Insecticide exposure can affect honey bees in agro-ecosystems, posing behavioral stresses that can lead to population decline. In this study, insecticide incidence, DNA damage, and antioxidant enzyme activity were studied in Apis florea and A. dorsata honey bee samples collected from insecticide-treated and insecticide-free areas of Punjab, Pakistan. Seven insecticides: chlorpyrifos, dimethoate, imidacloprid, phorate, emamectin, chlorfenapyr, and acetamiprid were detected in seven samples of A. florea and five samples of A. dorsata. In total, 12 samples (22.2%) of honey bees were found positive to insecticide presence out of 54 samples. The most frequently detected insecticide was chlorpyrifos, which was found in four samples (7.4%), with a concentration ranging from 0.01 to 0.05 μg/g and an average concentration 0.03 μg/g. The comet assay or single cell gel electrophoresis assay, a simple way to measure DNA strand breaks in eukaryotic cells, was used to microscopically find damage of DNA at the level of a single cell. Comet tail lengths of DNA in A. florea and A. dorsata samples from insecticidetreated areas were significantly higher (P < 0.05) than samples from insecticide-free areas. The highest comet tail length (19.28 ± 2.67 μm) was observed in DNA of A. dorsata from insecticidetreated areas, while the minimum one (3.18 ± 1.46 μm) was noted in A. dorsata from insecticidefree areas. Catalase (CAT) activity did not vary significantly between honey bee samples from insecticide-treated and insecticide-free areas, while glutathione S transferase (GST) activity showed a significant reduction in response to insecticide exposure. Significant positive correlations were detected between enzyme activity and insecticide concentration in honey bee species from insecticide-treated areas compared with control groups. Toxicity from pesticide exposure at sub-lethal levels after application or from exposure to pesticide residues should not be underestimated in honey bees, as it may induce physiological impairment that can decline honey bees'' health. Insecticides, essential for crop protection measures, leave behind several toxic residues that can result in a series of human health disorders. Therefore, the determination of residues and adverse effects of insecticides in blood samples of sprayers, pesticide-industry workers and controls by using blood parameters of these individuals as biomarkers, was also carried out. Optimized analytical methods using GC-MS and HPLC for the simultaneous detection of 22 currently used insecticides were adopted. Eight of twenty-seven (22.22%) sprayers’ blood samples were found positive for five different insecticides. Eleven of twentyseven (40.74%) pesticide industry workers were found positive for eight different insecticides. The blood samples of both the exposed groups, sprayers and industry workers had significantly (P < 0.001; Mann-Whitney U-tests) low hemoglobin-Hb concentrations (12.17 ± 2.13 and 12.22 ± 2.37 g/dl respectively) than the average value of the control group with 14.23 ± 2.37 g/dl. The erythrocyte sedimentation rates (ESRs) in sprayers and insecticide industry workers (28.78 ± 20.72 and 28.17 ± 25.14 mm/1st h respectively) were greater significantly (P <0.001; MannWhitney U test) than the control blood samples (9.53 ± 3.34 mm/1st h). Comet tail length in comet assay of the two exposed groups, i.e., pesticide operators and industry workers (16.33 ± 3.78 and 16.88 ± 4.57 µm) differed significantly (P < 0.01) from comet tail length of the control group (6.53 ± 2.75 µm).Values of serum cholinesterase (SChE) concentration were slightly lower (P > 0.05) in exposed individuals, whereas values of alanine aminotransferase (ALT), aspartate aminotransferase (AST), and alkaline phosphatase (ALP) concentration were significantly higher (P < 0.01) in exposed individuals compared with control group. Exposure duration and total insecticide concentration in blood samples were positively associated with comet length, ALT activity, AST activity, and ALP activity, but negatively with SChE. DNA damage was significantly higher in smokers. Also, a positive association was found between comet length and number of cigarettes per day. These results indicate that the exposed individuals have experienced significant genotoxic and hemotoxic effects during insecticide exposure. The study also predicts the risk to exposed individuals in developing countries like Pakistan and demands realization of safety measures to prevent such dangerous effects of pesticide exposures. Out of total of 143 samples analyzed (59 fruits & vegetables, 36 pollen, 36 nectar and 27 water samples), 50 samples (34.96%) were found positive for one or more insecticides. Fruits and vegetables 24(40.67%), pollen 8(22.22%) and nectar 6(16.66%) and water 18(66.67%) samples were found positive. Total 13 insecticides were detected in 27 watersamples of three zones of Punjab (Pakistan) ranging from 0.02 to 0.8 μg/L. Different insecticides including carbosulfan, profenofos, cypermethrin, endosulfan sulfate and chlorpyriphos-methyl were frequently detected in the fruit and vegetable samples. The results suggest that consumers of Punjab province are exposed to the lower concentrations of insecticides that can cause longlasting disorders.