Human Bocavirus: Detection,Quantification and Molecular Characterization in Sewage and Surface Waters in Uruguay

Human bocavirus (HBoV) infections are related to respiratory and gastroenteric diseases. The aim of this study was to investigate the presence of HBoV in both sewage and surface waters in Uruguay. Sixty-eight sewage samples from the cities of Salto, Paysandú, Bella Unión, Fray Bentos, Treinta y Tres and Melo and 36 surface water samples from the cities of Salto, Florida and Santa Lucía were studied. HBoV was screened by multiplex qPCR for the detection of the four subtypes, followed by monoplex qPCRs for the independent quantification of each subtype. A qualitative PCR followed by DNA sequencing and phylogenetic analysis was carried out for molecular characterization of HBoV strains. HBoV was present in a high frequency (69%) in sewage and only one positive sample (3%) was found in surface water. Concerning sewage samples, HBoV1 was detected in 11 (23%) out of the 47 positives samples, with a mean concentration of 8.2 × 10⁴ genomic copies/Liter (gc/L), HBoV3 was detected in 35 (74%) of the positive samples with a mean concentration of 4.1 × 10⁶ gc/L and subtypes 2 and/or 4 were detected in 39 (83%) of the positive samples with a mean concentration of 7.8 × 10⁶ gc/L. After the phylogenetic analysis performed by a Bayesian approach, the four HBoV subtypes were confirmed. This is the first study determining a high frequency of HBoV and the presence of the four HBoV subtypes in aquatic matrices in Latin America, mainly in sewage. Although HBoV was scarcely detected in surface water, a waterborne transmission is likely to occur if people enter in contact with polluted surface waters for recreational activities such as fishing or swimming since an elevated frequency of HBoV was detected in raw sewage which is usually directly discharged into surface waters.     

Relative Abundance of Human Bocaviruses in Urban Sewage in Greater Cairo,Egypt

Human bocavirus (HBoV) is predominantly found in the respiratory tract infections and in the stool of patients with gastroenteritis symptoms. However, data on the prevalence of HBoV genotypes in environmental samples are limited. Here we addressed the prevalence of HBoV in sewage collected from three different wastewater treatment plants in Egypt. HBoV-1, HBoV-2, and HBoV-3 were detected, whereas none of the samples were positive for HBoV-4. The median concentration of HBoV in influent samples was 8.5 × 10³ GC/l for HBoV-1, 3.0 × 10⁴ GC/l for HBoV-2, and 2.5 × 10⁴ GC/l for HBoV-3. The concentration was reduced but not completely removed in the effluent samples. The median concentration in the outlet samples was 2.9 × 10³ GC/l for HBoV-1, 4.1 × 10³ GC/l for HBoV-2, and 2.1 × 10³ GC/l for HBoV-3. Moreover, no seasonality pattern of HBoVs was found. The high incidence of HBoV in sewage samples provided an evidence of its circulation in the local population. Although the role of HBoV in respiratory or gastro-intestinal infections still remains to be fully elucidated, the risk of infection via contaminated water should be taken into consideration.     

Assessment of Gastroenteric Viruses from Wastewater Directly Discharged into Uruguay River,Uruguay

The aim of this study was to assess the viral contamination of group A rotavirus (RVA), norovirus (NoV), and human astrovirus (HAstV) in sewage directly discharged into Uruguay River and to characterize RVA genotypes circulating in Uruguay. For this purpose, sewage samples (n = 96) were collected biweekly from March 2011 to February 2012 in four Uruguayan cities: Bella Unión, Salto, Paysandú, and Fray Bentos. Each sample was concentrated by ultracentrifugation method. Qualitative and quantitative RT-PCR for RVA, NoV, and HAstV were performed. A wide dissemination of gastroenteric viruses was observed in the sewage samples analyzed with 80 % of positivity, being NoV (51 %) the most frequently detected followed by RVA with a frequency of 49 % and HAstV with 45 %. Genotypes of RVA were typed using multiplex semi-nested RT-PCR as follows: P[8] (n = 15), P[4] (n = 8), P[10] (n = 1), P[11] (n = 1), G2 (n = 29), and G3 (n = 2). The viral load ranged from 10³ to 10⁷ genomic copies/liter, and they were detected roughly with the same frequency in all participant cities. A peak of RVA and HAstV detection was observed in colder months (June to September), whereas no seasonality was observed for NoV. This study demonstrates for the first time, the high degree of gastroenteric viral contamination in the country; highlighting the importance of developing these analyses as a tool to determine the viral contamination in this hydrographic boundary region used by the local populations for recreation and consumption, establishing an elevated risk of gastroenteric diseases for human health.     

