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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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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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Combinations of plant essential oils and their major compositions inducing mortality and morphological abnormality of Aedes aegypti and Aedes albopictus
(2022-05-01)
Soonwera, Mayura
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Moungthipmalai, Tanapoom
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Aungtikun, Jirapon
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Sittichok, Sirawut
Extensive uses of synthetic insecticides to control mosquito's populations have induced the insects to develop resistance against them, rendering them ineffective today. Moreover, they cause serious impacts on human health and the ecosystem. Therefore, safe and effective natural alternatives are needed. This study evaluated the larvicidal and pupicidal activities of essential oils (EOs) from Illicium verum and Zanthoxylum limonella and the major constituents against Aedes aegypti and Aedes albopictus mosquitoes as well as recorded their morphological aberrations at death. The GC-MS analysis showed that trans-anethole was the major constituent of I. verum EO, and limonene was the major constituent of Z. limonella EO. Both were more effective against the larvae and pupae of Ae. aegypti than those of Ae. albopictus. A 2.5% I. verum EO + 2.5% trans-anethole combination showed the highest larvicidal and pupicidal effects against Ae. aegypti and Ae. albopictus with an LT<inf>50</inf> ranging from 0.2-6.9 h. Between the two tested constituents, trans-anethole exhibited stronger larvicidal and pupicidal activities (LC<inf>50</inf> ranging 2.4–3.4%) against the two tested mosquito species than d-limonene (LC<inf>50</inf> ranging 2.5–3.7%). Most importantly, 5% trans-anethole, 5% d-limonene, and 2.5% I. verum EO + 2.5% trans-anethole were more effective (LT<inf>50</inf> ranging 0.1–0.3 h) than 1% (w/w) temephos (LT<inf>50</inf> ranging 2.9–3.1 h). Morphological aberrations at death observed were such as color pigment and thorax shape abnormalities. To conclude, trans-anethole, d-limonene, and a combination of I. verum EO + trans-anethole, are natural compounds that not only are as effective as temephos at the time of this study, but should be also be much safer to human health.
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Adulticidal activities of Cymbopogon citratus (Stapf.) and Eucalyptus globulus (Labill.) essential oils and of their synergistic combinations against Aedes aegypti (L.), Aedes albopictus (Skuse), and Musca domestica (L.)
(2020-06-01)
Soonwera, Mayura
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Sittichok, Sirawut
The knockdown and adulticidal activities of individual Cymbopogon citratus and Eucalyptus globulus essential oils (EOs) and their combinations were evaluated against three medical insect pests (Aedes aegypti, Aedes albopictus, and Musca domestica) using a WHO susceptibility test. The knockdown and adulticidal activities against the three medical insect pests of combinations of C. citratus and E. globulus EOs were higher than those of individual EOs alone. Combinations of 7.5% C. citratus + 7.5% E. globulus EOs and 10% C. citratus + 10% E. globulus EOs exhibited the highest efficacy against females of the three species with 100% knockdown and mortality rates at 1 and 24 h after exposure, respectively. Their adulticidal activities were equivalent to that of 10% w/v cypermethrin. In contrast, 70% v/v ethyl alcohol negative control was not effective at all. The combinations of EOs showed a synergistic effect, i.e., their adulticidal activity was improved by 0.2 to 100%, with increased knockdown and mortality rates, compared to individual EOs. The highest synergistic effect on effective knockdown and adulticidal activities against females of the three species was achieved by a combination of 2.5% C. citratus + 2.5% E. globulus EOs, with 36.6 to 100% knockdown rate increase and 33.5 to 98.9% mortality rate increase. This study demonstrates that all tested combinations of C. citratus and E. globulus EOs were effective adulticidal agents against females of Ae. aegypti, Ae. albopictus, and M. domestica and have a high potential for development into a botanical insecticide for controlling populations of Aedes mosquitoes and houseflies.
