Qualitative and Quantitative Assessment of Hepatitis A Virus in Wastewaters in Tunisia

Hepatitis A causes substantial morbidity in both industrialized and non-industrialized countries and represents an important health problem in several southern Mediterranean countries. The objectives of the study were as follows: (a) to assess the occurrence of hepatitis A virus (HAV) in Tunisia through the monitoring of urban wastewaters collected at wastewater treatment plants (WTPs); (b) to characterize environmental strains; and (c) to estimate the viral load in raw and treated sewages, in order to evaluate the potential impact on superficial waters receiving discharges. A total of 150 raw and treated wastewaters were collected from three WTPs and analyzed by both qualitative (RT-PCR/nested) and quantitative (qRT-PCR) methods. Of these, 100 (66 %) were found to be positive for HAV by the qualitative assay: 68.3 % in influents and 64.7 % in effluents. The vast majority of HAV sequences belonged to sub-genotype IA, with 11 different strains detected found to be identical to clinical strains isolated from Tunisian patients with acute hepatitis. Five unique variants were also detected, not previously reported in clinical cases. Only two IB strains were found, confirming the rarity of this sub-genotype in this country. The results of the present study indicate a wide circulation of the pathogen in the population, most probably in the form of asymptomatic infections, a finding consistent with the classification of the country as having intermediate/high endemicity. Quantitative data showed high viral loads in influents (3.5E+05 genome copies/liter, mean value) as well as effluents (2.5E+05 genome copies/liter, mean value), suggesting that contaminated water could be a critical element in transmission.     

Heat Inactivation of Hepatitis A Virus in Shellfish Using Steam

Shellfish are an important cause of foodborne viral illness. Consumer-friendly cooking recommendations for shellfish could improve food safety and decrease the risk for infection from contaminated products. Thermal inactivation parameters were established for hepatitis A virus (HAV) in mussels and validated with cooking experiments. Steaming for only 2–5 min was not sufficient to inactivate HAV in mussels in all layers of a steamer. Steaming mussels for 6 min was sufficient to inactivate HAV in all layers. These cooking guidelines produce shellfish with a reduced risk for foodborne virus transmission.     

Hepatitis A Virus,Hepatitis E Virus,and Rotavirus in Foods of Animal Origin Traded at the Borders of Brazil,Argentina, and Uruguay

The aim of this study was to investigate hepatitis A virus (HAV), hepatitis E (HEV), and rotavirus (RV) in fresh and processed meat traded on the border of Brazil with Argentina and Uruguay. In total, 159 samples of raw and processed foods of animal origin were collected in Paso de los Libres, Argentina (n?=?53 raw meat, n?=?24 processed meat) and Rivera, Uruguay (n?=?55 raw meat, n?=?18 processed meat), or were seized by the Brazilian International Agricultural Surveillance System—VIGIAGRO (Brazil–Argentina border) (n?=?8 raw meat, n?=?1 bush meat). All samples were tested for the presence of HAV, HEV, and RV genomes. HAV genes were detected in 18.23% of samples and RV genes in 23.89%. No HEV-positive samples were detected. HAV was also detected in two of the VIGIAGRO samples. Processed meats from Argentina and Uruguay had a higher rate of HAV and RV than raw meat (P?>?0.05). The median HAV in the Argentinian and Uruguayan samples was 6.9?×?10⁴ and 3.5?×?10³ copies/g, respectively. The presence of RV viral genes in raw meats from Argentina was significant, and this was not observed in processed meats. The presence of HAV and RV genes in a significant portion of products from Argentina and Uruguay is a potential source of human infection. This also indicates precarious conditions of acquisition, processing, and manipulation, which could be improved by improved regulation of food across borders.     

