Study of the different evaluation areas in the pesticide risk assessment process: Focus on pesticides based on microorganisms.

Both chemical and microbial active substances can currently be approved as pesticides in the EU, the provisions of their approval being set under Regulation (EC) No 1107/2009. Although sharing the same legal framework, chemicals and microorganisms used as pesticides have different risk profiles especially because once released into the environment, microbial active substances may produce secondary metabolites, multiply, spread and possibly genetically adapt or transfer antimicrobial resistance genes to other microorganisms. Consequently, the risk assessment process must adjust to the specificities ensuing from the chemical or microbial nature of the active substance. This specific programme focused on the risk assessment of microorganisms used as pesticides, especially on the low-risk criteria linked to antimicrobial resistance and the risk assessment of secondary metabolites. The use of microorganisms in integrated pest management (IPM) programmes was also investigated. In 2020, the recently adopted Farm to Fork Strategy and the Biodiversity Strategy for 2030, two important action plans of the European Green Deal, called for a 50% reduction in the use of and risk from chemical and more hazardous pesticides. Many microorganisms are likely to be approved as low-risk active substances, thus representing important tools to achieve this goal. Given the central role that microbial active substances could play towards a more sustainable food system, a need for information regarding the actual production of secondary metabolites by the microorganisms of interest and projects investigating IPM programmes at national and EU levels was identified.

A study using QSAR/QSPR models focused on the possible occurrence and risk of alloxydim residues from chlorinated drinking water, according to the EU Regulation.

Any active substance with phytosanitary capacity intended to be marketed in Europe must pass exhaustive controls to assess its risk before being marketed and used in European agriculture. Since the implementation of Regulation (EC) No 1107/2009, agrochemical companies have been obliged to study the formation of pesticide transformation products (TPs) during the treatment of drinking water containing pesticide residues. However, there is no consensus on how to address this requirement. In this research work, the open literature collection on alloxydim was used to propose potential chlorination paths from alloxydim isomers. Furthermore, several QSAR/QSPR models have been used to fill the of knowledge gap relative to some key parameters in the physico-chemical, environmental and ecotoxicological areas of potential alloxydim TPs from chlorinated water for which little information exists. In this way, it has been possible to estimate the state of aggregation of these TPs (they exist mainly as liquids) as well as their ease of transit between the different phases, to predict their possible behaviour in the three environmental compartments (e.g., thermophysical properties point to a change in their evolution with respect to the parent alloxydim isomers) and to anticipate their potential risk to human and animal health (e.g., all of them cause developmental toxicity). These and other results highlight that the hazards of several TPs, i.e., both chlorinated and nonchlorinated from parent alloxydim or from those obtained after cleavage of the N - O bond and the subsequent reaction with chlorine, should be seriously considered. The obtained results reopen the debate on the implications of the use of QSAR/QSPR models for pesticide risk assessment in the legislative framework.

Analysis of Pesticide Residues by QuEChERS Method and LC-MS/MS for a New Extrapolation of Maximum Residue Levels in Persimmon Minor Crop.

According to EU guidance SANCO/7525/VI/95 Rev. 10.3, residue data extrapolation from a surrogate major crop to a minor crop can be used for setting maximum residue levels (MRLs) with a reduced number of residue trials and representative selected pesticides. In this work, a QuEChERS method (citrate-buffered version and PSA with MgSO4 clean-up) and LC-ESI-MS/MS for the determination of boscalid, pyraclostrobin, fludioxonil, fluopyram and tebuconazole in persimmon was developed and validated according to EU Commission guidelines and afterwards used for the determination of residues in four field trials. Residue levels at harvest for each pesticide ranged between 0.347 and 0.028 mg/kg. After comparing EFSA residue data on apples, as the surrogate major crop, and conducting a consumer risk assessment, a proposal of residue data extrapolation to set MRLs in persimmons was performed. The results showed that pesticide residues in persimmons at harvest were consistently lower than residues in apples when substances were applied according to the same critical GAP. MRLs were set at 0.5 mg/kg for fludioxonil, 0.6 mg/kg for boscalid, 0.3 mg/kg for tebuconazole, 0.4 mg/kg for fluopyran and 0.3 mg/kg for pyraclostrobin. The ratio of the MRLs for apple/persimmon varied between 2.5 for boscalid and 1.25 for fluopyram, suggesting that residue extrapolation can be feasible, promoting the process of pesticide registration for minor crops and the settlement of MRL.

Study of the different evaluation areas in the pesticide risk assessment process.

