Report on the fifth meeting of INRA’s national network of ecotoxicologists,ECOTOX

The fifth meeting of INRA’s national network of ecotoxicologists took place on 25 to 27 November 2014 in Biarritz, France. The main aim of the meeting was to bring together ecotoxicologists from INRA and associated partners, providing them ample opportunity to share and discuss their latest scientific results as well as the national policy of research in ecotoxicology and to precise perspectives for the network.     

Academic expertise in assisting private companies in the fields of environment and environmental toxicology: the role of individual expertise

Environmental Science and Pollution Research - The scientific knowledge produced by academic research can be valued in all sectors of human activity, including private sector. The ROVALTAIN...     

Fungal laccases: from structure-activity studies to environmental applications

Laccases are multicopper oxidases mainly secreted by filamentous fungi. Producing radical forms from organic substrates, they are involved in numerous reactions leading to the degradation and polymerization of xenobiotics. Our studies have led to a better knowledge of the structural, catalytic and genetic properties of laccases and allowed to develop a strategy for their evolution through genetic engineering. Here, we show that fungal laccases, wild or engineered, may be potent tools to develop bioremediation processes of soils polluted by organic compounds, and assays to assess the ecotoxicological impact of these pollutants on soil fungi.     

Oxidoreductases provide a more generic response to metallic stressors (Cu and Cd) than hydrolases in soil fungi: new ecotoxicological insights

The present study investigates the effect of metals on the secretion of enzymes from12 fungal strains maintained in liquid cultures. Hydrolases (acid phosphatase, β-glucosidase, β-galactosidase, and N-acetyl-β-glucosaminidase) and ligninolytic oxidoreductases (laccase, Mn, and lignin peroxidases) activities, as well as biomass production, were measured in culture fluids from fungi exposed to Cu or Cd. Our results showed that all fungi secreted most of the selected hydrolases and that about 50 % of them produced a partial oxidative system in the absence of metals. Then, exposure of fungi to metals led to the decrease in biomass production. At the enzymatic level, Cu and Cd modified the secretion profiles of soil fungi. The response of hydrolases to metals was contrasted and complex and depended on metal, enzyme, and fungal strain considered. By contrast, the metals always stimulated the activity of ligninolytic oxidoreductases in fungal strains. In some of them, oxidoreductases were specifically produced following metal exposure. Fungal oxidoreductases provide a more generic response than hydrolases, constituting thus a physiological basis for their use as biomarkers of metal exposure in soils.     

Incorporation of pesticides by soil micro-organisms as a way of bound residues formation

We studied the ¹⁴C-tagged biochemical compartments such as proteins, lipids and polysaccharides from bacterial and fungal cultures supplied with ¹⁴C-pesticides and ¹⁴C-glucose. We showed that ¹⁴C incorporation depends both on the substrate structure and on the nature of the micro-organism. Both bacterial and fungal cells incorporated glucose ¹⁴C mainly into cell wall proteins. By contrast, glyphosate ¹⁴C was mainly incorporated into cytoplasm carbohydrates by fungi. On the other hand, 2,4-dichlorophenoxyacetic acid ¹⁴C was mainly found in the carbohydrate fractions of the bacteria walls.     

Effect of the endectocide ivermectin on filamentous fungi

We investigated the effect of ivermectin, at environmental concentrations and above, on several strains of filamentous fungi. Ivermectin did not show significant short-term effect on biomass production and germination rate of the spores. By contrast, the spore production was stimulated twofold in Phanerochaete chrysosporium and Mucor racemosus cultures, whereas that endpoint was reduced in Fusarium cultures. A long-term exposure of F. oxysporum to ivermectin reduced both production and germination of fungal spores. Ivermectin was not an inducer of exocellular oxidases.     

Secretion profiles of fungi as potential tools for metal ecotoxicity assessment: a study of enzymatic system in Trametes versicolor

The relationship between the expression of extracellular enzymatic system and a metal stress is scarce in fungi, hence limiting the possible use of secretion profiles as tools for metal ecotoxicity assessment. In the present study, we investigated the effect of Zn, Cu, Pb and Cd, tested alone or in equimolar cocktail, on the secretion profiles at enzymatic and protein levels in Trametesversicolor. For that purpose, extracellular hydrolases (acid phosphatase, β-glucosidase, β-galactosidase and N-acetyl-β-glucosaminidase) and ligninolytic oxidases (laccase, Mn-peroxidase) were monitored in liquid cultures. Fungal secretome was analyzed by electrophoresis and laccase secretion was characterized by western-blot and mass spectrometry analyses. Our results showed that all hydrolase activities were inhibited by the metals tested alone or in cocktail, whereas oxidase activities were specifically stimulated by Cu, Cd and metal cocktail. At protein level, metal exposure modified the electrophoretic profiles of fungal secretome and affected the diversity of secreted proteins. Two laccase isoenzymes, LacA and LacB, identified by mass spectrometry were differentially glycosylated according to the metal exposure. The amount of secreted LacA and LacB was strongly correlated with the stimulation of laccase activity by Cu, Cd and metal cocktail. These modifications of extracellular enzymatic system suggest that fungal oxidases could be used as biomarkers of metal exposure.