Presence of Torque Teno Virus (TTV) in Tap Water in Public Schools from Southern Brazil

Torque teno virus (TTV) was surveyed in tap water collected in schools from three municipalities located in the south of Brazil. TTV genomes were found in 11.7 % (4/34) of the samples. TTV DNA was detected in 10.5 % (2/19) of the samples collected at the city of Caxias do Sul and in 25 % (2/8) of the samples from Pelotas. Those cities have a low rate of sewage treatment. All samples from Santa Cruz do Sul, which has nearly 92 % of its sewage treated, were negative. These results suggest that the amount of sewage treated may have an effect on the detection rates of TTV DNA in drinking water in a given urban area, showing a mild negative correlation (r = ?0.76), when comparing the percentage of sewage treatment to the detection of TTV genomes. The detection rate of TTV was also compared with Escherichia coli, showing a strong correlation (r = 0.97), indicating that TTV may be a suitable marker of fecal contamination.     

我国城市污水处理回用调查研究

在水利部主持下,开展了2007年全国城市污水处理回用现状的调查,取得了全国城市污水回用现状的基础数据.结果表明,2007年,全国城市污水回用总量为17.9×108m3,城市污水处理回用率为5.23%,主要用于工业、景观、农林牧业、城市非饮用水、地下水回灌.全国共有127座再生水厂,再生水管道总长1 421.78 km.再生水厂总投资为56.44×108元,中央财政投资、地方财政投资和其他投资分别占16%、26%和58%,再生水管网其他投资和地方财政投资分别为66%和32%,中央投资仅为2%.再生水发展状况具有较强的地域性,与当地的水资源状况、产业结构和经济状况有密切关系.     

GIV Noroviruses in Wastewaters and in Stool Specimens from Hospitalized Patients

Noroviruses (NoVs) are important human pathogens associated with foodborne and waterborne gastroenteritis. These viruses are genetically highly heterogeneous, with more than forty genotypes within three genogroups (GI, GII, and GIV) identified in humans. However, the vast majority of human infections are associated with variants of a unique genotype, GII.4. Aside from these NoV strains of epidemiological relevance, NoV strains of genogroup GIV (Alphatron-like) are reported in a sporadic fashion and their overall prevalence in the community is unknown and this likely reflects the lack of specific diagnostic tools. We analyzed raw sewages collected from 32 wastewater treatment plants distributed throughout Italy (307 samples) and stool specimens collected from hospitalized patients with clinical signs of diarrhea of unknown etiology (285 samples). By using specific qualitative and quantitative RT-PCR assays, 21.8 % of the sewage samples and 3.2 % of the stool specimens tested positive for GIV NoVs. The number of genome copies in fecal samples ranged from 5.08 × 10⁴ to 1.73× 10⁶/g of feces. Sequence analysis showed limited genetic variability in human GIV viruses. The presence of GIV NoV both in sewage and in clinical samples confirms that not only GI and GII NoVs but also GIV strains are circulating in humans. Monitoring of GIV NoV is recommended in order to understand the dynamics of circulation in human populations, environmental contamination, and potential health risks.     

Coxsackievirus B4 as a Causative Agent of Diabetes Mellitus Type 1: Is There a Role of Inefficiently Treated Drinking Water and Sewage in Virus Spreading?

