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Here, we review the state-of-the-art of limnological investigations in the large and deep lakes south of the Alps (DSLs), lakes Garda, Maggiore, Como, Iseo and Lugano. In the pre-industrial age, these lakes were ultra- or oligotrophic. Increasing anthropogenic pressure and the impact of global warming has led to an acceleration of eutrophication and a decrease in the frequency of full mixing episodes, which have induced a state of meromixis in lakes Lugano and Iseo. In the last two decades, other changes have been identified, including fundamental variations in the long-term dynamics and structure of phytoplankton communities, identification of new toxigenic cyanobacteria and cyanotoxins, increases in the introduction and establishment of allochthonous species, and continuous detection of new, emerging chemical pollutants. Overall, these fundamental changes are quickly transforming the features of the DSLs. The ability to document and reconstruct changes in lake aquatic biota and micropollutants was strongly dependent on both research efforts and the availability of technologies to appraise these changes. In this context, the ongoing adoption of new technological tools, such as high-frequency monitoring and high throughput sequencing, is opening the way to a new level of comprehension of physical, chemical and biological processes, and aquatic biodiversity. New outlooks and new conceptual frameworks are necessary to cope with the huge dimensions of high-throughput data and analysis of big data.

The taxonomic identification of organisms based on the amplification of specific genetic markers (metabarcoding) implicitly requires adequate discriminatory information and taxonomic coverage of environmental DNA sequences in taxonomic databases. These requirements were quantitatively examined by comparing the determination of cyanobacteria and microalgae obtained by metabarcoding and light microscopy. We used planktic and biofilm samples collected in 37 lakes and 22 rivers across the Alpine region. We focused on two of the most used and best represented genetic markers in the reference databases, namely the 16S rRNA and 18S rRNA genes. A sequence gap analysis using blastn showed that, in the identity range of 99-100%, approximately 30% (plankton) and 60% (biofilm) of the sequences did not find any close counterpart in the reference databases (NCBI GenBank). Similarly, a taxonomic gap analysis showed that approximately 50% of the cyanobacterial and eukaryotic microalgal species identified by light microscopy were not represented in the reference databases. In both cases, the magnitude of the gaps differed between the major taxonomic groups. Even considering the species determined under the microscope and represented in the reference databases, 22% and 26% were still not included in the results obtained by the blastn at percentage levels of identity ≥95% and ≥97%, respectively. The main causes were the absence of matching sequences due to amplification and/or sequencing failure and potential misidentification in the microscopy step. Our results quantitatively demonstrated that in metabarcoding the main obstacles in the classification of 16S rRNA and 18S rRNA sequences and interpretation of high-throughput sequencing biomonitoring data were due to the existence of important gaps in the taxonomic completeness of the reference databases and the short length of reads. The study focused on the Alpine region, but the extent of the gaps could be much greater in other less investigated geographic areas.

The formation of chemical and biological heterogeneity in lakes can be favored by physical and morphometric constrains. This study describes the results of four whole-lake field campaigns carried out in Lake Como (north Italy) during thermal stratification. The aim was to analyze the distribution of chemical-biological variables in a multi-basin lake as a result of internal and external physical drivers and constrains. Lake Como has a y-like shape encompassing three main sub-basins: northern, south-eastern, and south-western. Field data underlined: the presence of chemical-biological gradients between the south-western basin and the rest of the lake and the propagation of a fresher water-plume (formed by the two main northern inflows) into the northern basin and then into the south-western closed arm. The use of a three-dimensional hydrodynamic model showed a periodic movement of this plume that tends to enter the south-western basin first and then to return toward north, moving forward and back through the junction of the three arms. The entrance into the eastern basin, instead, occurs only secondarily. Wavelet transform analysis revealed common periodicity between the plume movement and the action of different external and internal lake-stressors, including: the discharge of the main inflow (period centered at 1, 3.3, and 6.5 day), the wind intensity (0.5 and 1 day) and the two main basin-scale internal wave motions: (3.3 day and 7.1 day). The periodic movement of the fresher water-plume enhances the water exchange and reduces the chemical-biological gradients between the western closed basin and the main lake, playing a crucial role in distributing both inorganic and organic materials at the lake basin-scale.

