Electroantennogram techniques (EAGs) were employed to record olfactory responses in the antennae of the adult female stable fly, Stomoxys calcitrans (Diptera: Muscidae), to Zanthoxylum piperitum pericarp steam distillate (ZP-SD), Z. armatum seed oil (ZA-SO) and their 29 volatile constituents alone as well as binary mixture of the ZP-SD and ZA-SO with 1-octen-3-ol. The electrophysiological responses of the test materials were compared with those of DEET and 1-octen-3-ol. At concentration of 10-1 (v/v) in mineral oil, ZP-SD, ZA-SO, and all volatiles elicited EAG responses in the fly antennae except for DEET, which is extremely low volatile. ZP-SD, ZA-SO, and some of the test volatiles elicited EAG responses rather lower than 1-octen-3-ol, which showed vapor phase repellency and toxicity to the fly in our previous behavioral research. This suggests that the stable fly possesses olfactory receptor neurons responding to given repellent compounds per se. In binary mixture with 1-octen-3-ol at concentration of 10-1, ZP-SD and ZA-SO elicited 16±0.55 and 18±0.63 mV while ZP-SD, ZA-SO, and 1-octen-3-ol alone elicited responses of 14±0.45, 15.8±0.37 and 16.2±0.58 mV, respectively. Among the volatile compounds, terpinen- 4-ol, β-myrcene, α-phellanderene, citronellal, and limonene oxide elicited 80 – 96% relative EAG amplitude compared to 1-octen-3-ol as a reference (100%). Based on structure-activity relationships, constituents having aldehyde moiety elicited bigger EAG responses than those of alcohol or ether one. The perception in the fly antenna to plant volatile compounds exhibited complicated patterns of olfactory responses. Current and future directions of this study on sensory processing mechanisms underlying repellent behaviors will be discussed.

The vapor phase repellency and toxicity of Zanthoxylum piperitum pericarp steam distillate (ZP-SD), Z. armatum seed oil (ZA-SO), and their 29 constituents to the adult female stable fly, Stomoxys calcitrans (Diptera: Muscidae), were examined using filter paper fumigation bioassay. Results were compared with those of the currently used repellent: DEET. Both of ZP-SD and ZA-SO exhibited vapor phase repellency and toxicity to female flies at 5 to 40 mg/filter paper (0.23 to 1.82 mg/cm3 air) during a 120-min exposure, whereas DEET exhibited neither repellency nor toxicity to the stable fly. At 5 mg/filter paper, vapor phase of ZP-SD and ZA-SO repelled 50 to 67% and 61 to 51% flies, respectively, to control area during 30 to 120 min. At 40 mg/filter paper, vapor phase of ZP-SD and ZA-SO caused 100% and 81% mortality, respectively, after 120 min of exposure. Among the tested volatile constituents, cuminaldehyde was the strongest repellent and toxic compound to the fly and was four times more toxic than ZP-SD and ZA-SO. Cuminaldehyde vapor phase repelled 62% flies to control area after 30 min at 2.5 mg/filter paper and caused 100% mortality after 120 min at 10 mg/filter paper. Based on the structure-activity relationships, the toxicity and repellency of constituents having aldehyde moiety were better than those of alcohol or ether one. Overall, ZP-SD, ZA-SO, and their bioactive constituents could be useful as potential vapor repellents to control stable fly population.

The repellency to female Stomoxys calcitrans (L.) of 21 essential oils alone or in combination with Calophyllum inophyllum nut oil (tamanu oil) was examined using a skin bioassay. Results were compared with those following treatment with the commonly used repellent DEET (N,N-diethyl-3-methylbenzamide). As judged by the protection time (PT) to first bite at 0.5 mg cm-2, patchouli (3.67 h) was the most effective essential oil, followed by clove bud, lovage root, clove leaf and thyme white essential oils (3.50-2.12 h). Thyme red, oregano and geranium essential oils exhibited moderate protection time (PT, 1.24-1.11 h). At 0.25 mg cm-2, effective protection time of clove bud, clove leaf and lovage root essential oils was about 1 h. The protection times of DEET were 4.47 and 2.17h at 0.5 and 0.25 mg cm-2 respectively. The remarkable increase in the protection time were produced by binary mixtures of five essential oils (clove bud, clove leaf, thyme white, patchouli and savory) and tamanu oil (0.25:2.0 mg cm-2) compared with those of either the constituted essential oil, tamanu oil or DEET alone, indicating the involvement in synergy. These essential oils, tamanu oil and binary mixtures did not cause any adverse effects on the human volunteers at 0.5 mg cm-2 except savory oil. Thus binary mixtures of essential oils and tamanu oil described merit further study as potential insect repellents for protection from humans and domestic animals from biting and nuisance caused by S. calcitrans.