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Synergistic Adulticidal Activity of Lemongrass (Cymbopogon citratus), Star Anise (Illicium verum), Nutmeg (Myristica fragrans) Essential Oil and Their Components Against the Housefly (Musca domestica) and Their Safety for Key Non-Target Organisms
(2026-04-01)
Passara, Hataichanok
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Laosinwattana, Chamroon
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Moungthipmalai, Tanapoom
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Murata, Kouhei
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Soonwera, Mayura
Botanical insecticides containing a mixture of plant essential oils (EOs) are considered suitable for the management of houseflies (M. domestica). The adulticidal efficacies of single EOs and mixtures of EOs, including lemongrass (C. citratus), star anise (I. verum), nutmeg (M. fragrans), and their components (geranial, trans-anethole, and α-pinene), against houseflies were determined in comparison to 2% (w/v) α-cypermethrin as the positive control and distilled water as the negative control. The mixture of star anise EO (1%) + geranial (1%) was the most effective adulticide, superseding single EOs, other combinations of EOs, and its active component, α-cypermethrin, and distilled water. This mixture was highly synergistic and was found to be over 74% more toxic than all single EOs and almost 2.6 times more toxic than α-cypermethrin. Furthermore, the tested EOs did not cause mortality in guppies (P. reticulata) or earthworms (E. fetida), and caused a maximum of 48% mortality in honeybees (A. mellifera) at 24 h; by contrast, α-cypermethrin led to 100% mortality in honeybees within 0.5 h and in guppies and earthworms within 24 h, although it had low toxicity toward houseflies. Thus, a mixture of star anise EO + geranial is a promising source of EO-derived insecticides for housefly control that is also safe for important non-target species.
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Terpenoid Mixtures as Repellents Against the American Cockroach: Their Synergy and Low Toxicity Against Non-Target Species
(2026-01-01)
Passara, Hataichanok
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Moungthipmalai, Tanapoom
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Laosinwattana, Chamroon
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Sittichok, Sirawut
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Murata, Kouhei
The repellent activities (Periplaneta americana) of lone and binary mixtures of terpenoids—geranial, trans-anethole, and trans-cinnamaldehyde—against adult American cockroaches were evaluated in this study. The respective efficacies of these mixtures were compared with that of 12% (w/w) DEET. Safety bioassays for all formulations on non-target species, namely, earthworms (Eudrilus eugeniae) and guppy fish (Poecilia reticulata), were conducted to identify natural compounds with repellent efficacy equal to or surpassing that of DEET while ensuring ecological safety for non-target organisms such as fish and earthworms. All mixtures (RC<inf>50</inf> of 0.3 to 1.6 µL/cm3) were more effective than all lone terpenoids (RC<inf>50</inf> of 6.2 to 9.1 µL/cm3) and DEET (RC<inf>50</inf> of 3.0 µL/cm3), demonstrating strong synergy, with an increased repellent value (IV) of 79 to 96%. The strongest repellency, 98.0% at 72 h and an RC<inf>50</inf> of 0.3 µL/cm3, was achieved using geranial + trans-cinnamaldehyde (1:1). This mixture was 5.9 to 10 times more effective as a repellent than DEET. The toxicity of every lone terpenoid and terpenoid mixture to non-target species was low; hence, these mixtures can be considered safe, whereas DEET was highly toxic to non-target species (100% mortality). The terpenoid mixture geranial + trans-cinnamaldehyde showed strong repellency against P. americana.
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Tick fauna in grasslands on Mt. Aso and the possibility of the tick reduction to cattle by the essential oil
(2025-12-08)
Shibuya, Hikaru
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Tamagawa, Yoshiki
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Murata, Kouhei
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Tanapoon, Moungthipmalai
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Puwanard, Cheepchanok
The objectives of this study were to evaluate the tick fauna in the Aso region and the repellent effects and acaricidal activity of three plant–derived essential oils against ticks. It was revealed that the most dominant species of tick in the region was Haemaphysalis longicornis, with large annual fluctuations in the number of juvenile stages. The essential oils also showed significant repellent effects against H. longicornis and were highly toxic to this species. These results suggest that spraying essential oils on cattle in spring and autumn, when juvenile stages emerge, may be useful in controlling this species by disrupting infestation.
