Ecotoxicological Evaluation of Diesel- Contaminated Soil Before and After a Bioremediation Process L. Molina-Barahona, 1 L. Vega-Loyo, 1 M. Guerrero, 2 S. Ramı ´rez, 3 I. Romero, 3 C. Vega-Jarquı ´n, 4 A. Albores 1 1 Seccio ´n Externa de Toxicologı´a, Centro de Investigacio ´ n y de Estudios Avanzados del Instituto Polite ´ cnico Nacional, Mexico City, Mexico 2 Departamento de Fı´sica, Centro de Investigacio ´ n y de Estudios Avanzados del Instituto Polite ´ cnico Nacional, Mexico City, Mexico 3 Centro Nacional de Investigacio ´ n y Capacitacio ´ n Ambiental (CENICA-INE), Mexico City, Mexico 4 Departamento de Biotecnologı´a y Bioingenierı´a, Centro de Investigacio ´ n y de Estudios Avanzados del Instituto Polite ´ cnico Nacional, Mexico City, Mexico Received 16 January 2004; revised 20 August 2004; accepted 13 October 2004 ABSTRACT: Evaluation of contaminated sites is usually performed by chemical analysis of pollutants in soil. This is not enough either to evaluate the environmental risk of contaminated soil nor to evaluate the efficiency of soil cleanup techniques. Information on the bioavailability of complex mixtures of xenobiotics and degra- dation products cannot be totally provided by chemical analytical data, but results from bioassays can integrate the effects of pollutants in complex mixtures. In the preservation of human health and environment quality, it is important to assess the ecotoxicological effects of contaminated soils to obtain a better evaluation of the healthiness of this system. The monitoring of a diesel-contaminated soil and the evaluation of a bioremediation technique conducted on a microcosm scale were performed by a battery of ecotoxicological tests including phytotoxicity, Daphnia magna, and nematode assays. In this study we biostimulated the native microflora of soil contaminated with diesel by adding nutrients and crop residue (corn straw) as a bulking agent and as a source of microorganisms and nutrients; in addition, moisture was adjusted to enhance diesel removal. The bioremediation process efficiency was evaluated directly by an innovative, simple phytotoxicity test system and the diesel extracts by Daphnia magna and nematode assays. Contaminated soil samples were revealed to have toxic effects on seed germination, seedling growth, and Daphnia survival. After biostimulation, the diesel concentration was reduced by 50.6%, and the soil samples showed a significant reduction in phytotoxicity (9%–15%) and Daphnia assays (3-fold), confirming the effectiveness of the bioremediation process. Results from our microcosm study suggest that in addition to the evaluation of the bioremediation processes efficiency, toxicity testing is different with organisms representative of diverse phylogenic levels. The integration of analytical, toxicological and bioremediation data is necessary to properly assess the ecological risk of bioremediation processes. © 2005 Wiley Periodicals, Inc. Environ Toxicol 20: 100 –109, 2005. Keywords: diesel contamination; ecotoxicology; ecotoxicological tests; phytotoxicity tests; microbial bioremediation; microcosms INTRODUCTION Around the world, large areas of soil and water reservoirs have been contaminated with oil products such as diesel fuel. Mexico is one of the world’s largest producers of oil Correspondence to: A. Albores; e-mail: aalbores@cinvestav.mx Published online in Wiley InterScience (www.interscience.wiley.com). DOI 10.1002/tox.20083 © 2005 Wiley Periodicals, Inc. 100