Exposure to organophosphate insecticides induces behavioral changes and acetylcholinesterase inhibition in Apis mellifera

European honey bee (Apis mellifera L.) is an essential pollinator that contributes significantly to the global ecosystem and agricultural productivity. However, their population has been facing unprecedented threats, primarily due to their exposure to various pesticides, including organophosphates. These pesticides are being widely used in agriculture to control insect pests due to their efficacy, but their non-selective nature raises concerns about their impact on honey bees. Insecticides viz., chlorpyriphos 20 Emulsifiable Concentrate, dimethoate 30 Emulsifiable Concentrate, and profenophos 50 Emulsifiable Concentrate at a range of 0.005–0.09 per cent concentration were evaluated through two modes of application viz., topical and oral. Acetylcholinesterase (AChE) activity was measured at various intervals (1 and 24 hours) to assess enzyme inhibition. Behavioral observations and statistical analyses, including factor analysis with Eigen values, were employed to evaluate the impact of exposure on bee behavior and physiological responses.The results revealed two factors with Eigen value > 0.95 in both topical as well as oral method which accounted for 88.28 and 88.80 per cent of the variation in behaviour, respectively. Insecticides applied to honey bee A. mellifera in both topical as well as oral methods resulted in significant inhibition of the Acetyl choline esterase enzyme (AChE) activity. Studies revealed higher AChE inhibition (%) in oral method as compared to topical method. AChE inhibition percentage increased from 25.15 (1 Hour after treatment) to 58.25 (24 Hours after treatment) with lower concentration (0.005) of chlorpyriphos in topical method while as it reached from 27.66 to 60.94 with same concentration and same time in oral method of application. AChE inhibition percentage increased from 35.81 (1 Hour after treatment) to 78.30 (24 Hours after treatment) with higher concentration (0.06) of chlorpyriphos in topical method while as it reached from 40.35 to 80.18 with same concentration and same time in oral method. Similar trend was observed in dimethoate, and profenophos where AChE inhibition increased from 17.30, 27.15 (1 Hour after treatment) to 57.18, 61.81 (24 Hours after treatment), respectively in topical method and 20.67, 28.80 (1 Hour after treatment) to 59.85 64.04 (24 Hours after treatment), respectively in oral method. Similarly, with higher concentrations of dimethoate (0.07), and profenophos (0.09), per cent inhibition increased from 34.54, 38.60 (1 Hour after treatment) to 75.68, 79.62 (24 Hours after treatment), in topical method and 37.25, 41.23 (1 Hours after treatment) to 77.86, 82.73 (24 Hours after treatment), in oral method, respectively. Thorough risk assessments are vital for evaluating the effects of agrochemicals on Apis mellifera. The findings highlight the necessity for updated pesticide regulations and broadened conservation strategies that take into account the diverse ways pollinators are exposed to agrochemicals in the environment. The study assessed the impact of different insecticides on Apis mellifera by comparing topical and oral exposure methods. The study also aimed to analyze the behavioral effects of insecticide exposure on Apis mellifera, assessing variations in response and enzyme inhibition.The research focused on evaluating Acetylcholinesterase (AChE) inhibition to better understand the risks posed to honeybees in agricultural environments.