PHAGOCYTIC ACTIVITY OF EARTHWORM COELOMYCYTES USED AS A SUBLETHAL BIOMARKER OF METAL TOXICITY Sauvé S, Hendawi M, Brousseau P, Fournier M Département de.

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PHAGOCYTIC ACTIVITY OF EARTHWORM COELOMYCYTES USED AS A SUBLETHAL BIOMARKER OF METAL TOXICITY Sauvé S, Hendawi M, Brousseau P, Fournier M Département de chimie – Université de Montréal INRS-Institut Armand Frappier-Santé Humaine

© Sauvé 2001 Earthworms Earthworms lead an essential ecological role for maintaining soil health, fertility, structure, nutrient cycling, … Standard worm assays evaluate lethality Sublethal markers such as reproduction and immunocompetence can be quantified

© Sauvé 2001 Worm species Terrestrial earthworms  Lumbricus terrestris  Eisenia fetida  Aporrectodea turgida Aquatic worm  Tubifex tubifex

© Sauvé 2001 Cell Extrusion By puncture with pasteur pipette or syringe Ethanol extrusion Electrical stimulation Ultrasound Extruded cells are suspended in a Lumbricus Basal Salt Solution (LBSS)

© Sauvé 2001 Species-specific parameters E. fetidaL. terrestrisA. turgidaT. tubifex Fresh weight (g) Total cells / animals (10 6 ) Viability (%) Trypan Blue Viability (%) PI

© Sauvé 2001 Phagocytosis Earthworm coelomocytes (Eisenia fetida)

© Sauvé 2001 Flow Cytometry

© Sauvé 2001 Phagocytosis SpeciesControl Toxic (10 -4 M HgCl 2 ) L. terrestris E. fetida

© Sauvé 2001 Silver and Arsenic

© Sauvé 2001 Cadmium and Copper

© Sauvé 2001 Mercury

Lead and Zinc

© Sauvé 2001 In vitro Toxicity thresholds Compound EC 50 (mg L -1 ) AgNO Na 2 HAsO 4 ND CdCl CoCl 2 ND KCr 2 O 7 ND CuCl HgCl Compound EC 50 (mg L -1 ) CH 3 HgCl0.4 MoCl * NiCl PbCl 2 134* NaSeO * ZnCl * ND = not detected * = extrapolated

© Sauvé 2001 Transfer in vitro to in vivo In vitro cellular exposure Petri dish, solution exposure Soil incubation (using the OECD reproduction protocol) Field, in situ exposure in mesocosm « worm bags » Some recent field applications to the evaluation of cement factory emissions

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