Molecular insight into endosulfan degradation by Ese protein from Arthrobacter: Evidence-based structural bioinformatics and quantum mechanical calculations.

Endosulfan is an organochlorine insecticide widely used for agricultural pest control. Many nations worldwide have restricted or completely banned it due to its extreme toxicity to fish and aquatic invertebrates. Arthrobacter sp. strain KW has the ability to degrade α, ß endosulfan and its intermediate metabolite endosulfate; this degradation is associated with Ese protein, a two-component flavin-dependent monooxygenase (TC-FDM). Employing in silico tools, we obtained the 3D model of Ese protein, and our results suggest that it belongs to the Luciferase Like Monooxygenase family (LLM). Docking studies showed that the residues V59, V315, D316, and T335 interact with α-endosulfan. The residues: V59, T60, V315, D316, and T335 are implicated in the interacting site with ß-endosulfan, and the residues: H17, V315, D316, T335, N364, and Q363 participate in the interaction with endosulfate. Topological analysis of the electron density by means of the Quantum Theory of Atoms in Molecules (QTAIM) and the Non-Covalent Interaction (NCI) index reveals that the Ese-ligands complexes are formed mainly by dispersive forces, where Cl atoms have a predominant role. As Ese is a monooxygenase member, we predict the homodimer formation. However, enzymatic studies must be developed to investigate the Ese protein's enzymatic and catalytic activity.

Environmental Behavior of Chlorpyrifos and Endosulfan in a Tropical Soil in Central Brazil.

The environmental behavior of chlorpyrifos and endosulfan in soil was studied in the central-western region of Brazil by means of a field experiment. Sorption was evaluated in laboratory batch experiments. Chlorpyrifos and endosulfan were applied to experimental plots on uncultivated soil and the following processes were studied: leaching, runoff, and dissipation in top soil. Field dissipation of chlorpyrifos and endosulfan was more rapid than reported in temperate climates. Despite the high Koc of the studied pesticides, the two endosulfan isomers and endosulfan sulfate as well as chlorpyrifos were detected in percolated water. In runoff water and sediment, both endosulfan isomers and endosulfan sulfate were detected throughout the period of study. Observed losses of endosulfan by leaching (below a depth of 50 cm) and runoff were 0.0013 and 1.04% of the applied amount, whereas chlorpyrifos losses were 0.003 and 0.032%, respectively. Leaching of these highly adsorbed pesticides was attributed to preferential flow.

Analytical methodology for the study of endosulfan bioremediation under controlled conditions with white rot fungi.

A general procedure to study the biodegradation of endosulfan under laboratory conditions by white rot fungi isolated from native sources growing in YNB (yeast nitrogen base) media with 1% of glucose is presented. The evaluation of endosulfan biodegradation as well as endosulfan sulfate, endosulfan ether and endosulfan alcohol production throughout the whole bioremedation process was performed using an original and straightforward validated analytical procedure with recoveries between 78 and 112% at all concentration levels studied except for endosulfan sulfate at 50 mg L(-1) that yielded 128% and RSDs<20%. Under the developed conditions, the basidiomycete Bjerkandera adusta was able to degrade 83% of (alpha+beta) endosulfan after 27 days, 6 mg kg(-1) of endosulfan diol were determined; endosulfan ether and endosulfan sulfate were produced below 1 mg kg(-1) (LOQ, limit of quantitation).

Evaluation of the QuEChERS method and gas chromatography-mass spectrometry for the analysis pesticide residues in water and sediment.

A method for the determination of pesticide residues in water and sediment was developed using the QuEChERS method followed by gas chromatography--mass spectrometry. The method was validated in terms of accuracy, specificity, linearity, detection and quantification limits. The recovery percentages obtained for the pesticides in water at different concentrations ranged from 63 to 116%, with relative standard deviations below 12%. The corresponding results from the sediment ranged from 48 to 115% with relative standard deviations below 16%. The limits of detection for the pesticides in water and sediment were below 0.003 mg L⁻¹ and 0.02 mg kg⁻¹, respectively.

Assessing pesticide leaching and desorption in soils with different agricultural activities from Argentina (Pampa and Patagonia).

