CYP4CS5-mediated thiamethoxam and clothianidin resistance is accompanied by fitness cost in the whitefly Bemisia tabaci.

BACKGROUND: Understanding the trade-offs between insecticide resistance and the associated fitness is of particular importance to sustainable pest control. One of the most devastating pest worldwide, the whitefly Bemisia tabaci, has developed resistance to various insecticides, especially the neonicotinoid group. Although neonicotinoid resistance often is conferred by P450s-mediated metabolic resistance, the relationship between such resistance and the associated fitness phenotype remains largely elusive. By gene cloning, quantitative reverse transcription (qRT)-PCR, RNA interference (RNAi), transgenic Drosophila melanogaster, metabolism capacity in vitro and 'two sex-age stage' life table study, this study aims to explore the molecular role of a P450 gene CYP4CS5 in neonicotinoid resistance and to investigate whether such resistance mechanism carries fitness costs in the whitefly. RESULTS: Our bioassay tests showed that a total of 13 field-collected populations of B. tabaci MED biotype displayed low-to-moderate resistance to thiamethoxam and clothianidin. Compared to the laboratory susceptible strain, we then found that an important P450 CYP4CS5 was remarkably upregulated in the field resistant populations. Such overexpression of CYP4CS5 had a good match with the resistance level among the whitefly samples. Further exposure to the two neonicotinoids resulted in an increase in CYP4CS5 expression. These results implicate that overexpression of CYP4CS5 is closely correlated with thiamethoxam and clothianidin resistance. RNAi knockdown of CYP4CS5 increased mortality of the resistant and susceptible populations after treatment with thiamethoxam and clothianidin in bioassay, but obtained an opposite result when using a transgenic line of D. melanogaster expressing CYP4CS5. Metabolic assays in vitro revealed that CYP4CS5 exhibited certain capacity of metabolizing thiamethoxam and clothianidin. These in vivo and in vitro assays indicate an essential role of CYP4CS5 in conferring thiamethoxam and clothianidin resistance in whitefly. Additionally, our life-table analysis demonstrate that the field resistant whitefly exhibited a prolonged development time, shortened longevity and reduced fecundity compared to the susceptible, suggesting an existing fitness cost as a result of the resistance. CONCLUSION: Collectively, in addition to the important role of CYP4CS5 in conferring thiamethoxam and clothianidin resistance, this resistance mechanism is associated with fitness costs in the whitefly. These findings not only contribute to the development of neonicotinoids resistance management strategies, but also provide a new target for sustainable whitefly control. © 2023 Society of Chemical Industry.

Single and joint toxicity assessment of acetamiprid and thiamethoxam neonicotinoids pesticides on biochemical indices and antioxidant enzyme activities of a freshwater fish Catla catla.

Neonicotinoids pesticides are extensively used in many countries due to their high insect selectivity. Acetamiprid and thiamethoxam are the neonicotinoids most commonly detected in the aquatic environment. This work examined the single and joint toxicity of acetamiprid and thiamethoxam in a freshwater fish Catla catla. Fish were exposed to acetamiprid (0.5 mg/L and 1 mg/L), thiamethoxam (0.01 mg/L and 0.5 mg/L) and their binary mixtures (0.5 mg/L of acetamiprid and 0.01 mg/L of thiamethoxam) for 96 h. The stress biomarkers such as glucose, protein, electrolytes, Na+/K+ -ATPase and oxidative stress were evaluated. Among the biochemical parameters, plasma protein, electrolytes (sodium, potassium and chloride) and gill ATPase activity were decreased in response to individual and binary mixtures treatments. In contrast, blood glucose level showed significant increase in all the treatments. Exposure to various concentrations of acetamiprid and thiamethoxam resulted in significant decrease in superoxide dismutase (SOD) activity in the gill tissue. However, SOD activity was significantly elevated during binary mixtures treatment. Glutathione peroxidase (GPx), catalase (CAT), glutathione-S-transferase (GST) and reduced glutathione (GSH) levels in gills were decreased significantly after individual and binary mixtures treatments. Fish exposed at individual and binary mixtures significantly elevated the level of LPO in gill tissue. Our findings suggest that multi-biomarker approach can be effectively used to assess the effects of joint toxicity of pesticides and to monitor the neonicotinoids pesticides in the aquatic environment.

Monitoring residue levels and dietary risk assessment of thiamethoxam and its metabolite clothianidin for Chinese consumption of Chinese kale.

BACKGROUND: Thiamethoxam is widely used to control pests in Chinese kale, popularly consumed leafy vegetables. The potential risk to the environment and human health has aroused much public concern. Therefore, it is important to investigate the degradation behavior, residue distribution and dietary risk assessment of thiamethoxam in Chinese kale. RESULTS: A sensitive analytical method for determination of thiamethoxam and its metabolite clothianidin residue in Chinese kale was established and validated through a quick, easy, cheap, effective, rugged, and safe (QuEChERS) technique with ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS). The recoveries were 85.4-101.2% for thiamethoxam and 79.5-108.1% for clothianidin, with the relative standard deviations (RSDs) of 0.9-10.2% and 1.8-6.0%, respectively. For the dissipation kinetics, the data showed that thiamethoxam in Chinese kale was degraded with the half-lives of 4.1 to 4.5 days. In the terminal residue experiments, the residues of thiamethoxam were 0.017-0.357 mg kg-1 after application 2-3 times with a preharvest interval (PHI) of 7 days under the designed dosages. The chronic and acute dietary exposure assessment risk quotient (RQ) values of thiamethoxam in Chinese kale for different Chinese consumers were 0.08-0.19% and 0.05-0.12%, respectively, and those of clothianidin were 0.01-0.04% and 0.02-0.04%, respectively, all of the RQ values were lower than 100%. CONCLUSION: Thiamethoxam in Chinese kale was rapidly degraded following first-order kinetics models. The dietary risk of thiamethoxam and clothianidin through Chinese kale was negligible to consumers. The results from this study are important reference for Chinese governments to developing criteria for the safe and rational use of thiamethoxam, setting maximum residue levels (MRLs), monitoring the quality safety of agricultural products and protecting consumer health. © 2021 Society of Chemical Industry.

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.