Metabolic Detoxification Of Insecticides Research Articles

Knockdown of CYP301B1 and CYP6AX1v2 increases the susceptibility of the brown planthopper to beta-asarone, a potential plant-derived insecticide

The cytochrome P450 monooxygenases of insects play crucial roles in the metabolic detoxification of insecticides. Our previous finding showed that two cytochrome P450 genes, both CYP301B1 and CYP6AX1v2, in the BPH underwent overexpression due to β-asarone. In this study, we investigated the molecular characteristics, expression patterns and functions of these two cytochrome P450 genes. The results showed that CYP301B1 had the highest expression level in the eggs, while CYP6AX1v2 was expressed in macropterous female adults. Moreover, the expression level of CYP301B1 in the head was higher than that in the integument, fat body and gut. The expression level of CYP6AX1v2 in the fat body and gut was higher than that in head and integument. Importantly, silencing CYP301B1 and CYP6AX1v2 separately could increase the sensitivity, resulting in significant higher mortality of BPH following treatment with β-asarone. Our findings indicated that CYP301B1 and CYP6AX1v2 could contribute to the resistance of BPH to β-asarone, and these two genes may be involved in the detoxification metabolism of β-asarone in BPH.

Energy Reserve Compensating for Trade-Off Between Metabolic Resistance and Life History Traits in the Brown Planthopper (Hemiptera: Delphacidae).

Development of insecticide resistance often changes life history traits of insect pests, because metabolic detoxification of insecticides in insect bodies requires huge energetic reserves. The brown planthopper, Nilaparvata lugens (Stål), an important insect pest of rice crop in East and Southeast Asia, has developed strong resistance to imidacloprid from mid-2000s. The aim of this study was to examine the costs of life history traits and reveal changes in energy reserves with developing imidacloprid resistance. We compared the life history traits (survival time, fecundity, developmental time, and hatchability) and total lipid content between imidacloprid-resistant and imidacloprid-susceptible (control) brown planthopper strains. As compared to the control strains, adults' survival time of the resistant females was shorter, and their fecundity was lower; the other life history traits did not differ significantly between the resistant and control strains. As the results, net reproductive rates (R0) were lower in the resistant strains than in the susceptible strains. However, the amount of stored lipids was larger in resistant females than control ones. Our findings demonstrated a physiological trade-off between the development of imidacloprid resistance and the reproductive traits of brown planthopper. The imidacloprid-resistant strains are likely to store lipids for metabolic detoxification rather than consume them for reproduction.

Deciphering pyrethroid resistance in Cx. pipiens (L): Implications of cytochrome P450; expression profiling and regulatory microRNA

Over the past decades, the extensive use of pyrethroids insecticides for vector control has resulted in the development of insecticide resistance. Cytochrome P450 has been recognized to play a critical role in the metabolic detoxification of insecticides. In the current study, Culex pipiens mosquitoes were collected from Giza Governorate in Egypt and tested for insecticide susceptibility against deltamethrin. First detection of Knockdown resistance gene (Kdr) mutations in field collected mosquitoes was performed. Activities of cytochrome oxidase P450 detoxification enzyme that synchronized with the resistance development, was assessed. Expression profiles of cytochrome P450s and their putative corresponding regulating miRNAs, which was previously reported in Cx. pipiens pallens were evaluated in pyrethroid resistant field-collected Cx. pipiens using RT-qPCR and stem-loop RT-qPCR, respectively. Specific stem-loop reverse transcription primers and forward primers were designed for miRNAs profiling. Our results elucidated the pyrethroid resistance development and revealed its relation to the metabolic and target site modification mechanisms with a first report of L1014F-kdr mutation detection. RT-qPCR results have showed an up-regulation in the expression of the studied P450 transcripts. Negative correlations were found between the expression of P450s and their regulatory miRNAs except for CYP9J35, where positive correlation was found with its corresponding miR-13. Interestingly, our data was the first to detect negative correlation between miR-285 and its putative CYP6Cp1 target gene. These findings highlighted the significance of identifying P450 gene along with regulatory miRNAs as a key mechanism implicated in pyrethroid resistance in field Culex vector population. The elucidation of this mechanism would shed light on the development of insecticide resistance and would help in shaping strategies to combat such vectors.

Identification of a novel cytochrome P450 CYP3356A1 linked with insecticide detoxification in Bradysia odoriphaga.

Cytochrome P450 monooxygenases play an important role in the metabolic detoxification of insecticides in insect pests. However, little is known about the role of a specific P450 gene and its responses to insecticide exposure in Bradysia odoriphaga, a major pest in Chinese chive production. In this study, a novel P450 gene, CYP3356A1, was cloned from Bradysia odoriphaga. The full-length cDNA sequence of CYP3356A1 is 2153 bp and its open reading frame (ORF) encodes 508 amino acids. Quantitative real time PCR(qRT-PCR) analyses in different tissues showed that CYP3356A1 expression was the highest in the Malpighian tubule. Moreover, among the different developmental stages of the insect, the highest expression of CYP3356A1 was found in fourth-instar larvae. Expression of CYP3356A1 was upregulated by treatment with imidacloprid, thiamethoxam, and β-cypermethrin at median lethal concentrations (LC50 ). RNA interference (RNAi)-mediated silencing of CYP3356A1 significantly increased mortality by 36.90%, 25.17%, and 36.73 when fourth-instar B. odoriphaga larvae were exposed to imidacloprid, thiamethoxam, and β-cypermethrin, respectively, at the LC50 dose. These results demonstrate that CYP3356A1 is related to the detoxification of imidacloprid, thiamethoxam, and β-cypermethrin in B. odoriphaga. Moreover, the study also increased our understanding of the molecular mechanisms of insecticide detoxification in this pest insect. © 2018 Society of Chemical Industry.

