Simultaneous removal of neonicotinoid insecticides by a microbial degrading consortium: Detoxification at reactor scale.

Neonicotinoid insecticides show high persistence in the environment, and standard biological approaches such as biopurification systems have shown mostly inefficient removal of such compounds. In this work, soil pre-exposed to imidacloprid was used to obtain presumptive imidacloprid-degrading consortia. Cometabolic enrichment yielded a microbial consortium composed of eight bacterial and one yeast strains, capable of degrading not only this compound, but also thiamethoxam and acetamiprid, as demonstrated in cross-degradation assays. The biological removal process was scaled-up to batch stirred tank bioreactors (STBR); this configuration was able to simultaneously remove mixtures of imidacloprid + thiamethoxam or imidacloprid + thiamethoxam + acetamiprid, reaching elimination of 95.8% and 94.4% of total neonicotinoids, respectively. Removal rates in the bioreactors followed the pattern imidacloprid > acetamiprid > thiamethoxam, including >99% elimination of imidacloprid in 6 d and 17 d (binary and ternary mixtures, respectively). A comprehensive evaluation of the detoxification in the STBR was performed using different biomarkers: seed germination (Lactuca sativa), bioluminescence inhibition (Vibrio fischeri), and acute oral tests in honeybees. Overall, ecotoxicological tests revealed partial detoxification of the matrix, with clearer detoxification patterns in the binary mixture. This biological approach represents a promising option for the removal of neonicotinoids from agricultural wastewater; however, optimization of the process should be performed before application in farms.

Removal of triazines, triazoles and organophophates in biomixtures and application of a biopurification system for the treatment of laboratory wastewaters.

Biopurification systems (BPS) have been barely explored for removing complex mixtures of pesticides. In this study, the potential of a biomixture to remove simultaneously a mixture of herbicides (triazines), fungicides (triazoles) and insecticides (organophosphates) is presented. Also, a BPS using the same biomixture was used for treating a pesticide testing laboratory wastewater containing a mixture of 38 compounds. Ecotoxicological assays were conducted on the BPS elutriates to investigate the mixture detoxification. A mixture (concentrations of 4-8 mg kg-1) run in small-scale biomixture systems (SSB) for 128 d showed 59.3% removal of triazines, 68.5% of organophosphates and no elimination of triazoles. The treatment of the laboratory wastewater (individual concentrations range: 0.0036-0.25 mg kg-1) in the pilot-scale BPS for 281 d resulted in the elimination pattern of organophosphates (90.0%) > triazoles (73.4%) > carbamates (71.3%) > triazines (54.3%). Complete detoxification towards Daphnia magna and partial detoxification in Lactuca sativa seeds germination occurred in the BPS. Although the pesticide mixture complexity is higher in the BPS, the lower concentrations found in this matrix, could explain removal differences between SSB and BPS and the apparent inhibition in the elimination of carbamates and some triazines observed in the latter. These findings suggest that disposal of pesticide-containing laboratory-wastewater should be done in separate containers, according to chemical groups before their treatment in separate BPS, in order to reduce treatment periods. Monitoring the treatment process in the BPS with a battery of ecotoxicological tests is strongly recommended.

Removal of herbicides in a biopurification system is not negatively affected by oxytetracycline or fungally pretreated oxytetracycline.

The disposal of agricultural antibiotic-containing wastewater in biopurification systems (BPS) employed in the treatment of pesticides, may negatively affect the removal capacity of these devices. This work aimed to employ a fungal pretreatment of oxytetracycline (OTC)-rich wastewater, before its disposal in a BPS used for the treatment of two pesticides. The fungal treatment at reactor scale (stirred tank reactor, 3L) with biomass of Trametes versicolor efficiently removed 100 mg L-1 OTC in only 60 h. However, ecotoxicity tests on seed germination with Lactuca sativa revealed that antibiotic elimination did not correlate with a decrease in toxicity. After the pretreatment, treated OTC was discarded in biomixtures used for the elimination of the herbicides ametryn and terbutryn. The co-application of treated or untreated OTC did not inhibit the removal of the herbicides; moreover, in both cases their removal seemed to be slightly enhanced in the presence of OTC or its residues, with respect to antibiotic-free biomixtures. Estimated half-lives ranged from 28.4 to 34.8 d for ametryn, and 34.0-51.0 d for terbutryn. In addition, the biomixture was also able to remove OTC in the presence of the herbicides, with an estimated half-life of 38 d. Remarkably, the toxicity of the wastewater containing OTC or treated OTC was mostly eliminated after its disposal in the biomixture. Overall results suggest that, given the high efficiency of the biomixture, the fungal pretreatment of OTC-containing wastewater is not mandatory before its disposal in the BPS.