For effective lakes’ management, high-frequent water quality data on a synoptic scale are essential. The aim of this study is to test the suitability of the latest generation of satellite sensors to provide information on lake water quality parameters for the five largest Italian subalpine lakes. In situ data of phytoplankton composition, chlorophyll-a (chl-a) concentration and water reflectance were used in synergy with satellite observations to map some algal blooms in 2016. Chl-a concentration maps were derived from satellite data by applying a bio-optical model to satellite data, previously corrected for atmospheric effects. Results were compared with in situ data, showing good agreement. The shape and magnitude of water reflectance from different satellite data were consistent. Output chl-a concentration maps, show the distribution within each lake during blooming events, suggesting a synoptic view is required for these events monitoring. Maps show the dynamic of bloom events with concentration increasing from 2 up to 7 mg m−3 and dropping again to initial value in less than 20 days. Latest generation sensors were shown to be valuable tools for lakes monitoring, thanks to frequent, free of charge data availability over long time periods.

A first synoptic and trans-domain overview of plankton dynamics was conducted across the aquatic sites belonging to the Italian Long-Term Ecological Research Network (LTER-Italy). Based on published studies, checked and complemented with unpublished information, we investigated phytoplankton and zooplankton annual dynamics and long-term changes across domains: from the large subalpine lakes to mountain lakes and artificial lakes, from lagoons to marine coastal ecosystems. This study permitted identifying common and unique environmental drivers and ecological functional processes controlling seasonal and long-term temporal course. The most relevant patterns of plankton seasonal succession were revealed, showing that the driving factors were nutrient availability, stratification regime, and freshwater inflow. Phytoplankton and mesozooplankton displayed a wide interannual variability at most sites. Unidirectional or linear long-term trends were rarely detected but all sites were impacted across the years by at least one, but in many case several major stressor(s): nutrient inputs, meteo-climatic variability at the local and regional scale, and direct human activities at specific sites. Different climatic and anthropic forcings frequently co-occurred, whereby the responses of plankton communities were the result of this environmental complexity. Overall, the LTER investigations are providing an unparalleled framework of knowledge to evaluate changes in the aquatic pelagic systems and management options.

We estimated the external phosphorus load (EPL) between 1960 and 2015 in Lake Pusiano, a mid-sized subalpine lake that reached its maximum trophic state in the mid-1980s. Using historical data we also estimated the internal phosphorus load (IPL) between 1972 and 2015. EPL reached its maximum in the mid-1980s with phosphorus (P) values ~21 t yr−1 compared to the current value close to 6 t yr−1. IPL was one order of magnitude less and ranged between 0.25 t yr−1 (2015) and 3.6 (1985) t yr−1. The strong reduction of the P load determined a marked decrease of both P (from 200 to 23 μg L−1) and chlorophyll a (from 18 to 8 μg L−1) in-lake concentrations. The process of eutrophication and subsequent recovery, however, showed hysteresis between P load and in-lake P concentrations. In recent years, in particular, the P concentrations at winter overturn seem to be independent from the P load, related to modifications in the hydrological management of the lake that favored P flush-out in October–November, when the EPL is maximal. This process led to a marked decrease of the total P concentrations at winter overturn (23 μg L−1), which recently resulted in lower-than-target concentrations (30 μg L−1) established by the Lombardy Region. The lake, nevertheless, suffers from the presence of the toxic cyanobacterium Planktothrix rubescens, and eradicating this species and further improving water quality will likely require an additional abatement of the EPL.

The availability of more than thirty years of historical satellite data is a valuable source which could be used as an alternative to the sparse in-situ data. We developed a new homogenised time series of daily day time Lake Surface Water Temperature (LSWT) over the last thirty years (1986–2015) at a spatial resolution of 1Âkm from thirteen polar orbiting satellites. The new homogenisation procedure implemented in this study corrects for the different acquisition times of the satellites standardizing the derived LSWT to 12:00 UTC. In this study, we developed new time series of LSWT for five large lakes in Italy and evaluated the product with in-situ data from the respective lakes. Furthermore, we estimated the long-term annual and summer trends, the temporal coherence of mean LSWT between the lakes, and studied the intra-annual variations and long-term trends from the newly developed LSWT time series. We found a regional warming trend at a rate of 0.017°CÂyrÂ−1annually and 0.032°CÂyrÂ−1during summer. Mean annual and summer LSWT temporal patterns in these lakes were found to be highly coherent. Amidst the reported rapid warming of lakes globally, it is important to understand the long-term variations of surface temperature at a regional scale. This study contributes a new method to derive long-term accurate LSWT for lakes with sparse in-situ data thereby facilitating understanding of regional level changes in lake's surface temperature.

