Metal concentrations in transitional and coastal waters measured by passive (Diffusive Gradients in Thin-films) and spot sampling: MONITOOL Project Dataset.

The MONITOOL project (2017-2023) was carried out to describe the relationships between total dissolved and labile metal concentrations measured in spot water samples and in concurrently deployed Diffusive Gradients in Thin-films (DGTs) passive samplers, respectively. The ultimate aim was to adapt existing marine metal Environmental Quality Standards (EQS marine water) for DGTs, enabling their use in the context of the European Directives (the Water Framework Directive (WFD) and the Marine Strategy Framework Directive (MSFD)). Time-integrated metal concentrations provided by DGTs, representing several days, are an advantage compared to conventional spot sampling, especially in highly dynamic systems, such as transitional waters. Hence, the MONITOOL project aimed to provide a robust database of dissolved and labile metal concentrations in transitional and coastal waters, based upon co-deployments of DGTs and collection of spot water samples at several sampling sites (England, France, Ireland, Italy, Northern Ireland, Portugal, Scotland and Spain), followed subsequently by DGT and water metal analysis. Samplings were carried out in 2018 and 2022, following agreed protocols developed in the framework of the project. The MONITOOL dataset includes metal concentrations from DGTs, measured with Inductively Coupled Plasma Mass Spectrometry (ICP-MS: Cd, Co, Cu, Fe, Mn, Ni, Pb, Zn) and in concurrently collected spot water samples by ICP-MS (Al, Cd, Co, Cu, Mn, Ni, Pb, Zn) and Anodic/Cathodic Stripping Voltammetry (ASV/CSV: Cd, Pb, Ni). Moreover, data on seawater physical-chemical parameters (salinity, temperature, dissolved oxygen, pH, turbidity, total suspended solids, dissolved organic carbon, and total organic carbon) is provided. This database presents the results obtained using, concurrently, different forms of sampling and analytical techniques, enabling the comparison of the results obtained by these strategies and allowing the adaptation of EQS in marine water (EQS marine water) to DGTs (EQS DGT), in the context of the WFD. Moreover, due to the large number of sampling sites, it could also be used for other types of research, such as those dealing with metal speciation or the determination of baseline levels.

An international intercomparison exercise on passive samplers (DGT) for monitoring metals in marine waters under a regulatory context.

In order to move forward in the acceptance of a novel contaminant monitoring technique (Diffusive Gradients in Thin-films: DGT) for assessment of marine water bodies, sensu the WFD, an Inter-Laboratories Comparison (ILC) exercise (nine Europeans laboratories) was organized in the framework of the Interreg Atlantic Area MONITOOL project, which focused on the use of the DGT technique for the measurement of WFD priority metals (Cd, Ni and Pb). Reproducible results were obtained for each metal by several laboratories, supporting the assertion that DGT analysis can be performed satisfactorily by laboratories experienced in measuring metals at trace levels in marine environments, even if they have limited practice in DGT analysis. According to the Z-score analysis, among the 9 participating laboratories, 3 had 100 % of satisfactory results for Cd, Ni, and Pb, 3 had >80 % satisfactory results and 2 had about 60 % satisfactory results. This work highlights the need to clearly describe the DGT method in order to control sources of contamination during analytical steps, in particular the resin gel retrieval and the elution steps. Such international intercomparison exercise is an important step to develop the laboratory network involved in DGT analysis and contributes to the improvement of data quality.

Metals concentrations in transitional and coastal waters by ICPMS and voltammetry analysis of spot samples and passive samplers (DGT).

This study investigates the relationships among Ni, Cd and Pb's different chemical forms determined by different methodologies in coastal and transitional waters across a broad geographical scale. Concentrations were measured in spot samples and through passive sampling (DGT). High variability of metal concentrations was found among sampling sites and methodologies due to natural water fluctuations rather than to a given metal or method. Total dissolved metal concentrations in spot samples were lower than the EQS-WFD values. The labile fractions of Cd and Pb, measured in spot samples by Anodic Stripping Voltammetry and by DGT-ICPMS, were highly correlated. Similar labilities were found for Cd, while for Pb, the ASV labile fraction was ≈50% lower. These results reflect the pool of mobile and labile species available towards each technique kinetic window, and they seem not to be affected by discrete sampling flaws.

