Detection of Norovirus GII.17 Kawasaki 2014 in Shellfish,Marine Water and Underwater Sewage Discharges in Italy

Norovirus (NoV) is a major cause of non-bacterial acute gastroenteritis worldwide, and the variants of genotype GII.4 are currently the predominant human strains. Recently, a novel variant of NoV GII.17 (GII.P17_GII.17 NoV), termed Kawasaki 2014, has been reported as the cause of gastroenteritis outbreaks in Asia, replacing the pandemic strain GII.4 Sydney 2012. The GII.17 Kawasaki 2014 variant has also been reported sporadically in patients with gastroenteritis outside of Asia, including Italy. In this study, 384 shellfish samples were subjected to screening for human NoVs using real-time PCR and 259 (67.4%) tested positive for Genogroup II (GII) NoV. Of these, 52 samples, selected as representative of different areas and sampling dates, were further amplified by conventional PCR targeting the capsid gene, using broad-range primers. Forty shellfish samples were characterized by amplicon sequencing as GII.4 (n = 29), GII.2 (n = 4), GII.6 (n = 2), GII.12 (n = 2), and GII.17 (n = 3). Sixty-eight water samples (39 seawater samples from the corresponding shellfish production areas and 29 water samples from nearby underwater sewage discharge points) were also tested using the above broad-range assay: eight NoV-positive samples were characterized as GII.1 (n = 3), GII.2 (n = 1), GII.4 (n = 2), and GII.6 (n = 2). Based on full genome sequences available in public databases, a novel RT-PCR nested assay specific for GII.17 NoVs was designed and used to re-test the characterized shellfish (40) and water (8) samples. In this second screening, the RNA of GII.17 NoV was identified in 17 additional shellfish samples and in one water sample. Upon phylogenetic analysis, these GII.17 NoV isolates were closely related to the novel GII.17 Kawasaki 2014. Interestingly, our findings chronologically matched the emergence of the Kawasaki 2014 variant in the Italian population (early 2015), as reported by hospital-based NoV surveillance. These results, showing GII.17 NoV strains to be widespread in shellfish samples collected in 2015 in Italy, provide indirect evidence that this strain has started circulating in the Italian population. Notably, using a specific assay, we were able to detect many more samples positive for GII.17 NoV, indicating that, in food and water matrices, broad-range assays for NoV may grossly underestimate the prevalence of some, less common, NoVs. The detection of the GII.17 strain Kawasaki 2014 in clinical, water and food samples in Italy highlights the need for more systematic surveillance for future disease control and prevention.     

水环境容量研究在环境管理中的实施过程初探

本文在地表水环境容量测算分析的基础上，分析测算中需要重视的关键问题，提出几个对测算结论影响较大的关键参数，并分析了环境容量在管理中应用的技术方法与手段，提出了水环境容量资源分配与管理中的解决方案。     

Chorionic mosaicism: Association with fetal loss but not with adverse perinatal outcome

Cytogenetic data from the United States NICHD collaborative study of chorionic villus sampling (CVS) were used to evaluate the clinical significance of chorionic mosaicism. The 10 754 patients with normal cytogenetic results were compared wtih 108 patients (1.0 per cent) with placental mosaicism and 181 patients (1.6 per cent) with pseudomosaicism. Of the pregnancies intended to continue, the pregnancy loss rate was significantly greater in patients with placental mosaicism than in the cytogenetically normal cohort (8.6 vs. 3.4 per cent, p <0.05). However, there was no difference in the frequencies of abruptio placenta, preterm labour or delivery, small-for-gestational-age newborns, pregnancy-induced hypertension, or neonates with Apgar scores less than 7.     

Vermicomposting of winery wastes: a laboratory study

In Mediterranean countries, millions of tons of wastes from viticulture and winery industries are produced every year. This study describes the ability of the earthworm Eisenia andrei to compost different winery wastes (spent grape marc, vinasse biosolids, lees cakes, and vine shoots) into valuable agricultural products. The evolution of earthworm biomass and enzyme activities was tracked for 16 weeks of vermicomposting, on a laboratory scale. Increases in earthworm biomass for all winery wastes proved lower than in manure. Changes in hydrolytic enzymes and overall microbial activities during the vermicomposting process indicated the biodegradation of the winery wastes. Vermicomposting improved the agronomic value of the winery wastes by reducing the C:N ratio, conductivity and phytotoxicity, while increasing the humic materials, nutrient contents, and pH in all cases. Thus, winery wastes show potential as raw substrates in vermicomposting, although further research is needed to evaluate the feasibility of such wastes in large-scale vermicomposting systems.     

Effect of operational parameters on the removal of particulate chemical oxygen demand in the activated sludge process.