This study proposed to detect the enterovirus (EV) infection in children with type 1 diabetes mellitus (T1D) and to assess the role of insufficiently treated water and sewage as sources of viral spreading. Three hundred and eighty-two serum specimens of children with T1D, one hundred serum specimens of children who did not suffer from T1D as control, and forty-eight water and sewage samples were screened for EV RNA using nested RT-PCR. The number of genome copies and infectious units of EVs in raw and treated sewage and water samples were investigated using real-time (RT)-PCR and plaque assay, respectively. T1D markers [Fasting blood glucose (FBG), HbA1c, and C-peptide], in addition to anti-Coxsackie A & B viruses (CVs A & B) IgG, were measured in control, T1D-negative EV (T1D–EV^?), and T1D-positive EV (T1D–EV⁺) children specimens. The prevalence of EV genome was significantly higher in diabetic children (26.2%, 100 out of 382) than the control children (0%, 0 out of 100). FBG and HbA1c in T1D–EV^? and T1D–EV⁺ children specimens were significantly higher than those in the control group, while c-peptide in T1D–EV^? and T1D–EV⁺ children specimens was significantly lower than that in the control (n = 100; p < 0.001). Positivity of anti-CVs A & B IgG was 70.7, 6.7, and 22.9% in T1D–EV⁺, T1D–EV^?, and control children specimens, respectively. The prevalence of EV genome in drinking water and treated sewage samples was 25 and 33.3%, respectively. The prevalence of EV infectious units in drinking water and treated sewage samples was 8.5 and 25%, respectively. Quantification assays were performed to assess the capabilities of both wastewater treatment plants (WWTPs) and water treatment plants (WTPs) to remove EV. The reduction of EV genome in Zenin WWTP ranged from 2 to 4 log₁₀, while the reduction of EV infectious units ranged from 1 to 4 log₁₀. The reduction of EV genome in El-Giza WTP ranged from 1 to 3 log₁₀, while the reduction of EV infectious units ranged from 1 to 2 log₁₀. This capability of reduction did not prevent the appearance of infectious EV in treated sewage and drinking water. Plaque purification was performed for isolation of separate EV isolates from treated and untreated water and sewage samples. Characterization of the EV amplicons by RT-PCR followed by sequencing of these isolates revealed high homology (97%) with human coxsackievirus B4 (CV B4) in 60% of the isolates, while the rest of the isolates belonged to poliovirus type 1 and type 2 vaccine strains. On the other hand, characterization of the EV amplicons by RT-PCR followed by sequencing for T1D–EV⁺ children specimens indicated that all samples contained CV B4 with the same sequence characterized in the environmental samples. CV B4-contaminated drinking water or treated sewage may play a role as a causative agent of T1D in children.     

Pepper Mild Mottle Virus as Indicator of Pollution: Assessment of Prevalence and Concentration in Different Water Environments in Italy

Pepper mild mottle virus (PMMoV), a plant pathogenic virus belonging to the family Virgoviridae, has been proposed as a potential viral indicator for human faecal pollution in aquatic environments. The present study investigated the occurrence, amount and diversity of PMMoV in water environments in Italy. A total of 254 water samples, collected between 2017 and 2019 from different types of water, were analysed. In detail, 92 raw sewage, 32 treated sewage, 16 river samples, 9 estuarine waters, 20 bathing waters, 67 groundwater samples and 18 drinking waters were tested. PMMoV was detected in 79% and 75% of untreated and treated sewage samples, respectively, 75% of river samples, 67% and 25% of estuarine and bathing waters and 13% of groundwater samples. No positive was detected in drinking water. The geometric mean of viral concentrations (genome copies/L) was ranked as follows: raw sewage (2.2 × 10⁶) > treated sewage (2.9 × 10⁵) > river waters (6.1 × 10²) > estuarine waters (4.8 × 10²) > bathing waters (8.5 × 10¹) > groundwater (5.9 × 10¹). A statistically significant variation of viral loads could be observed between raw and treated sewage and between these and all the other water matrices. PMMoV occurrence and viral loads did not display seasonal variation in raw sewage nor correlation with faecal indicator bacteria in marine waters and groundwater. This study represents the first report on the occurrence and quantification PMMoV in different water environments in Italy. Further studies are required to evaluate the suitability of PMMoV as a viral indicator for human faecal pollution and for viral pathogens in waters.