This work investigated the combined effects of nutrient availability and temperature on phytoplankton in large and deep lakes south of the Alps (lakes Garda, Iseo, Como, Lugano and Maggiore). The more eutrophic basins (Lugano and Iseo) showed a higher presence of cyanobacteria, green algae (Chlorophyta and Charophyta) and dinoflagellates (Dinophyta). Besides these two water bodies, high biomasses of cyanobacteria were recorded also in the oligo-mesotrophic Lake Garda. The development of these algal groups during the growing season showed a strong dependence on the surface spring availability of SRP, which, in turn, was related to winter climatic oscillations, deep mixing dynamics, and trophic status. A specific analysis carried out by applying additive mixed modelling, generalized least squares and mixed modelling, allowed investigation of the direct, seasonal effects of water temperature variations and trophic status on different algal groups. The dominant cyanobacteria (Oscillatoriales) showed only a partial relationship with temperature, while Nostocales and Chroococcales, which did not appear to have a close relationship with the trophic status of the lakes, were characterised by abrupt increases during the warmer months. High positive relationships with temperature were found for a few other algal groups (e.g., Chlorophyta, Charophyta and Dinophyta). Overall, the results indicated a positive relationship between the seasonal development of the more abundant and eutrophic-sensitive algal groups and the concurrent effect of trophic status and water temperature. Nevertheless, it was stressed that specific differences could be interpreted taking into account the different autoecological characteristics and susceptibilities of different species and functional groups to other stressing factors favouring losses, including, e.g., vertical sinking and grazing.

The concept of ¿reference conditions¿ describes the benchmark against which current conditions are compared when assessing the status of water bodies. In this paper we focus on the establishment of reference conditions for European lakes according to a phytoplankton biomass indicator - the concentration of chlorophyll-a. A mostly spatial approach (selection of existing lakes with no or minor human impact) was used to set the reference conditions for chlorophyll-a values, supplemented by historical data, paleolimnological investigations and modelling. The work resulted in definition of reference conditions and the boundary between ¿high¿ and ¿good¿ status for 15 main lake types and five ecoregions of Europe: Alpine, Atlantic, Central/Baltic, Mediterranean, and Northern. Additionally, empirical models were developed for estimating site-specific reference chlorophyll-a concentrations from a set of potential predictor variables. The results were recently formulated into the EU legislation, marking the first attempt in international water policy to move from chemical quality standards to ecological quality targets., JRC.H.1-Water Resources

A growing concern exists about the effects of chloride (Cl) on freshwater systems. Increasing Cl concentrations have been observed in the last few decades in several rivers and lakes, mainly in northern countries. In Italy, present levels and temporal changes of sodium (Na) and Cl in water bodies have rarely been assessed. Based on long-term data for the lakes of the subalpine district in Italy (Maggiore, Lugano, Como, Iseo, Garda), we analyzed trends affecting Cl and Na concentrations during the last 25years, with the aim of identifying temporal changes and assessing possible causes. An in-depth analysis is presented for Lake Maggiore. Positive temporal Na and Cl trends were evident in all studied lakes, with the trends increasing since early 2000s. Data for Lake Maggiore tributaries showed a clear seasonality (higher values in winter and early spring). The NaCl used as road de-icing agent, together with Cl discharge from wastewater treatment plants, were identified as the main causes for the observed trends. Chloride concentrations in the lakes are below the threshold limit for reduced water quality and below concentrations known to harm aquatic biota. However, considering the relevance of deep subalpine lakes, representing almost 80% of the total freshwater volume in Italy, these trends indicate an important chemical change, which warrants further analysis.