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Binary Mixtures of Essential Oils: Potent Housefly Adulticides That Are Safe Against Non-Target Species
(2025-08-01)
Passara, Hataichanok
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Sittichok, Sirawut
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Moungthipmalai, Tanapoom
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Laosinwattana, Chamroon
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Murata, Kouhei
In this study, we investigated the insecticidal potential of Eucalyptus globulus Labill. and Cymbopogon citratus Stapf essential oils (EOs), both alone and in synergistic blends with their primary active compounds, against adult houseflies (Musca domestica L.). Toxicity assessments were also conducted on non-target organisms—dwarf honeybees (Apis florea Fabricius) and guppies (Poecilia reticulata Peters)—to evaluate environmental safety. All binary EO mixtures demonstrated superior efficacy compared to individual EOs and the synthetic pyrethroid α-cypermethrin (1% positive control). The most potent formulation, combining 2.5% (v/v) geranial with 2.5% (v/v) E. globulus EO, exhibited a synergistic effect, achieving complete fly mortality (LT<inf>50</inf>: 0.06 h). This mixture’s mortality index significantly exceeded those of single-component formulations, with a mortality index of 0.22, confirming greater toxicity to flies than α-cypermethrin. Importantly, all the tested EOs and their blends were non-toxic to honeybees and guppies; in comparison, α-cypermethrin caused significant harm. These findings highlight the 2.5% (v/v) geranial + 2.5% (v/v) E. globulus EO blend as a highly effective and environmentally friendly alternative to conventional insecticides. Further research is recommended to optimize its formulation for practical use in sustainable fly management.
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Synergistic Larvicidal and Pupicidal Effects of Monoterpene Mixtures Against Aedes aegypti with Low Toxicity to Guppies and Honeybees
(2025-07-01)
Sittichok, Sirawut
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Passara, Hataichanok
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Moungthipmalai, Tanapoom
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Sinthusiri, Jirisuda
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Murata, Kouhei
The present study evaluated the larvicidal and pupicidal activities of pure and mixed monoterpene formulations—eucalyptol, geranial, trans-anethole, and trans-cinnamaldehyde—against Aedes aegypti and compared them with 1% (w/w) temephos. Safety bioassays of all formulations on non-target species confirmed their safety. The combined mixture of eucalyptol + trans-anethole at 400 µg/mL exhibited stronger larvicidal activity, with an LC<inf>50</inf> of 176 µg/mL, while the combination of trans-anethole + geranial at 400 µg/mL exhibited stronger pupicidal activity with an LC<inf>50</inf> of 167 µg/mL. Both formulations were more effective than a 1% temephos. All the mixture formulations were more strongly synergistic compared to pure formulations, with an increased mortality value (IMV) of 25% to 95%. External morphological aberrations observed at death included swelling of the respiratory system. Importantly, all the formulations were safe for two non-target species: guppies (Poecilia reticulata) and honeybees (Apis mellifera). The combination formulations are strong larvicides and pupicides for controlling Ae. Aegypti, which will help reduce the spread of viruses carried by this vector.