Approval of active substances and authorisation of plant protection products in the EU is made based on a strict risk assessment of the agronomic use of the plant protection products. Regulation 1107/2009 regulates the procedure in the EU with complex procedures involving many actors. 'The Farm to Fork strategy' and 'The Biodiversity for 2030 strategy', that are the heart of the 'European Green Deal', aiming to make food systems fair, healthy, environmentally friendly and put Europe's biodiversity on the path to recovery by 2030, for the benefit of people, climate and the planet. Therefore, 'The Farm to Fork strategy' and 'The Biodiversity for 2030 strategy' represents a challenge for the evaluation and authorisation of plant protection products in which the risk management will constitute a key element on the approval of active substances and authorisation of plant protection products. The aim of the work was to get knowledge of the large body of EU legislation and guidelines in the plant production products, identifying the most critical points of the pesticide evaluation in each of its areas, analysing the complexity and the interaction between these different areas. This study allowed to have a global and clearer vision of these procedures, with the focus on highlighting inconsistency and to propose speed up alternatives. Finally, this work will also facilitate not only the risk assessment but also the decision-making on the approval of active substances and the authorisation of plant protection products.

Computational-Based Study of QuEChERS Extraction of Cyclohexanedione Herbicide Residues in Soil by Chemometric Modeling.

Assessment of two buffered QuEChERS (quick, easy, cheap, effective, rugged, and safe) versions (i.e., citrate and acetate) modified by including methanol to recover the residues of three cyclohexanedione oxime (CHD) herbicides and three of their byproducts from agricultural soil was performed. In this context, a full second-order face-centered factorial experimental design was developed to quantify the influences of the main five variables (i.e., extraction time, water content, soil weight, and extraction solvent volume and composition) on the target compound recoveries. The fitting equations satisfactorily described the extraction process behavior. The mathematical models also showed the most influencing independent variables (i.e., extraction solvent composition and soil weight). Handling simpler expressions was possible with the acetate QuEChERS but not with the citrate QuEChERS. The recoveries of the CHD residues were close to 100% after performing the extraction under suitable conditions. Furthermore, dispersive solid-phase extraction (dSPE) clean-up steps were assessed to reduce the matrix effect in mass spectrometry. In this sense, the citrate QuEChERS in combination with the PSA + C18 clean-up step was the best option for the extraction of CHD residues.

Considerations of nano-QSAR/QSPR models for nanopesticide risk assessment within the European legislative framework.

The European market for pesticides is currently legislated through the well-developed Regulation (EC) No. 1107/2009. This regulation promotes the competitiveness of European agriculture, recognizing the necessity of safe pesticides for human and animal health and the environment to protect crops against pests, diseases and weeds. In this sense, nanotechnology can provide a tremendous opportunity to achieve a more rational use of pesticides. However, the lack of information regarding nanopesticides and their fate and behavior in the environment and their effects on human and animal health is inhibiting rapid nanopesticide incorporation into European Union agriculture. This review analyzes the recent state of knowledge on nanopesticide risk assessment, highlighting the challenges that need to be overcame to accelerate the arrival of these new tools for plant protection to European agricultural professionals. Novel nano-Quantitative Structure-Activity/Structure-Property Relationship (nano-QSAR/QSPR) tools for risk assessment are analyzed, including modeling methods and validation procedures towards the potential of these computational instruments to meet the current requirements for authorization of nanoformulations. Future trends on these issues, of pressing importance within the context of the current European pesticide legislative framework, are also discussed. Standard protocols to make high-quality and well-described datasets for the series of related but differently sized nanoparticles/nanopesticides are required.

The impact of restrictions on neonicotinoid and fipronil insecticides on pest management in maize, oilseed rape and sunflower in eight European Union regions.

BACKGROUND: In 2013, the European Commission restricted the use of three neonicotinoids (clothianidin, imidacloprid and thiamethoxam) and the pyrazole fipronil, which are widely used to control early-season pests. Here, we used original farm survey data to examine the impact of the restrictions on pest management practices in eight regional case studies including maize, oilseed rape and sunflower in seven European Union (EU) countries. RESULTS: In four case studies, farmers switched to using untreated seeds as no alternative seed treatments were available. In three case studies, farmers switched to using unrestricted neonicotinoid- or pyrethroid-treated seeds. In five case studies, farmers increased the use of soil or foliar treatments, with pyrethroids as the principal insecticide class. Other changes in pest management practices ranged from increased sowing density to more frequent scouting for pests. Many farmers perceived that the time, cost and amount of insecticides required to protect crops increased, along with pest pressure. Alternative seed treatments were mostly perceived as being less effective than the restricted seed treatments. CONCLUSION: Farmers generally relied on alternative seed treatments or more soil/foliar treatments in the first growing season after the restrictions took effect. Further study is required to assess the effectiveness and sustainability of these alternatives compared with the restricted insecticides. © 2017 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Photolysis of clethodim herbicide and a formulation in aquatic environments: Fate and ecotoxicity assessment of photoproducts by QSAR models.