This paper offers a synoptic account of studies on the phytoplankton communities in the deep southern subalpine lakes (DSL) Garda, Iseo, Como, Lugano and Maggiore. The main cause of the degradation of the water quality in the DSL is eutrophication. The euphotic layers of these lakes are trophically different, ranging from the oligo-mesotrophy of lakes Maggiore and Garda to the meso-eutrophy of lakes Iseo and Lugano. The trophic status as estimated by using total phosphorus and chlorophyll a has provided consistent results in agreement with the models proposed by OECD (1982. Eutrophication of Waters. Monitoring, Assessment and Control, OECD, Paris). Though related with chlorophyll a and TP, the Secchi disk depths have significantly underestimated the trophic status of the DSL. Two trophic indices using the algal orders (PTIorders) and species (PTIspecies) were drawn up on the basis of the distribution of phytoplankton along a trophic gradient defined by the application of multivariate methods; the scores emerging from these indices were used to make a definitive ecological classification of water bodies on a scale from 1 to 5, in accordance with the Water Framework Directive. A third index (PTIOE) was computed as the ratio between the annual mean values of the cumulative biovolumes of two groups of algal orders with opposite trophic characteristics. The three PTI indices were highly correlated, providing a consistent classification of the water bodies. The indices proposed in this work were specifically adopted for use in the DSL. However, the criteria for their implementation constitute a robust and impartial tool for assessing similar indices in other lake typologies and for evaluating the degree of specificity of the trophic indicator values assigned to the single phytoplankton orders and species.

This paper presents a synoptic account of the most important results emerging from studies on the phytoplankton communities in the deep southern subalpine lakes Garda, Iseo, Como, Lugano and Maggiore (DSL) in the second half of the 1990s. At present, the trophogenic layers of these lakes are trophically different, ranging from the oligo-mesotrophy of lakes Maggiore and Garda to the meso-eutrophy of lakes Iseo and Lugano. The research confirmed the existence of a common pool of species developing in the DSL, as already suggested by early studies conducted on a seasonal basis from the end of the 1970s to the first half of the 1980s. However, multivariate analyses (Correspondence Analysis, CA, and a subsequent application of Non Metric Multidimensional Scaling) demonstrated that the species in this common pool were developing differently or exclusively along a geographic and a trophic gradient. The major differences in the geographic distribution were found between the easternmost lakes (Garda and Iseo) and those farthest to the West (Lugano and, partly, Maggiore), with intermediate characteristics in Lake Como. These differences were due mainly to changes in the dominance relationships and only secondarily to compositional changes. The detection of the ultimate causes of these differences should take into account other factors not considered in the paper (i.e. the specific analysis of the food webs, local climatic conditions, hydrology and seasonal input of nutrients). Despite the observed differences, common patterns in the sequence of seasonal assemblages in the DSL could be recognised and defined. The second gradient in the species distribution identified by CA was strongly correlated with the principal trophic descriptors (algal biomass and total phosphorus); this meant that the phytoplankton taxa could be ranked along a trophic spectrum, from oligotrophy to eutrophy. A brief examination of the main differences which have historically arisen with the progress of eutrophication in the DSL showed that the species identified in this study as being indicative of more eutrophic conditions were increasing in importance in some of the lakes.

Si propone il protocollo nazionale di campionamento dei macroinvertebrati in ambiente lacustre, standardizzato ed approvato dal Gruppo di Lavoro delle agenzie ambientali creato appositamente da ISPRA ambiente. Tale protocollo, inserito nel Decreto sul Monitoraggio, è quindi quello che ogni agenzia deve adottare per il campionamento dei laghi per ottemperare alle richieste della Direttiva Quadro sulle Acque.

Collection of methods to evaluate lake quality using biological elements

Winter air and spring surface water temperatures, spring epilimnetic phosphorus, and hypolimnetic oxygen in the deep lakes south of the Alps (Garda, Iseo, Como, Lugano, and Maggiore) showed a high degree of temporal coherence. The common temporal patterns were originating from the effects caused by winter climate, and from corresponding synchronisms in the interannual variations in the extent of the spring water renewal and replenishment of nutrients. In turn, the sequence of linked causal events was triggered by two atmospheric modes of variability relevant for the Mediterranean region, i.e. the East Atlantic pattern (EA) and the Eastern Mediterranean Pattern (EMP). In contrast, there were no significant relationships of air and water temperatures with the North Atlantic Oscillation. In oligotrophic lakes, which were characterised by weak vertical nutrient gradients, the spring replenishment of P was negligible and difficult to detect (Maggiore), or detectable but of minor importance (Garda and Como) compared to that measured in more enriched lakes (Iseo and Lugano). The applicability of EA and EMP in the study of the impact of climate on aquatic ecosystems will require to be tested by expanding the number, typology, and geographical location of water bodies in the Alpine and Mediterranean regions.

