Snejana Moncheva

Intrinsic Tryptophan Fluorescence of Human Serum Proteins and Related Conformational Changes

The unfolding of human serum proteins (HSP) was studied by measuring the intrinsic fluorescence intensity at a wavelength of excitation corresponding to tryptophans or typosines fluorescence and surface hydrophobicity. The maxima emission wavelengths (λmax) of human serum albumin (HSA) and human serum globulin (HSG) before beer consumption (BC) were 336.0 and 337.0 nm and after BC shifted to 335.0 and 334.0 nm, respectively. The surface hydrophobicity slightly increased after BC. In a solution of 8 M urea the λmax of BSA shifted to 346.4 and that of BSG to 342.5 nm. In contrast, in the same solution but after BC the λmax positions of HSA and HSG shifted to 355.9 and 357.7 nm, respectively. A decrease in fluorescence intensity, a shift in the maximum of emission, and an increase in surface hydrophobicity which reflected unfolding of proteins were observed. Here we provide evidence that the loosening of the HSP structure takes place primarily in various concentrations of urea before and after beer consumption. Differences in the fluorescence behavior of the proteins are attributed to disruption of the structure of proteins by denaturants as well as by the change in their compactability as a result of ethanol consumption.

Implementing and Innovating Marine Monitoring Approaches for Assessing Marine Environmental Status

Marine environmental monitoring has tended to focus on site-specific methods of investigation. These traditional methods have low spatial and temporal resolution and are relatively labor intensive per unit area/time that they cover. To implement the Marine Strategy Framework Directive (MSFD), European Member States are required to improve marine monitoring and design monitoring networks. This can be achieved by developing and testing innovative and cost-effective monitoring systems, as well as indicators of environmental status. Here, we present several recently developed methodologies and technologies to improve marine biodiversity indicators and monitoring methods. The innovative tools are discussed concerning the technologies presently utilized as well as the advantages and disadvantages of their use in routine monitoring. In particular, the present analysis focuses on: (i) molecular approaches, including microarray, Real Time quantitative PCR (qPCR), and metagenetic (metabarcoding) tools; (ii) optical (remote) sensing and acoustic methods; and (iii) in situ monitoring instruments. We also discuss their applications in marine monitoring within the MSFD through the analysis of case studies in order to evaluate their potential utilization in future routine marine monitoring. We show that these recently-developed technologies can present clear advantages in accuracy, efficiency and cost.

European Marine Biodiversity Monitoring Networks: Strengths, Weaknesses, Opportunities and Threats

By 2020, European Union Member States should achieve Good Environmental Status (GES) for eleven environmental quality descriptors for their marine waters to fulfill the Marine Strategy Framework Directive (MSFD). By the end of 2015, in coordination with the Regional Seas Conventions, each EU Member States was required to develop a marine strategy for their waters, together with other countries within the same marine region or sub-region. Coherent monitoring programs, submitted in 2014, form a key component of this strategy, which then aimed to lead to a Program of Measures (submitted in 2015). The European DEVOTES FP7 project has produced and interrogated a catalogue of EU marine monitoring related to MSFD descriptors 1 (biological diversity), 2 (non-indigenous species), 4 (food webs) and 6 (seafloor integrity). Here we detail the monitoring activity at the regional and sub-regional level for these descriptors, as well as for 11 biodiversity components, 22 habitats and the 37 anthropogenic pressures addressed. The metadata collated for existing European monitoring networks were subject to a SWOT (strengths, weaknesses, opportunities and threats) analysis. This interrogation has indicated case studies to address the following questions: a) what are the types of monitoring currently in place?; b) who does what and how?; c) is the monitoring fit-for-purpose for addressing the MSFD requirements?, and d) what are the impediments to better monitoring (e.g. costs, shared responsibilities between countries, overlaps, co-ordination)? We recommend the future means, to overcome the identified impediments and develop more robust monitoring strategies and as such the results are especially relevant to implementing coordinated monitoring networks throughout Europe, for marine policy makers, government agencies and regulatory bodies. It is emphasized that while many of the recommendations given here require better, more extensive and perhaps more costly monitoring, this is required to avoid any legal challenges to the assessments or to bodies and industries accused of causing a deterioration in marine quality. More importantly the monitoring is required to demonstrate the efficacy of management measures employed. Furthermore, given the similarity in marine management approaches in other developed systems, we consider that the recommendations are also of relevance to other regimes worldwide.

