Identification of plant-derived inhibitors of acetylcholinesterase and glutathione S-transferase in Rhipicephalus microplus and Anopheles stephensi

Hematophagous insects, especially mosquitoes and ticks, are responsible for transmitting various pathogens in humans and animals. Due to global warming, there has been a rise in their population, and they are spreading to new geographical territories. Therefore, vector control measures seem essential for preventing the spread of insect-borne diseases. To address this, the current study investigated the insecticidal properties of twenty-four medicinal plants against Rhipicephalus microplus and Anopheles stephensi . A soxhlet extracts of these plants were prepared using three solvents: hexane, chloroform, and methanol. All the 72 extracts were screened for acaricidal and mosquitocidal activities. Nine plant extracts exhibited adulticidal activity, while seven showed larvicidal effects against R. microplus . In the case of A. stephensi , the bed net assay revealed no significant adulticidal activity from any extracts. However, five plant extracts demonstrated larvicidal activity and also inhibited larval-to-pupal molting. The hexane extracts of Annona muricata and Cinnamomum zeylanicum showed a notable inhibition of acetylcholinesterase (AChE) and glutathione S-transferase (GST) activities in R. microplus and A. stephensi . Further, GC-MS analysis, molecular docking, and molecular dynamics simulation studies identified 2-tert-butyl-4,6-bis(3,5-di-tert-butyl-4-hydroxybenzyl)phenol, as one of the potential molecules in these extracts, which can bind and form a stable complex with AChE and GST of R. microplus and A. stephensi. • Screening of plant extracts led to the identification of potent plant extracts with acaricidal and mosquitocidal activity. • Extracts of C. zeylanicum and A. muricata inhibited acetylcholinesterase (AChE) and glutathione S-transferase (GST). • Phytochemical analysis of active extracts revealed diverse bioactive compounds with potential insecticidal properties • Insilico studies suggested that 2-tert-Butyl-4,6-bis(3,5-di-tert-butyl-4-hydroxybenzyl)phenol could be a possible inhibitor.