Integrated Risk Assessment for WFD Ecological Status classification applied to Llobregat river basin (Spain). Part II - Evaluation process applied to five environmental Lines of Evidence.

Many indicators and indices related to a variety of biological, physico-chemical, chemical, and hydromorphological water conditions have been recently developed or adapted by scientists in order to support water managers in the Water Framework Directive (WFD) implementation. In this context, the achievement of a comprehensive and reliable Ecological Status classification of water bodies across Europe is hampered by the lack of harmonised procedures for selecting an appropriate set of indicators and integrating heterogeneous information in a flexible way. To this purpose, an Integrated Risk Assessment (IRA)(2) methodology was developed based on the Weight of Evidence approach. This method analyses and combines a set of environmental indicators grouped into five Lines of Evidence (LoE), i.e. Biology, Chemistry, Ecotoxicology, Physico-chemistry and Hydromorphology. The whole IRA methodology has been implemented as a specific module into a freeware GIS (Geographic Information System)-based Decision Support System, named MODELKEY DSS. This paper focuses on the evaluation of the four supporting LoE (i.e. Chemistry, Ecotoxicology, Physico-chemistry and Hydromorphology), and includes a procedure for a comparison of each indicator with proper thresholds and a subsequent integration process to combine the obtained output with the LoE Biology results in order to provide a single score expressing the Ecological Status classification. The approach supports the identification of the most prominent stressors, which are responsible for the observed alterations in the river basin under investigation. The results provided by the preliminary testing of the IRA methodology through application of the MODELKEY DSS to the Llobregat case study are finally reported and discussed.

Integrated risk assessment for WFD ecological status classification applied to Llobregat river basin (Spain). Part I-Fuzzy approach to aggregate biological indicators.

Water Framework Directive (WFD) requirements and recommendations for Ecological Status (ES) classification of surface water bodies do not address all issues that Member States have to face in the implementation process, such as selection of appropriate stressor-specific environmental indicators, definition of class boundaries, aggregation of heterogeneous data and information and uncertainty evaluation. In this context the "One-Out, All-Out" (OOAO) principle is the suggested approach to lead the entire classification procedure and ensure conservative results. In order to support water managers in achieving a more comprehensive and realistic evaluation of ES, an Integrated Risk Assessment (IRA) methodology was developed. It is based on the Weight of Evidence approach and implements a Fuzzy Inference System in order to hierarchically aggregate a set of environmental indicators, which are grouped into five Lines of Evidence (i.e. Biology, Chemistry, Ecotoxicology, Physico-chemistry and Hydromorphology). The whole IRA methodology has been implemented as an individual module into a freeware GIS (Geographic Information System)-based Decision Support System (DSS), named MODELKEY DSS. The paper focuses on the conceptual and mathematical procedure underlying the evaluation of the most complex Line of Evidence, i.e. Biology, which identifies the biological communities that are potentially at risk and the stressors that are most likely responsible for the observed alterations. The results obtained from testing the procedure through application of the MODELKEY DSS to the Llobregat case study are reported and discussed.

Environmental quality of transitional waters: the lagoon of Venice case study.

The health status of European aquatic environments, including transitional waters such as coastal lagoons, is regulated by the Water Framework Directive (WFD), which requires the classification of the water bodies' environmental quality and the achievement of a good ecological status by 2015. In the Venice lagoon, a transitional water body located in the northeastern part of Italy, the achievement of a good ecological status is hampered by several anthropogenic and natural pressures, such as sediment and water chemical contamination, and sediment erosion. In order to evaluate the lagoon's environmental quality according to the WFD (i.e. 5 quality classes, from High to Bad), an integrated Weight-of-Evidence methodology was developed and applied to classify the quality of the lagoon water bodies, integrating biological, physico-chemical, chemical, ecotoxicological, and hydromorphological data (i.e. Lines of Evidence, LOE). The quality assessment was carried out in two lagoon habitat typologies (previously defined on the basis of morphological, sediment, and hydrodynamic characteristics) which were selected taking into account the ecological gradient from sea to land, and the differences in anthropogenic pressure and contamination levels. The LOE classification was carried out by using indicators scored by comparing site specific conditions to reference conditions measured in lagoon reference sites, or provided by local, national or European regulations (e.g. Environmental Quality Standards, EQS, for chemicals). Finally, the overall quality status for each water body was calculated by a probabilistic approach, i.e. by reporting the final result as the frequency distribution of quality classes. The developed procedure was applied by using data and information concerning selected LOE and collected from monitoring programs and research studies carried out in the last 15 years in the lagoon of Venice. A set of sampling stations characterized by spatially and temporally coherent information for each LOE was selected, and among these stations, potential reference sites for each water body typology were identified. The quality assessment highlighted that there are specific lagoon areas, especially those located near the industrially developed area, which are highly affected by anthropogenic activities, and that chemical contamination is one of the main pressures affecting ecological status (e.g. macro-benthonic biodiversity) in the Venice lagoon. The integrated quality assessment procedure that was developed provided a new tool supporting decision making, as well as lagoon assessment and management.