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Anise and Fennel Essential Oils and Their Combination as Natural and Safe Housefly Repellents
(2025-01-01)
Passara, Hataichanok
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Sittichok, Sirawut
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Puwanard, Cheepchanok
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Sinthusiri, Jirisuda
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Moungthipmalai, Tanapoom
Essential oils (EOs) are effective repellents and eco-friendly alternatives. We tested single and combination EOs of fennel and anise for repellency and stability against houseflies. All treatments were stored at 27 °C for up to 360 days. Efficacies were compared against α-cypermethrin as a reference. Safety bioassay used on two representative non-target species (guppies and earthworms) confirmed safety. The strongest repellency and stability, 100% repellency and RC<inf>50</inf> of 0.4 mL/m3 on day 1, was achieved by a fennel + anise EO combination (1:1). After 360 days, these combinations remained effective (95% repellency), and RC<inf>50</inf> = 0.8 mL/m3. The EO combinations were more effective than all single EOs and α-cypermethrin at all times, showing strong synergy with a synergistic repellent index of 2.4 to 3.3. This fennel + anise EO combination was more than 24 times more effective as a repellent than α-cypermethrin. Morphological damage included damaged antennae with twisted flagella and abnormal aristae. All single and combination EOs were not toxic to the non-targets and could be considered safe, whereas α-cypermethrin was highly toxic to them. Thus, the fennel + anise EO combination has great potential to be developed as a safe, natural repellent for managing housefly populations.
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Synergistic Larvicidal and Pupicidal Toxicity and the Morphological Impact of the Dengue Vector (Aedes aegypti) Induced by Geranial and trans-Cinnamaldehyde
(2024-09-01)
Sittichok, Sirawut
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Passara, Hataichanok
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Sinthusiri, Jirisuda
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Moungthipmalai, Tanapoom
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Puwanard, Cheepchanok
Monoterpenes are effective and eco-friendly alternatives to conventional chemical larvicides. We tested single and binary mixtures of monoterpenes—geranial and trans-cinnamaldehyde—for their larvicidal and pupicidal activities against Aedes aegypti L. and for non-target toxicity on guppies (Poecilia reticulata Peters), using 1% (w/w) temephos as a reference. Geranial and trans-cinnamaldehyde at 250 ppm showed stronger larvicidal and pupicidal activities with a 100% mortality rate and an LT<inf>50</inf> ranging from 0.3 to 0.6 h. All combinations were strongly synergistic against larvae and pupae compared to single formulations, with an increased mortality value (IMV) of 6% to 93%. The combination of geranial + trans-cinnamaldehyde (1:1) at 200 ppm showed the highest impact, with an IMV of 93%. The strongest larvicidal and pupicidal activities, a 100% mortality rate, and an LT<inf>50</inf> of 0.2 h were achieved by geranial + trans-cinnamaldehyde (1:1) 500 ppm. They were thirty times more effective than a 1% temephos solution (LT<inf>50</inf> ranging from 6.7 to 96 h) and caused obviously shriveled cuticles and a swollen respiratory system. All single and binary mixtures were not toxic to the guppies. Thus, the combination of geranial + trans-cinnamaldehyde has great potential as a safe insecticide for controlling mosquito larvae and pupae.
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Ovicidal Toxicity and Morphological Changes in Housefly Eggs Induced by the Essential Oils of Star Anise and Lemongrass and Their Main Constituents
(2024-07-01)
Passara, Hataichanok
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Sittichok, Sirawut
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Sinthusiri, Jirisuda
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Moungthipmalai, Tanapoom
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Puwanard, Cheepchanok
This study attempted to evaluate the ovicidal activity of single-component formulations and combination formulations of lemongrass and star anise essential oils (EOs) and their main constituents against housefly eggs. The efficacies of the combinations were compared with those of single-component formulations and α-cypermethrin. Safety bioassays of all treatments and α-cypermethrin on non-target predators—guppy and molly—were conducted. Two combinations: 1% lemongrass EO + 1% trans-anethole and 1% star anise EO + 1% geranial, exhibited a strong ovicidal activity with an inhibition rate of 94.4 to 96.2%. They were 1.1 times as effective as α-cypermethrin. The two combinations also showed high synergistic activity compared to single-component formulations, with a high synergistic index and a high increased inhibition value of 37.4 to 57.7%. All EO treatments were benign for all non-target aquatic species with a high 50% lethal time (LT<inf>50</inf>) and safety index. In contrast, α-cypermethrin was highly toxic to them with a low LT<inf>50</inf>. The morphological abnormalities observed in housefly eggs at death were those such as the shrivelling of the eggs, aberrations and damage to the eggshells, hatching lines, aeropyles, plastron, and micropyle. The potential of these two combinations as a cypermethrin replacement is compelling.
