Valutazione del rischio ecologico e sviluppo di strategie innovative per il monitoraggio della qualita’ dei corpi idrici: applicazione di un nuovo approccio integrato in un bacino pilota.

Freshwater is extremely precious; but even more precious than freshwater is clean freshwater. From the time that 2/3 of our planet is covered in water, we have contaminated our globe with chemicals that have been used by industrial activities over the last century in a unprecedented way causing harm to humans and wildlife. We have to adopt a new scientific mindset in order to face this problem so to protect this important resource. The Water Framework Directive (European Parliament and the Council, 2000) is a milestone legislative document that transformed the way that water quality monitoring is undertaken across all Member States by introducing the Ecological and Chemical Status. A “good or higher” Ecological Status is expected to be achieved for all waterbodies in Europe by 2015. Yet, most of the European waterbodies, which are determined to be at risk, or of moderate to bad quality, further information will be required so that adequate remediation strategies can be implemented. To date, water quality evaluation is based on five biological components (phytoplankton, macrophytes and benthic algae, macroinvertebrates and fishes) and various hydromorphological and physicochemical elements. The evaluation of the chemical status is principally based on 33 priority substances and on 12 xenobiotics, considered as dangerous for the environment. This approach takes into account only a part of the numerous xenobiotics that can be present in surface waters and could not evidence all the possible causes of ecotoxicological stress that can act in a water section. The mixtures of toxic chemicals may constitute an ecological risk not predictable on the basis of the single component concentration. To improve water quality, sources of contamination and causes of ecological alterations need to be identified. On the other hand, the analysis of the community structure, which is the result of multiple processes, including hydrological constrains and physico-chemical stress, give back only a “photograph” of the actual status of a site without revealing causes and sources of the perturbation. A multidisciplinary approach, able to integrate the information obtained by different methods, such as community structure analysis and eco-genotoxicological studies, could help overcome some of the difficulties in properly identifying the different causes of stress in risk assessment. In synthesis, the river ecological status is the result of a combination of multiple pressures that, for management purposes and quality improvement, have to be disentangled from each other. To reduce actual uncertainty in risk assessment, methods that establish quantitative links between levels of contamination and community alterations are needed. The analysis of macrobenthic invertebrate community structure has been widely used to identify sites subjected to perturbation. Trait-based descriptors of community structure constitute a useful method in ecological risk assessment. The diagnostic capacity of freshwater biomonitoring could be improved by chronic sublethal toxicity testing of water and sediment samples. Requiring an exposure time that covers most of the species’ life cycle, chronic toxicity tests are able to reveal negative effects on life-history traits at contaminant concentrations well below the acute toxicity level. Furthermore, the responses of high-level endpoints (growth, fecundity, mortality) can be integrated in order to evaluate the impact on population’s dynamics, a highly relevant endpoint from the ecological point of view. To gain more accurate information about potential causes and consequences of environmental contamination, the evaluation of adverse effects at physiological, biochemical and genetic level is also needed. The use of different biomarkers and toxicity tests can give information about the sub-lethal and toxic load of environmental compartments. Biomarkers give essential information about the exposure to toxicants, such as endocrine disruptor compounds and genotoxic substances whose negative effects cannot be evidenced by using only high-level toxicological endpoints. The increasing presence of genotoxic pollutants in the environment has caused concern regarding the potential harmful effects of xenobiotics on human health, and interest on the development of new and more sensitive methods for the assessment of mutagenic and cancerogenic risk. Within the WFD, biomarkers and bioassays are regarded as important tools to gain lines of evidence for cause-effect relationship in ecological quality assessment. Despite the scientific community clearly addresses the advantages and necessity of an ecotoxicological approach within the ecological quality assessment, a recent review reports that, more than one decade after the publication of the WFD, only few studies have attempted to integrate ecological water status assessment and biological methods (namely biomarkers or bioassays). None of the fifteen reviewed studies included both biomarkers and bioassays. The integrated approach developed in this PhD Thesis comprises a set of laboratory bioassays (Daphnia magna acute and chronic toxicity tests, Comet Assay and FPG-Comet) newly-developed, modified tacking a cue from standardized existing protocols or applied for freshwater quality testing (ecotoxicological, genotoxicological and toxicogenomic assays), coupled with field investigations on macrobenthic community structures (SPEAR and EBI indexes). Together with the development of new bioassays with Daphnia magna, the feasibility of eco-genotoxicological testing of freshwater and sediment quality with Heterocypris incongruens was evaluated (Comet Assay and a protocol for chronic toxicity). However, the Comet Assay, although standardized, was not applied to freshwater samples due to the lack of sensitivity of this species observed after 24h of exposure to relatively high (and not environmentally relevant) concentrations of reference genotoxicants. Furthermore, this species demonstrated to be unsuitable also for chronic toxicity testing due to the difficult evaluation of fecundity as sub-lethal endpoint of exposure and complications due to its biology and behaviour. The study was applied to a pilot hydrographic sub-Basin, by selecting section subjected to different levels of anthropogenic pressure: this allowed us to establish the reference conditions, to select the most significant endpoints and to evaluate the coherence of the responses of the different lines of evidence (alteration of community structure, eco-genotoxicological responses, alteration of gene expression profiles) and, finally, the diagnostic capacity of the monitoring strategy. Significant correlations were found between the genotoxicological parameter Tail Intensity % (TI%) and macrobenthic community descriptors SPEAR (p<0.001) and EBI (p<0.05), between the genotoxicological parameter describing DNA oxidative stress (ΔTI%) and mean levels of nitrates (p<0.01) and between reproductive impairment (Failed Development % from D. magna chronic bioassays) and TI% (p<0.001) as well as EBI (p<0.001). While correlation among parameters demonstrates a general coherence in the response to increasing impacts, the concomitant ability of each single endpoint to be responsive to specific sources of stress is at the basis of the diagnostic capacity of the integrated approach as demonstrated by stations presenting a mismatch among the different lines of evidence. The chosen set of bioassays, as well as the selected endpoints, are not providing redundant indications on the water quality status but, on the contrary, are contributing with complementary pieces of information about the several stressors that insist simultaneously on a waterbody section providing this monitoring strategy with a solid diagnostic capacity. Our approach should provide opportunities for the integration of biological effects into monitoring programmes for surface water, especially in investigative monitoring. Moreover, it should provide a more realistic assessment of impact and exposure of aquatic organisms to contaminants. Finally this approach should provide an evaluation of drivers of change in biodiversity and its causalities on ecosystem function/services provision, that is the direct and indirect contributions to human well-being.

