A previously developed potential cleanup tool for atrazine contaminated soils was evaluated in larger open soil microcosms for optimization under more realistic conditions, using a natural crop soil spiked with an atrazine commercial formulation (Atrazerba FL). The doses used were 20x or 200x higher than the recommended dose (RD) for an agricultural application, mimicking over-use or spill situations. Pseudomonas sp. strain ADP was used for bioaugmentation (around 10(7) or 10(8) viable cells g(-1) of soil) and citrate for biostimulation (up to 4.8 mg g(-1) of soil). Bioremediation treatments providing fastest and higher atrazine biodegradation proved to differ according to the initial level of soil contamination. For 20x RD of Atrazerba FL, a unique inoculation with Pseudomonas sp. ADP (9 +/- 1 x 10(7) CFU g(-1)) resulted in rapid atrazine removal (99% of the initial 7.2 +/- 1.6 microg g(-1) after 8d), independent of citrate. For 200x RD, an inoculation with the atrazine-degrading bacteria (8.5 +/- 0.5 x 10(7) CFU g(-1)) supplemented with citrate amendment (2.4 mg g(-1)) resulted in improved biodegradation (87%) compared with bioaugmentation alone (79%), even though 7.8 +/- 2.1 microg of atrazine g(-1) still remained in the soil after 1 wk. However, the same amount of inoculum, distributed over three successive inoculations and combined with citrate, increased Pseudomonas sp. ADP survival and atrazine biodegradation (to 98%, in 1 wk). We suggest that this bioremediation tool may be valuable for efficient removal of atrazine from contaminated field soils thus minimizing atrazine and its chlorinated derivatives from reaching water compartments.

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The aim of the present study is to contribute an ecologically relevant assessment of the ecotoxicological effects of pesticide applications in agricultural areas in the tropics, using an integrated approach with information gathered from soil and aquatic compartments. Carbofuran, an insecticide/nematicide used widely on sugarcane crops, was selected as a model substance. To evaluate the toxic effects of pesticide spraying for soil biota, as well as the potential indirect effects on aquatic biota resulting from surface runoff and/or leaching, field and laboratory (using a cost-effective simulator of pesticide applications) trials were performed. Standard ecotoxicological tests were performed with soil (Eisenia andrei, Folsomia candida, and Enchytraeus crypticus) and aquatic (Ceriodaphnia silvestrii) organisms, using serial dilutions of soil, eluate, leachate, and runoff samples. Among soil organisms, sensitivity was found to be E. crypticus < E. andrei < F. candida. Among the aqueous extracts, mortality of C. silvestrii was extreme in runoff samples, whereas eluates were by far the least toxic samples. A generally higher toxicity was found in the bioassays performed with samples from the field trial, indicating the need for improvements in the laboratory simulator. However, the tool developed proved to be valuable in evaluating the toxic effects of pesticide spraying in soils and the potential risks for aquatic compartments.

Purpose To mitigate the environmental effects of atrazine, one of the cleanup strategies available is based on the use of atrazine-degrading bacteria. This work aimed to evaluate the efficacy of a previously developed bioremediation tool for atrazine-contaminated soils (combining bioaugmentation with Pseudomonas sp. ADP, hereafter designated as P. ADP, and biostimulation with citrate) on both soil habitat and retention functions, by performing ecotoxicological tests with standard soil and aquatic species. Materials and methods Soil microcosms (incorporating earthworms, collembolans, and plants) were spiked with three doses of Atrazerba FL, an atrazine commercial formulation: the recommended dose (RD; 2 L/ha), 10×RD and 20×RD to simulate overuse/accidental spills scenarios. The experiment included two main groups of treatments:(1) microcosms sprayed solely with Atrazerba, i.e., nonbioremediated soils (NB) and (2) microcosms sprayed with both Atrazerba and the bioremediation tool (addition of P. ADP plus citrate), i.e., bioremediated soils (B). Control microcosms with no herbicide or P. ADP plus citrate addition were also set up. Besides soil chemical analysis, the following ecotoxicological endpoints were assessed to monitor bioremediation: plant biomass production, earthworm reproduction, microalgae growth (in eluatescollected 5 and 10 days after the bioremediation treatmentand leachates-collected on day seven), and cladoceran reproduction (in soil eluates). Results In NB soils, all Atrazerba doses induced a severe reduction in plant biomass production, and no effects were found for earthworm's reproduction. Eluates and leachates obtained from the NB soils caused deleterious effects on both microalgae growth and cladoceran reproduction. Chemical analysis showed that atrazine degradation was faster in B soils than in the correspondent NB soils. Data from toxicity tests indicated that test organism performance was enhanced in B soils and respective eluates and leachates, compared to the NB samples. In fact, for soils contaminated with 10 and 20×RD Atrazerba doses, plant biomass production was significantly higher in the B soils than in the correspondent NB soils. Regarding the effects of soil bioremediation on the toxicity of soil eluates and leachates, for the soil contaminated with 10 ×RD of Atrazerba, over a 5-day treatment period, both microalgae growth and cladoceran reproduction were significantly higher in water extracts obtained from the B soils when compared with the NB extracts and also similar to the control. By the contrary, for the highest Atrazerba dose tested (20×RD), no significant differences were found on the toxicity of B and NB eluates toward both aquatic test organisms. However, for this same dose, after 7 days, microalgae growth was higher in B than in the NB leachates and similar to the control. Yet, after a longer bioremediation period of 10 days, eluates were also no longer toxic to both aquatic organisms. Discussion Based on atrazine soil chemical analysis, one can state that the addition ...

A current challenge in soil ecotoxicology is the use of natural soils as test substrates to increase ecological relevance of data. Despite the existence of six natural reference soils (the Euro-soils), some parallel projects showed that these soils do not accurately represent the diversity of European soils. Particularly, Mediterranean soils are not properly represented. To fill this gap, 12 natural soils from the Mediterranean regions of Alentejo, Portugal; Cataluña, Spain; and Liguria, Italy, were selected and used in reproduction and avoidance tests to evaluate the soil habitat function for earthworms (Eisenia andrei) and enchytraeids (Enchytraeus crypticus). Predictive models on the influence of soil properties on the responses of these organisms were developed using generalized linear models. Results indicate that the selected soils can impact reproduction and avoidance behavior of both Oligochaete species. Reproduction of enchytraeids was affected by different soil properties, but the test validity criteria were fulfilled. The avoidance response of enchytraeids was highly variable, but significant effects of texture and pH were found. Earthworms were more sensitive to soil properties. They did not reproduce successfully in three of the 10 soils, and a positive influence of moisture, fine sand, pH, and organic matter and a negative influence of clay were found. Moreover, they strongly avoided soils with extreme textures. Despite these limitations, most of the selected soils are suitable substrates for ecotoxicological evaluations.