Trace metal levels in the edible tissues of sea cucumbers (Holothuria tubulosa and Holothuria polii) from Sardinia (Western Mediterranean).

Sea cucumbers represent an important part of the diet in Asian and Pacific regions and are also used in traditional medicine. These habits have led to the overexploitation of local sea cucumber populations in these areas, driving the pursuit of new stock regions, such as Mediterranean areas. In Italy, contrarily to that observed for other Mediterranean countries, the exploitation of sea cucumber stocks is not extensive, which opens a new market opportunity. Thus, from a food safety perspective, this work aims at reporting the first assessment of trace metal concentrations (As, Cd, Cr, Cu, Hg, Ni, Pb) in the edible tissues of Holothuria polii and Holoturia tubulosa collected in Sardinia, the second-largest island in the Mediterranean Sea. Metal concentrations found in H. polii were generally higher than in H. tubulosa. However, in both species, they were lower than those reported for other areas of the Western Mediterranean. Cd, Hg, and Pb were below the limits established for seafood in Europe. As concentrations were in the range of those measured in other commercial seafood species in the Mediterranean. Thus, these species may be harvested and traded to fulfil the demands of local and international markets.

Concurrent sampling of transitional and coastal waters by Diffusive Gradient in Thin-films (DGT) and spot sampling for trace metals analysis.

This protocol was developed based on the knowledge acquired in the framework of the Interreg MONITOOL project (EAPA_565/2016) where extensive sampling campaigns were performed in transitional and coastal waters covering eight European countries. It provides detailed procedures and guidelines for the sampling of these waterbodies by concurrent collection of discrete water samples and the deployment of Diffusive Gradient in Thin-films (DGT) passive samplers for the measurement of trace metal concentrations. In order to facilitate the application of this protocol by end-users, it presents steps to follow in the laboratory prior to sampling campaigns, explains the procedures for field campaigns (including in situ measurement of supporting parameters) and subsequent sample processing in the laboratory in preparation for trace metal analyze by inductively coupled plasma-mass spectrometry (ICP-MS) and voltammetry. The protocol provides a systematic, coherent field sampling and sample preparation strategy that was developed in order to ensure comparability and reproducibility of the data obtained from each project Partner in different regions. • Standardization of the concurrent sampling of transitional and coastal waters by DGT passive samplers and spot sampling. • Robust procedures and tips based on existing international standards and comprehensive practical experience. • Links to demonstration videos produced within the MONITOOL project.

Assessing variability in the ratio of metal concentrations measured by DGT-type passive samplers and spot sampling in European seawaters.

The current study evaluates the effect of seawater physico-chemical characteristics on the relationship between the concentration of metals measured by Diffusive Gradients in Thin films (DGT) passive samplers (i.e., DGT-labile concentration) and the concentrations measured in discrete water samples. Accordingly, Inductively Coupled Plasma Mass Spectrometry (ICP-MS) was used to measure the total dissolved metal concentrations in the discrete water samples and the labile metal concentrations obtained by DGT samplers; additionally, lead and cadmium conditional labile fractions were determined by Anodic Stripping Voltammetry (ASV) and total dissolved nickel was measured by Cathodic Stripping Voltammetry (CSV). It can be concluded that, in general, the median ratios of DGT/ICP and DGT/ASV(CSV) were lower than 1, except for Ni (median ratio close to 1) and Zn (higher than 1). This indicates the importance of speciation and time-integrated concentrations measured using passive sampling techniques, which is in line with the WFD suggestions for improving the chemical assessment of waterbodies. It is the variability in metal content in waters rather than environmental conditions to which the variability of the ratios can be attributed. The ratios were not significantly affected by the temperature, salinity, pH, oxygen, DOC or SPM, giving a great confidence for all the techniques used. Within a regulatory context such as the EU Water Framework Directive this is a great advantage, since the simplicity of not needing to use corrections to minimize the effects of environmental variables could help in implementing DGTs within monitoring networks.

