Modeling flow and nitrate fate at catchment scale in Brittany (France).

In the intensive pig-farming (Sus scrofa) area of Brittany (western France), many surface and subsurface water resources are contaminated by nitrate (NO3) with concentrations that chronically exceed the European Community 50 mg L(-1) drinking standard. To ensure sustainable water supply, the fate of NO3 must be considered in both surface water and ground water. The fate of N was investigated in a Britain catchment, the Coët-Dan watershed, with an integrated management tool: the hydrological SWAT model coupled with the ground water model MODFLOW, and its companion contaminant and solute transport model MT3DMS. The model was validated with respect to water quantity during a 6-yr period and for the NO3 concentration during a 44-mo period, at two gauging stations in the catchment. The coupled models reproduced accurately the measurements. At the basin outlet, the Nash-Sutcliffe coefficients were 0.88 for monthly flow for the entire period and 0.87 for monthly N load. Alternative scenarios were simulated and showed potential benefits of decreasing manure application from 210 to 170 kg N ha(-1) as required by the European Commission Nitrates Directive.

Comprehensive approach for the detection of antifungal compounds using a susceptible strain of Candida albicans and confirmation of in vivo activity with the Galleria mellonella model.

An efficient screening strategy for the identification of potentially interesting low-abundance antifungal natural products in crude extracts that combines both a sensitive bioautography assay and high performance liquid chromatography (HPLC) microfractionation was developed. This method relies on high performance thin layer chromatography (HPTLC) bioautography with a hypersusceptible engineered strain of Candida albicans (DSY2621) for bioactivity detection, followed by the evaluation of wild type strains in standard microdilution antifungal assays. Active extracts were microfractionated by HPLC in 96-well plates, and the fractions were subsequently submitted to the bioassay. This procedure enabled precise localisation of the antifungal compounds directly in the HPLC chromatograms of the crude extracts. HPLC-PDA-mass spectrometry (MS) data obtained in parallel to the HPLC antifungal profiles provided a first chemical screening about the bioactive constituents. Transposition of the HPLC analytical conditions to medium-pressure liquid chromatography (MPLC) allowed the efficient isolation of the active constituents in mg amounts for structure confirmation and more extensive characterisation of their biological activities. The antifungal properties of the isolated natural products were evaluated by their minimum inhibitory concentration (MIC) in a dilution assay against both wild type and engineered strains of C. albicans. The biological activity of the most promising agents was further evaluated in vitro by electron microscopy and in vivo in a Galleria mellonella model of C. albicans infection. The overall procedure represents a rational and comprehensive means of evaluating antifungal activity from various perspectives for the selection of initial hits that can be explored in more in-depth mode-of-action studies. This strategy is illustrated by the identification and bioactivity evaluation of a series of antifungal compounds from the methanolic extract of a Rubiaceae plant, Morinda tomentosa, which was used as a model in these studies.