Elimination of fungicides in biopurification systems: Effect of fungal bioaugmentation on removal performance and microbial community structure.

Bioaugmentation with ligninolytic fungi represents a potential way to improve the performance of biomixtures used in biopurification systems for the treatment of pesticide-containing agricultural wastewater. The fungus Trametes versicolor was employed in the bioaugmentation of a biomixture to be used in the simultaneous removal of seven fungicides. Liquid cultures of the fungus were able to remove tebuconazole, while no evidence of carbendazim, metalaxyl and triadimenol depletion was found. When applied in the biomixture, the bioaugmented matrix failed to remove all the triazole fungicides (including tebuconazole) under the assayed conditions, but was efficient to eliminate carbendazim, edifenphos and metalaxyl (the latter only after a second pesticide application). The re-addition of pesticides markedly increased the elimination of carbendazim and metalaxyl; nonetheless, no clear enhancement of the biomixture performance could be ascribed to fungal bioaugmentation, not even after the re-inoculation of fungal biomass. Detoxification efficiently took place in the biomixture (9 d after pesticide applications) according to acute tests on Daphnia magna. DGGE-analysis revealed only moderate time-divergence in bacterial and fungal communities, and a weak establishment of T. versicolor in the matrix. Data suggest that the non-bioaugmented biomixture is useful for the treatment of fungicides other than triazoles.

Removal of pesticides and ecotoxicological changes during the simultaneous treatment of triazines and chlorpyrifos in biomixtures.

Biopurification systems constitute a biological approach for the treatment of pesticide-containing wastewaters produced in agricultural activities, and contain an active core called biomixture. This work evaluated the performance of a biomixture to remove and detoxify a combination of three triazine herbicides (atrazine/terbuthylazine/terbutryn) and one insecticide (chlorpyrifos), and this efficiency was compared with dissipation in soil alone. The potential enhancement of the process was also assayed by bioaugmentation with the ligninolytic fungi Trametes versicolor. Globally, the non-bioaugmented biomixture exhibited faster pesticide removal than soil, but only in the first stages of the treatment. After 20 d, the largest pesticide removal was achieved in the biomixture, while significant removal was detected only for chlorpyrifos in soil. However, after 60 d the removal values in soil matched those achieved in the biomixture for all the pesticides. The bioaugmentation failed to enhance, and even significantly decreased the biomixture removal capacity. Final removal values were 82.8% (non-bioaugmented biomixture), 43.8% (fungal bioaugmented biomixture), and 84.7% (soil). The ecotoxicological analysis revealed rapid detoxification (from 100 to 170 TU to <1 TU in 20 d) towards Daphnia magna in the biomixture and soil, and slower in the bioaugmented biomixture, coinciding with pesticide removal. On the contrary, despite important herbicide elimination, no clear detoxification patterns were observed in the phytotoxicity towards Lactuca sativa. Findings suggest that the proposed biomixture is useful for fast removal of the target pesticides; even though soil also removes the agrochemicals, longer periods would be required. On the other hand, the use of fungal bioaugmentation is discouraged in this matrix.

Simultaneous removal of structurally different pesticides in a biomixture: Detoxification and effect of oxytetracycline.

The biopurification systems (BPS) used for the treatment of pesticide-containing wastewater must present a versatile degrading ability, in order to remove different active ingredients according to the crop protection programs. This work aimed to assay the simultaneous removal of several pesticides (combinations of herbicides/insecticides/fungicides, or insecticides/fungicides) in a biomixture used in a BPS over a period of 115 d, and in the presence of oxytetracycline (OTC), an antibiotic of agricultural use that could be present in wastewater from agricultural pesticide application practices. The biomixture was able to mostly remove the herbicides during the treatment (removal rates: atrazine ≈ linuron > ametryn), and suffered no inhibition by OTC (only slightly for ametryn). Two fungicides (carbendazim and metalaxyl) were removed, nonetheless, in the systems containing only fungicides and insecticides, a clear increase in their half-lives was obtained in the treatments containing OTC. The neonicotinoid insecticides (imidacloprid and thiamethoxam) and the triazole fungicides (tebuconazole and triadimenol) were not significantly eliminated in the biomixture. Globally, the total removal of active ingredients ranged from 40.9% to 61.2% depending on the system, following the pattern: herbicides > fungicides > insecticides. The ecotoxicological analysis of the process revealed no detoxification towards the microcrustacean Daphnia magna, but a significant decay in the phytotoxicity towards Lactuca sativa in some cases, according to seed germination tests; in this case, OTC proved to be partially responsible for the phytotoxicity. The patterns of pesticide removal and detoxification provide inputs for the improvement of BPS use and their relevance as devices for wastewater treatment according to specific pesticide application programs.