Maize weevils, Sitophilus zeamais, are stored product pests mostly found in warm and humid regions around the globe. In the present study, acute toxicity via contact and residual bioassay and fumigant bioassay of 28 essential oils as well as their attraction–inhibitory activity against the adults of S. zeamais were evaluated. Chemical composition of the essential oils was analyzed by gas chromatography-mass spectrometry, and a compound elimination assay was conducted on the four most active oils (cinnamon, tea tree, ylang ylang, and marjoram oils) to identify major active constituents. Amongst the oils examined, cinnamon oil was the most active in both contact/residual and fumigant bioassays, and exhibited strong behavioral inhibitory activity. Based on the compound elimination assay and chemical analyses, trans-cinnamaldehyde in cinnamon oil, and terpinen-4-ol in tea tree and marjoram oils were identified as the major active components. Although cinnamon oil seemed promising in the lab-scale bioassay without rice grains, it failed to exhibit strong insecticidal activity when the container was filled with rice. When a cinnamon oil-based product was applied both in an empty glass jar and a rice-filled container, all weevils in the empty jar were killed, whereas fewer than 15% died in the rice-filled container.

Many plant essential oils and their terpenoid constituents possess bioactivities including insecticidal activity, and they sometimes act synergistically when mixed. Although several hypotheses for this have been proposed, the underlying mechanism has not been fully elucidated thus far. In the present study, we report that in larvae of the cabbage looper, Trichoplusia ni, most synergistic or antagonistic insecticidal activities among mixtures of plant essential oil constituents are pharmacokinetic effects, owing to changes in solubility as well as spreadability on a wax layer. Among the major constituents of rosemary (Rosmarinus officinalis) oil, in vitro analysis revealed up to a 19-fold increase in penetration of camphor in a binary mixture with 1,8-cineole through the larval integument, suggesting increased penetration as the major mechanism for synergy. A total of 138 synergistic or antagonistic interactions among 39 compounds were identified in binary mixtures via topical application, and these were highly correlated to changes in surface tension as measured by contact angle of the mixtures on a beeswax layer. Among compounds tested, trans-anethole alone showed evidence of internal synergy, whereas most of remaining synergistic or antagonistic combinations among the three most active compounds were identified as penetration-related interactions, confirmed via a divided-application bioassay.

Plant essential oils are intricate blends comprising predominantly of monoterpenes and some sesquiterpenes. These oils display diverse bioactivities against targeted organisms, often arising from complex interactions among their constituents, which may demonstrate synergistic or antagonistic effects. Despite their wide use as botanical insecticides, the mechanisms behind these interactions and their effects on bioactivity are poorly understood. This study investigated the synergistic interaction of thymol and p-cymene, two major constituents of Thymus vulgaris essential oil, on the larvae and adults of the housefly, Musca domestica. The results showed that p-cymene synergized the insecticidal activity of thymol in adult houseflies, but not in larvae. GC–MS analyses and bioassays indicated the increased cuticular penetration of thymol by p-cymene was the mechanism of synergy, which was observed only in the adults. Two potential routes were proposed: the expansion of the wetting area, or the disruption of cuticular integrity through dissolving the wax layer. The sequential application and large-volume treatment bioassay results suggested that the former was the more likely mechanism. Also, the hydrophobicity of the cuticle seemed critical for this stage-specific synergy. Wax-devoid adults failed to show synergistic toxicity, whereas artificially wax-coated larvae gained a synergistic effect. Overall, the findings provide insights into the synergistic mechanism of insecticidal activity of plant essential oils and suggest potential applications in developing effective strategies using penetration-enhancing synergists.