The ecology of algae in relation to river water equality surveillance

The broad objectives of the work were to develop standard methods for the routine biological surveillance of river water quality, using the non-planktonic algae. Studies on sampling methodology indicated that natural substrata should be sampled directly wherever possible, but for routine purposes, only a semi-quantitative approach was found to be feasible. Artificial substrata were considered to be useful for sample collection in deeper waters, and of three different types tested, Polythene strips were selected for further investigation essentially on grounds of practicality. These were tested in the deeper reaches of a wide range of river types and water qualities: 26 pool sites in 14 different rivers were studied over a period of 9 months. At each site, the assemblages developing on 3 strips following a 4, or less commonly, an 3 week immersion period were analysed quantitatively. Where possible, the natural substrata were also sampled semi-quantitatively at each site, and at a nearby riffle. The results of this survey were very fragmentary: many strips failed to yield useful data, and the results were often difficult to interpret, and of limited value for water quality surveillance purposes. In one river, the Churnet, the natural substrata at 14 riffle sites were sampled semi-quantitatively on 14 occasions at intervals of 4 weeks. In this survey, the results were more readily interpreted in relation to water quality, and no special data processing was found to be necessary or helpful. Further studies carried out on the filamentous green alga Cladophora showed that this alga may have some value as a bioaccumulation indicator for metals, and as a bioassay organism for the assessment of the algal growth promoting potential of natural river waters.

Ecological studies on:(a) the use of colonisation samplers in relation to biological surveillance of river water quality (b) the requirements of freshwater gastropoda

