Laura Mancini

The European technical report on aquatic effect-based monitoring tools under the water framework directive

The Water Framework Directive (WFD), 2000/60/EC, requires an integrated approach to the monitoring and assessment of the quality of surface water bodies. The chemical status assessment is based on compliance with legally binding Environmental Quality Standards (EQSs) for selected chemical pollutants (priority substances) of EU-wide concern. In the context of the mandate for the period 2010 to 2012 of the subgroup Chemical Monitoring and Emerging Pollutants (CMEP) under the Common Implementation Strategy (CIS) for the WFD, a specific task was established for the elaboration of a technical report on aquatic effect-based monitoring tools. The activity was chaired by Sweden and co-chaired by Italy and progressively involved several Member States and stakeholders in an EU-wide drafting group. The main aim of this technical report was to identify potential effect-based tools (e.g. biomarkers and bioassays) that could be used in the context of the different monitoring programmes (surveillance, operational and investigative) linking chemical and ecological status assessment. The present paper summarizes the major technical contents and findings of the report.

Least Disturbed Condition for European Mediterranean rivers.

The present report describes a three-step approach that was used to characterize and define thresholds for the Least Disturbed Condition in Mediterranean streams of four different types, regarding organic pollution and nutrients, hydrological and morphological alterations, and land use. For this purpose, a common database composed of national reference sites (929 records) from seven countries, sampled for invertebrates, diatoms and macrophytes was used. The analyses of reference sites showed that small (catchment <100 km(2)) siliceous and non-siliceous streams were mainly affected by channelization, bank alteration and hydropeaking. Medium-sized siliceous rivers were the most affected by stressors: 25-43% of the samples showed at least slight alterations regarding channelization, connectivity, upstream dam influence, hydropeaking and degradation of riparian vegetation. Temporary streams were the least affected by hydromorphological changes, but they were nevertheless affected by alterations in riparian vegetation. There were no major differences between all permanent stream types regarding water quality, but temporary streams showed lower values for oxygenation (DO) and wider ranges for other variables, such as nitrates. A lower threshold value for DO (60%) was determined for this stream type and can be attributed to the streams natural characteristics. For all other river types, common limits were found for the remaining variables (ammonium, nitrate, phosphate, total P, % of artificial areas, % of intensive and extensive agriculture, % of semi-natural areas in the catchment). These values were then used to select the list of reference sites. The biological communities were characterized, revealing the existence of nine groups of Mediterranean invertebrate communities, six for diatoms and five for macrophytes: each group was characterized by specific indicator taxa that highlighted the differences between groups.

Biological quality of running waters in protected areas: the influence of size and land use

There are still substantial questions about whether protected areas affect the quality and biodiversity of surface waters within their borders. In this study, the size and land use of 19 protected areas of Latium Region (central Italy) were related to the biological quality of 32 streams running inside them. Additionally, the biological quality of 18 out of the 32 streams was compared with the quality recorded on the same streams outside the boundaries of the protected areas. The biological quality was assessed using the Extended Biotic Index, which indicates the macroinvertebrate community health. The quality of 32 study streams running through the protected areas was not related to the size of these areas, but it did reflect land use. On average, the 18 study sites inside protected areas had biological quality similar to external control sites. In the protected areas, the biological quality of streams was higher than for the same streams in the surrounding territory provided that anthropogenic changes were fewer. These data indicate that the creation of protected areas per se does not increase freshwater biodiversity and that land use has a major impact on the biological quality of the stream in a protected area. As a consequence, a higher number of reserves or landscape designations specifically created for aquatic conservation is necessary and recovery programs aimed at restoring physical habitats and reducing sources of impact to aquatic life have to be pursued. Also, where the anthropogenic impact is high (e.g., as in the case of strongly urbanised areas), the creation of effective protected areas might improve the biological quality of water courses.

The presence of a river bird, the dipper, in relation to water quality and biotic indices in central Italy

A good water quality bio-indicator should be sensitive to different forms of water degradation in any geographical area, and effectively track water quality changes in time. We investigated whether the presence of a bird, the dipper (Cinclus cinclus), in 47 reaches of 35 streams of central Italy was affected by stream pollution as assessed by the Extended Biotic Index. Data from the literature and interviews with ornithologists furnished information about past dipper presence and water quality in the study streams. In 93.3% of cases dippers occurred in unpolluted reaches. The dipper was absent from 93.7% of polluted or strongly polluted streams. Dippers disappeared from 50% of the study streams for which past information was available. In a few streams dippers disappeared after the quality of water had degraded. Water quality degradation in the study area, especially in the hill streams, is causing dipper areale to contract to higher altitudes. Dipper is a good water quality bio-indicator for Mediterranean streams.