     

Quantification and Genotyping of Aichi Virus 1 in Water Samples in the Kathmandu Valley,Nepal

Aichi virus 1 genomes were detected by quantitative PCR in groundwater from shallow dug (10/22) and tube wells (1/15), river water (14/14), and sewage (1/1), with the maximum concentration of 4.0 × 10⁹ copies/l. Nucleotide sequencing analysis demonstrated the prevalence of genotype B in the virus positive samples.     

Environmental Surveillance for Noroviruses in Selected South African Wastewaters 2015–2016: Emergence of the Novel GII.17

Norovirus (NoV) GII.4 is the predominant genotype associated with gastroenteritis pandemics and new strains emerge every 2–3 years. Between 2008 and 2011, environmental studies in South Africa (SA) reported NoVs in 63% of the sewage-polluted river water samples. The aim of this study was to assess whether wastewater samples could be used for routine surveillance of NoVs, including GII.4 variants. From April 2015 to March 2016, raw sewage and effluent water samples were collected monthly from five wastewater treatment plants in SA. A total of 108 samples were screened for NoV GI and GII using real-time RT-qPCR. Overall 72.2% (78/108) of samples tested positive for NoVs with 4.6% (5/108) GI, 31.5% (34/108) GII and 36.1% (39/108) GI + GII strains being detected. Norovirus concentrations ranged from 1.02 × 10² to 3.41 × 10⁶ genome copies/litre for GI and 5.00 × 10³ to 1.31 × 10⁶ genome copies/litre for GII. Sixteen NoV genotypes (GI.2, GI.3, GI.4, GI.5, GI.6, GII.2, GII.3, GII.4, GII.7, GII.9, GII.10, GII.14, GII.16, GII.17, GII.20, and GII.21) were identified. Norovirus GII.2 and GII.17 co-dominated and the majority of GII.17 strains clustered with the novel Kawasaki 2014 variant. Sewage surveillance facilitated detection of Kawasaki 2014 in SA, which to date has not been detected with surveillance in children with gastroenteritis <5 years of age. Combined surveillance in the clinical setting and environment appears to be a valuable strategy to monitor emergence of NoV strains in countries that lack NoV outbreak surveillance.     

Effects of Al3+ on the microstructure and bioflocculation of anoxic sludge

The aluminum ions generated from mining aluminum,electrolytic aluminum and the industrial production of aluminum-based coagulants(such as AlCl_3 and Al_2(SO_4)_3) enter sewage treatment plants and interact with activated sludges.An anaerobic/anoxic/oxic(A~2 O) process was used to reveal the effects of Al~(3+) on the pollutant removal efficiencies,bioflocculation and the micro structure of sludge.The results showed that a low concentration of Al~(3+) improved the pollutant removal efficiencies and increased the sludge particle size.However,a high concentration of Al~(3+) hindered microbial flocculation and reduced the pollutant removal efficiencies.With a 10 mg/L Al~(3+) addition,the chemical oxygen demand(COD),total nitrogen(TN) and NH_4~+-N increased by 3%,16% and 27%,and reached as high as 68%,60% and 87%,respectively.At the same time,the dehydrogenase activity,flocculation ability(FA) and contact angle of the sludge reached their maximum levels at 41.3 mg/L/hr,45% and 79.63°,respectively.The specific surface area of the sludge decreased to 7.084 m2/g and the sludge pore size distribution shifted to concentrate in the me soporous range.Most of Al~(3+) was adsorbed on the surface of sludge,changing the physicochemical properties and physical structure of the sludge.     