Occurrence and Trend of Hepatitis A Virus in Bivalve Molluscs Production Areas Following a Contamination Event

The aim of this study was to assess the trend of hepatitis A virus (HAV) in a coastal zone impacted by a contamination event, providing data for the development of management strategies. A total of 352 samples, including four bivalve mollusc species (Mytilus galloprovincialis, Solen vagina, Venus gallina and Donax trunculus), were taken over a period of 6 months from 27 production areas of the coast and analysis were performed according to ISO/TS 15216-1:2013. HAV presence was detected in 77 samples from 11 production areas and all positive results were related to samples collected in the first 3 months of the surveillance, during which HAV prevalence was 39.9% and values as high as 5096 genome copies/g were detected. A progressive reduction of viral contamination was evident during the first trimester of the monitoring, with prevalence decreasing from 78.8% in the first month, to 37.8% in the second and 3.9% in the third and quantitative levels reduced from an average value of 672 genome copies/g to 255 genome copies/g over a period of 4 weeks (virus half-life: 21.5 days). A regression analysis showed that, during the decreasing phase of the contamination, the data fitted a reciprocal quadratic model (Ra² = 0.921) and, based on the model, a residual presence of HAV could be estimated after negativization of the production areas. The statistical analysis of the results per shellfish species and per production area showed that there were limited differences in contamination prevalence and levels among diverse bivalve species, while a statistically significant difference was present in quantitative levels of one production area. These data could be useful for the development of both risk assessment models and code of practice for the management of viral contamination in primary production.     

不同淋洗剂对土壤中多氯联苯的洗脱

应用批量平衡振荡法研究了4种化学表面活性剂〔TX-100(曲拉通X-100)、TW-80(吐温80)、SDBS(十二烷基苯磺酸钠)、CTMAB(十六烷基三甲基溴化铵)〕、2种生物表面活性剂〔Saponin(皂素)、RL(鼠李糖脂发酵液)〕和1种生物产品(β-环糊精)对污染土壤中PCBs(以Aroclor1254为例)的洗脱效果. 结果表明:在设定的质量浓度范围内,洗脱效果总体表现为TX-100≈TW-80＞SDBS＞β-环糊精＞Saponin＞RL＞CTMAB;ρ(PCBs)为10g/L时,TX-100、TW-80、SDBS、β-环糊精、Saponin、RL、CTMAB对污染土壤中PCBs的洗脱率分别为54.99%、53.12%、46.99%、25.35%、14.76%、13.70%、0%. 通过对洗脱效果、土壤吸附量、淋洗剂成本和环境友好性4个方面的综合考量,筛选出TX-100和β-环糊精2种相对较好的淋洗剂,对其洗脱时间、洗脱次数和淋洗剂pH的影响进行研究,结果表明,TX-100和β-环糊精对土壤中PCBs的洗脱率在洗脱时间为12h时出现拐点,之后洗脱率提高不大;2种淋洗剂的3次洗脱均以第1次为主,第2次和第3次洗脱对洗脱效果没有显著贡献;相同体积的淋洗剂分2次洗脱与1次洗脱效果相当;pH为4~10时,对2种淋洗剂的洗脱效果影响很小.      

砷污染土壤磷酸盐淋洗修复技术研究

淋洗修复技术是治理砷污染土壤的一种高效快速方法.本研究通过砷污染土壤批量振荡淋洗实验,筛选出环境友好且高效的淋洗剂磷酸钾,系统分析液固比、淋洗时间、淋洗液浓度、p H对土壤中砷的去除效果的影响,确定磷酸钾的最佳淋洗条件.进一步以磷酸钾为基本淋洗剂,采用不同试剂和磷酸钾进行组合二步淋洗,探究最佳复合淋洗组合.结果表明,磷酸钾在浓度为0.5 mol·L-1、液固比为4 m L·g-1、淋洗时间为8 h及p H为4.3的淋洗条件下,单独使用磷酸钾进行淋洗时,土壤砷的去除率达到74.03%.当采用0.5 mol·L-1Na OH+0.5 mol·L-1KH2PO4进行复合二步淋洗时,可使土壤中砷的去除率提高到82.60%.     