Pesticide distribution in the soil profile depends on soil and pesticide properties as well as on the composition of irrigation water. Water containing surfactants, acids or solvents, may alter pesticide desorption from soil. The distribution of organochlorine pesticides (OCPs) in two Argentinean agricultural areas, Pampa and Patagonia, was evaluated. Furthermore, pesticide desorption from aged and freshly spiked soils was performed by the batch technique, using solutions of sodium oxalate and citrate, dissolved organic carbon (DOC), wastewater and surfactants. Patagonian soil showed the highest OCP levels (46.5-38.1 µg g(-1) OC) from 0 to 30 cm depth and the predominance of p,p'-DDE residues reflected an extensive and past use of DDT. Pampean soil with lower levels (0.039-0.07 µg g(-1) OC) was mainly polluted by the currently used insecticide endosulfan. Sodium citrate and oxalate, at levels usually exuded by plant roots, effectively enhanced desorption of p,p'-DDT, p,p'-DDE and α-cypermethrin, while no effects were observed for α-endosulfan and endosulfan sulfate. The non-ionic surfactant Tween 80 behaved similarly to the acids, whereas the anionic sodium dodecyl sulfate enhanced desorption of all pesticides. Increased desorption of the hydrophobic pesticides also occurred when DOC from humic acids but not from sewage sludge or wastewater were used. Soil profile distribution of pesticides was in accordance with results from desorption studies. Data suggest pesticide leaching in Pampean and Patagonian soils, with risk of endosulfan to reach groundwater and that some organic components of wastewaters may enhance the solubilisation and leaching of recalcitrant compounds such as p,p'-DDT and p,p'-DDE.

Kinetic studies of endosulfan photochemical degradations by ultraviolet light irradiation in aqueous medium.

Kinetic studies of endosulfan photochemical degradation in controlled aqueous systems were carried out by ultraviolet light irradiation at lambda = 254 nm. The photolysis of (alpha + beta: 2 + 1) endosulfan, alpha-endosulfan and beta-endosulfan were first-order kinetics. The observed rate constants obtained from linear least-squares analysis of the data were 1 x 10(-4) s(-1); 1 x 10(-4) s(-1); and 2 x 10(-5) s(-1), respectively, and the calculated quantum yields (phi) were 1, 1 and 1.6, respectively. Preliminary differential pulse polarographic (DPP) analysis allowed to observe the possible endosulfan photochemical degradation pathway. This degradation route involves the formation of the endosulfan diol, its transformation to endosulfan ether and finally the ether's complete degradation by observing the potential shifts.

Endosulfan induces oxidative stress and changes on detoxication enzymes in the aquatic macrophyte Myriophyllum quitense.

Endosulfan (1) is a chlorinated insecticide still in use in both developed and emerging countries. Although its toxicity on animals has been studied in the last years, scarce information is available on its effects on plants. In this study, we exposed the aquatic macrophyte Myriophyllum quitense to environmentally relevant concentrations of endosulfan (microg/L) (1) for a short time, simulating exposures that might occur after either accidental spills or toxic run-off from agricultural areas. The main goal was to evaluate changes in both detoxication and antioxidant enzymatic systems of this plant upon exposure to endosulfan (1). Thus, we measured the activities of catalase (CAT), soluble and membrane associated glutathione-S-transferases (s- and m-GSTs) and glutathione reductase (GR), as well as the hydrogen peroxide (H2O2) content. Results showed that endosulfan (1) exerts oxidative stress on M. quitense, which was evidenced by the increase of CAT activity and the H2O2 content in exposed plants. At 5 microg/L endosulfan (1), we found a generalized induction of activities of tested enzymes, indicating that this xenobiotic activates the protection system of this plant, increasing its capacity to scavenge reactive oxygen species. On the other hand, we did not find significant changes at 0.02 microg/L endosulfan (1), which is the maximal concentration allowed for freshwater. We conclude that runoff events, which can produce significant amounts of endosulfan (1) in aquatic environments during short time, can result in oxidative stress on M. quitense, and probably on similar macrophytes.

Fate of (14)C-labeled soybean and corn pesticides in tropical soils of Brazil under laboratory conditions.

