Assessing the effectiveness of a three-stage on-farm biobed in treating pesticide contaminated wastewater

Agricultural point source pesticide pollution arising from contaminated machinery washings and accidental spillages pose a significant threat to river water and groundwater quality. In this study, we assess the effectiveness of a three-stage on-farm biobed for treating pesticide contaminated waste water from a large (20 km2) commercial arable estate. The facility consisted of an enclosed machinery wash-down unit (stage 1), a 49 m2 lined compost-straw-topsoil biobed (stage 2), and a 200 m2 drainage field with a trickle irrigation system (stage 3). Pesticide concentrations were analysed in water samples collected fortnightly between November 2013 and November 2015 from the biobed input and output sumps and from 20 porous pots buried at 45 cm and 90 cm depth within the drainage field. The results revealed that the biobed removed 68–98% of individual pesticides within the contaminated washings, with mean total pesticide concentrations reducing by 91.6% between the biobed input and output sumps. Drainage field irrigation removed a further 68–99% of individual pesticides, with total mean pesticide concentrations reducing by 98.4% and 97.2% in the 45 cm and 90 cm depth porous pots, respectively. The average total pesticide concentration at 45 cm depth in the drainage field (57 µg L-1) was 760 times lower than the mean concentration recorded in the input sump (43,334 µg L-1). There was no evidence of seasonality in the efficiency of biobed pesticide removal, nor was there evidence of a decline in removal efficiency over the two-year monitoring period. However, higher mean total pesticide concentrations at 90 cm (102 µg L-1) relative to 45 cm (57 µg L-1) depth indicated an accumulation of pesticide residues deeper within the soil profile. Overall, the results presented here demonstrate that a three-stage biobed can successfully reduce pesticide pollution risk from contaminated machinery washings on a commercial farm

Dissipation of pure and broccoli-released glucosinolates in soil under high and low moisture content

Glucosinolates (GSLs) are secondary metabolites found in Brassica species. Upon tissue disruption GSLs are hydrolyzed by myrosinase enzymes to isothiocyanates (ITCs) which are highly toxic to microbes. Therefore the incorporation of fresh Brassicaceae tissues into soil for the control of soil-born plant pathogens is viewed as a biofumigation process. Its efficacy relies on the rate of GSL conversion to ITCs, but also on environmental factors controlling GSL availability in the soil matrix. We studied the dissipation of GSLs, applied either by broccoli leaves or as pure compounds, in a clay loam soil at two soil moisture content levels. Regardless of their mode of application, GSLs were rapidly dissipated in soil with half-life values ranging from 3.2 to 15.5 h. Increasing moisture from 20 to 90% of the soil water holding capacity significantly accelerated their dissipation. Indolyl broccoli-derived GSLs dissipated faster than aliphatic GSLs at high moisture levels, while at low moisture levels a three-fold reduction in their dissipation rates was observed. The dissipation of aliphatic GSLs was less affected by soil moisture levels. Application of pure GSLs resulted in increased soil metabolic quotients (qCO(2)), suggesting a decline is microbial metabolic efficiency. Their dissipation was related to myrosinase activity apparently derived from soil microbes. (c) 2013 Published by Elsevier Masson SAS

Isolation of a bacterial consortium able to degrade the fungicide thiabendazole: the key role of a Sphingomonas phylotype