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Effect of Cymbopogon citratus (lemongrass) and Syzygium aromaticum (clove) oils on the morphology and mortality of Aedes aegypti and Anopheles dirus larvae
(2016-04-01)
Soonwera, Mayura
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Phasomkusolsil, Siriporn
Cymbopogon citratus (lemongrass) and Syzygium aromaticum (clove) oils were evaluated to determine mortality rates, morphological aberrations, and persistence when used against third and fourth larval instars of Aedes aegypti and Anopheles dirus. The oils were evaluated at 1, 5, and 10 % concentrations in mixtures with soybean oil. Persistence of higher concentrations was measured over a period of 10 days. For Ae. aegypti, both plant oils caused various morphological aberrations to include deformed larvae, incomplete eclosion, white pupae, deformed pupae, dead normal pupae, and incomplete pupal eclosion. All of these aberrations led to larval mortality. In Ae. aegypti larvae, there were no significant differences in mortality at days 1, 5, and 10 or between third and fourth larval instar exposure. In An. dirus, morphological aberrations were rare and S. aromaticum oil was more effective in causing mortality among all larval stages. Both oils were equally effective at producing mortality on days 1, 5, and 10. Both oils had slightly increased LT<inf>50</inf> rates from day 1 to day 10. In conclusion, both lemongrass and clove oils have significant effects on the immature stages of Ae. aegypti and An. dirus and could potentially be developed for use as larvicides.
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Efficacy of Thai herbal essential oils as green repellent against mosquito vectors
(2015-02-01)
Soonwera, Mayura
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Phasomkusolsil, Siriporn
Repellency activity of Thai essential oils derived from ylang ylang (Cananga odorata (Lamk.) Hook.f. & Thomson: Annonaceae) and lemongrass (Cymbopogon citratus (DC.) Stapf: Poaceae) were tested against two mosquito vectors, Aedes aegypti (L.) and Culex quinquefasciatus (Say). There were compared with two chemical repellents (DEET 20% w/w; Sketolene Shield® and IR3535, ethyl butylacetylaminopropionate 12.5% w/w; Johnson's Baby Clear Lotion Anti-Mosquito®). Each herbal repellent was applied in three diluents; coconut oil, soybean oil and olive oil at 0.33μl/cm2 on the forearm of volunteers. All herbal repellent exhibited higher repellent activity than IR3535 12.5% w/w, but lower repellent activity than DEET 20% w/w. The C. odorata oil in coconut oil exhibited excellent activity with 98.9% protection from bites of A. aegypti for 88.7±10.4min. In addition, C. citratus in olive oil showed excellent activity with 98.8% protection from bites of C. quinquefasciatus for 170.0±9.0min. While, DEET 20% w/w gave protection for 155.0±7.1-182.0±12.2min and 98.5% protection from bites of two mosquito species. However, all herbal repellent provided lower repellency activity (97.4-98.9% protection for 10.5-88.7min) against A. aegypti than C. quinquefasciatus (98.3-99.2% protection for 60-170min). Our data exhibited that C. odorata oil and C. citratus oil are suitable to be used as green repellents for mosquito control, which are safe for humans, domestic animals and environmental friendly.
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Repellent activity of herbal essential oils against Aedes aegypti (Linn.) and Culex quinquefasciatus (Say.)
(2013-08-01)
Sritabutra, Duangkamon
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Soonwera, Mayura
Objective: To determine the mosquito repellent activity of herbal essential oils against female Aedes aegypti and Culex quinquefasciatus. Methods: On a volunteer's forearm, 0.1 mL of each essential oil was applied to 3 cm×10 cm of exposed skin. The protection time was recorded for 3 min after every 30 min. Results: Essential oil from clove oil in olive oil and in coconut oil gave the longest lasting period of 76.50 min and 96.00 min respectively against Aedes aegypti. The citronella grass oil in olive oil, citronella grass oil in coconut oil and lemongrass oil in coconut oil exhibited protection against Culex quinquefasciatus at 165.00, 105.00, and 112.50 min respectively. Conclusions: The results clearly indicated that clove, citronella and lemongrass oil were the most promising for repellency against mosquito species. These oils could be used to develop a new formulation to control mosquitoes. © 2013 Asian Pacific Tropical Medicine Press.