Some of the factors affecting colonisation of a colonisation sampler, the Standard Aufwuchs Unit (S. Auf. U.) were investigated, namely immersion period, whether anchored on the bottom or suspended, and the influence of riffles. It was concluded that a four-week immersion period was best. S. Auf. U. anchored on the bottom collected both more taxa and individuals than suspended ones. Fewer taxa but more individuals colonised S. Auf. U. in the potamon zone compared to the rhithron zone with a consequent reduction in the values of pollution indexes and diversity. It was concluded that a completely different scoring system was necessary for lowland rivers. Macroinvertebrates colonising S. Auf. U. in simulated streams, lowland rivers and the R. Churnet reflected water quality. A variety of pollution and diversity indexes were applied to results from lowland river sites. Instead of these, it was recommended that an abbreviated species - relative abundance list be used to summarise biological data for use in lowland river surveillance. An intensive study of gastropod populations was made in simulated streams. Lynnaea peregra increased in abundance whereas Potamopyrgas jenkinsi decreased with increasing sewage effluent concentration. No clear-cut differences in reproduction were observed. The presence/absence of eight gastropod taxa was compared with concentrations of various pollutants in lowland rivers. On the basis of all field work it appeared that ammonia, nitrite, copper and zinc were the toxicants most likely to be detrimental to gastropods and that P. jenkinsi and Theodoxus fluviatilis were the least tolerant taxa. 96h acute toxicity tests of P. jenkinsi using ammonia and copper were carried out in a flow-through system after a variety of static range finding tests. P. jenkinsi was intolerant to both toxicants compared to reports on other taxa and the results suggested that these toxicants would affect distribution of this species in the field.

The application of artificial neural networks to the interpretation and classification of freshwater benthic invertebrate communities

This thesis presents a thorough and principled investigation into the application of artificial neural networks to the biological monitoring of freshwater. It contains original ideas on the classification and interpretation of benthic macroinvertebrates, and aims to demonstrate their superiority over the biotic systems currently used in the UK to report river water quality. The conceptual basis of a new biological classification system is described, and a full review and analysis of a number of river data sets is presented. The biological classification is compared to the common biotic systems using data from the Upper Trent catchment. This data contained 292 expertly classified invertebrate samples identified to mixed taxonomic levels. The neural network experimental work concentrates on the classification of the invertebrate samples into biological class, where only a subset of the sample is used to form the classification. Other experimentation is conducted into the identification of novel input samples, the classification of samples from different biotopes and the use of prior information in the neural network models. The biological classification is shown to provide an intuitive interpretation of a graphical representation, generated without reference to the class labels, of the Upper Trent data. The selection of key indicator taxa is considered using three different approaches; one novel, one from information theory and one from classical statistical methods. Good indicators of quality class based on these analyses are found to be in good agreement with those chosen by a domain expert. The change in information associated with different levels of identification and enumeration of taxa is quantified. The feasibility of using neural network classifiers and predictors to develop numeric criteria for the biological assessment of sediment contamination in the Great Lakes is also investigated.

Sedimentology, palaeontology and diagenesis of the Much Wenlock limestone formation

Lithofacies distribution indicates that the Much Wenlock Limestone Formation of England and South Wales was desposited on a shelf which was flat and gently subsiding in the north, but topographically variable in the south. Limestone deposition in the north began with 12m of alga-rich limestone, which formed an upward shoaling sequence. Deepening then led to deposition of calcareous silty mudstones on the northern shelf. The remainder of the formation in this area formed during a shelf-wide regression, culminating in the production of an E to W younging sandbody. Lithofacies distribution on the southern shelf was primarily controlled by local subsidence. Six bedded lithofacies are recognised which contain 14 brachiopod/bryozoan dominated assemblages, of which 11 are in situ and three consist of reworked fossils. Microfacies analysis is necessary to distinguish assemblages which reflect original communities from those which reflect sedimentary processes. Turbulence, substrate-type, ease of feeding and other organisms in the environment controlled faunal distribution. Reefs were built dominantly by corals, stromatoporoids, algae and crinoids. Coral/stromatoporoid (Type A) reefs are common, particularly on the northern shelf, where they formed in response to shallowing, ultimately growing in front of the advancing carbonate sandbody. Algae dominate Type B and Type C reefs, reflecting growth in areas of poor water circulation. Lithification of the formation began in the marine-phreatic environment with precipitation of aragonite and high Mg calcite, which was subsequently altered to turbid low Mg calcite. Younger clear spars post-date secondary void formation. The pre-compactional clear spars have features which resemble the products of meteoric water diagenesis, but freshwater did not enter the formation at this time. The pre-compactional spars were precipitated by waters forced from the surrounding silty mudstones at shallow burial depths. Late diagenetic products are stylolites, compaction fractures and burial cements.