Water quality assessment of rivers using diatom metrics across Mediterranean Europe: A methods intercalibration exercise

The European Water Framework Directive establishes a framework for the protection of water resources. However, common water management tools demand common understanding of assessment methods, so quality goals are equally met. Intercalibration of methods ensures the comparability of biological elements across similar geographical areas. Many aspects can influence the outcome of intercalibration: data sampling, treatment methods, taxonomic reliability of databases, choice of metrics for ecological quality status classification, and criteria for selecting reference sites. This study describes the potentials and constraints of the intercalibration of indices using diatoms for assessment of Mediterranean rivers. Harmonisation of diatom taxonomy and nomenclature was based on a previous ring test which took place at the European level. Four diatom indices (Indice de Polluosensibilité Spécifique-IPS, Indice Biologique Diatomées-IBD 2007, Intercalibration Common Metric Italy-ICMi and Slovenian Ecological Status assessment system) were intercalibrated using data from six European Mediterranean countries (Cyprus, France, Italy, Portugal, Slovenia and Spain). Boundaries between High/Good and Good/Moderate quality classes were harmonised by means of the Intercalibration Common Metric (ICM). Comparability between countries was assured through boundary bias and class agreement. The national boundaries were adjusted when they deviated more than a quarter of a class equivalent (0.25) from the global mean. All national methods correlated well with the ICM, which was sensitive to water quality (negatively correlated to nutrients). Achnanthidium minutissimum sensu lato was the most discriminative species of Good ecological status class. Planothidium frequentissimum, Gomphonema parvulum and Nitzschia palea were the most contributive to Moderate ecological status class. Some taxa were discriminative for both Good and Moderate ecological status classes due to low indication and ecological discriminative power but also due to differences in taxonomy between countries. This intercalibration exercise allowed establishment of common water quality goals across Mediterranean Europe, which is substantiated with the ICM.

Detection of Emerging and Re-Emerging Pathogens in Surface Waters Close to an Urban Area

Current knowledge about the spread of pathogens in aquatic environments is scarce probably because bacteria, viruses, algae and their toxins tend to occur at low concentrations in water, making them very difficult to measure directly. The purpose of this study was the development and validation of tools to detect pathogens in freshwater systems close to an urban area. In order to evaluate anthropogenic impacts on water microbiological quality, a phylogenetic microarray was developed in the context of the EU project µAQUA to detect simultaneously numerous pathogens and applied to samples from two different locations close to an urban area located upstream and downstream of Rome in the Tiber River. Furthermore, human enteric viruses were also detected. Fifty liters of water were collected and concentrated using a hollow-fiber ultrafiltration approach. The resultant concentrate was further size-fractionated through a series of decreasing pore size filters. RNA was extracted from pooled filters and hybridized to the newly designed microarray to detect pathogenic bacteria, protozoa and toxic cyanobacteria. Diatoms as indicators of the water quality status, were also included in the microarray to evaluate water quality. The microarray results gave positive signals for bacteria, diatoms, cyanobacteria and protozoa. Cross validation of the microarray was performed using standard microbiological methods for the bacteria. The presence of oral-fecal transmitted human enteric-viruses were detected using q-PCR. Significant concentrations of Salmonella, Clostridium, Campylobacter and Staphylococcus as well as Hepatitis E Virus (HEV), noroviruses GI (NoGGI) and GII (NoGII) and human adenovirus 41 (ADV 41) were found in the Mezzocammino site, whereas lower concentrations of other bacteria and only the ADV41 virus was recovered at the Castel Giubileo site. This study revealed that the pollution level in the Tiber River was considerably higher downstream rather than upstream of Rome and the downstream location was contaminated by emerging and re-emerging pathogens.

Weak Concordance between Fish and Macroinvertebrates in Mediterranean Streams

Although anthropogenic degradation of riverine systems stimulated a multi-taxon bioassessment of their ecological integrity in EU countries, specific responses of different taxonomic groups to human pressure are poorly investigated in Mediterranean rivers. Here, we assess if richness and composition of macroinvertebrate and fish assemblages show concordant variation along a gradient of anthropogenic pressure in 31 reaches across 13 wadeable streams in central Italy. Fish and invertebrate taxonomic richness was not correlated across sites. However, Mantel test showed that the two groups were significantly, albeit weakly, correlated even after statistically controlling for the effect of environmental variables and site proximity. Variance partitioning with partial Canonical Correspondence Analysis showed that the assemblages of the two groups were influenced by different set of environmental drivers: invertebrates were influenced by water organic content, channel and substratum features, while fish were related to stream temperature (mirroring elevation) and local land-use. Variance partitioning revealed the importance of biotic interactions between the two groups as a possible mechanisms determining concordance. Although significant, the congruence between the groups was weak, indicating that they should not be used as surrogate of each other for environmental assessments in these Mediterranean catchments. Indeed, both richness and patterns in nestedness (i.e. where depauperate locations host only a subset of taxa found in richer locations) appeared influenced by different environmental drivers suggesting that the observed concordance did not result from a co-loss of taxa along similar environmental gradients. As fish and macroinvertebrates appeared sensitive to different environmental factors, we argue that monitoring programmes should consider a multi-assemblage assessment, as also required by the Water Framework Directive.