The photochemical fate of the herbicide clethodim in natural waters was investigated under simulated and natural sunlight radiation. This herbicide exhibited a rapid degradation rate in simulated aquatic environment with half-lives ranged from 27.9min to 4.6h. The commercial formulation of clethodim showed a faster degradation with half-lives from 19.3min to 1.4h. It has also been demonstrated that the photolytic behavior of clethodim was affected by the water composition and the radiation intensity. Nine major photoproducts were identified and their distribution was dependent on the experimental conditions. Photodegraded solutions of clethodim were shown to be more toxic to the bacteria Vibrio fischeri than the herbicide itself, reaching the maximum toxicity when the herbicide is completely degraded. QSAR analysis of the fate, ecotoxicological and physicochemical endpoints of the degradation products provided positive alerts for several identified by-products. Environmental fate and transport estimates showed that all photoproducts, unlike the active substance, are potential leachers. Moreover, predicted vapor pressures suggested that dermal contact and ingestion are the most probable exposure routes for workers and general population to both clethodim and its photoproducts. These results highlight the importance of the degradation products in attaining a complete knowledge of the fate and behavior of an herbicide in the environment. To our knowledge, this is the first study to report a detailed QSAR study on clethodim photoproducts under environmental conditions. These results provide a very valuable information that will guide further experimental studies leading to a better pesticide risk assessment.

Development of a rapid and direct method for the determination of organic acids in peach fruit using LC-ESI-MS.

An accurate, simple and rapid liquid chromatography mass spectrometry method for the determination of organic acids in peach fruit has been developed. Direct injection and sample clean-up with a mixed-mode sorbent was compared. The best results for the determination of gluconic, oxalic, malic, citric and fumaric acids were obtained with only a simple dilution and filtration step, and nylon filters should be avoided since some organic acids are retained by them. It is the first time that gluconic acid has been determined in peach fruit. Different parameters involved in the separation and detection process have been optimized. Since matrix effects were observed in the peach commodity, organic acids were quantified by the standard addition method. All validation parameters of the method were found acceptable of all organic acids. Finally, the method was successfully applied to the analysis of samples of peach from two cultivars.

Aqueous photodegradation of sethoxydim herbicide: Qtof elucidation of its by-products, mechanism and degradation pathway.

The photochemical fate of sethoxydim herbicide in water was studied under simulated sunlight radiation (Suntest apparatus). This compound exhibited a rapid degradation rate in water with a half-life of approximately 1h, which is in accordance with the high calculated value of the quantum yield (Φ=0.26). Consequently, the photolysis of sethoxydim should be considered as an efficient route of sethoxydim removal in aqueous media. During the photodegradation studies, ten by-products were detected, and their identification was accomplished using high-performance liquid chromatography (HPLC) coupled with quadrupole time-of-flight mass spectrometry (Qtof) and electrospray ionization in positive mode (ESI+). A detailed and exhaustive study of the mass spectra of the precursor ions and their MS/MS fragmentation patterns allowed for their identification. The photodegradation products resulted from the cleavage and isomerization of the NO bond of the oxime, oxidation of the sulfur atom, oxidative CS bond cleavage and Beckmann rearrangement followed by intramolecular cyclization. Additionally, the reactive species involved and a plausible mechanism for the photodegradation of sethoxydim in water are discussed. Finally, based on the identified products, a rational pathway for the photodegradation of sethoxydim in water is proposed.

Field-incurred fenitrothion residues in kakis: comparison of individual fruits, composite samples, and peeled and cooked fruits.

Field trials have been carried out to determine the variability of residue levels of fenitrothion and its main metabolites fenitrothion-oxon and 3-methyl-4-nitrophenol in individual kaki fruits versus composite samples, in peel versus flesh, and in whole uncooked versus whole cooked fruits. Residue levels have been determined by gas chromatography with thermionic specific detection after extraction with ethyl acetate and without further cleanup. At harvest, residue levels of fenitrothion were below maximum residue levels (MRLs) and the two metabolites 3-methyl-4-nitrophenol and fenitrothion-oxon could be quantified with average amounts of 0.080 and 0.012 mg/kg, respectively. Levels of fenitrothion decreased 88% after peeling, whereas temperature did not result in a high variation. The ratios of the highest residue level in the individual fruits to the corresponding mean of residue levels in the composite samples for fenitrothion were <3. This value is lower than that recommended by the World Health Organization as default value for consumer risk assessment.

Occurrence, Distribution, and Relative Incidence of Mosaic Viruses Infecting Field-Grown Melon in Spain.

The main areas for field-grown melon (Cucumis melo) production in Spain were surveyed for the occurrence and relative incidence of cucumber mosaic virus (CMV), papaya ringspot virus-watermelon strain (PRSV-W), watermelon mosaic virus-2 (WMV-2), and zucchini yellow mosaic virus (ZYMV) during the growing seasons of 1995 and 1996. Samples from 1,152 plants showing symptoms of virus infection were collected from commercial melon fields and analyzed by enzyme-linked immunosorbent assay (ELISA). CMV and WMV-2 were the most frequently found viruses, both by the number of locations and by their incidence in each location. In contrast, PRSV-W and ZYMV were detected in fewer sites and at lower incidences. PRSV-W was not found in 1996. In 79% of the samples, only one virus was detected; 15% of the samples were doubly infected. Both the incidence of plants showing symptoms of viral infection and the relative incidence of each of the four viruses varied according to the region, while the main trends of virus distribution were similar for 1995 and 1996.