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The Campos de Palmas Wildlife Refuge (CPWR) (Brazil) is a full protection Conservation Unit (CU) formed by private properties, where the use of land and natural resources are allowed sustainably according to the rules of the CU Management Plan. Inadequate practices of land use and occupation within this CU can affect its quality and the organisms that depend on them. Considering the above, the objective of the present study was to biomonitor different land uses and occupations within the CPWR and its surroundings, during the four seasons of a year, using the bioindicators Allium cepa (cytotoxicity and genotoxicity tests) and Eisenia fetida (leakage test). The soil samples were collected in areas of silviculture, agriculture, and native grasslands within the CU and agriculture outside the limits of the CPWR. The use by silviculture (spring) and native grassland (summer) showed cytotoxic effects for A. cepa. The use by agriculture outside the CPWR (spring) showed genotoxic effect and stimulation of mitotic cell division. For the animal bioindicator, the use by agriculture within the CPWR (winter), native grassland (autumn and spring), silviculture (autumn and spring), and agriculture outside the CU (spring) showed a toxic effect. A PCA analysis showed a correlation between the results of toxicity, cytotoxicity, and genotoxicity with the presence of macronutrients and metals in the evaluated soil samples. Possibly influenced by their soil composition characteristics (silviculture and native grassland) or the management of agricultural areas. In addition, samples from the rainy season (spring) showed higher ecotoxicity. These results show that biomonitoring through different organisms is important for evaluating environmental quality and indicate the need for the implementation of preventive measures in the CPWR to avoid the toxic, cytotoxic, and genotoxic effects found. They also ensure the integrity of this CU and the active protection of the environment and biodiversity., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)