Wave transport in random systems: multiple resonance character of necklace modes and their statistical behavior.

We present the experimental observation of multiple resonance transport of light waves, due to necklace states, in disordered one-dimensional systems. Transmission phase measurements allow us to identify these states unambiguously and investigate their statistical properties. A theoretical model is developed to describe the resonance statistics and the frequency dependance of the localization length.

Effects of transient cerebral ischemia on glial fibrillary acidic protein phosphorylation and immunocontent in rat hippocampus.

Transient global cerebral ischemia induced in rats by four-vessel occlusion for 20 min produced an increase in the immunocontent of glial fibrillary acidic protein and a protein phosphorylation response that was different in the CA1 and dentate gyrus areas of the hippocampus. We studied different times of reperfusion (one, four, seven, 14 and 30 days) and observed that the immunocontent and in vitro rate of phosphorylation of glial fibrillary acidic protein in the CA1 region was significantly increased at all intervals after the ischemic insult, indicating that the astrocytic response was maintained for at least 30 days. After reperfusion for 14 days a significant increase in the ratio "in vitro phosphorylation rate/immunocontent" in the CA1 region was observed when compared to control values, to other intervals and to the dentate gyrus, suggesting a hyperphosphorylation of this intermediate filament protein at this interval. In the dentate gyrus, an area less vulnerable to the insult, labelling and immunocontent of glial fibrillary acidic protein were equally increased from four days of reperfusion and the increase remained significant until 30 days, confirming that neuronal death is not the only determining factor for gliosis to occur. In control sham-operated animals, neither the CA1 region nor the dentate gyrus showed significant increases in labelling or immunocontent. Changes in the phosphorylation of glial fibrillary acidic protein may be essential for the plastic response of astrocytes to neuronal damage, as neurons and astrocytes can act as functional units involved in homeostasis, plasticity and neurotransmission.

Temporal profiles of the in vitro phosphorylation rate and immunocontent of glial fibrillary acidic protein (GFAP) after kainic acid-induced lesions in area CA1 of the rat hippocampus: demonstration of a novel phosphoprotein associated with gliosis.

The in vitro phosphorylation rate and immunocontent of glial fibrillary acidic protein was studied in slices of area CA1 of the rat hippocampus after stereotaxic injection of 1 nmol of kainic acid. For controls the contralateral hippocampus was injected with saline. Hippocampal tissue was incubated with [32P]phosphate and analysed by two-dimensional electrophoresis for phosphorylation rate and by immunoblotting for immunocontent. Both these parameters decreased during the first 4 days after injection and then started to increase at 10 days and continued to increase until at least 84 days. Except for a small excess of phosphorylation rate at 28 days, the relationship between immunocontent and in vitro phosphorylation rate of glial fibrillary acidic protein remained constant, indicating that the reactive gliosis was not associated with hypo- or a major hyperphosphorylation of this protein. Histology showed a pronounced loss of CA1 pyramidal cells 1 day after injection. At 28 days after injection the pyramidal cells had disappeared and only a few abnormal neurones were present. In contrast, immunocytochemistry after 28 days showed a marked increase in astrocytes reacting positive to the antibody in the strata radiatum and lacunosum moleculare. Besides glial fibrillary acidic protein the expression of several other proteins was upregulated as a result of the injection of kainic acid. These included phosphovimentin and an unknown phosphoprotein designated pp25 which co-migrated on 2-D gels with a prominent phosphoprotein expressed in primary cultures of astrocytes. Pp25 was expressed in lesioned tissue more frequently than phosphovimentin and with a time course that started earlier. Of particular interest was the expression of pp25 in the contralateral saline-injected hippocampus 1 day after injection of kainic acid. It is possible that pp25 will prove to be a sensitive marker of gliosis.