微生物营养剂浓度对生物沥浸法促进城市污泥脱水性能的影响

通过实验室摇瓶试验和实际工程应用试验研究了不同污泥浓度(2%～5%)下,营养剂加入量对生物沥浸促进城市污泥脱水性能的影响,同时分析了pH值和营养剂利用率的变化.结果表明,生物沥浸过程中,不同浓度污泥各处理下pH值均呈直线下降后趋于稳定的趋势;生物沥浸2 d后,各处理中营养剂几乎都被微生物完全利用;2%、3%、4%、5%浓度污泥比阻随着沥浸时间的延长先迅速减小后逐渐回升,且浓度越高回升幅度越大,各浓度污泥选择最佳营养剂浓度分别为3.0、4.5、8.3和12.8 g.L-1,此时污泥最低比阻分别为0.61×1012、1.22×1012、3.09×1012和4.83×1012m.kg-1.通过工程应用试验表明,将5%浓度城市污泥稀释成3%浓度再生物沥浸的方法不仅能够改善生物沥浸污泥脱水性能,表现在比阻从3.29×1012m.kg-1下降到1.10×1012m.kg-1,同时还可以缩短污泥停留时间(从4 d缩短为2.35 d)及降低运行成本.这为生物沥浸工艺处理高浓度污泥运行参数的优化提供了科学依据.     

Source-specific risk assessment for cadmium in wheat and maize: Towards an enrichment model for China

Cadmium (Cd) pollution of agricultural soil is of public concern due to its high potential toxicity and mobility. This study aimed to reveal the risk of Cd accumulation in soil and wheat/maize systems, with a specific focus on the source-specific ecological risk, human health risk and Cd enrichment model. For this we investigated more than 6100 paired soil and grain samples with 216 datasets including soil Cd contents, soil pH and grain Cd contents of 85 sites from China. The results showed that mining activities, sewage irrigation, industrial activities and agricultural practices were the critical factors causing Cd accumulation in wheat and maize cultivated sites. Thereinto, mining activities contributed to a higher Cd accumulation risk in the southwest China and Middle Yellow River regions; sewage irrigation influenced the Cd accumulation in the North China Plain. In addition, the investigated sites were classified into different categories by comparing their soil and grain Cd contents with the Chinese soil screening values and food safety values, respectively. Cd enrichment models were developed to predict the Cd levels in wheat and maize grains. The results showed that the models exhibited a good performance for predicting the grain Cd contents among safe and warning sites of wheat (R² = 0.61 and 0.72, respectively); while the well-fitted model for maize was prone to the overestimated sites (R² = 0.77). This study will provide national viewpoints for the risk assessments and prediction of Cd accumulation in soil and wheat/maize systems.     

First Detection of Hepatitis E Virus in Shellfish and in Seawater from Production Areas in Southern Italy

Shellfish samples (n = 384) from production areas, water samples from the same areas (n = 39) and from nearby sewage discharge points (n = 29) were analyzed for hepatitis E virus (HEV) by real-time and nested RT-PCR. Ten shellfish samples (2.6%) and five seawater samples (12.8%) tested positive for HEV; all characterized strains were G3 and showed high degree of sequence identity. An integrated surveillance in seafood and waters is relevant to reduce the risk of shellfish-associated illnesses.     

不同处理水芹浮床对城市河道黑臭污水的脱氮效果及其机理研究

通过对冬季不同留茬高度水芹（Oenanthe Javanica）浮床对城市河道污水氮循环细菌数量和脱氮效果的影响研究,试图阐明浮床植物去除污染水体氮素的可能途径.研究结果表明：不同处理水芹浮床对城市河道黑臭污水硝氮和氨氮的去除效果分别达到了52.5%～69.8%和68.1%～76.7%,19cm留茬高度水芹浮床对脱氮效...     

Spatial and Temporal Distribution of Norovirus and <Emphasis Type="Italic">E. coli</Emphasis> in Sydney Rock Oysters Following a Sewage Overflow into an Estuary

This paper reports a study of norovirus (NoV) GII distribution and persistence in Sydney rock oysters (SRO) (Saccostrea glomerata) located in an estuary after a pump station sewage overflow. SRO were strategically placed at six sites spanning the length of the estuary from the pump station to the sea. The spatial and temporal distribution of NoV, hepatitis A virus (HAV) and Escherichia coli (E. coli) in oysters was mapped after the contamination event. NoV GI and GII, HAV and E. coli were quantified for up to 48 days in oysters placed at six sites ranging from 0.05 to 8.20 km from the sewage overflow. NoV GII was detected up to 5.29 km downstream and persisted in oysters for 42 days at the site closest to the overflow. NoV GII concentrations decreased significantly over time; a reduction rate of 8.5% per day was observed in oysters (p < 0.001). NoV GII concentrations decreased significantly as a function of distance at a rate of 5.8% per km (p < 0.001) and the decline in E. coli concentration with distance was 20.1% per km (p < 0.001). HAV and NoV GI were not detected. A comparison of NoV GII reduction rates from oysters over time, as observed in this study and other published research, collectively suggest that GII reduction rates from oysters may be broadly similar, regardless of environmental conditions, oyster species and genotype.     