Identifying Current and Missing Knowledge in the Control of Pyrethroid-Resistant Triatoma infestans, Vector of Chagas Disease

Triatomines are blood-sucking bugs that occur mainly in Latin America. They are vectors of Trypanosoma cruzi, the parasite that causes Chagas disease. Chemical control of Chagas disease´s vectors by using pyrethroid insecticides has been highly successful for the elimination of domestic infestation and consequently the reduction of the vector transmission. However, at the beginning of the 2000s a decrease in the effectiveness of the chemical control of triatomines was detected in several areas from Argentina and Bolivia, particularly in the Gran Chaco eco-region.During the last 15 years, several studies demonstrated the evolution of insecticide resistance in Triatoma infestans and established the presence of different toxicological profiles, the autosomal inherence of resistance, the biological costs of deltamethrin resistance, the expression of deltamethrin resistance thorough the embryonic development, and the main mechanisms of resistance (target-site insensitivity and metabolic detoxification of insecticides).The emergence of pyrethroid resistance coupled with the usual difficulties in sustaining adequate rates of insecticide applications emphasize the need of incorporating other tools for integrated vector and disease control, such as the proposal of the organo-phosphorus insecticide fenitrothion as an alternative chemical strategy for the management of the resistance because it was effective against pyrethroid-resistant populations in laboratory and semi-field trials.New studies on the current situation of presence and spread of resistant populations of triatomines and the acceptance of the use of alternative insecticides are critical requirements in the implementation of strategies for the management of resistance and for the rational design of campaigns oriented to reducing the vector transmission of Chagas’ disease.

Induction and activity of glutathione S-transferases extracted from Zonocerus variegatus (Orthoptera: Pyrgomorphidae) exposed to insecticides

Glutathione S-transferases (GSTs) have been recognized as important metabolic detoxifying enzymes in many phytophagous insects. However, the contribution of GST to insecticide resistance in Zonocerus variegatus (L.) has not been studied. Therefore, we carried out an initial study on the induction and kinetics of GST in Z. variegatus exposed to pyrethroids (PYRs) and Ocimum gratissimum leaf extract. Fifth-instar nymphs of Z. variegatus collected from cassava farms (with no history of insecticide exposure) on the campus of the University of Ibadan were reared to adult stage. Adult insects were divided into four groups of two replicates, each consisting of 30 insects. The groups were exposed to 20 ml of PYR insecticide and 25 mg/dl and 250 mg/dl of O. gratissimum leaf extract, respectively, while the last group served as the control. GST was extracted from the body tissues of the insects using the Bradford method and the kinetics of the enzyme was evaluated using the Lineweaver-Burk plots. No mortality was recorded in insects exposed to 25 mg/dl of O. gratissimum leaf extract, while mortality rates of 93.3 and 43.3% were recorded in insects exposed to PYR insecticide and 250 mg/dl of O. gratissimum leaf extract, respectively. The activity of GST was higher in insects exposed to the insecticides than in the control insects, while a higher enzyme activity was recorded in insects that died after exposure to the insecticides than in insects that were alive after exposure. The GST extracted from insects that were alive after exposure to PYR insecticide and O. gratissimum leaf extract exhibited a high affinity for the glutathione substrate compared with that extracted from insects that died after exposure. Higher GST activity in insects exposed to insecticides is indicative of the role of this enzyme in the metabolic detoxification of insecticides by Z. variegatus.

Using Drosophila melanogaster to validate metabolism-based insecticide resistance from insect pests

Identifying molecular mechanisms of insecticide resistance is important for preserving insecticide efficacy, developing new insecticides and implementing insect control. The metabolic detoxification of insecticides is a widespread resistance mechanism. Enzymes with the potential to detoxify insecticides are commonly encoded by members of the large cytochrome P450, glutathione S-transferase and carboxylesterase gene families, all rapidly evolving in insects. Here, we demonstrate that the model insect Drosophila melanogaster is useful for functionally validating the role of metabolic enzymes in conferring metabolism-based insecticide resistance. Alleles of three well-characterized genes from different pest insects were expressed in transgenic D. melanogaster : a carboxylesterase gene (αE7) from the Australian sheep blowfly Lucilia cuprina, a glutathione S-transferase gene (GstE2) from the mosquito Anopheles gambiae and a cytochrome P450 gene (Cyp6cm1) from the whitefly Bemisia tabaci. For all genes, expression in D. melanogaster resulted in insecticide resistance phenotypes mirroring those observed in resistant populations of the pest species. Using D. melanogaster to assess the potential for novel metabolic resistance mechanisms to evolve in pest species is discussed.