Fish farm effluent control and the development of an expert system

In recent years, freshwater fish farmers have come under increasing pressure from the Water Authorities to control the quality of their farm effluents. This project aimed to investigate methods of treating aquacultural effluent in an efficient and cost-effective manner, and to incorporate the knowledge gained into an Expert System which could then be used in an advice service to farmers. From the results of this research it was established that sedimentation and the use of low pollution diets are the only cost effective methods of controlling the quality of fish farm effluents. Settlement has been extensively investigated and it was found that the removal of suspended solids in a settlement pond is only likely to be effective if the inlet solids concentration is in excess of 8 mg/litre. The probability of good settlement can be enhanced by keeping the ratio of length/retention time (a form of mean fluid velocity) below 4.0 metres/minute. The removal of BOD requires inlet solids concentrations in excess of 20 mg/litre to be effective, and this is seldom attained on commercial fish farms. Settlement, generally, does not remove appreciable quantities of ammonia from effluents, but algae can absorb ammonia by nutrient uptake under certain conditions. The use of low pollution, high performance diets gives pollutant yields which are low when compared with published figures obtained by many previous workers. Two Expert Systems were constructed, both of which diagnose possible causes of poor effluent quality on fish farms and suggest solutions. The first system uses knowledge gained from a literature review and the second employs the knowledge obtained from this project's experimental work. Consent details for over 100 fish farms were obtained from the public registers kept by the Water Authorities. Large variations in policy from one Authority to the next were found. These data have been compiled in a computer file for ease of comparison.

A solar-powered reverse osmosis system for high recovery of freshwater from saline groundwater

Desalination of groundwater is essential in arid regions that are remote from both seawater and freshwater resources. Desirable features of a groundwater desalination system include a high recovery ratio, operation from a sustainable energy source such as solar, and high water output per unit of energy and land. Here we propose a new system that uses a solar-Rankine cycle to drive reverse osmosis (RO). The working fluid such as steam is expanded against a power piston that actuates a pump piston which in turn pressurises the saline water thus passing it through RO membranes. A reciprocating crank mechanism is used to equalise the forces between the two pistons. The choice of batch mode in preference to continuous flow permits maximum energy recovery and minimal concentration polarisation in the vicinity of the RO membrane. This study analyses the sizing and efficiency of the crank mechanism, quantifies energy losses in the RO separation and predicts the overall performance. For example, a system using a field of linear Fresnel collectors occupying 1000 m2 of land and raising steam at 200 °C and 15.5 bar could desalinate 350 m3/day from saline water containing 5000 ppm of sodium chloride with a recovery ratio of 0.7.

Liquid desiccant dehumidification and regeneration process to meet cooling and freshwater needs of desert greenhouses

Agriculture accounts for ~70% of freshwater usage worldwide. Seawater desalination alone cannot meet the growing needs for irrigation and food production, particularly in hot, desert environments. Greenhouse cultivation of high-value crops uses just a fraction of freshwater per unit of food produced when compared with open field cultivation. However, desert greenhouse producers face three main challenges: freshwater supply, plant nutrient supply, and cooling of the greenhouse. The common practice of evaporative cooling for greenhouses consumes large amounts of fresh water. In Saudi Arabia, the most common greenhouse cooling schemes are fresh water-based evaporative cooling, often using fossil groundwater or energy-intensive desalinated water, and traditional refrigeration-based direct expansion cooling, largely powered by the burning of fossil fuels. The coastal deserts have ambient conditions that are seasonally too humid to support adequate evaporative cooling, necessitating additional energy consumption in the dehumidification process of refrigeration-based cooling. This project evaluates the use of a combined-system liquid desiccant dehumidifier and membrane distillation unit that can meet the dual needs of cooling and freshwater supply for a greenhouse in a hot and humid environment.

Case-based reasoning enabling database mining for cryo-preserving algae applications

Case-based Reasoning's (CBR) origins were stimulated by a desire to understand how people remember information and are in turn reminded of information, and that subsequently it was recognized that people commonly solve problems by remembering how they solved similar problems in the past. Thus CBR became an appropriate way to find out the most suitable solution method for a new problem based on the old methods for the same or even similar problems. The research highlights how to use CBR to aid biologists in finding the best method to cryo preserve algae. The study found CBR could be used successfully to find the similarity percentage between the new algae and old cases in the case base. The prediction result showed approximately 93.75% accuracy, which proves the CBR system can offer appropriate recommendations for most situations. © 2011 IEEE.

Climate change and freshwater zooplankton: what does it boil down to?

