Min Liao

Insecticidal Activity of Melaleuca alternifolia Essential Oil and RNA-Seq Analysis of Sitophilus zeamais Transcriptome in Response to Oil Fumigation.

Background The cereal weevil, Sitophilus zeamais is one of the most destructive pests of stored cereals worldwide. Frequent use of fumigants for managing stored-product insects has led to the development of resistance in insects. Essential oils from aromatic plants including the tea oil plant, Melaleuca alternifolia may provide environmentally friendly alternatives to currently used pest control agents. However, little is known about molecular events involved in stored-product insects in response to plant essential oil fumigation. Results M. alternifolia essential oil was shown to possess the fumigant toxicity against S. zeamais. The constituent, terpinen-4-ol was the most effective compound for fumigant toxicity. M. alternifolia essential oil significantly inhibited the activity of three enzymes in S. zeamais, including two detoxifying enzymes, glutathione S-transferase (GST), and carboxylesterase (CarE), as well as a nerve conduction enzyme, acetylcholinesterase (AChE). Comparative transcriptome analysis of S. zeamais through RNA-Seq identified a total of 3,562 differentially expressed genes (DEGs), of which 2,836 and 726 were up-regulated and down-regulated in response to M. alternifolia essential oil fumigation, respectively. Based on gene ontology (GO) analysis, the majority of DEGs were involved in insecticide detoxification and mitochondrial function. Furthermore, an abundance of DEGs mapped into the metabolism pathway in the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway database were associated with respiration and metabolism of xenobiotics, including cytochrome P450s, CarEs, GSTs, and ATP-binding cassette transporters (ABC transporters). Some DEGs mapped into the proteasome and phagosome pathway were found to be significantly enriched. These results led us to propose a model of insecticide action that M. alternifolia essential oil likely directly affects the hydrogen carrier to block the electron flow and interfere energy synthesis in mitochondrial respiratory chain. Conclusion This is the first study to perform a comparative transcriptome analysis of S. zeamais in response to M. alternifolia essential oil fumigation. Our results provide new insights into the insecticidal mechanism of M. alternifolia essential oil fumigation against S. zeamais and eventually contribute to the management of this important agricultural pest.

Design, Synthesis and Anti-Tobacco Mosaic Virus (TMV) Activity of 5-Chloro-N-(4-cyano-1-aryl-1H-pyrazol-5-yl)-1-aryl-3-methyl-1H-pyrazole-4-carboxamide Derivatives

A series of novel pyrazole amide derivatives 3a–3p which take TMV PC protein as the target has been designed and synthesized by the reactions of 5-chloro-1-aryl-3-methyl-1H-pyrazole-4-carboxylic acids with 5-amino-1-aryl-1H-pyrazole-4-carbonitriles. All the compounds were characterized by 1H-NMR, mass spectroscopy and elemental analysis. Preliminary bioassays indicated that all the compounds acted against the tobacco mosaic virus (TMV) with different in vivo and in vitro modes at 500 μg/mL and were found to possess promising activity. Especially, compound 3p showed the most potent biological activity against tobacco mosaic virus (TMV) compared to ningnanmycin, and a molecular docking study was performed and the binding model revealed that the pyrazole amide moiety was tightly embedded in the binding sites of TMV PC (PDB code: 2OM3).

Degradation Dynamics and Dietary Risk Assessments of Two Neonicotinoid Insecticides during Lonicera japonica Planting, Drying, and Tea Brewing Processes

The degradation dynamics and dietary risk assessments of thiamethoxam and thiacloprid during Lonicera japonica planting, drying, and tea brewing processes were systematically investigated using high-performance liquid chromatography. The half-lives of thiamethoxam and thiacloprid were 1.0-4.1 d in the honeysuckle flowers and leaves, with degradation rate constants k ranging from -0.169 to -0.696. The safety interval time was 7 d. The sun- and oven-drying (70 °C) percent digestions were 59.4-81.0% for the residues, which were higher than the shade- and oven-drying percentages at lower temperatures (30, 40, 50, and 60 °C, which ranged from 37.7% to 57.0%). The percent transfers of thiamethoxam and thiacloprid were 0-48.4% and 0-25.2%, respectively, for the different tea brewing conditions. On the basis of the results of this study, abiding by the safety interval time is important, and using reasonable drying methods and tea brewing conditions can reduce the transfer of thiamethoxam and thiacloprid to humans.

Factors Affecting Transfer of Pyrethroid Residues from Herbal Teas to Infusion and Influence of Physicochemical Properties of Pesticides

The transfer of pesticide residues from herbal teas to their infusion is a subject of particular interest. In this study, a multi-residue analytical method for the determination of pyrethroids (fenpropathrin, beta-cypermethrin, lambda-cyhalothrin, and fenvalerate) in honeysuckle, chrysanthemum, wolfberry, and licorice and their infusion samples was validated. The transfer of pyrethroid residues from tea to infusion was investigated at different water temperatures, tea/water ratios, and infusion intervals/times. The results show that low amounts (0–6.70%) of pyrethroids were transferred under the different tea brewing conditions examined, indicating that the infusion process reduced the pyrethroid content in the extracted liquid by over 90%. Similar results were obtained for the different tea varieties, and pesticides with high water solubility and low octanol–water partition coefficients (log Kow) exhibited high transfer rates. Moreover, the estimated values of the exposure risk to the pyrethroids were in the range of 0.0022–0.33, indicating that the daily intake of the four pyrethroid residues from herbal tea can be regarded as safe. The present results can support the identification of suitable tea brewing conditions for significantly reducing the pesticide residue levels in the infusion.