Thiabendazole (TBZ) is a fungicide used in fruit-packaging plants. Its application leads to the production of wastewaters requiring detoxification. In the absence of efficient treatment methods, biological depuration of these effluents could be a viable alternative. However, nothing is known regarding the microbial degradation of the recalcitrant and toxic to aquatics TBZ. We report the isolation, via enrichment cultures from a polluted soil, of the first bacterial consortium able to rapidly degrade TBZ and use it as a carbon source. Repeated efforts using various culture-dependent approaches failed to isolate TBZ-degrading bacteria in axenic cultures. Denaturating gradient gel electrophoresis (DGGE) and cloning showed that the consortium was composed of α-, β- and γ-Proteobacteria. Culture-independent methods including antibiotics-driven selection with DNA/RNA-DGGE, q-PCR and stable isotope probing (SIP)-DGGE identified a Sphingomonas phylotype (B13) as the key degrading member. Cross-feeding studies with structurally related chemicals showed that ring substituents of the benzimidazole moiety (thiazole or furan rings) favoured the cleavage of the imidazole moiety. LC-MS/MS analysis verified that TBZ degradation proceeds via cleavage of the imidazole moiety releasing thiazole-4-carboxamidine, which was not further transformed, and the benzoyl moiety, possibly as catechol, which was eventually consumed by the bacterial consortium as suggested by SIP-DGGE. © 2017, Springer-Verlag Berlin Heidelberg

Exploring the potential of biobeds for the depuration of pesticide-contaminated wastewaters from the citrus production chain: Laboratory, column and field studies

The high wastewater volumes produced during citrus production at pre- and post-harvest level presents serious pesticide point-source pollution for groundwater bodies. Biobeds are used for preventing such point-source pollution occurring at farm level. We explored the potential of biobeds for the depuration of wastewaters produced through the citrus production chain following a lab-to-field experimentation. The dissipation of pesticides used pre- or post-harvest was studied in compost-based biomixtures, soil, and a straw-soil mixture. A biomixture of composted grape seeds and skins (GSS-1) showed the highest dissipation capacity. In subsequent column studies, GSS-1 restricted pesticides leaching even at the highest water load (462 L m(-3)). Ortho-phenylphenol was the most mobile compound. Studies in an on-farm biobed filled with GSS-1 showed that pesticides were fully retained and partially or fully dissipated. Overall biobeds could be a valuable solution for the depuration of wastewaters produced at pre- and post-harvest level by citrus fruit industries. (C) 2012 Elsevier Ltd. All rights reserved

Bioaugmentation of thiabendazole-contaminated soils from a wastewater disposal site: Factors driving the efficacy of this strategy and the diversity of the indigenous soil bacterial community

The application of the fungicide thiabendazole (TBZ) in fruit packaging plants (FPP) results in the production of effluents which are often disposed in adjacent field sites. These require remediation to prevent further environmental dispersal of TBZ. We assessed the bioaugmentation potential of a newly isolated TBZ-degrading bacterial consortium in a naturally contaminated soil (NCS) exhibiting a natural gradient of TBZ levels (12000, 400, 250 and 12 mg kg−1). The effect of aging on bioaugmentation efficacy was comparatively tested in a soil with similar physicochemical properties and soil microbiota, which was artificially, contaminated with the same TBZ levels (ACS). The impact of bioaugmentation and TBZ on the bacterial diversity in the NCS was explored via amplicon sequencing. Bioaugmentation effectively removed TBZ from both soils at levels up to 400 mg kg−1 but failed at the highest contamination level (12000 mg kg−1). Dissipation of TBZ in bioaugmented samples showed a concentration-dependent pattern, while aging of TBZ had a slight effect on bioaugmentation efficiency. Bioaugmentation had no impact on the soil bacterial diversity, in contrast to TBZ contamination. Soils from the hotspots of TBZ contamination (12000 mg kg−1) showed a drastically lower α-diversity driven by the dominance of β- and γ-proteobacteria at the expense of all other bacterial phyla, especially Actinobacteria. Overall, bioaugmentation with specialized microbial inocula could be an effective solution for the recovery of disposal sites contaminated with persistent chemicals like TBZ. Bioaugmentation with a specialized bacterial consortium was effective in the remediation of soils naturally contaminated with the persistent fungicide thiabendazole, and did not affect the soil bacterial diversity, the latter being heavily affected by high fungicide levels. © 2017 Elsevier Lt

Impact of Nitrogen and Sulfur Fertilization on the Composition of Glucosinolates in Relation to Sulfur Assimilation in Different Plant Organs of Broccoli

