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The framework integrated the quantification of a group of 51 pesticides and ecotoxicological endpoints with organisms from different trophic categories (the bacterium Aliivibrio fischeri, the microalga Pseudokirchneriella subcapitata, and the crustaceans Daphnia magna and Thamnocephalus platyurus) at two sampling points in the reservoir (Lage (L) and Lage S (LS)) between 2018 and 2020. Over the three-year study, we quantified 36 of the 51 pesticides analyzed in the Lage reservoir. Total concentrations increased successively from 0.95 \u00b5g L\u22121 to 1.99 and 2.66 \u00b5g L\u22121. Among these, the pesticides most frequently detected were terbuthylazine (100% of detection) and metolachlor (83% of detection), with maximum concentrations of 115.6 and 85.5 \u00b5g L\u22121, respectively. Samples from the LS site showed higher toxicity, where A. fischeri presented 30 min EC50 values of 39\u201351%. Microalgae growth was consistently inhibited, correlating with agricultural activity, mainly the application of herbicides and insecticides, while D. magna feeding rates revealed no inhibitory effects in the Lage samples. The results highlight that although the detected pesticide levels were below regulatory limits, they still induced toxic effects in the tested organisms. The potential ecological status of the reservoir was classified as moderate, and the integration of the proposal toolbox allowed refinement of the classification of water status. The results demonstrated that this integrated approach, combining multiple assessment methods, establishes a more robust water quality evaluation methodology, allowing it to be used as a tool complementary to the WFD methodology. This proposal not only identified existing pollution impacts but also enabled (1) early detection of the toxic effects of emerging contaminants to prevent ecological damage; (2) proactive management through specific actions to restore water status; and (3) improved sustainable water use.<\/jats:p>","DOI":"10.3390\/w17172642","type":"journal-article","created":{"date-parts":[[2025,9,8]],"date-time":"2025-09-08T08:06:32Z","timestamp":1757318792000},"page":"2642","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Integrating Ecotoxicological Assessment to Evaluate Agricultural Impacts on Aquatic Ecosystems: A Case Study of the Lage Reservoir (Mediterranean Region)"],"prefix":"10.3390","volume":"17","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-4792-3633","authenticated-orcid":false,"given":"Adriana","family":"Catarino","sequence":"first","affiliation":[{"name":"Instituto Polit\u00e9cnico de Beja, Escola Superior Agr\u00e1ria de Beja, Rua Pedro Soares, 7800-295 Beja, Portugal"},{"name":"Center for Sci-Tech Research in Earth System and Energy (CREATE)\u2014Pole of the Polytechnic Institute of Beja, 7800-295 Beja, Portugal"},{"name":"MARE\u2014Marine and Environmental Sciences Centre, ARNET\u2014Aquatic Research Network, Department of Landscape, Environment and Planning, University of \u00c9vora, Rua Rom\u00e3o Ramalho 59, 7000-671 \u00c9vora, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8813-3315","authenticated-orcid":false,"given":"Clarisse","family":"Mourinha","sequence":"additional","affiliation":[{"name":"Instituto Polit\u00e9cnico de Beja, Escola Superior Agr\u00e1ria de Beja, Rua Pedro Soares, 7800-295 Beja, Portugal"},{"name":"Center for Sci-Tech Research in Earth System and Energy (CREATE)\u2014Pole of the Polytechnic Institute of Beja, 7800-295 Beja, Portugal"}]},{"given":"Mariana","family":"Cust\u00f3dio","sequence":"additional","affiliation":[{"name":"Instituto Polit\u00e9cnico de Beja, Escola Superior Agr\u00e1ria de Beja, Rua Pedro Soares, 7800-295 Beja, Portugal"},{"name":"Center for Sci-Tech Research in Earth System and Energy (CREATE)\u2014Pole of the Polytechnic Institute of Beja, 7800-295 Beja, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1808-3847","authenticated-orcid":false,"given":"Pedro","family":"Anast\u00e1cio","sequence":"additional","affiliation":[{"name":"MARE\u2014Marine and Environmental Sciences Centre, ARNET\u2014Aquatic Research Network, Department of Landscape, Environment and Planning, University of \u00c9vora, Rua Rom\u00e3o Ramalho 59, 7000-671 \u00c9vora, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9719-079X","authenticated-orcid":false,"given":"Patr\u00edcia","family":"Palma","sequence":"additional","affiliation":[{"name":"Instituto Polit\u00e9cnico de Beja, Escola Superior Agr\u00e1ria de Beja, Rua Pedro Soares, 7800-295 Beja, Portugal"},{"name":"Center for Sci-Tech Research in Earth System and Energy (CREATE)\u2014Pole of the Polytechnic Institute of Beja, 7800-295 Beja, Portugal"},{"name":"GeoBioTec, NOVA School of Sciences and Technology, Campus da Caparica, 2829-516 Caparica, Portugal"},{"name":"\u00c1guas P\u00fablicas do Alentejo, Grupo \u00c1guas de Portugal, Rua Dr. Aresta Branco n\u00ba51, 7800-310 Beja, Portugal"}]}],"member":"1968","published-online":{"date-parts":[[2025,9,6]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Derossi, F.N., Ribeiro, P.F., and Santos, J.L. 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