Assessment of the eutrophication status at Mediterranean sub-basin scale, within the European Marine Strategy Framework Directive.

The aim of this work is to define harmonized reference conditions and assessment thresholds for selected criteria elements of the Marine Strategy Framework Directive (MSFD) Descriptor 5 (Eutrophication) in the Eastern Mediterranean Sea and to test if a tool for integrated assessment of the status of marine systems can be used as a common methodological approach. In this frame, we tested two statistical approaches in order to set threshold values for four criteria of Descriptor 5: nutrients, chlorophyll a, transparency and dissolved oxygen in the bottom waters. It is noteworthy that this work revealed the need to apply common procedures in data treatment and assessment evaluation. This is the first attempt to set common methods for the assessment of eutrophication in the Eastern Mediterranean, which is essential in marine environments, especially those shared by several countries. To this end, we have applied common criteria and metrics and established thresholds "Good" and "Moderate" for nutrients, chlorophyll a, transparency and dissolved oxygen in the bottom waters for the different Water Types of the Adriatic and Aegean Seas (I, II, IIIW, IIIE), based on datasets provided by Italy, Slovenia, Croatia and Greece. The selected criteria elements were common for all countries, providing a unified approach to GES assessment of two case study areas: the Adriatic Sea and the Saronikos Gulf. Dissolved Inorganic Nitrogen (DIN) threshold values of 15.6, 6.85, 1.61 and 2.11 µmol L-1 were set for the Water Types I, II, IIIW and IIIE, respectively. We also tested if an aggregation tool for GES assessment, such as Nested Environmental status Assessment Tool (NEAT), could be used as a common methodological approach. The comparison of NEAT with TRIX showed good comparability. In this end, NEAT can be used as a useful and much needed assessment tool for assessing eutrophication status of the marine.

The Influence of the Toxic Dinoflagellate Alexandrium minutum, Grown under Different N:P Ratios, on the Marine Copepod Acartia tonsa.

HABs pose a threat to coastal ecosystems, the economic sector and human health, and are expanding globally. However, their influence on copepods, a major connector between primary producers and upper trophic levels, remains essentially unknown. Microalgal toxins can eventually control copepod survival and reproduction by deterring grazing and hence reducing food availability. We present several 24-h experiments in which the globally distributed marine copepod, Acartia tonsa, was exposed to different concentrations of the toxic dinoflagellate, Alexandrium minutum, grown under three N:P ratios (4:1, 16:1 and 80:1), with the simultaneous presence of non-toxic food (the dinoflagellate Prorocentrum micans). The different N:P ratios did not affect the toxicity of A. minutum, probably due to the low toxicity of the tested strain. Production of eggs and pellets as well as ingested carbon appeared to be affected by food toxicity. Toxicity levels in A. minutum also had an effect on hatching success and on the toxin excreted in pellets. Overall, A. minutum toxicity affected the reproduction, toxin excretion and, to an extent, the feeding behavior of A. tonsa. This work indicates that even short-term exposure to toxic A. minutum can impact the vital functions of A. tonsa and might ultimately pose serious threats to copepod recruitment and survival. Still, further investigation is required for identifying and understanding, in particular, the long-term effects of harmful microalgae on marine copepods.

Sympatric occurrence of two Azadinium poporum ribotypes in the Eastern Mediterranean Sea.

The marine dinoflagellate Azadinium poporum produce azaspiracids (AZA) and has been recorded widely in the world. However, information on its biogeography is still limited, especially in view of the fact that A. poporum comprises several genetically differentiated groups. A total of 18 strains of A. poporum were obtained from the Eastern Mediterranean area by incubating surface sediment collected from Ionian Sea of Greece. The morphology of these strains was examined with light microscopy and scanning electron microscopy. Small subunit ribosomal DNA (SSU rDNA), large subunit ribosomal DNA (LSU rDNA) and internal transcribed spacer (ITS) sequences were obtained from all cultured strains. Molecular phylogeny based on concatenated SSU, LSU and ITS sequences confirmed three ribotypes within A. poporum and revealed two subclades within ribotypes A and C. Greek strains of A. poporum ribotype A were nested within ribotype A2 together with strains from Western Mediterranean Sea and French Atlantic, and Greek strains of A. poporum ribotype C were nested within ribotype C2 together with a strain from the Gulf of Mexico. Growth experiments on four selected strains revealed that ribotypes A and C from Greece differed in their growth at higher temperatures, indicating that they are physiologically differentiated. Azaspiracid profiles were analyzed for 15 cultured A. poporum strains using LCMS/MS and demonstrate that the A. poporum ribotype A from Greece produce low level or no AZA and A. poporum ribotype C from Greece produces predominantly AZA-40 (9.6-30.2 fg cell-1) followed by AZA-2 (2.1-2.6 fg cell-1). The first record of AZA-40 producing A. poporum from the Mediterranean suggests that this species is a potential source for azaspiracid contaminations in shellfish from the Eastern Mediterranean Sea.

One Step forward: Benthic Pelagic Coupling and Indicators for Environmental Status.

A large data set from the Eastern Mediterranean was analyzed to explore the relationship between seawater column variables and benthic community status. Our results showed a strong quantitative link between the seawater column variables (Chlorophyll a and Eutrophication Index) and various indicators describing benthic diversity and community composition. The percentage of benthic opportunistic species increased significantly in the stations with high trophic status of the seawater column and so did the strength of the coupling between values of seawater column and benthic indicators. The Eutrophication Index threshold level of 0.85, separating the "Bad and Poor" from "Moderate to High" conditions could serve as an acceptable critical value above which there is a readily observable change in benthic community composition.