to other viruses or bacteria, no information is available on the stability of polyomavirus toward UV irradiation. Here, the inactivation of dsDNA (HAdV2 and JCPyV) and ssRNA (MS2 bacteriophage) viruses was analyzed at increasing UVC fluences. A minor decay of 2-logs was achieved for both infectious JC polyomaviruses (JCPyV) and human adenoviruses 2 (HAdV2) exposed to a UVC fluence of 1,400 J/m², while a decay of 4-log was observed for MS2 bacteriophages (ssRNA). The present study reveals the high UVC resistance of dsDNA viruses, and the UV fluences needed to efficiently inactivate JCPyV and HAdV2 are predicted. Furthermore, we show that in conjunction with appropriate mathematical models, qPCR data may be used to accurately estimate virus infectivity.     

Oncogenic Papillomavirus and Polyomavirus in Water Environments: Is There a Potential for Waterborne Transmission?

Waterborne exposure to human viruses through contact with sewage-contaminated water environments can result in infections associated with a wide range of illnesses. Gastrointestinal symptoms are the most commonly encountered manifestations of waterborne viral illness. Respiratory diseases, neurological diseases and paralysis can also occur. Whether viral infections resulting in health outcomes like cancer might also be transmitted by the waterborne route is unknown. Recently, viruses belonging to two oncogenic groups—Human Papillomaviruses (HPVs) and Human Polyomaviruses (HPyVs)—have been detected in urban sewages worldwide. The latter have also been identified in other water environments. HPVs are epitheliotropic viruses responsible for several diseases of skin and mucosae, from common warts to squamous intraepithelial lesions that can either heal or progress to invasive carcinoma of the cervix, vulva, vagina, penis, anus or oropharynx. Human PyVs infect different tissues and organs, causing infections that are usually subclinical in immunocompetent individuals but can be serious in immunocompromised hosts. These pathogens belong to a family of DNA tumour viruses. Merkel cell polyomavirus, a HPyV identified in recent years, has attracted much attention due to its link with a rare and aggressive form of human cancer. Merkel cell carcinoma, the incidence of which has tripled over the past two decades. JC polyomavirus and BK polyomavirus are also potentially oncogenic. The observed abundance and wide dissemination of HPVs and HPyVs in water environments strongly suggest the need to shed light on the fate of these viruses in water environments and to elucidate their potential for waterborne transmission. Such information is essential for the improvement of wastewater management programs in terms of both sewage treatment and water quality surveillance.