Indicator-Based Assessment of Marine Biological Diversity–Lessons from 10 Case Studies across the European Seas

The Marine Strategy Framework Directive requires the environmental status of European marine waters to be assessed using biodiversity as one out of 11 descriptors, but the complexity of marine biodiversity and its large span across latitudinal and salinity gradients have been a challenge to the scientific community aiming to produce approaches for integrating information from a broad range of indicators. The Nested Environmental status Assessment Tool (NEAT), developed for the integrated assessment of the status of marine waters, was applied to ten marine ecosystems to test its applicability and compare biodiversity assessments across the four European regional seas. We evaluate the assessment results as well as the assessment designs of the ten cases, and how the assessment design, particularly the choices made regarding the area and indicator selection, affected the results. The results show that only 2 out of the 10 case study areas show more than 50 % probability of being in good status in respect of biodiversity. No strong pattern among the ecosystem components across the case study areas could be detected, but marine mammals, birds, and benthic vegetation indicators tended to indicate poor status while zooplankton indicators indicated good status when included into the assessment. The analysis shows that the assessment design, including the selection of indicators, their target values, geographical resolution and habitats to be assessed, has potentially a high impact on the result, and the assessment structure needs to be understood in order to make an informed assessment. Moreover, recommendations are provided for the best practice of using NEAT for marine status assessments.

Sources of uncertainty in assessment of marine phytoplankton communities

Characterisation of phytoplankton communities is important for classification of the ecological status of marine waters. In order to design a monitoring programme, it is important to know what degree of variation in the measurements occur at each level (water body, station and sample), so that resources can be spent in a way that maximise the precision of the measured parameters. Seven European water bodies were sampled to assess the variation in pigment concentrations and population densities attributed to water body, station and sample levels. It was found that the main proportion of the variation between pigment measurements was explained by the variation between stations (12–91% of variation) followed by the variation between water bodies (0–89% of variation). For measurements of population density, the main proportion of the variation between densities of cells recorded was explained by the variation between the taxonomists counting the samples (61%), whilst the main proportion of the variation between numbers of taxa recorded was explained by the variation between water bodies (83%). When the cell density of the nine dominant classes were analysed separately, the main proportion of variation was explained at the water body level for all but two class.

A Catalogue of Marine Biodiversity Indicators

A Catalogue of Marine Biodiversity Indicators was developed with the aim of providing the basis for assessing the environmental status of the marine ecosystems. Useful for the implementation of the Marine Strategy Framework Directive (MSFD), this catalogue allows the navigation of a database of indicators mostly related to biological diversity, non-indigenous species, food webs, and seafloor integrity. Over 600 indicators were compiled, which were developed and used in the framework of different initiatives (e.g. EU policies, research projects) and in national and international contexts (e.g. Regional Seas Conventions, and assessments in non-European seas). The catalogue reflects the current scientific capability to address environmental assessment needs by providing a broad coverage of the most relevant indicators for marine biodiversity and ecosystem integrity. The available indicators are reviewed according to their typology, data requirements, development status, geographical coverage, relevance to habitats or biodiversity components, and related human pressures. Through this comprehensive overview, we discuss the potential of the current set of indicators in a wide range of contexts, from large-scale to local environmental programs, and we also address shortcomings in light of current needs. Developed by the DEVOTES Project, the catalogue is freely available through the DEVOTool software application, which provides browsing and query options for the associated metadata. The tool allows extraction of ranked indicator lists best fulfilling selected criteria, enabling users to search for suitable indicators to address a particular biodiversity component, ecosystem feature, habitat or pressure in a marine area of interest. This tool is useful for EU Member States, Regional Sea Conventions, the European Commission, non-governmental organizations, managers, scientists and any person interested in marine environmental assessment. It allows users to build, complement or adjust monitoring programs and has the potential to improve comparability and foster transfer of knowledge across marine regions.