Evaluation of Rhodotorula spp. contamination in hospital environments.

Microbiological monitoring of air and surfaces is a valuable tool for assessing the level of environmental contamination by bacteria and fungi in hospitals, especially in high-risk wards. The presence of these microorganisms requires strict control in view of their role as opportunistic pathogens. As regard yeasts, Candida albicans is one of the main opportunistic pathogens. However, in recent years, many other "non-Albicans" species have been identified that are implicated in the onset of both localised infections in immunocompetent patients and systemic infections in immunocompromised patients. The aim of the work was to evaluate the frequency of isolation of Rhodotorula spp. in environmental samples of hospital origin in order to highlight how its presence may represent an increased risk for hospitalised patients. The samples were taken in at-risk wards, divided into six types of environments, in three hospitals in southern Sardinia between 2016 and 2019. The presence of moulds and yeasts was higher in the air (55.7%) than on surfaces (44.3%) in the hospital environments (p < 0.001). Similarly, the isolation frequency of Rhodotorula spp. was found to be greater (p < 0.001) in the air (14.2%) than on surfaces (9.2%). This study made it possible to evaluate the presence of pathogenic fungi in hospital environments, paying particular attention to Rhodotorula spp. since it is an emerging opportunistic yeast. It also highlights the importance of control measures especially in those where immunocompromised patients are present.

Potential issues of well water in domestic use in a retrospective study from 2014 to 2018.

Well water requires chemical, physical-chemical, and microbiological analysis to ensure that the water used for irrigation or for human consumption presents no specific risks. The aim of the present work was to determine the microbiological risk of samples taken from wells in south-central Sardinia. In this survey, 55 water samples were taken from private wells in the period from 2014 to 2018. Quality was assessed through the detection of the parameters required by national law. The analysis of the data showed that only 36.4% of the samples (20 wells) complied with legal limits for water for human consumption. The most isolated species was P. aeruginosa, present in 29.1%; its high concentration and recorded Coliforms suggest that the disinfection of the wells was insufficient or not performed properly if implemented. Such data suggest the need to pay more attention to the waters used not only for human consumption but also for irrigation, since the cultivated vegetables could become a vehicle for potentially pathogenic microorganisms. National legislation sets contamination limits only for water intended for human consumption and not for irrigation water. However, based on these considerations, it appears necessary to develop reference standards for irrigation water.

Ten-Year Retrospective Analysis of Legionella Diffusion in Hospital Water Systems and Its Serogroup Seasonal Variation.

INTRODUCTION: Legionella spp. are ubiquitous aquatic organisms found to be associated with community-acquired pneumoniae (CAP) as well as hospital-acquired pneumonia (HAP). Direct inhalation of aerosols from environmental colonisation is typically the source of infection. The aim of this study was to determine the level of colonisation in hospital water supply systems in order to assess the criticality of the water distribution network and strengthen preventive measures. METHODS: From 2009 to 2018, 769 water samples were collected and then analysed according to the standard methods indicated in ISO11731-2:2004 and ISO11731:2017 for Legionella detection. RESULTS: The samples were positive in 37.1% cases (n. 285) and negative in 62.9% cases (n. 484). The threshold of 10,000 CFU/L was exceeded in 15.1% cases and led to decolonisation as indicated by Italian and European ECDC guidelines. In the autumn-winter period SG1 showed a positivity of 41.2% (n. 40) with a decrease in the spring-summer period with 9.6% (n. 18) of positivity. In contrast, SG2-15 showed a positivity of 30.9% (n. 30) in autumn-winter, which tends to increase to 56.9% (n. 112) in spring-summer (p < 0.001). CONCLUSION: Surprisingly, besides showing a seasonal trend already described previously in the literature, the positivity of our sample was not balanced even for serogroups in the two periods. This could be due to genetic differences and ecological niches to be further investigated that could also have links with the greater pathogenicity of SG1. Environmental microbiological surveillance and risk assessment should be performed more frequently and disinfection must be carried out, especially in health facilities where people are more susceptible to infections.