Discrimination of Infectious Hepatitis A Viruses by Propidium Monoazide Real-Time RT-PCR

The discrimination of infectious and inactivated viruses remains a key obstacle when using quantitative RT-PCR (RT-qPCR) to quantify enteric viruses. In this study, propidium monoazide (PMA) and RNase pretreatments were evaluated for the detection and quantification of infectious hepatitis A virus (HAV). For thermally inactivated HAV, PMA treatment was more effective than RNase treatment for differentiating infectious and inactivated viruses, with HAV titers reduced by more than 2.4 log₁₀ units. Results showed that combining 50 μM of PMA and RT-qPCR selectively quantify infectious HAV in media suspensions. Therefore, PMA treatment previous to RT-qPCR detection is a promising alternative to assess HAV infectivity.     

Detection and Characterization of Hepatitis A Virus and Norovirus in Mussels from Galicia (NW Spain)

Shellfish are recognized as a potential vehicle of viral disease and despite the control measures for shellfish safety there is periodic emergence of viral outbreaks associated with shellfish consumption. In this study a total of 81 mussel samples from Ría do Burgo, A Coruña (NW Spain) were analysed. Samples were collected in seven different harvesting areas with the aim to establish a correlation between the prevalence of norovirus (NoV) and hepatitis A virus (HAV) in mussel samples and the water quality. In addition, the genogroup of the detected HAV and NoV strains was also determined. The HAV presence was detected in 18.5 % of the samples. Contamination levels for this virus ranged from 1.1 × 10² to 4.1 × 10⁶ RNA copies/g digestive tissue. NoV were detected in 49.4 % of the cases reaching contamination levels from 5.9 × 10³ to 1.6 × 10⁹ RNA copies/g digestive tissue for NoV GI and from 6.1 × 10³ to 5.4 × 10⁶ RNA copies/g digestive tissue for NoV GII. The χ²-test showed no statistical correlation between the number of positive samples and the classification of molluscan harvesting area based on the E. coli number. All the detected HAV strains belong to genogroup IB. NoV strains were assigned to genotype I.4, II.4 and II.6.     

铅蓄电池厂污染土壤中重金属铅的清洗及形态变化分析

以我国西南某铅蓄电池厂区内不同污染负荷的铅污染土壤为研究对象,对其中铅含量和形态进行分析,并利用不同清洗剂清洗进行筛选.对不同pH条件下铅的清洗效率和铅形态进行分析,并对不同粒径土壤进行不同时间清洗以确定最佳清洗时间.结果表明,厂区内A和B点土壤污染严重,分别达到15 703.22 mg.kg-1和1 747.78 mg.kg-1,活动态铅比例较大,残渣态仅占17.32%、11.64%、14.6%和10.2%.EDTA、盐酸、柠檬酸、鼠李糖脂和SDS这5种清洗剂中EDTA和盐酸的提取效果最好.酸性条件下的清洗不仅能有效地提取铅总量并能有效地减小铅活动态的环境风险,建议pH 4～7最为合适.粗沙粒和细沙粒的清洗效果较好,清洗粉黏粒建议改进工艺,清洗时间定为240 min效果最好.     

北京市4种不同污水处理系统中病原菌变化研究

采用定量PCR技术,对北京市4种不同污水处理系统中大肠杆菌、军团菌和沙门氏菌随工艺及四季的数量变化进行了追踪研究,以评估病原菌的去除效果及污水回用的健康风险.结果发现,大肠杆菌在夏季进水和出水中的浓度分别在107~108copies·m L-1和105copies·m L-1左右;G-A/O系统对大肠杆菌的去除率最高,平均去除率达99.88%.军团菌在4个污水处理系统中进水浓度为104~105copies·m L-1,出水浓度约为104copies·m L-1,其在剩余污泥样品中浓度较高,达到105~106copies·m L-1.沙门氏菌进水浓度较低,为102~103copies·m L-1,且其在多个工艺段样品中未检出.季节变化对于病原菌的去除具有较大影响.研究表明,大肠杆菌在各污水处理系统中均可检出,且其分布具有一定的季节性,夏季的进出水中浓度相对较高;而军团菌和沙门氏菌浓度在各工艺中则并未呈现出明显的季节性变化.G-A/O系统对3种细菌的整体去除效果最为稳定,去除率较高.大肠杆菌在污水处理厂的出水及剩余污泥中浓度仍然较高,此外,冬季出水中也能检测到沙门氏菌的存在,因此,污水处理厂的出水和污泥排放仍存在一定的生态和健康风险.     

