Detection of Enteroviruses in Influent and Effluent Flow Samples from Wastewater Treatment Plants in Italy

This study evaluated the presence and seasonal distribution of polio and other enteroviruses in four wastewater treatment plants in three cities in Italy, using different treatment systems. Detection of enteroviruses was carried out by virus isolation in cell cultures after concentration of water samples collected at both inlet and outlet of the treatment plants, following the methods described in the WHO guidelines. Viral serotypes isolated before and after water treatment were compared. Forty-eight non-polio enteroviruses were isolated from 312 samples collected at the inlet of the four wastewater treatment plants, 35 of which were Coxsackievirus type B (72.9 %) and 13 Echovirus (27.1 %). After treatment, 2 CVB3, 1 CVB5, and 1 Echo 6 were isolated. CVB3 and Echo 6 serotypes were also detected in samples collected at the inlet of the TP, in the same month and year. The high rate of detection of infectious enteroviruses in inlet sewage samples (30.1 %) indicates wide diffusion of these viruses in the populations linked to the collectors. The incomplete removal of infectious viruses following sewage treatment highlights possible risks for public health relate to treated waters discharge into the environment.     

Identification and Genotyping of Human Sapoviruses Collected from Sewage Water in Naples and Palermo, Italy, in 2011

Human sapoviruses were identified in 15 (12.4 %) of 121 inlet sewage samples collected from wastewater treatment plants in Naples and Palermo, Italy, in 2011. All strains, except one GI.1, were genotyped as GI.2 by sequencing a capsid gene fragment. This is the first detection of sapovirus in wastewaters in Italy.     

Loud calls as a mechanism of social coordination in a fission–fusion taxon, the white-bellied spider monkey (Ateles belzebuth)

Spider monkeys (Ateles spp.) live in social groups that exhibit high levels of fission–fusion dynamics, in which group members form subgroups of varying sizes and compositions. Within these fluid societies, how individuals establish contact with dispersed group members with whom they might choose to associate remains unclear. Long-range vocalizations might facilitate interactions between group members and provide a means of social coordination in fission–fusion societies. We evaluated this possibility for one spider monkey vocalization, the loud call, by examining calling behavior, the relationship between loud calls and changes in subgroup size, and the response of individuals to distant calls and playback experiments in a single study group. We found that 82 % of loud calls were emitted within 30 min of a call from a different location, suggesting that individuals frequently emit loud calls in response to the calls of distant group members. Subgroups that emitted loud calls, especially those that responded to distant calls, were much more likely to experience an increase in subgroup size within an hour after calling than those that did not. Animals also approached distant loud calls more than they avoided or ignored these calls. Finally, playbacks of male calls demonstrated that females respond preferentially to the calls of some individuals over others. Taken together, these results provide support for the hypothesis that spider monkey loud calls function to facilitate and initiate interactions between dispersed group members and suggest that vocal signals can play an important role in influencing social interactions in fission–fusion societies.     

Historical maps and satellite images as tools for shoreline variations and territorial changes assessment: the case study of Volturno Coastal Plain (Southern Italy)

Anthropic pressure has caused severe variations of Mediterranean coastal areas currently hosting about 480 million people. The replacement of natural land covers with crops and urban environment coupled with the reduction of sediment supply to the coast, subsidence, Relative Sea Level Rise and the high frequency of storm events, have caused severe shoreline erosion. In this paper, we stress the key role of historical maps, topographic maps and free satellite images to forecast the rates of coastline changes and to recognize the main features of past landscapes as tools for risk reduction. This data was recorded into a Geographical Information System dedicated to coastal management that allows to compare different coastal zones and elaborate maps. The analysis was applied to the case study of Volturno Coastal Plain (VCP), extending from the town of Mondragone to Patria Lake (Campania Region, Southern Italy). Indeed, the intense territorial modification that occurred between the seventies and eighties, coupled with the exposure to coastal erosion, make the VCP a good test area. The spatial analysis algorithms allowed to outline the main features of past landscapes and how they changed from roman times to present while the coastal evolution (erosion, accretion) and possible future coastal trend was assessed with the Digital Shoreline Analysis System (DSAS) software. Starting from the Bourbon domain, the reclamation caused the first great territorial change (e.g. wetlands were transformed into agricultural lands, regimentation of surperficial water) but the negative effects of antrophic pressure, as the intense urbanization of the coastal belt, emerged in the seventies of the last century. The shoreline position was defined for 9 time intervals (from 1817 to 2012) as the ratio of the distance between two shorelines and the relative elapsed time. Moreover, for the 1957–1998 and 1998–2012 time windows, the shoreline trends were calculated with the weighted linear regression method. The first trend pointed out an intensive erosional phase (mean value: 5 m/yr) for a wide sector close to the Volturno River mouth, the eroded sediment nourished the beaches of other coastal sectors. This phase was related to the reduction of River sediment supply mainly due to the construction of the Ponte Annibale dam on the Volturno River. The second (1998–2012) showed an alternation of erosion and accretion sectors due to a sediment starved condition to deltaic zone and to a sequence of 52 sea protection works in the Gaeta Gulf. Furthermore, the regression values of more recent time interval, was assumed as a scenario to draw the probable shoreline position in 2022. The overlay of this shoreline on the Technical Maps of Campania Region at 1:5000 scale highlighted the urban area that could be exposed to damages.     