雄安新区唐河污水库残留污染物对地下水水化学动态的作用机制

唐河污水库位于雄安新区白洋淀西侧,污水库渗漏和污水灌溉历时长达40 a,包气带中累积大量污染物.识别污水渗漏和灌溉复合系统残留污染物对地下水水化学动态的作用机制,对雄安新区的水环境保护至关重要.利用水化学离子示踪及改进的氯碱指数等水文地球化学方法,解析浅层地下水水化学时空特征及演化机制.结果表明,污水库区地下水水化学类型为SO₄ ·HCO₃-Na型,平均污水混合比例为48.4%,阳离子交替吸附作用和岩盐溶解作用分别为地下水提供了29.9%和8.6%的钠离子;污灌区地下水水化学类型为SO₄ ·HCO₃-Na ·Mg型,平均污水混合比例为58.3%,阳离子交替吸附作用降低了地下水中8.1%的钠离子;降水和灌溉水的混合稀释作用导致残留污水混合比例和矿物饱和指数降低,促进地下水中钠离子与土壤胶体吸附的钙镁离子的交换作用.含水层反硝化作用能有效减少地下水硝酸盐污染.此外,污水库修复前后污水比例分别为61.5%和49.3%.污水渗漏和灌溉复合系统中长期滞留的污染物将在降雨和灌溉模式驱动下以不同程度的污水物理混合和水岩相互作用持续影响浅层地下水水质.     

重力出流式膜生物反应器的膜通量及膜污染控制研究

采用新型的重力出流式膜生物反应器(MBR)处理生活污水和垃圾渗滤液,考察了其在长期运行过程中膜通量的变化规律及其影响因素.结果表明,该MBR能够在较低液位水头(8.5～15.0 kPa)的作用下连续出水,并获得较高的膜通量.处理生活污水时,平均膜通量为11.2 L·(m²·h)^-1;处理垃圾渗滤液时,平均膜通量为6.4 L·(m²·h)^-1.研究发现,污泥浓度对膜通量影响大小与曝气强度有关.当曝气强度小于400 m³·(m²·h)^-1时,膜通量随着污泥浓度的升高显著下降;当曝气强度大于400 m³·(m²·h)^-1时,膜通量几乎不受污泥浓度和曝气强度的影响.对膜的化学清洗试验表明,NaOH+NaClO溶液清洗效果最佳,膜通量可恢复至初始通量的85%以上.进一步研究表明,混合液中高浓度的溶解性胞外聚合物是MBR处理垃圾渗滤液时膜通量较低的主要原因.     

Persistence of MS-2 Bacteriophage Within Eastern Oysters

Male-specific bacteriophages have been proposed as human enteric virus indicators for shellfish. In this study, Eastern oysters (Crassostrea virginica) were individually exposed to 5.6 × 10¹⁰ PFU of MS-2 for 48 h at 15 °C followed by collective maintenance in continuously UV-sterilized seawater for 0–6 weeks at either 7, 15, or 24 °C. Initial contamination levels of MS-2 were >6 log PFU. Assessment of weekly declines of viable MS-2 indicated that cooler temperatures dramatically enhanced the persistence of MS-2 within oyster tissues. At 3 weeks, the average log PFU reductions for MS-2 within oysters were 2.28, 2.90, and 4.57 for oysters held at 7, 15, and 24 °C, respectively. Fitting temporal survival data with linear and nonlinear Weibull models indicated that the Weibull model best fit the observed reductions. In total, these data can serve as a guideline for regulatory agencies regarding the influence of water temperature on indicator phage after episodic sewage exposure.