The dissipation rate of seven currently used soybean and corn pesticides in two tropical soils (Ustox and Psamments) of Brazil was studied in a laboratory incubation experiment. Dissipation half-lives of pesticides ranged between 2 (monocrotofos) and 90 days (endosulfan-beta). The contrasting clay contents of the studied tropical soils (130 versus 470 g of clay kg(-1) of soil) did not influence the dissipation dynamics of pesticides substantially. Mineralization to CO(2) was high [up to 78% of the applied radioactivity (AR)] for the studied organophosphorus compounds and deltamethrin, which also formed considerable amounts of bound residues (>20% of AR) during the 80 days of incubation. The highest portion of nonextractable residues was found for alachlor and simazine (55-60% of AR). In contrast, the nonpolar trifluralin and endosulfan formed only small amounts of bound residues (mostly <20% of AR) but showed the highest dissipation half-lives (>14 days) in the studied soils, also due to a low mineralization rate. When endosulfan-sulfate, as the main metabolite of endosulfan, was considered, the half-life time of endosulfan compounds (sum of -alpha, -beta, and -sulfate) was enhanced to >160 days in both soils. In comparison with the laboratory experiments, dissipation half-life times of chlorpyrifos, endosulfan-alpha, and trifluralin were shortened by a factor of 10-30 in field trials with the same soils, which was related to the volatilization potential of pesticides from soils.

Leaching and degradation of corn and soybean pesticides in an Oxisol of the Brazilian Cerrados.

Pesticide pollution of ground and surface water is of growing concern in tropical countries. The objective of this pilot study was to evaluate the leaching potential of eight pesticides in a Brazilian Oxisol. In a field experiment near Cuiabá, Mato Grosso, atrazine, chlorpyrifos, lambda-cyhalothrin, endosulfane alpha, metolachlor, monocrotofos, simazine, and trifluraline were applied onto a Typic Haplustox. Dissipation in the topsoil, mobility within the soil profile and leaching of pesticides were studied for a period of 28 days after application. The dissipation half-life of pesticides in the topsoil ranged from 0.9 to 14 d for trifluraline and metolachlor, respectively. Dissipation curves were described by exponential functions for polar pesticides (atrazine, metolachlor, monocrotofos, simazine) and bi-exponential ones for apolar substances (chlorpyrifos, lambda-cyhalothrin, endosulfane alpha, trifluraline). Atrazine, simazine and metolachlor were moderately leached beyond 15 cm soil depth, whereas all other compounds remained within the top 15 cm of the soil. In lysimeter percolates (at 35 cm soil depth), 0.8-2.0% of the applied amounts of atrazine, simazine, and metolachlor were measured within 28 days after application. Of the other compounds less than 0.03% of the applied amounts was detected in the soil water percolates. The relative contamination potentials of pesticides, according to the lysimeter study, were ranked as follows: metolachlor > atrazine = simazine >> monocrotofos > endsulfane alpha > chlorpyrifos > trifluraline > lambda-cyhalothrin. This order of the pesticides was also achieved by ranking them according to their effective sorption coefficient Ke, which is the ratio of Koc to field-dissipation half-life.

The use of the mitochondrial transmembrane electric potential as an effective biosensor in ecotoxicological research.

In this work, the mitochondrial transmembrane electric potential (delta psi) of isolated mitochondria was used to evaluate the toxicity of some chemicals (endosulfan, 3,4-dichloroaniline, parathion, tributyltin and cadmium) and wastewater. Mitochondria were isolated from rat liver, and the delta psi measured in a suitable assay medium, using a sensitive tetraphenylphosphonium (TPP+) electrode. The test substance was pre-incubated in a rotenone-containing medium during 3 min with 1.0 mg of mitochondrial protein. Mitochondria were energised with succinate and after the establishment of a constant maximal potential ADP was added to induce the phosphorylative cycle. Chosen endpoints were the membrane potential from mitochondria oxidising succinate and the depolarisation induced by ADP. After the appropriate transformations the EC50 (effective concentration) was calculated for each toxicant. Even very low concentrations of a toxicant were able to affect the delta psi, thus showing its suitability as a biosensor in ecotoxicology and results were reproducible between tests. The utilisation of delta psi in screening tests of pure substances and wastewater seems to be very effective and can be carried out routinely.