Heavy metal bioavailability and chelate mobilization efficiency in an assisted phytoextraction process

The heavy metal bioavailable fraction of a soil is a core parameter to verify the potential risks of contaminant exposure to organisms or plants. The purpose of the present work is to identify the bioavailable metal fraction in soils treated with chelates. This fraction was evaluated directly by analyzing metal concentrations in soil solution and indirectly using sequential extraction procedures. The metal bioavailable fraction was compared with metal accumulated in plant leaves, grown in both untreated and chelate-treated reactors. In order to verify the effect of the readily and slowly biodegradable chelates [S,S]-ethylenediaminedisuccinic acid (EDDS), methylglycine diacetic acid (MGDA), and ethylenediaminetetraacetic acid (EDTA) on metal speciation in soils, a simulation of chelate treatment was made and metal concentrations in different soil compartments before and after the simulation were compared. Lead concentration in the soil solution was positively correlated with metal concentration in the test plants. The soluble fraction showed the best correlation with metal concentration in soil solution. The simulation of the chelate treatment demonstrated that EDTA and EDDS were able to extract part of the organic- and sulfide-bound fraction, which are less available to plants.     

Bioprospecting at former mining sites across Europe: microbial and functional diversity in soils

The planetary importance of microbial function requires urgently that our knowledge and our exploitation ability is extended, therefore every occasion of bioprospecting is welcome. In this work, bioprospecting is presented from the perspective of the UMBRELLA project, whose main goal was to develop an integral approach for remediation of soil influenced by mining activity, by using microorganisms in association with plants. Accordingly, this work relies on the cultivable fraction of microbial biodiversity, native to six mining sites across Europe, different for geographical, climatic and geochemical characteristics but similar for suffering from chronic stress. The comparative analysis of the soil functional diversity, resulting from the metabolic profiling at community level (BIOLOG ECOPlates) and confirmed by the multivariate analysis, separates the six soils in two clusters, identifying soils characterised by low functional diversity and low metabolic activity. The microbial biodiversity falls into four major bacterial phyla: Actinobacteria, Proteobacteria, Firmicutes and Bacteroidetes, including a total of 47 genera and 99 species. In each soil, despite harsh conditions, metabolic capacity of nitrogen fixation and plant growth promotion were quite widespread, and most of the strains showed multiple resistances to heavy metals. At species-level, Shannon’s index (alpha diversity) and Sørensen's Similarity (beta diversity) indicates the sites are indeed diverse. Multivariate analysis of soil chemical factors and biodiversity identifies for each soil well-discriminating chemical factors and species, supporting the assumption that cultured biodiversity from the six mining sites presents, at phylum level, a convergence correlated to soil factors rather than to geographical factors while, at species level, reflects a remarkable local characterisation.     

Bacterial cell monitoring in wastewater treatment plants by flow cytometry.

The activated sludge process is performed by a variable and mixed community of microorganisms in an aerobic aquatic environment, in which bacteria constitute the majority and represent the main microorganisms responsible for the degradation process in a plant. In this work, we monitored bacterial charge in different wastewater treatment plants by flow cytometry, also evaluating chlorination effects on bacterial viability, both by flow cytometry and traditional plate counts. Maximum values of bacterial charge were registered in the aeration tank of all plants monitored. Cell viability did not show significant differences (p > 0.05) in samples collected in "before chlorination" and "wastewater effluent" treatment steps; this suggests that the chlorination was not able to decrease total viable bacterial charge. In this work, we discuss the need to improve microbiological analyses, both in terms of measuring other potential pathogens and of using new methodological approaches in the traditional evaluation of the microbiological quality of effluents.     

Poliovirus and Other Enteroviruses from Environmental Surveillance in Italy, 2009–2015

Within the initiatives for poliomyelitis eradication by WHO, Italy activated an environmental surveillance (ES) in 2005. ES complements clinical Acute Flaccid Paralysis (AFP) surveillance for possible polio cases, detects poliovirus circulation in environmental sewage, and is used to monitor transmission in communities. In addition to polioviruses, the analyses comprised: (i) the monitoring of the presence of non-polio enteroviruses in sewage samples and (ii) the temporal and geographical distribution of the detected viruses. From 2009 to 2015, 2880 sewage samples were collected from eight cities participating in the surveillance. Overall, 1479 samples resulted positive for enteroviruses. No wild-type polioviruses were found, although four Sabin-like polioviruses were detected. The low degree of mutation found in the genomes of these four isolates suggests that these viruses have had a limited circulation in the population. All non-polio enteroviruses belonged to species B and the most frequent serotype was CV-B5, followed by CV-B4, E-11, E-6, E-7, CV-B3, and CV-B2. Variations in the frequency of different serotypes were also observed in different seasons and/or Italian areas. Environmental surveillance in Italy, as part of the ‘WHO global polio eradication program’, is a powerful tool to augment the polio surveillance and to investigate the silent circulation or the re-emergence of enteroviruses in the population.     

Large-Scale Survey of Human Enteroviruses in Wastewater Treatment Plants of a Metropolitan Area of Southern Italy

Human enteroviruses (HEVs) occur in high concentrations in wastewater and can contaminate receiving environmental waters, constituting a major cause of acute waterborne disease worldwide. In this study, we investigated the relative abundance, occurrence, and seasonal distribution of polio and other enteroviruses at three wastewater treatment plants (WWTPs) in Naples, Southern Italy, from January 2010 to December 2014. Influent and effluent samples from the three WWTPs were collected monthly. One hundred and sixty-one of the 731 wastewater samples collected (22.0%) before and after water treatment were CPE positive on RD cells; while no samples were positive on L20B cells from any WWTPs. Among the 140 non-polio enterovirus isolated from inlet sewage, 69.3% were Coxsackieviruses type B and 30.7% were Echoviruses. Among these, CVB3 and CVB5 were most prevalent, followed by CVB4 and Echo6. The twenty-one samples tested after treatment contained 6 CVB4, 5 CVB3, 3 Echo11, and 2 Echo6; while other serotypes were isolated less frequently. Data on viral detection in treated effluents of WWTPs confirmed the potential environmental contamination by HEVs and could be useful to establish standards for policies on wastewater management.