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Adulticidal synergy of two plant essential oils and their major constituents against the housefly Musca domestica and bioassay on non-target species
(2024-03-15)
Soonwera, Mayura
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Moungthipmalai, Tanapoom
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Puwanard, Cheepchanok
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Sittichok, Sirawut
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Sinthusiri, Jirisuda
Single and mixture formulations of lemongrass (Cymbopogon citratus (DC.) Stapf.) and star anise (Illicium verum (J. Presl.)) essential oils (EOs) and their major constituents were assayed for their adulticidal activities against housefly, Musca domestica L., and two non-target species, stingless bee (Tetragonula pegdeni Schwarz) and guppy (Poecilia reticulata Peters). The efficacies of the mixture formulations were compared against those of the single formulations and 1.0% α-cypermethrin, a common synthetic insecticide. GC-MS analysis found that the major constituent of lemongrass EO was geranial (45.23%), and that of star anise EO was trans-anethole (93.23%). Almost all mixture formulations were more effective in adulticidal activity against housefly adults than single formulations and 1.0% α-cypermethrin. A mixture of 1.0% lemongrass EO + 1.0% trans-anethole exhibited the strongest synergistic insecticidal activity with a 100% mortality rate (KT<inf>50</inf> of 3.2 min and LT<inf>50</inf> of 0.07 h). The relative percentage increase in mortality rate over single formulations was between 1.6 and 91.9%. In addition, it was three times more effective than 1.0% α-cypermethrin. To find the mechanism of adulticidal action, scanning electron microscopy (SEM) was done to find morphological aberrations, such as antennal and mouthpart aberrations, after the houseflies were treated with 1.0% lemongrass EO + 1.0% trans-anethole. The aberrations included deformed and abnormal shape of arista and flagellum, change in labellum pigmentation, and damage to pseudotracheae. Regarding toxicity against non-target species, all single and mixture formulations were not toxic to the two non-target species, while 1.0% α-cypermethrin was highly toxic. To conclude, a mixture of 1.0% lemongrass EO + 1.0% trans-anethole can be an excellent, natural, sustainable housefly adulticidal agent.
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Combinations of Lemongrass and Star Anise Essential Oils and Their Main Constituent: Synergistic Housefly Repellency and Safety against Non-Target Organisms
(2024-03-01)
Soonwera, Mayura
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Sinthusiri, Jirisuda
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Passara, Hataichanok
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Moungthipmalai, Tanapoom
;
Puwanard, Cheepchanok
The present study evaluated the housefly repellency of single-component formulations and combinations of lemongrass and star anise essential oils (EOs) and their main constituents. The efficacies of the combinations were compared against those of single-component formulations and DEET. Safety bioassays of all formulations and DEET on non-target species—guppy, molly, dwarf honeybee, and stingless bee—were conducted. GC–MS analysis showed that the main constituent of lemongrass EO was geranial (46.83%) and that of star anise EO was trans-anethole (92.88%). All combinations were highly synergistic compared to single-component formulations, with an increased repellent value (IR) of 34.6 to 51.2%. The greatest synergistic effect was achieved by 1.0% lemongrass EO + 1.0% trans-anethole combination, with an IR of 51.2%. The strongest, 100% repellent rate at 6 h was achieved by 1.0% geranial + 1.0% trans-anethole. They were twice as effective as DEET and caused obvious damage to housefly antennae under microscopic observation. All single-component formulations and combinations were benign to the four tested non-target species. In contrast, DEET was highly toxic to them. The synergistic repellency and biosafety of these two combinations are compellingly strong support for developing them into an effective green repellent.

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