Transfer and risk assessment of fipronil in laying hen tissues and eggs.

Fipronil is a persistent insecticide known to transfer into hen eggs from exposure from animal drinking water and feed, but some questions remain regarding its transfer behavior and distribution characteristics. Therefore, the dynamic metabolism, residue distribution and transfer factor (TF) of fipronil were investigated in 11 edible tissues of laying hens and eggs over 21 days. After a continuous low-dose drinking water exposure scenario, the sum of fipronil and all its metabolites (defined as fipronilT) quickly transferred to each edible tissue and gradually increased with exposure time. FipronilT residue in eggs first appeared at 3 days and then gradually increased. After a single high-dose feed exposure scenario, fipronilT residue in edible tissues first appeared after 2 h, quickly peaked at 1 day, and then gradually decreased. In eggs, fipronilT residue first appeared at 2 days, peaked 6-7 days and then gradually decreased. The TF values followed the order of the skin (0.30-0.73) > egg yolk (0.30-0.71) > bottom (0.21-0.59) after drinking water exposure, and the order of the skin (1.01-1.59) > bottom (0.75-1.1) > egg yolk (0.58-1.10) for feed exposure. Fipronil sulfone, a more toxic compound, was the predominant metabolite with higher levels distributed in the skin and bottom for both exposure pathways. FipronilT was distributed in egg yolks rather than in albumen owing to its lipophilicity, and the ratio of egg yolk to albumen may potentially reflect the time of exposure. The distinction is that the residues after feed exposure were much higher than that after drinking water exposure in edible tissues and eggs. The study highlights the residual characteristics of two exposure pathways, which would contribute to the tracing of contamination sources and risk assessment.

Enantioselective fate and risk assessment of chiral fungicide pydiflumetofen in rice-fish and wheat farming systems.

Fungicides have been widely used for reducing the losses caused by plant diseases. Rice and wheat are the most basic food crops, and the potential risks after applying fungicides are worthy of attention. Especially rice-fish farming system is an ecological symbiosis system that is beneficial to both environmental and ecological protection. However, the application of pesticides will stress the ecosystem, and the pesticide residues in rice and fish would be transmitted along the food chain, which is harmful to human health. Here, the enantioselective behaviors of chiral pydiflumetofen in rice-fish and wheat farming systems were clarified. In the rice-fish farming system, pydiflumetofen enantiomers were preferentially attached to the plants, entering the paddy water and settling into the paddy soil, and then accumulating and dissipating in the fish. With the growth of rice, it was transported to rice fruits. The wheat farming system was similar. Enantioselective dissipation occurred in carp (Cyprinus carpio), brown rice and wheat soil, and S-(+)-pydiflumetofen was preferentially dissipated. In other words, R-(-)-pydiflumetofen showed higher concentrations, especially in carp, which meant R-(-)-pydiflumetofen was more easily accumulated in the environment, and posed a greater potential risk to the farming system. The pydiflumetofen residues in brown rice and wheat were lower than MRLs from the EFSA (0.02 mg/kg) and eCFR (0.3 mg/kg), respectively. What deserves attention is that the MRL of pydiflumetofen in fish is not clear. Meanwhile, pydiflumetofen in paddy soil and wheat soil had a persistent residual effect, and the risks could not be ignored. Combined with the previous research, developing S-(+)-pydiflumetofen products will help to reduce the dosage and reduce the risks to environment and people. This study evaluated the environmental fate and risk of chiral pydiflumetofen from the perspective of farming system, and would provide data support for its rational use and risk assessment.

Stereoselective study on chiral fungicide metconazole in four kinds of fruits: Absolute configuration, SFC-MS/MS enantioseparation, degradation and risk assessment.