The removal of particulate material in the aeration basin of the activated sludge process is mainly attributed to bioflocculation and hydrolysis of particulate substrate. The bioflocculation process in the aeration tank of the activated sludge process occurs only under favorable conditions in the system, and several common operational parameters affect its performance. The principal objective of this research was to observe the effect of mixed liquor suspended solids, solids retention time (SRT), and extracellular polymer substances on the removal of particulate substrate by bioflocculation. A first-order particulate removal expression, based on flocculation, accurately described the removal rates for supernatant suspended solids and colloidal chemical oxygen demand. Based on the results presented in this investigation, a mixed liquor concentration of approximately 2200 mg/L, an SRT of at least 3 days, and a contact time of 30 minutes are needed for relatively complete removal of the particulate substrate in a plug-flow reactor.     

Waterborne and sediment toxicity of fluoxetine to select organisms

Ecological risk assessments of pharmaceuticals are currently difficult because little-to-no aquatic hazard and exposure information exists in the peer-reviewed literature for most therapeutics. Recently several studies have identified fluoxetine, a widely prescribed antidepressant, in municipal effluents. To evaluate the potential aquatic toxicity of fluoxetine, single species laboratory toxicity tests were performed to assess hazard to aquatic biota. Average LC(50) values for Ceriodaphnia dubia, Daphnia magna, and Pimephales promelas were 0.756 (234 microg/l), 2.65 (820 microg/l), and 2.28 microM (705 microg/l), respectively. Pseudokirchneriella subcapitata growth and C. dubia fecundity were decreased by 0.044 (14 microg/l) and 0.72 microM (223 microg/l) fluoxetine treatments, respectively. Oryias latipes survival was not affected by fluoxteine exposure up to a concentration of 28.9 microM (8.9 mg/l). An LC(50) of 15.2 mg/kg was estimated for Chironomus tentans. Hyalella azteca survival was not affected up to 43 mg/kg fluoxetine sediment exposure. Growth lowest observed effect concentrations for C. tentans and H. azteca were 1.3 and 5.6 mg/kg, respectively. Our findings indicate that lowest measured fluoxetine effect levels are an order of magnitude higher than highest reported municipal effluent concentrations.     

Occurrence of polybrominated diphenylethers, polychlorinated dibenzo-p-dioxins, dibenzofurans and biphenyls in coastal sediments from Spain

Fifteen sediment samples were analysed in order to determine their content of polybrominated diphenylethers (PBDEs), polychlorinated dibenzo-p-dioxins (PCDDs), dibenzofurans (PCDFs) and biphenyls (PCBs). Samples were collected from several hot spots on the Spanish coast, such as the harbours of Almeria and Tarragona, and the mouths of the Besos and Llobregat rivers in Barcelona. A generic analytical procedure based on Soxhlet extraction followed by an automated cleanup system and gas chromatography-ion trap-mass spectrometry was employed for determining the toxic congeners of PCDDs and PCDFs, as well as dioxin-like PCBs. As regards PBDE determinations, a rapid method based on the use of selective pressurized liquid extraction followed by gas chromatography-negative ion chemical ionization-mass spectrometry was applied. Total toxicity equivalent (WHO-TEQ) values were calculated using the toxicity equivalent factors proposed by WHO for dioxin-like PCBs, PCDDs and PCDFs. WHO-TEQ values ranged from 0.3 to 75 pg/g dry weight (dw), with PCB contribution on the toxicity of the samples between 1 and 84%. Total PBDE levels ranged from 2.7 to 134 ng/g dw, with BDE-209 contribution on the total PBDE contamination between 50 and 99%.     

Effect of different radiotracer labelling techniques on radionuclide excretion from marine organisms

Experiments were designed to assess the effect of different techniques of radiotracer labelling on subsequent radioisotope excretion rates in marine crustaceans. A small amphipod (Gammarus locusta) that accumulated ⁶⁵Zn under a close approximation of natural conditions excreted the radiotracer during a 3-month period at a markedly different rate than those of comparable amphipods labelled with ⁶⁵Zn in the laboratory via different combinations of radioactive food and seawater. Shrimp (Lysmata seticauda) administered ⁶⁵Zn by three different methods in the laboratory displayed different excretion kinetics during the first 2 months of loss. The difference between excretion rates most likely was a reflection of the degree to which the various zinc pools within the shrimp had equilibrated with the radiotracer. During the next several months all ⁶⁵Zn loss rates were quite similar, probably indicating that radiotracer excretion was taking place from similar zinc pools within the shrimp. The importance of adequate radiotracer labelling techniques as well as proper design of subsequent excretion experiments, used to gain reliable information on flux rates of the corresponding stable metal, is discussed. It is concluded that laboratory radiotracer experiments which are intended to supply information on actual situations in the sea require careful design and execution.