Operational DGT threshold values for metals in seawater from protected coastal areas in Sardinia (Western Mediterranean).

Diffusive gradients in thin films (DGTs) were used for monitoring metal (Cd, Cu, Ni, and Pb) concentrations in protected and non-protected coastal areas in Sardinia (Western Mediterranean). The deployment of DGTs in relatively undisturbed areas enabled calculation of operational DGT threshold values, which can be used for assessments of the environmental quality of coastal areas. The DGT thresholds were defined as the median metal concentrations that were found in protected areas, which ensured consideration of the natural variability of the different study sites. The calculated DGT thresholds were 11.6 ng L-1 for Pb, 5.1 ng L-1 for Cd, 63 ng L-1 for Cu and 152 ng L-1 for Ni. A comparison of the calculated DGT thresholds with previous DGT studies in the area demonstrated their suitability for identifying sites of environmental concern in the Western Mediterranean.

Passive sampling monitoring of PAHs and trace metals in seawater during the salvaging of the Costa Concordia wreck (Parbuckling Project).

Passive sampling techniques were used for monitoring trace metals and polycyclic aromatic hydrocarbons (PAHs) in the seawater surrounding the Costa Concordia shipwreck (Isola del Giglio, Italy). The monitoring lasted two and a half years (2012-2014) and considered all four phases of the "parbuckling project": stabilisation of the wreckage, installation of steel caissons on both sides of the wreck, parbuckling, and refloating. Dissolved trace metals (Cd, Cr, Cu, Hg, Ni, Pb, and V) were measured with diffusive gradients in thin films (DGT), while freely dissolved PAHs were measured with semi-permeable membrane devices (SPMDs). Passive sampling allowed to detect very low concentrations of contaminants, and indicated significant differences among the sampling stations and among the different steps for salvaging the wreck. The results suggested that the main source of contamination was the heavy working vessel traffic at the disaster site, rather than the release of contaminants from the wreck.

Presence of Trihalomethanes in ready-to-eat vegetables disinfected with chlorine.

Trihalomethanes (THMs) - CHCl3, CHCl2Br, CHClBr2 and CHBr3 - are drinking water disinfection by-products (DBPs). These compounds can also be absorbed by different types of foods, including ready-to-eat (RTE) fresh vegetables. The potential absorption of THMs during washing of RTE vegetables could pose a potential risk to consumers' health. The concentration of THMs in the water used in the manufacturing process of these products shall not exceed the limit of 100 or 80 µgL-1 according to European Union (EU) and United States legislation, respectively. By contrast, there is little information about the presence of such compounds in the final product. This study evaluated the concentration of THMs in different types of RTE vegetables (carrots, iceberg lettuce, lettuce, mixed salad, parsley, parsley and garlic, rocket salad, valerian) after washing with chlorinated water. In the 115 samples analysed, the average value of total THMs was equal to 76.7 ng g-1. Chloroform was the THM present in the largest percentage in all the RTE vegetables. These results show that the process of washing RTE vegetables should be optimised in order to reduce the risk for consumers associated with the presence of DBPs.

Exploring the bacterial gut microbiota of supralittoral talitrid amphipods.

Talitrid amphipods (sandhoppers and beach fleas) are typical of the supralittoral zone. They are known to thrive on stranded materials, including detrital marine angiosperms and macroalgae, as well as occasional dead animals. In this work, the gut microbiota of five species of talitrid amphipods (Talitrus saltator, Talorchestia ugolinii, Sardorchestia pelecaniformis, Orchestia montagui and Orchestia stephenseni) collected in Sardinia (Italy) has been investigated through: i) metabarcoding analysis of the amplified 16S rRNA V4 region; and ii) quantification of family 48 glycosyl hydrolase genes (GHF48), involved in cellulose degradation. Results indicate that, though talitrid gut biodiversity is not directly related to taxon or sampling locality, the animals' digestive tracts may host species-specific bacterial communities. In particular, gut microbiota of O. montagui, an inhabitant of Posidonia banquettes and macro-algae mat, showed the greatest differences in taxonomic composition and the highest proportion of GHF48 genes with respect to 16S rRNA genes. These results suggest that the different talitrid species may host species-specific bacterial communities whose function could partially reflect the different microhabitats and food preferences of their host.