Microbiological and 16S rRNA analysis of sulphite-reducing clostridia from river sediments in central Italy

BackgroundMicrobiological indicators are commonly used in the assessment of public health risks associated with fecal contamination of freshwater ecosystems. Sediments are a reservoir of microorganisms, and can thus provide information on past pollution events, not obtainable through the testing of surface water. Moreover, pathogens present in sediment may represent future threats to human health. Clostridium perfringens, a typical colonizer of sediments, has been suggested as an alternative indicator of fecal pollution. In order to be suitable for such purpose, the microorganism should be widely distributed in contaminated environments. The objective of this study was thus to determine the composition of the anaerobic community in sediment samples of the lower Tiber basin, in central Italy, through a combined approach involving granulometric analysis of sediment samples, as well as a microbiological and molecular (16S rRNA) analysis of strains.ResultsGranulometry showed a similar, clayey sediment composition, in most sampling sites. The microbiological method, employing, an adaptation of the standard method, proved to be effective in isolating anaerobic bacteria from the environmental matrix for the purpose of genetic analysis. Eighty-three strains of bacteria were isolated and the partial 16S rRNA gene sequenced. While biochemical analysis detected only C. perfringens strains, phylogenetic analysis indicated the presence of three clusters: C. perfringens, C. bifermentans and B. cereus, comprising eight taxa. C. perfringens, the commonest in almost all sediment sampling sites, was present in all sites, and in both seasons (seasonal sampling was carried out only along the Tiber and Aniene rivers). None of the described genetic profiles showed complete similarity with GenBank sequences.ConclusionThe study underlines the value of C. perfringens as an alternative microbial indicator of fecal contamination in river sediments. This is supported by the bacteriums presence in all sampling sites, and in both seasons, coupled with its detectability using commercial diagnostic kits.The study also illustrates the presence of an anaerobic community of considerable biodiversity in the lower Tiber basin, with C. perfringens as its main component. The 16S rRNA analysis, while confirming the phylogenetic relationships among isolated species, also showed haplotype patterns different from those present in the NCBI database.

Mercury (Hg) and methyl mercury (MeHg) concentrations in fish from the coastal lagoon of Orbetello, central Italy

Total mercury (Hg tot) and methyl mercury (MeHg) were quantified in several specimens of Dicentrarchus labrax and Sparus aurata from the east basin of the Orbetello lagoon, central Italy. The size of each specimen was recorded to estimate body burdens (BBs); =Hg tot and MeHg were measured in fillets of both species. Hg tot and MeHg in S. aurata ranged between 0.355-1.58 and 0.341-1.53 μg/g wet weight (ww), respectively; in D. labrax, their ranges were 0.284-2.54 and 0.214-2.35 μg/g ww. Approximately 90% of the concentrations measured exceeded Hg tot regulatory maximum level of 0.5 μg/g ww; however, exceedance rate was different in the two species studied. No correlations between specimen size and Hg tot or MeHg BBs were detected in this study.

Monitoring of freshwater toxins in European environmental waters by using novel multi-detection methods

Monitoring the quality of freshwater is an important issue for public health. In the context of the European project μAqua, 150 samples were collected from several waters in France, Germany, Ireland, Italy, and Turkey for 2 yr. These samples were analyzed using 2 multitoxin detection methods previously developed: a microsphere-based method coupled to flow-cytometry, and an ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method. The presence of microcystins, nodularin, domoic acid, cylindrospermopsin, and several analogues of anatoxin-a (ATX-a) was monitored. No traces of cylindrospermopsin or domoic acid were found in any of the environmental samples. Microcystin-LR and microcystin-RR were detected in 2 samples from Turkey and Germany. In the case of ATX-a derivatives, 75% of samples contained mainly H2 -ATX-a and small amounts of H2 -homoanatoxin-a, whereas ATX-a and homoanatoxin-a were found in only 1 sample. These results confirm the presence and wide distribution of dihydro derivatives of ATX-a toxins in European freshwaters. Environ Toxicol Chem 2017;36:645-654.