Metconazole is a novel chiral fungicide with two chiral carbon atoms, but the research on its stereoselective behavior is limited. Therefore, the stereoselective behaviors of metconazole in four fruits, including grape, peach, pear and jujube, were summarized in this study. After determining the absolute configuration of metconazole stereoisomers, a chiral separation method through supercritical fluid chromatography/tandem triple quadrupole mass spectrometry was first developed, which combined an improved QuEChERS method obtained the recoveries of 71.6-113 % with RSD ≤ 19.8 %. The LOD and LOQ were 4.30-95.9 and 10.5-143.2 ng/kg, respectively. Different stereoselective and diastereoselective behaviors were observed in four fruits. Dietary risk assessments of rac-metconazole were performed in populations with different ages and genders. Both acute (RQa, 0.0124-0.140 %) and chronic (HQ, 0.0234-0.0794 %) intake risks were acceptable. The results of this study would contribute to more complete risk assessments of metconazole and provide data for chiral studies.

Stereoselective, Diastereoselective Dissipation and Risk Assessment of Chiral Metconazole in Soybean, Peanut, Cabbage, Celery, Tomato, and Soil.

The stereoselective behaviors and dietary risks of metconazole (MZE) in soil and five vegetables were investigated. The results showed that there was species-specific stereoselective and diastereoselective dissipation, and the half-lives ranged from 0.69 to 8.17 days. cis-(+)-1S,5R-MZE was preferentially dissipated in soybean pods, cabbages, celeries, and tomatoes, which was contrary to soybean plants and soil. trans-(+)-1R,5R-MZE was preferentially dissipated in peanut plants, peanut shells, celeries, and tomatoes, while trans-(-)-1S,5S-MZE was preferentially dissipated in soybean plants. cis-MZE was preferentially dissipated in the test vegetables and soil, except celery. The stereoisomeric excess changes were higher than 10%, indicating that the stereoselectivity and diastereoselectivity should be considered in the risk assessment of MZE in soybean plants, pods, and peanut plants. The acute and chronic dietary intake risks of rac-MZE for different groups of people were acceptable. The preferentially dissipated and high activity cis-(+)-1S,5R-MZE with lower toxicity might be suitable for application as monocase.

Evaluation of Chiral Fungicide Penflufen in Legume Vegetables: Enantioseparation and Its Mechanism, Enantioselective Behaviors, and Risk Assessment.

Illustrating the enantioselective behaviors of the novel chiral fungicide penflufen was extremely important for ecological safety and human health. For penflufen enantiomers, an excellent separation method including a short analysis time (4 min), a high sensitivity (2 ng/g), and lesser consumption of an organic solvent was first established through supercritical fluid chromatography-tandem mass spectrometry. The enantioseparation mechanism was explained by computational chemistry, and the stronger binding ability of S-(+)-penflufen with cellulose tris-(3-chloro-4-methylphenylcarbamate) (the chiral stationary phase OZ-3 column) contributed to the posterior elution. In legume vegetables, penflufen dissipation was the fastest in Pisum sativum Linn plants (half-life, 1 day) and the slowest in Glycine max plants (half-lives, 11.3-12.9 days). After 30, 50, and 40 days, the rac-penflufen residues were lower than the maximum residue level value in the Electronic Code of Federal Regulations (10 ng/g) in G. max, P. sativum Linn, and Vigna unguiculata, respectively. Abundant S-(+)-penflufen was found in these plants with stereoisomeric excess (se) changes being >10% in the initial stage, so the risk assessment might be driven by S-(+)-penflufen. However, the se changes were <10% in V. unguiculata plants, and the risk assessment might be calculated based on rac-penflufen. Moreover, penflufen enantiomers could be transferred from legume vegetables to soils, and the concentrations increased with time. The high persistence and medium mobility of penflufen in soils might lead to potential groundwater contamination, which was noteworthy. These results could contribute to a more accurate risk assessment of penflufen in legume vegetables.

Evaluation of chiral triticonazole in three kinds of fruits: enantioseparation, degradation, and dietary risk assessment.