Heavy metal accumulation in surface sediments at the port of Cagliari (Sardinia, western Mediterranean): Environmental assessment using sequential extractions and benthic foraminifera.

Superficial sediments were taken at the port of Cagliari (Sardinia, Italy), which includes the oil terminal of one of the largest oil refineries in the Mediterranean. Significant trace metal concentrations were found in the whole port area. Sequential extraction of metals from the different sediment fractions (BCR method) showed a higher risk of remobilisation for Cd, which is mostly bound to the exchangeable fraction. Foraminiferal density and richness of species were variable across the study area. The living assemblages were characterized by low diversity in samples collected close to the port areas. Ammonia tepida and bolivinids, which were positively correlated with concentrations of heavy metals and organic matter content, appeared to show tolerance to the environmental disturbance. The sampling sites characterized by the highest values of biotic indices were located far from the port areas and present an epiphytic and epifaunal biocoenosis.

Interpretation of coastal sediment quality based on trace metal and PAH analysis, benthic foraminifera, and toxicity tests (Sardinia, Western Mediterranean).

An integrated approach for the assessment of coastal sediment quality was utilised in three areas of Sardinia (Western Mediterranean, Italy). Sediments were analysed for trace metals and polycyclic aromatic hydrocarbons (PAHs), while benthic foraminifera were used as bioindicators. Furthermore, the embryo-toxicity test was used to provide ecologically relevant information using rapid and cost-effective screening tools. The aim was to evaluate the usefulness of coupling different analytical tools. The results revealed the presence of polluted sediments in areas exposed to petrochemical industries, smelters or military settlements. However, while foraminifera have presented similar indications for chemical analysis of contamination levels in the different areas, the toxicity test exhibited a poor relationship with the contaminants measured individually. The results raise questions concerning the bioavailability of contaminants released by sediments in the water column. Overall, the toxicity rate was significant in many samples in comparison with other sites studied in other Mediterranean regions.

Using SPMDs to monitor the seawater concentrations of PAHs and PCBs in marine protected areas (Western Mediterranean).

Aqueous concentrations of individual polycyclic aromatic hydrocarbons (PAHs) and polychlorinated biphenyls (PCBs) were determined in coastal sites of two marine protected areas (MPAs), that is, Asinara and the La Maddalena Archipelago, in Sardinia (Western Mediterranean Sea). The use of semi-permeable membrane devices (SPMDs) enabled the detection of dissolved PAHs and PCBs, even in very low concentrations of (pgL(-1)), in seawater. The results reveal significant differences between the two sampling areas relative to the concentration of the individual PAHs, which provide information concerning the pollution sources affecting relatively pristine environments. The PCBs were generally observed at levels below the detection limits of the utilised method.

Macroalgae and DGT as indicators of available trace metals in marine coastal waters near a lead-zinc smelter.

The levels of Cd, Cu, Pb, and Zn were determined in the commonest species of green, red, and brown algae collected from five coastal sites in south-western Sardinia (Italy), an area with a long history of mining and smelting. The usefulness of employing Enteromorpha sp. and Padina pavonica (L.) Thivy to monitor metal pollution was evaluated, while diffusive gradients in thin films (DGT) devices were used to measure dissolved metals in seawater. Levels of Cd and Pb were high enough to be of environmental concern in the whole study area. A significant relationship was found between the content of Pb in P. pavonica and DGT-labile Pb in seawater, suggesting that gross elemental concentrations of nonessential metals such as Pb in algal tissues are apparently controlled by the abundance of dissolved metal species in the ambient seawater. The results pointed out the usefulness of using both DGT and algal methods for a better understanding of trace metal availability in coastal waters.