The author, underlined the general importance of the information towards the persons who receive hospital assistance and recalled also the historical bases and the international inquiry upon this matter, precises the reasons that need particular information procedure regarding the hospital infections, because the problems raised by these infections and the safety measures against them request to involve also all people entering the hospital as visitors. On the basis of some specific items fixed by the Joint Commission International Accreditation in order of the duties of the hospital directions, well applicable on this matter, the author suggests that the material impossibility to zeroing occurrence of the hospital infections, in case of litigations between hospitals and patients needs alternative dispute solutions. In this respect the author mentions the opportunities created by law in France and, very recently, in Italy too. Finally, the author points out the pretentions of the Italian Supreme Court about the completeness and the precision that must caractherize the procedure of informed consent about all risks of every medical activity, otherwise the liability of the hospitals and the members of their care staffs is quite presumed--even from the point of view of the penal aforethought--while the medical performance has been proper.

The intercalibration (IC) exercise is a key element in the implementation of the Water Framework Directive (WFD) in Europe. Its focus lies on the harmonization of national classification methods to guarantee a common understanding of ‘Good Ecological Status’ in surface waters. This article defines reference conditions and sets class boundaries for deep (mean depth >15 m, IC lake type L-AL3) and moderately deep (mean depth 3–15 m, IC lake type L-AL4) Alpine lakes >0.5 km2. Data were collated from each of the five EU member states included in the Alpine Geographical Intercalibration Group (Alpine GIG: Austria, France, Germany, Italy and Slovenia). Hydro-morphological, chemical and biological data from 161 sites (sampling stations) in 144 Alpine lakes over a period of seven decades were collated in a database. Based on a set of reference criteria, 18 L-AL3 and 13 L-AL4 reference sites were selected. Reference conditions were defined using a combined approach, based on historical, paleolimnological and monitoring data in conjunction with trophic modelling and expert judgement. Reference values and class boundaries were set for annual mean total biomass (biovolume), and then derived for annual mean chlorophyll-a using a regression between the two parameters. In order to allow for geographical differences within the Alpine GIG and to facilitate the inclusion of the broadly defined common IC types and national lake types, ranges were defined for each reference value. Range of reference values are 0.2–0.3 mg l−1 (L-AL3) and 0.5–0.7 mg l−1 (L-AL4) for total biovolume and 1.5–1.9 μg l−1 (L-AL3) and 2.7–3.3 μg l−1 (L-AL4) for chlorophyll-a. Depending on lake type and variable, the ecological quality ratios (EQR) for setting the class boundaries lie between 0.60 and 0.75 for the high/good class boundary and between 0.25 and 0.41 for the good/moderate class boundary. The response of sensitive phytoplankton taxa along a nutrient gradient and the occurrence of ‘undesirable conditions and secondary effects’ as defined in the WFD was used to validate the class boundary values, which are thus considered to be compliant with the requirements of the WFD.

We describe the larva of Protanypus sp. A from the Italian Alps. All the larval characteristics fit the diagnosis of the genus, but it is impossible to assign the specimens examined to one of the known species. The low number of labral scales (12-14) and the serrated median lamellae of the medioventral appendix of the prementum exclude the identity of the species with P. morio or with the East Palaearctic P. pseudomorio. The antennal ratio (2.3) excludes the identity with P. caudatus or P. forcipatus, which are the other two Protanypus species known from the Alpine region. In Sæther’s key (1975) the larva fits with the Nearctic P. ramosus, but identification of the species needs to be supported by pupal and adult material. In the Southern Alps, the genus is restricted to cold lakes at high altitude and is confirmed as an indicator of oligotrophic lakes.