Transcriptome profiling reveals differential gene expression of detoxification enzymes in Sitophilus zeamais responding to terpinen-4-ol fumigation

Plant essential oils with high bioactivity provide environmental friendly alternatives for synthetic pesticides. Melaleuca alternifolia essential oil and its main constituent terpinen-4-ol have high insecticidal activity to Sitophilus zeamais Motschulsky. Terpinen-4-ol may be metabolized by human and insect cytochrome P450s. However, little is known about how insects systemically respond to terpinen-4-ol. In this study, we used an RNA-seq approach to evaluate the global gene expression of S. zeamais after terpinen-4-ol fumigation. Duplicates of fumigated and control groups, for a total of four libraries, were collected for sequencing. A total of 36,117 unigenes with an average length of 1036 bp were generated in the de novo assembled transcriptome. Comparative analysis of S. zeamais libraries identified 592 differentially expressed genes (DEGs), of which 308 and 284 genes were up- and down-regulated in response to terpinen-4-ol fumigation, respectively. GO and KEGG functional analyses were performed for up and downregulated DEGs separately, showing these DEGs were enriched for terms related to catalytic activity, carbohydrate metabolism, and xenobiotics biodegradation and metabolism. DEGs encoding enzymes for detoxification were detected, including sixteen cytochrome P450s (P450s), eight glutathione S-transferase (GSTs), fourteen esterase (ESTs), ten UDP-glucuronosyltransferase (UGTs), and two ATP-binding cassette transporter (ABC transporter) genes. Real-time quantitative PCR confirmed that ten P450s, three GSTs and one EST were up-regulated dramatically after exposure to terpinen-4-ol at different concentrations and over a time course. The results provided a transcriptional overview of the changes in a stored-grain pest in response to terpinen-4-ol fumigation. The analysis revealed the expression levels of detoxification genes were altered, especially for P450s, and provided candidate genes for understanding systemic metabolic responses to terpinen-4-ol in insects.

Study of Factors Influencing the Bioaccessibility of Triazolone in Cherry Tomatoes Using a Static SHIME Model

Estimating the influence of bioaccessibility of pesticide residues in fruits and vegetables on dietary exposure is a challenge for human health risk assessment. This study investigated the bioaccessibility of pesticide residues in cherry tomatoes and contributing factors (digestion time, pH, solid/liquid ratio, and dietary nutrition) using an in vitro test simulating the human gastrointestinal tract. pH had the largest effect on triazolone precipitation in the simulated gastric intestinal juice, which had a significant impact on the bioaccessibility. The bioaccessibility of triazolone in the intestinal stage was slightly higher than that in the stomach stage, owing to bile salts and pancreatic enzymes present in the intestinal juice. The bioaccessibility of triazolone did not change significantly with digestion time. In the gastric stage, there was a logarithmic relationship between the bioaccessibility and solid/liquid ratio (R2 = 0.9941). The addition of oil significantly changed the bioaccessibility in the gastrointestinal stage. Protein and dietary fiber only affected bioaccessibility in the stomach stage. Dietary nutrition can reduce the release of pesticides from fruits and vegetables into the stomach, sharply reducing the bioaccessibility, and the dietary exposure of pesticide residues in fruits and vegetables can be properly evaluated.

Factors Affecting the Bioaccessibility and Intestinal Transport of Difenoconazole, Hexaconazole, and Spirodiclofen in Human Caco-2 Cells Following in Vitro Digestion

This study examined how gastrointestinal conditions affect pesticide bioaccessibility and intestinal transepithelial transport of pesticides (difenoconazole, hexaconazole, and spirodiclofen) in humans. We used an in vitro model combining human gastric and intestinal digestion, followed with Caco-2 cell model for human intestinal absorption. Bioaccessibility of three tested pesticides ranged from 25.2 to 76.3% and 10.6 to 79.63% in the gastric and intestinal phases, respectively. A marked trend similar to the normal distribution was observed between bioaccessibility and pH, with highest values observed at pH 2.12 in gastric juice. No significant differences were observed with increasing digestion time; however, a significant negative correlation was observed with the solid-liquid (S/L) ratio, following a logarithmic equation. R2 ranged from 0.9198 to 0.9848 and 0.9526 to 0.9951 in the simulated gastric and intestinal juices, respectively, suggesting that the S/L ratio is also a major factor affecting bioaccessibility. Moreover, significant dose- and time-response effects were subsequently observed for intestinal membrane permeability of difenoconazole, but not for hexaconazole or spirodiclofen. This is the first study to demonstrate the uptake of pesticides by human intestinal cells, aiding quantification of the likely effects on human health and highlighting the importance of considering bioaccessibility in studies of dietary exposure to pesticide residues.