Surveillance of Enteric Viruses and Thermotolerant Coliforms in Surface Water and Bivalves from a Mangrove Estuary in Southeastern Brazil

This study was conducted to evaluate the microbiological quality of a mangrove estuary in the Vitória Bay region, Espírito Santo, Brazil. We analyzed the presence and concentration of enteric viruses and thermotolerant coliforms in water, mussels (Mytella charruana and Mytella guyanensis), and oysters (Crassostrea rhizophorae), collected over a 13-month period. Human adenovirus, rotavirus A (RVA), and norovirus genogroup II were analyzed by quantitative PCR. The highest viral load was found in RVA-positive samples with a concentration of 3.0 × 10⁴ genome copies (GC) L⁻¹ in water samples and 1.3 × 10⁵ GC g⁻¹ in bivalves. RVA was the most prevalent virus in all matrices. Thermotolerant coliforms were quantified as colony-forming units (CFU) by the membrane filtration method. The concentration of these bacteria in water was in accordance with the Brazilian standard for recreational waters (< 250 CFU 100 mL⁻¹) during most of the monitoring period (12 out of 13 months). However, thermotolerant coliform concentrations of 3.0, 3.1, and 2.6 log CFU 100 g⁻¹ were detected in M. charruana, M. guyanensis, and C. rhizophorae, respectively. The presence of human-specific viruses in water and bivalves reflects the strong anthropogenic impact on the mangrove and serves as an early warning of waterborne and foodborne disease outbreaks resulting from the consumption of shellfish and the practice of water recreational activities in the region.

     

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.     

Identification of Enteric Viruses in Foods from Mexico City

Foodborne viruses are a common and, probably, the most under-recognized cause of outbreaks of gastroenteritis. Among the main foods involved in the transmission of human enteric viruses are mollusks, and fruits and vegetables irrigated with wastewater and/or washed with non-potable water or contaminated by contact with surfaces or hands of the infected personnel during its preparation. In this study, 134 food samples were analyzed for the detection of Norovirus, Rotavirus, and Hepatitis A virus (HAV) by amplification of conserved regions of these viruses. From the 134 analyzed samples, 14 were positive for HAV, 6 for Norovirus, and 11 for Rotavirus. This is the first report in Mexico where emphasis is given to the presence of HAV and Norovirus on perishable foods and food from fisheries, as well as Rotavirus on frozen vegetables, confirming the role of vegetables and bivalve mollusks as transmitting vehicles of enteric viruses.     

表面活性剂对土壤中多氯联苯的洗脱研究

采用实验室配置的PCBs污染土壤,以曲拉通X-100、吐温-80、SDS、SDBS 4种表面活性剂进行洗脱试验研究。结果表明:对于单一表面活性剂,SDS对污染土壤中多氯联苯的洗脱效果最好;采用1:20的固液比,进行2~3次洗脱,洗脱率可达50%以上;洗脱时间对洗脱效果影响不大;SDS—吐温-80混合表面活性剂对多氯联苯有协同增溶作用,且在土壤上的吸附损失较小,SDS和吐温-80的最佳质量比为5:5,在此条件下,浓度为7 000 mg/L的混合表面活性剂对多氯联苯污染土壤进行3次洗脱,洗脱率可高达97.89%。     

吐温80降解菌的分离及其性能的初步研究

从生活污水污染的土壤中分离纯化得到一株高效降解非离子表面活性剂吐温80的菌株,经鉴定为假单胞菌(Pseudomonassp.),其降解吐温80的最适温度和pH值为30℃和6.5。该菌株在吐温80浓度高于5000mg/L情况下仍然可以旺盛生长。72h内可以将5000m g/L吐温80溶液降解98.6%,使其剩余浓度<70mg/L。