The enantioselective behaviors of chiral pesticides would affect the accuracy of risk assessment. This study evaluated the enantioselectivity of chiral triticonazole (a widely used fungicide) in three kinds of fruits. Firstly, the enantioseparation of triticonazole enantiomers was carried out within 1.2 min utilizing CHIRALPAK OJ-3 column with a mixture of CO2 and methanol (93:7, v/v) using SFC-MS/MS. Secondly, field trials were conducted to clarify the enantioselective degradation and residue of S-( +)-triticonazole and R-(-)-triticonazole in fruits. The initial concentrations of rac-triticonazole were 25.1-93.1 ng/g, and enantioselective degradation was observed in pear, peach, and jujube after 2 h, 10 days, and 3 days, respectively. The degradation of S-( +)-triticonazole was fastest in pear (T1/2, 2.01 days), while the T1/2 of R-(-)-triticonazole was 5.02 days. The residue concentrations of rac-triticonazole were less than the MRL set by EU (10 ng/g) on the 3rd and 21st day in pear and peach, respectively, which were lower than 10 ng/g in jujube on the 30th day (no MRL). Finally, we found that the dietary intake risks of rac-triticonazole in fruits were low for 2-7 age, 20-50 age/female, and 20-50 age/male. The current study could provide complimentary references for the rational usage, MRL formulation, and risk assessment of chiral triticonazole.

Enantioselective Dissipation, Residue, and Risk Assessment of Diniconazole Enantiomers in Four Kinds of Fruits.

Chiral diniconazole is a widely used triazole fungicide, while its enantioselective behaviors in fruits have not been reported. In this article, the absolute configuration was confirmed. A fast supercritical fluid chromatography-tandem mass spectrometry (SFC-MS/MS) method was developed for the chiral separation and enantioselective study of diniconazole in four kinds of fruits. The residual concentrations gradually decreased with time in four kinds of fruits after applying diniconazole. The dissipation half-lives of R-diniconazole and S-diniconazole were in the range of 5.3-7.9 and 2.5-7.1 days respectively, and S-diniconazole was degraded preferentially. The residue concentrations were lower than the EU's MRL (0.01 mg/kg) on the 40th (harvest time), 30th, and 10th day in pear, jujube, and apple, respectively. But, in peach, residue concentrations were still higher than the MRL after 60 days and the ratio of R/S was 2.2. These results could be helpful for the reasonable use and risk assessment of chiral diniconazole.

Dissipation, accumulation and risk assessment of fungicides after repeated spraying on greenhouse strawberry.

Repeated spraying of fungicides is a common phenomenon in greenhouse strawberry cultivation, and the continuous harvest of strawberries makes them prone to contamination by accumulated fungicides. Despite this, very few residue safety assessments of fungicides on greenhouse strawberries are conducted after repeated spraying of fungicides, and no research exists on fungicide dissipation and accumulation mechanism. Therefore, the present study investigated the dissipation and accumulation of four fungicides (pyraclostrobin, pyrimethanil, procymidone, and cyprodinil) after two typical repeated sprayings (a single fungicide repeated spraying and two fungicides sprayed using an alternate repeated technique). The half-life of pyraclostrobin after three single repeated sprayings was 18 d; however, its average half-life decreased to 9 d after alternate repeated spraying with cyprodinil. The shortened half-life may be attributed to cyprodinil water solution washing during alternate repeated spraying. The other three fungicides showed similar half-lives after single and alternate repeated spraying, following the order of cyprodinil (12 d and 10 d) > procymidone (11 d and 10 d) > pyrimethanil (6 d and 7 d). The octanol-water partition coefficient was a more efficient indicator of the half-life order of the fungicides than vapor pressure and water solubility. Pyraclostrobin showed the highest deposition efficiency but negligible residue accumulation; further, the residue accumulation of the four fungicides followed the order of procymidone > cyprodinil > pyrimethanil > pyraclostrobin after both single and alternate repeated spraying. A safety assessment demonstrated that the maximum number of times cyprodinil could be sprayed after single spraying was one; however, this number doubled after alternate spraying. The risk of exceeding the maximum residue limits of the fungicides on greenhouse strawberries decreased; however, the combined dietary risks of fungicides after alternate spraying might be high. Alternate repeated spraying of procymidone and pyrimethanil may be the optimal repeated spraying combination for greenhouse strawberries.