Recently, major advances in the climate–zooplankton interface have been made some of which appeared to receive much attention in a broader audience of ecologists as well. In contrast to the marine realm, however, we still lack a more holistic summary of recent knowledge in freshwater. We discuss climate change-related variation in physical and biological attributes of lakes and running waters, high-order ecological functions, and subsequent alteration in zooplankton abundance, phenology, distribution, body size, community structure, life history parameters, and behavior by focusing on community level responses. The adequacy of large-scale climatic indices in ecology has received considerable support and provided a framework for the interpretation of community and species level responses in freshwater zooplankton. Modeling perspectives deserve particular consideration, since this promising stream of ecology is of particular applicability in climate change research owing to the inherently predictive nature of this field. In the future, ecologists should expand their research on species beyond daphnids, should address questions as to how different intrinsic and extrinsic drivers interact, should move beyond correlative approaches toward more mechanistic explanations, and last but not least, should facilitate transfer of biological data both across space and time.

Climate change and plankton phenology in freshwater: current trends and future commitments

A solid body of empirical, experimental and theoretical evidence accumulated over recent years indicated that freshwater plankton experienced advance in phenology in response to climate change. Despite rapidly growing evidence for phenological changes, we still lack a comprehensive understanding of how climate change alters plankton phenology in freshwater. To overcome current limitations, we need to shed some light on trends and constraints in current research. The goal of this study is to identify current trends and gaps based on analysis of selected papers, by the help of which we can facilitate further advance in the field. We searched the literature for plankton phenology and confined our search to studies where climate change has been proposed to alter plankton phenology and rates of changes were quantified. We did not restrict our search for empirical ontributions; experimental and theoretical studies were considered as well. In the following we discuss the spatio-temporal setting of selected studies, contributions of different taxonomic groups, emerging methodological constraints, measures of phenological trends; and finally give a list of recommendations on how to improve our understanding in the field. The majority of studies were confined to deep lakes with a skewed geographical distribution toward Central Europe, where scientists have long been engaged in limnology. Despite these findings, recent studies suggest that plankton in running waters may experience change in phenology with similar magnitude. Average rate of advancement in phenology of freshwater plankton exceeded those of the marine plankton and the global average. Increasing study duration was not coupled either with increasing contribution of discontinuous data or with increasing rates of phenological changes. Future studies may benefit from i) delivering longterm data across scientific and political boundaries; ii) extending study sites to broader geographical areas with a more explicit consideration of running waters; iii) applying plankton functional groups; iv) increasing the application of satellite data to quantify phytoplankton bloom phenology; v) extending analyses of time series beyond the spring period; vi) using various metrics to quantify variation in phenology; vii) combining empirical, experimental and theoretical approaches; and last but not least viii) paying more attention to emergence dynamics, nonresponding species and trophic mismatch.

Quantifying nitrogen cycling rates in freshwater marshes of the southern Everglades using nitrogen-15 tracer techniques

This dissertation research project addressed the question of how hydrologic restoration of the Everglades is impacting the nutrient dynamics of marsh ecosystems in the southern Everglades. These effects were analyzed by quantifying nitrogen (N) cycle dynamics in the region. I utilized stable isotope tracer techniques to investigate nitrogen uptake and cycling between the major ecosystem components of the freshwater marsh system. I recorded the natural isotopic signatures (δ15N and δ 13C) for major ecosystem components from the three major watersheds of the Everglades: Shark River Slough, Taylor Slough, and C-111 basin. Analysis of δ15 N and δ13C natural abundance data were used to demonstrate the spatial extent to which nitrogen from anthropogenic or naturally enriched sources is entering the marshes of the Everglades. In addition, I measured the fluxes on N between various ecosystem components at both near-canal and estuarine ecotone locations. Lastly, I investigated the effect of three phosphorus load treatments (0.00 mg P m-2, 6.66 mg P m-2, and 66.6 mg P m-2) on the rate and magnitude of ecosystem N-uptake and N-cycling. The δ15N and δ13C natural abundance data supported the hypothesis that ecosystem components from near-canal sites have heavier, more enriched δ 15N isotopic signatures than downstream sites. The natural abundance data also showed that the marshes of the southern Everglades are acting as a sink for isotopically heavier, canal-borne dissolved inorganic nitrogen (DIN) and a source for "new" marsh derived dissolved organic nitrogen (DON). In addition, the 15N mesocosm data showed the rapid assimilation of the 15N tracer by the periphyton component and the delayed N uptake by soil and macrophyte components in the southern Everglades.