iTRAQ-based quantitative proteome revealed metabolic changes of Sitophilus zeamais in response to terpinen-4-ol fumigation: Proteomic analysis of Sitophilus zeamais exposure to terpinen-4-ol

BACKGROUND Terpinen-4-ol has high insecticidal activity towards the stored-grain pest Sitophilus zeamais, a key pest of cereals worldwide. Comparative proteomic analysis can identify proteins related to the response to terpinen-4-ol fumigation in S. zeamais, improving our understanding of the mechanisms behind the insecticidal activity and the detoxification of terpinen-4-ol in insects. RESULTS Using an isobaric tags for relative and absolute quantification (iTRAQ)-based strategy, 2761 proteins were obtained from S. zeamais adults. Comparative proteomic analysis showed that 215 proteins were upregulated and 129 were downregulated after exposure to terpinen-4-ol. Based on functional classifications, differentially expressed proteins (DEPs) were enriched in the carbohydrate, energy and xenobiotics metabolism pathways. DEPs associated with lactose, sucrose and starch metabolism were identified, including alcohol dehydrogenase, aldose reductase, β-galactosidase, maltase, and myrosinase. Among detoxification-related proteins, the levels of 14 cytochrome P450s, seven glutathione S-transferases, and three UDP-glucuronosyltransferases were altered, most of which were upregulated after exposure to terpinen-4-ol. CONCLUSION Metabolic changes indicated that terpinen-4-ol could affect the energy supply and potentially be metabolized and detoxified by various enzymes in S. zeamais. The results provide a foundation for further functional studies of key proteins mediated by terpinen-4-ol.

Dissipation and Migration of Pyrethroids in Auricularia polytricha Mont. from Cultivation to Postharvest Processing and Dietary Risk

In order to ensure raw consumption safety the dissipation behavior, migration, postharvest processing, and dietary risk assessment of five pyrethroids in mushroom (Auricularia polytricha Mont.) cultivated under Chinese greenhouse-field conditions. Half-lives (t1/2) of pyrethroids in fruiting body and substrate samples were 3.10–5.26 and 17.46–40.06 d, respectively. Fenpropathrin dissipated rapidly in fruiting bodies (t1/2 3.10 d); bifenthrin had the longest t1/2. At harvest, pyrethroid residues in A. polytricha (except fenpropathrin) were above the respective maximum residue limits (MRLs). Some migration of lambda-cyhalothrin was observed in the substrate-fruit body system. In postharvest-processing, sun-drying and soaking reduced pyrethroid residues by 25–83%. We therefore recommend that consumers soak these mushrooms in 0.5% NaHCO3 at 50 °C for 90 min. Pyrethroids exhibit a particularly low PF value of 0.08–0.13%, resulting in a negligible exposure risk upon mushroom consumption. This study provides guidance for the safe application of pyrethroids to edible fungi, and for the establishment of MRLs in mushrooms to reduce pesticide exposure in humans.

Behavior of pesticides and their metabolites in traditional Chinese medicine Paeoniae Radix Alba during processing and associated health risk

Graphical abstract TCM processing could be useful for the partial removal of several pesticide residues, with removal rate reaching 98%. A drying time of 1–9 h and temperature of 60 °C are recommended for the post‐harvest processing of P. Radix. Figure. No Caption available. HighlightsDetermination of pesticides and their metabolites in traditional Chinese medicine Paeoniae Radix Alba by QuEChERs‐UPLC–MS/MS.Residue behavior of pesticides was investigated during Chinese medicine processing.The metabolic transformation of chlorpyrifos and thiamethoxam in boiling and drying was investigated.PCA showed that polarity was the dominant variables affected the removal effectiveness of pesticides.Calculated PFs for pesticides were used to perform a risk assessment of dietary exposure. ABSTRACT In Traditional Chinese Medicine (TCM), crude herbs are processed to obtain the medicinal parts of the plant, from which extracts are prepared for people to take as medicine. However, there is no report on the pesticide residual behavior in Chinese medicines during processing at present, and thus, a systematic study of the effects of different processing factors (PF) on pesticide removal in Chinese medicines and the associated risk of dietary exposure is urgently needed. This study main investigated the dissipation and metabolism of pesticides in Chinese medicine Paeoniae Radix Alba during processing, and the calculated pesticide PFs were also used to assess the risk of dietary exposure. For analyze samples, a simple and high‐throughput multiresidue pesticide analysis method was developed and validated for pesticides and their metabolites in P. Radix. based on QuEChERS procedure combined with HPLC‐MS/MS. Recoveries at three concentration levels were within 61.37%–117.82% with an associated precision RSD < 15% indicating satisfactory accuracy. TCM processing could be useful for the partial removal of several pesticide residues, with removal rate reaching 98%. The polarity is the dominant variable, which with a high contribution was the effectiveness of the treatment and the concentration factors for pesticides. The hazard quotients of all pesticides were much lower than the safety level, indicating low risk of dietary exposure. Results are of great theoretical and practical value for the scientific evaluation of the safety of Chinese medicines, improvement of the quality and safety level of Chinese medicine.