Assessment of Carbendazim Residues and Safety in Celery Under Different Cultivation Conditions.

Although the carbendazim is widely used to manage spot blight in celery cultivation, information on residues identified is of interest. In this study, we examined the dissipation and residual amounts of carbendazim in celery and soil under different cultivation methods when using the suggested dose and ten times of that and the bioconcentration factor of carbendazim for celery. The results showed that when celery leaves were sprayed with the suggested dose, the half-lives in a celery field and greenhouse were 2.75 days and 3.29 days, respectively. When the soil matrix was sprayed with the recommended dose before cultivation, the half-lives of carbendazim residues were 16.86 days and 11.97 days. We also conducted a long-term dietary risk assessment using the corresponding criteria. The results showed that, in China, the use of carbendazim at a dose of 0.022 g/m2 is safer and more reasonable when the harvest interval is 28 days.

Combined toxicity assessment of myclobutanil and thiamethoxam to zebrafish embryos employing multi-endpoints.

It is necessary to understand the interactions between different pesticides in ecotoxicology because pesticides never appear as individual compounds but rather in combinations with other compounds. In this study, we planned to explicate the combined toxic effect of myclobutanil (MYC) and thiamethoxam (THI) on the zebrafish (Danio rerio) by adopting multiple biomarkers. Results unraveled that the 96-h LC50 values of MYC to D. rerio at various life phases ranged from 5.2 to 10.3 mg L-1, which were lower than those of THI ranging from 147 to 246 mg L-1. Combinations of MYC and THI exhibited synergetic toxicity to zebrafish embryos. The activities of antioxidative enzymes (T-SOD, Cu/Zn-SOD and POD) and detoxification enzyme (GST) were obviously varied in most of the MYC, THI and combined exposures compared to the control. The mRNA expressions of eight genes (Cu-sod, cas3, il-8, cxcl, erα, crh, cyp17 and dio1) involved in antioxidation, apoptosis, immunity and endocrine were obviously altered in the combined exposure of MYC and THI compared to their individual exposures. Our findings hinted the threats when YMC and THI co-existed, which would be beneficial for the risk assessments of pesticide mixtures.

Evaluation of cyromazine transferred from feed to chicken products and subsequent assessment of dietary risks to Chinese consumers.

In this work, levels of cyromazine and its metabolite melamine in chicken products and its transfer factors (TFs) and dietary risks assessment for Chinese consumers are presented. Cyromazine was added to chicken feed at doses of 5, 25, and 50 mg/kg for 42 days. Cyromazine residues were found in frequently consumed chicken products (meat, heart, liver, gizzard, and eggs), with liver and eggs displaying the highest concentrations (0.935 and 1.281 mg/kg, respectively). Low levels of melamine residues (<0.029 mg/kg) were detected in chicken products from chickens treated with 25 and 50 mg/kg cyromazine, but not in eggs from chickens treated with 5 mg/kg cyromazine. The TFs for the investigated chicken products varied from 0.0074 to 0.0229 across the dosage levels. The chronic exposure assessment showed that 0.001 to 0.190% of the acceptable daily intake (0.06 mg/kg body weight [b.w.]/day) of cyromazine was consumed through chicken products among the various age and gender groups of Chinese consumers. The acute exposure assessments of different age and gender groups were 0.0004% to 0.178% of the acute reference dose (0.1 mg/kg b.w.). Although the results suggest that the risk associated with cyromazine residues in chicken products was low in China, the possible risk associated with cyromazine residues in chicken products should not be ignored while cyromazine remains a legal feed additive. PRACTICAL APPLICATION: The present analytical method could be used for cyromazine and melamine detection in different chicken products, and dietary risk assessments of cyromazine provided a support for the work of regulatory bodies to conduct surveillance programs regarding food safety evaluation of cyromazine.

Residue behavior and risk assessment of imidacloprid applied on greenhouse-cultivated strawberries under different application conditions.

