Surface and ground waters Vineyards s u m m a r y A multi-residual analytical method based on solid phase extraction (SPE) followed by liquid chromatography-electrospray ionisation-mass spectrometry (LC-MS) was developed to monitor pesticides in natural waters. Fifty-eight compounds, including herbicides, fungicides, insecticides and some of their degradation products, were surveyed to evaluate the quality of natural waters throughout the winegrowing region of La Rioja (Rioja DOCa). Ninety-two sampling points were selected, including surface and ground waters that could be affected by agricultural activities covering the region's three sub-areas. Different parameters that may affect the efficiency of the SPE procedure were optimised (sorbent type, elution solvent and sample volume), and matrix-matched standards were used to eliminate the variable matrix effect and ensure good quantification. The developed method allows the determination of target compounds below the level established by the European Union for waters for human use with suitable precision (relative standard deviations lower than 18%) and accuracy (with recoveries over 61%). Forty compounds included in this study (six insecticides, 12 herbicides, 16 fungicides and six degradation products) were detected in one or more samples. The herbicides terbuthylazine, its metabolite desethyl terbuthylazine, fluometuron and ethofumesate and the fungicides pyrimethanil and tebuconazole were the compounds most frequently detected in water samples (present in more than 60% of the samples). Concentrations above 0.1 lg L À1 were detected for 37 of the compounds studied, and in several cases recorded values of over 18 lg L À1 . The results reveal the presence of pesticides in most of the samples investigated. In 64% of groundwaters and 62% of surface waters, the sum of compounds detected was higher than 0.5 lg L À1 (the limit established by EU legislation for the sum of all pesticides detected in waters for human use).
Pesticides are needed to maintain high production in the vineyard area of La Rioja (Spain), and monitoring their spatial distribution is a priority for preserving the quality of natural resources. Accordingly, the purpose of this work was to conduct a study to evaluate the presence and seasonal distribution of herbicide and insecticide residues in ground and surface waters in this region. The monitoring network comprised 12 surface waters and 78 groundwaters, covering the three subareas (63,593ha) into which the vineyard region is divided. The quality of natural waters was examined through the analysis of twenty-two herbicides, eight of their main degradation products, and eight insecticides. Pesticides were extracted by solid-phase extraction, and analysed by gas chromatography-mass spectrometry or by liquid-chromatography-mass spectrometry. The results reveal the presence of most of the herbicides and insecticides included in the study in one or more of the samples collected during the four campaigns. The herbicide terbuthylazine and its metabolite desethylterbuthylazine were the compounds more frequently detected (present in >65% of the samples across all the campaigns). Other compounds detected in >50% of the samples in one sampling campaign were the herbicides fluometuron, metolachlor, alachlor and ethofumesate. Insecticides were present in a small number of samples, with only pirimicarb being detected in >25% of the samples in March and June campaigns. The results reveal that the sum of compounds detected (mainly herbicides) was higher than 0.5μgL in >50% of the samples, especially in the campaigns with the highest application of these compounds. A possible recovery of the quality of the waters was detected outside the periods of crop cultivation, although more monitoring programmes are needed to confirm this trend with a view to preventing and/or maintaining the sustainability of natural resources.
The abundance of libA, encoding a hydrolase that initiates linuron degradation in the linuron-metabolizing Variovorax sp. strain SRS16, was previously found to correlate well with linuron mineralization, but not in all tested environments. Recently, an alternative linuron hydrolase, HylA, was identified in Variovorax sp. strain WDL1, a strain that initiates linuron degradation in a linuron-mineralizing commensal bacterial consortium. The discovery of alternative linuron hydrolases poses questions about the respective contribution and competitive character of hylA-and libA-carrying bacteria as well as the role of linuron-mineralizing consortia versus single strains in linuron-exposed settings. Therefore, dynamics of hylA as well as dcaQ as a marker for downstream catabolic functions involved in linuron mineralization, in response to linuron treatment in agricultural soil and on-farm biopurification systems (BPS), were compared with previously reported libA dynamics. The results suggest that (i) organisms containing either libA or hylA contribute simultaneously to linuron biodegradation in the same environment, albeit to various extents, (ii) environmental linuron mineralization depends on multispecies bacterial food webs, and (iii) initiation of linuron mineralization can be governed by currently unidentified enzymes. IMPORTANCEA limited set of different isofunctional catabolic gene functions is known for the bacterial degradation of the phenylurea herbicide linuron, but the role of this redundancy in linuron degradation in environmental settings is not known. In this study, the simultaneous involvement of bacteria carrying one of two isofunctional linuron hydrolysis genes in the degradation of linuron was shown in agricultural soil and on-farm biopurification systems, as was the involvement of other bacterial populations that mineralize the downstream metabolites of linuron hydrolysis. This study illustrates the importance of the synergistic metabolism of pesticides in environmental settings.
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