Rapana venosa as a bioindicator of environmental pollution

The aim of this investigation was to examine biochemical differences in the gastropod Rapana venosa (R. venosa) from polluted (RvP) and nonpolluted (RvN) sites of the Black Seas Bulgarian coast that may serve as bioindicators of environmental quality. Mussels Mytilus galloprovincialis (MMg) were collected from polluted (MMgP) and nonpolluted (MMgN) sites at the same time for comparison. Bioactive compounds and the antioxidant activity of dry matter (DM) methanol extracts from RvP and RvN were determined. Three-dimensional fluorometry (3D-FL) and Fourier transform infrared (FTIR) analyses were used to evaluate polyphenols and proteins. DM methanol extracts from RvP, and to lesser extent RvN, showed high amounts of total phenolics (23.22±2.9 and 15.60±1.8 mg GAE · g−1 DW) and exhibited high levels of antioxidant activity in some radical scavenging assays (μ MTE · g−1 DW): 56.38±5.2 and 33.79±3.3 by 2,2azino-bis (3-ethyl-benzothiazoline-6-sulfonic acid) diamonium salt (ABTS·+); 54.79±5.6 and 33.7±2.6 by cupric reducing antioxidant capacity (CUPRAC); 62.34±6.8 and 30.31±3.9 by the 1diphenyl-2-picrylhydrazyl method (DPPH). A correlation in the above indices was found between R. venosa and M. galloprovincialis, but all results were relatively higher for M. galloprovincialis than for R. venosa. The obtained indices of R. venosa may serve as a bioindicator of environmental ecological quality.

Long-term changes in the Black Sea surface chlorophyll a according to in situ and modern satellite data

Analysis of temporal variability of extensive data sets on surface in situ chl a for the period 1964 to 1996 in the deep open Black Sea revealed pronounced features in its seasonal dynamics and long-term variability. Comparison (verification and patterns of seasonal and interannual changes) of satellite (Level-2 SeaWiFS) chl a for 1997–2002 and in situ chl a data did not show satisfactory results. A newly developed local algorithm (first preliminary version) to accurately transform SeaWiFS optical data, however, resulted in good correspondence of in situ and satellite data, the preliminary results of which are presented.

Characterization of Rapana thomasiana as an indicator of environmental quality of the Black Sea coast of Bulgaria

The aim of this investigation was to determine the contents of polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs), minerals, trace elements and bioactivity in the gastropod Rapana thomasiana, which can be used as an environmental bioindicator organism. The chemical differences between Rapana thomasiana from polluted (RapaPol) and non-polluted (RapaNPol) sites of the Black Sea coast in Bulgarian were investigated. Chromatography and high-resolution inductively coupled plasma mass spectrometry (HR-ICP-MS) were used for evaluation of PAHs, PCBs, minerals and trace elements. Methanol extracts from RapaPol and RapaNPol (to a lesser degree) contained relatively high amounts of free phenolics (2.50±0.3 and 1.57±0.18 mg GAE/g DW, respectively) and exhibited the following respective levels of antioxidant activities determined by two radical-scavenging assays (μMTE/g DW): 1.8±0.2 and 0.98±0.08 by 1-diphenyl-2-picrylhydrazyl method (DPPH); 1.74±0.17 and 1.04±0.12 by cupric reducing antioxidant capacity (CUPRAC). The total amounts of elements, PAHs and PCBs were higher in RapaPol than in RapaNPol. The obtained indices of Rapana thomasiana can serve as a bioindicator of the environmental ecological quality.

Biomass, protein- and carbohydrate-composition of phytoplankton in Varna Bay, Black Sea

It was shown that phytoplankton from the Varna Bay, Black Sea, has significantly more suspended carbohydrates, proteins and biomass in July than in April. The dominant species were Bacillariophyceae and Dinophyceae. Electrophoretic and fluorescent spectra have shown the main differences in molecular weight and stability of phytoplankton proteins. Phytoplankton included specific proteins distributed over a limited range of molecular weights between 14 and 72 kilodaltons (kDa). The most abundant protein constituents in phytoplankton samples collected in April were around 45–55 kDa. The seasonal variations of the environment influence the quantitative and qualitative changes in phytoplankton.