A risk assessment for imidacloprid applied on strawberries under different conditions was performed after residue determination using the quick, cheap, effective, rugged, and safe (QuEChERS) method. The application conditions were varied according to the applied dosage, addition of a plant oil or organosilicon surfactant, water volume, and sprayer type. The degradation dynamics of imidacloprid on strawberries followed first-order kinetics. At applied doses of 30-60 g a.i. ha-1, the half-lives of imidacloprid were 2.89-3.46, 1.98-3.65, and 2.57-2.77 days after application without a surfactant or with a plant oil or organosilicon surfactant, respectively. For water volumes of 112.5, 225, 450, 675, and 900 L ha-1, the half-lives of imidacloprid applied in the presence of the plant oil surfactant were 3.30, 7.70, 5.33, 7.70, and 6.30 days, respectively. The half-lives after application with a knapsack mist duster, electric sprayer, and manual sprayer were 2.16, 5.77, and 7.70 days, respectively. The health risk assessment revealed risk quotients less than 1 in all cases, indicating that the application of imidacloprid poses a low health risk to humans after a pre-harvest interval of 10 days under our application conditions. The risk assessment results can provide reference data for setting a reasonable maximum residue limit for imidacloprid on strawberries in China.

Conformational Sampling and Binding Site Assessment of Suppression of Tumorigenicity 2 Ectodomain.

Suppression of Tumorigenicity 2 (ST2), a member of the interleukin-1 receptor (IL-1R) family, activates type 2 immune responses to pathogens and tissue damage via binding to IL-33. Dysregulated responses contribute to asthma, graft-versus-host and autoinflammatory diseases and disorders. To study ST2 structure for inhibitor development, we performed the principal component (PC) analysis on the crystal structures of IL1-1R1, IL1-1R2, ST2 and the refined ST2 ectodomain (ST2ECD) models, constructed from previously reported small-angle X-ray scattering data. The analysis facilitates mapping of the ST2ECD conformations to PC subspace for characterizing structural changes. Extensive coverage of ST2ECD conformations was then obtained using the accelerated molecular dynamics simulations started with the IL-33 bound ST2ECD structure as instructed by their projected locations on the PC subspace. Cluster analysis of all conformations further determined representative conformations of ST2ECD ensemble in solution. Alignment of the representative conformations with the ST2/IL-33 structure showed that the D3 domain of ST2ECD (containing D1-D3 domains) in most conformations exhibits no clashes with IL-33 in the crystal structure. Our experimental binding data informed that the D1-D2 domain of ST2ECD contributes predominantly to the interaction between ST2ECD and IL-33 underscoring the importance of the D1-D2 domain in binding. Computational binding site assessment revealed one third of the total detected binding sites in the representative conformations may be suitable for binding to potent small molecules. Locations of these sites include the D1-D2 domain ST2ECD and modulation sites conformed to ST2ECD conformations. Our study provides structural models and analyses of ST2ECD that could be useful for inhibitor discovery.

Degradation of three fungicides following application on strawberry and a risk assessment of their toxicity under greenhouse conditions.

The health risk to humans of pesticide application on minor crops, such as strawberry, requires quantification. Here, the dissipation and residual levels of three fungicides (pyraclostrobin, myclobutanil, and difenoconazole) were studied for strawberry under greenhouse conditions using high-performance liquid chromatography (HPLC)-tandem mass spectrometry after Quick, Easy, Cheap, Effective, Rugged, and Safe extraction. This method was validated using blank samples, with all mean recoveries of these three fungicides exceeding 80%. The residues of all three fungicides dissipated following first-order kinetics. The half-lives of pyraclostrobin, myclobutanil, and difenoconazole were 1.69, 3.30, and 3.65 days following one time application and 1.73, 5.78, and 6.30 days following two times applications, respectively. Fungicide residue was determined by comparing the estimated daily intake of the three fungicides against the acceptable daily intake. The results indicate that the potential health risk of the three fungicides was not significant in strawberry when following good agricultural practices (GAP) under greenhouse conditions.