Broccoli (Brassica oleracea var. italica) is one of the most important winter season vegetables and a rich source of chemoprotective molecules, including glucosinolates (GSL). The aim of this study was to investigate the impact of nitrogen (N) and sulfur (S) fertilization on GSL concentration and composition in different parts of broccoli plants. A greenhouse experiment was performed, with four different treatments of sulfur (10, 30, 70, and 150 kg/ha) and three treatments of nitrogen (50, 250, and 600 kg/ha). GSL concentrations and plant growth responded to the N supply, but this was not observed above the 250 kg.N/ha dose. On the contrary, plant growth did not respond to the S supply, whereas GSL concentrations showed a sharp response to the whole range of S applications (from 10 to 150 kg/ha). Glucosinolate composition was altered differentially in the examined plant parts. Aliphatic GSL were more abundant in the florets and leaves, whereas indolyl GSLs were dominant in roots, in which aromatic GSL were also observed. High nitrogen fertilization had a higher impact on indolyl compared to aliphatic GSLs concentration. More importantly, a high concentration of aliphatic GSL, >2.4 mu mol/g dry weight (dw), and high S assimilation into aliphatic GSL were consistently observed in the florets compared to other broccoli parts, indicating adaptable processes for nitrogen and sulfur regarding synthesis and transport of aliphatic GSL for these organs

Relationships between nitrogen, dry matter accumulation and glucosinolates in Eruca sativa Mills. The applicability of the critical NO3-N levels approach

Rocket salad (Eruca sativa Mills) is one of the major leafy vegetables produced worldwide and has been characterized as a rich source of chemoprotective glucosinolates (GSL). The relationship between N fertilization and the resulting plant biomass and N status with GSL quantity and quality in rocket leaves was examined. A pot experiment was conducted, applying ten different N-rates and destructive sampling was carried out 15, 30 and 45 days after transplanting (DAT). The Mitscherlich equation was used to establish NO3-N critical levels at each growth stage and as an indicator of N demand for relative maximum dry matter accumulation and glucosinolate content and composition was determined. Glucosinolate content was significantly influenced by N rate, growth stage and their interaction. Different GSL types showed dissimilar responses to N fertilization: aliphatic GSLs were significantly reduced under increased N rates whereas indole GSL showed the reverse. Under excess N fertilization (> 1.04 g/plant), dry matter accumulation remained constant, NO3-N was significantly increased and total GSL content was significantly reduced, factors that could lead to an anticipated product quality decline. The application of the critical NO3-N level approach used to identify optimal N fertilization rates for plant growth could serve as means to obtain optimized GSL content in the edible plant parts

Impact of a beneficial and of a pathogenic Fusarium strain on the fingerprinting-based structure of microbial communities in tomato (Lycopersicon esculentum Milll.) rhizosphere

Fusarium solani strain FsK (FsK), isolated from a plant pathogen-suppressive compost, grows endophytically in tomato roots and controls infestations by Fusarium oxysporum f.sp. radicis-lycopersici (FORL). The effect of root colonization by the two fungi on the diversity of rhizosphere microbial community was studied. Tomato plants were inoculated with FsK and/or FURL and rhizosphere soil was collected 8, 15 and 30 days post inoculation (dpi) and analyzed by denaturating gradient gel electrophoresis (DGGE) of PCR-amplified internal transcribed spacer (ITS) sequences of fungi and 16S rRNA gene sequences of common rhizosphere bacterial guilds like alpha-proteobacteria and pseudomonads. Cluster analysis of DGGE fingerprints showed that FsK had a transient impact on the fungal and alpha-proteobacterial community only during its endophytic stage (15 dpi), while FURL had a readily distinguished and persistent effect on the fungal community. The changes observed in the rhizosphere fungal and bacterial communities may depict the interactions of the two fungal inocula with the plant. Cloning of selected DGGE bands stimulated by FsK showed that the responsive bacteria were closely related to species known to include biological control agents (BCA). Overall, the inoculation of FsK in tomato rhizosphere did not appear to have a significant impact on the diversity of non-target microbial groups inhabiting plant rhizosphere. (C) 2011 Elsevier Masson SAS. All rights reserved