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Faecal contamination of the coastal sea poses widespread hazard to human and environmental health and is predicted to rise in response to global change and human pressure. For better management and risk reduction it is thus imperative to clarify and predict trends of faecal pollution over spatial and temporal scales, and to assess links with climate and other variables. Here, we investigated the spatio-temporal variation in the Faecal Indicator Bacteria (FIB) Escherichia coli and enterococci, over a time frame spanning 11 years (2011-2021) along a coastal area covering approximately 40 km and 59 bathing sites in the Marche region (Adriatic Sea, Italy), characterized by intense beach tourism, high riverine inputs, resident population, maritime traffic and industrial activities. Our analysis, that considers 5,183 measurements during the bathing season (April to October), shows that FIB abundance varied significantly among years. A general, although not significant, increase over time of both FIB was observed, mainly due to a general reduction of structural zeros (i.e., zeros originated from the actual absence of the response variable) over the examined time period. FIB abundances displayed their maxima and minima in different years according to the municipality, with overall peaks recorded in different months (May-June or September), whereas the lowest values were always observed in October. FIB levels were not significantly related neither to rainfalls nor to river discharge, but the activation of combined sewer overflows (CSOs), typically occurring after intense rainfall events, appeared as a necessary condition for the high faecal contamination levels. Considering climate change scenarios predicting significant increases in extreme weather events, our findings support the usefulness of analysing long-term trends to identify pollution sources, and the prioritization of control strategies to better manage the release of microbial pollutants from combined sewer overflows in coastal waters to reduce human risks., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)

Microbial pollutants are a serious threat to human and environmental health in coastal areas. Based on the hypothesis that pollution from multiple sources may produce a distinct microbial signature and that microbial pollutants seem to distribute between a free-living and a particle-attached fraction, we investigated the occurrence, partitioning and sources of microbial pollutants in water samples collected in the Venice Lagoon (Italy). The area was taken as a case study of an environment characterized by a long history of industrial pollution and by growing human pressure. We found a variety of pollutants from several sources, with sewage-associated and faecal bacteria accounting for up to 5.98% of microbial communities. Sewage-associated pollutants were most abundant close to the city centre. Faecal pollution was highest in the area of the industrial port and was dominated by human inputs, whereas contamination from animal faeces was mainly detected at the interface with the mainland. Microbial pollutants were almost exclusively associated with the particle-attached fraction. The samples also contained other potential pathogens. Our findings stress the need for monitoring and managing microbial pollution in highly urbanized lagoon and semi-enclosed systems and suggest that management plans to reduce microbial inputs to the waterways should include measures to reduce particulate matter inputs to the lagoon. Finally, High-Throughput Sequencing combined with computational approaches proved critical to assess water quality and appears to be a valuable tool to support the monitoring of waterborne diseases., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 Elsevier B.V. All rights reserved.)

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 × 10 6 ) > treated sewage (2.9 × 10 5 ) > river waters (6.1 × 10 2 ) > estuarine waters (4.8 × 10 2 ) > bathing waters (8.5 × 10 1 ) > groundwater (5.9 × 10 1 ). 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.

High-Throughput Sequencing technologies are providing unprecedented inventories of microbial communities in aquatic samples, offering an invaluable tool to estimate the impact of anthropogenic pressure on marine communities. In this case study, the Mediterranean touristic site of Aci Castello (Italy) was investigated by High-Throughput Sequencing of 16S and 18S rRNA genes. The sampling area falls within a Marine Protected Area and, notwithstanding, features an untreated urban wastewater discharge. Seawater samples were collected close to the wastewater output (COL) and at a second station about 400 m further off (PAN), before and after a summer increase in population. Prokaryotic communities clustered according to stations, rather than to seasons. While PAN showed a typical, not impacted, marine microbial composition, COL was consistently enriched in Epsilonproteobacteria and Firmicutes. Protist communities showed a peculiar clustering, as COL at springtime stood alone and was dominated by Ciliophora, while the other samples were enriched in Dinophyta. Analysis of alternative, detectable by High-Throughput Sequencing, microbial indicators, including both faecal- and sewage-associated, allowed uncovering the different sources of pollution in coastal and anthropogenically impacted marine ecosystems, underpinning the relevance of High-Throughput Sequencing-based screening as rapid and precise method for water quality management., (© FEMS 2019.)