Future water quality monitoring--adapting tools to deal with mixtures of pollutants in water resource management.

Environmental quality monitoring of water resources is challenged with providing the basis for safeguarding the environment against adverse biological effects of anthropogenic chemical contamination from diffuse and point sources. While current regulatory efforts focus on monitoring and assessing a few legacy chemicals, many more anthropogenic chemicals can be detected simultaneously in our aquatic resources. However, exposure to chemical mixtures does not necessarily translate into adverse biological effects nor clearly shows whether mitigation measures are needed. Thus, the question which mixtures are present and which have associated combined effects becomes central for defining adequate monitoring and assessment strategies. Here we describe the vision of the international, EU-funded project SOLUTIONS, where three routes are explored to link the occurrence of chemical mixtures at specific sites to the assessment of adverse biological combination effects. First of all, multi-residue target and non-target screening techniques covering a broader range of anticipated chemicals co-occurring in the environment are being developed. By improving sensitivity and detection limits for known bioactive compounds of concern, new analytical chemistry data for multiple components can be obtained and used to characterise priority mixtures. This information on chemical occurrence will be used to predict mixture toxicity and to derive combined effect estimates suitable for advancing environmental quality standards. Secondly, bioanalytical tools will be explored to provide aggregate bioactivity measures integrating all components that produce common (adverse) outcomes even for mixtures of varying compositions. The ambition is to provide comprehensive arrays of effect-based tools and trait-based field observations that link multiple chemical exposures to various environmental protection goals more directly and to provide improved in situ observations for impact assessment of mixtures. Thirdly, effect-directed analysis (EDA) will be applied to identify major drivers of mixture toxicity. Refinements of EDA include the use of statistical approaches with monitoring information for guidance of experimental EDA studies. These three approaches will be explored using case studies at the Danube and Rhine river basins as well as rivers of the Iberian Peninsula. The synthesis of findings will be organised to provide guidance for future solution-oriented environmental monitoring and explore more systematic ways to assess mixture exposures and combination effects in future water quality monitoring.

An ultrasensitive NanoLuc-based luminescence system for monitoring Plasmodium berghei throughout its life cycle

BACKGROUND: Bioluminescence imaging is widely used for cell-based assays and animal imaging studies, both in biomedical research and drug development. Its main advantages include its high-throughput applicability, affordability, high sensitivity, operational simplicity, and quantitative outputs. In malaria research, bioluminescence has been used for drug discovery in vivo and in vitro, exploring host-pathogen interactions, and studying multiple aspects of Plasmodium biology. While the number of fluorescent proteins available for imaging has undergone a great expansion over the last two decades, enabling simultaneous visualization of multiple molecular and cellular events, expansion of available luciferases has lagged behind. The most widely used bioluminescent probe in malaria research is the Photinus pyralis firefly luciferase, followed by the more recently introduced Click-beetle and Renilla luciferases. Ultra-sensitive imaging of Plasmodium at low parasite densities has not been previously achieved. With the purpose of overcoming these challenges, a Plasmodium berghei line expressing the novel ultra-bright luciferase enzyme NanoLuc, called PbNLuc has been generated, and is presented in this work. RESULTS: NanoLuc shows at least 150 times brighter signal than firefly luciferase in vitro, allowing single parasite detection in mosquito, liver, and sexual and asexual blood stages. As a proof-of-concept, the PbNLuc parasites were used to image parasite development in the mosquito, liver and blood stages of infection, and to specifically explore parasite liver stage egress, and pre-patency period in vivo. CONCLUSIONS: PbNLuc is a suitable parasite line for sensitive imaging of the entire Plasmodium life cycle. Its sensitivity makes it a promising line to be used as a reference for drug candidate testing, as well as the characterization of mutant parasites to explore the function of parasite proteins, host-parasite interactions, and the better understanding of Plasmodium biology. Since the substrate requirements of NanoLuc are different from those of firefly luciferase, dual bioluminescence imaging for the simultaneous characterization of two lines, or two separate biological processes, is possible, as demonstrated in this work.

Water chemistry, and biological traits and mercury content of Salvelinus alpinus and S. namaycush in Laflamme and Pingualuit lake

Pingualuk Lake fills a deep crater in the Parc National des Pingualuit on the Ungava Peninsula (Nunavik, Canada) and is isolated from nearby surface waters. The main objectives of this study were to determine and compare the concentrations of two atmospherically derived contaminants, mercury and perfluorinated chemicals (PFCs), in the lake water column and fish of Pingualuk Lake and to assess the physical and biological factors influencing contaminant concentrations. Mercury concentrations in arctic char muscle tissue were comparable to those of char in other Arctic lakes, while the total amount of PFCs was below reported levels for remote lakes in the Arctic and elsewhere. Stable isotope and stomach content analyses were made to investigate the feeding ecology of the Pingualuk Lake arctic char population and indicated the possibility of multiple feeding groups. Genetics characteristics (MH and mtDNA) of fish from Pingualuk Lake revealed that this population is likely distinct from that of nearby Laflamme Lake. However, both arctic char populations exhibit differential variation of their allele families. Physical characteristics determined for Lake Pingualuk revealed that the water column was inversely stratified beneath the ice and extremely transparent to visible and ultraviolet radiation. The highest mercury concentrations (3- 6 pg/mL THg) occurred just beneath the ice surface in each lake. Pingualuk Lake, given its near pristine state and exceptional limnological features, may serve as a most valuable reference ecosystem for monitoring environmental stressors, such as contaminants, in the Arctic.

Spatiotemporal monitoring of water quality in coral reefs of Tayrona National Natural Park, Colombian Caribbean, 2011-2013

Tayrona National Natural Park (TNNP; 11°17' - 11°22' N and 73°53' - 74°12' W) is a hotspot of coral reef biodiversity in the Colombian Caribbean, located between the city of Santa Marta (>455,000 inhabitants) and several smaller river mouths (Rio Piedras, Mendihuaca, Guachaca). The region experiences a strong seasonal variation in physical parameters (temperature, salinity, wind, and water currents) due to alternating dry seasons with coastal upwelling and rainy seasons. Here, a range of water quality parameters relevant for coral reef functioning is provided. Water quality was measured directly above local coral reefs (~10 m water depth) by a monthly monitoring for up to 25 months in the four TNNP bays (Chengue, Gayraca, Neguanje, and Cinto) and at sites with different degree of exposition to winds, waves and water currents (exposed vs. sheltered sites) within each bay. The water quality parameters include: inorganic nutrient (nitrate, nitrite and soluble reactive phosphorus), chlorophyll a, particulate organic carbon and nitrogen concentrations (with a replication of n=3) as well as oxygen availability, biological oxygen demand, seawater pH, and water clarity (with a replication of n=4). This is by far the most comprehensive coral reefs water quality dataset for the region. A detailed description of the methods can be found within the referenced publications.