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Larvicidal and pupicidal activities of essential oils from Zingiberaceae plants against Aedes aegypti (Linn.) and Culex quinquefasciatus say mosquitoes
(2013-01-01)
Phukerd, Ubol
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Soonwera, Mayura
We conducted this study to investigate the efficacy of herbal essential oils from 12 species of Zingiberaceae plants to determine their larvicidal and pupicidal activity against fourth instar larvae and pupae of Aedes aegypti and Culex quinquefasciatus mosquitoes. Probit analysis was used to analyze the data. Larval mortality was recorded at 1, 5, 10, 15, 30 and 60 minutes and 24 hours. Pupal mortality was recorded at 15 and 30 minutes and 1, 3, 6, 12, 24 and 48 hours. All the essential oils tested showed larvicidal activity. Zingiber cassumunar and Amomum biflorum oils proved to have the greatest activity against Ae. aegypti larvae with LT<inf>50</inf> of 1.4 minutes and 100% mortality at 5 and 10 minutes, respectively. Boesenbergia rotunda, Curcuma zedoaria and Hedychium coronarium essential oils had activity against Cx. quinquefasciatus larvae with LT<inf>50</inf> of 1.7 minutes and 100% mortality at 10 minutes, 5 minutes and 15 minutes, respectively. All the herbal essential oils tested resulted in 100% mortality against Ae. aegypti and Cx. quinquefasciatus larvae at 60 minutes and 30 minutes, respectively. Ae. aegypti and Cx. quinquefasciatus pupae were susceptible to Z. ottensii oil (LT<inf>50</inf> of 0.2 hour) and Z. zerumbet oil (LT<inf>50</inf> of 0.6 hour) and had pupicidal activity with 100% mortality at 6 and 3 hours, respectively. All the essential oils test had pupicidal activity against Ae. aegypti and Cx. quinquefasciatus by inducing 100% mortality at 48 hours.
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Evaluation of herbal essential oil as repellents against Aedes aegypti (L.) and Anopheles dirus Peyton & Harrion
(2011-01-01)
Sritabutra, Duangkamon
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Soonwera, Mayura
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Waltanachanobon, Sirirat
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Poungjai, Supaporn
Objective: To investigate the repellent activity of herbal essential oils from garlic (Allium sativum), clove (Syzygium aromaticum), lemon grass (Cybopogon citratus), citronella grass (Cymbopogon nardus), peppermint (Mentha piperita), eucalyptus (Eucalyptus globulus), orange (Citrus sinensis) and sweet basil (Ocimum basilicum) and their combinations against Aedes aegypti (Ae. aegypti) (L.) and Anopheles dirus (An. dirus) Peyton & Harrion under laboratory conditions. Methods: In laboratory condition, 0.1 mL of each essential oil was applied to 3-10 cm of exposed area on a volunteer's forearm. The test was carried out every 30 min until fewer than two mosquitoes bit or land during the 3 min study period and then the repellency test was stopped. Results: Essential oil from lemon grass exhibited protection against biting from two mosquito species, for Ae. aegypti [(98.66±11.56) min protection time and 0.97% biting rate] and for An. dirus [(98.00±15.28) min protection time and 0.80% biting rate]. The combinations from eucalyptus oil and sweet basil oil were effective as repellents and feeding deterrents against Ae. aegypti [(98.87±10.28) min protection time and 0.90% biting rate] and An. dirus [(210±10.70) min protection time and 0.93% biting rate]. All herbal repellents exhibited the period of protection time against Ae. aegypti which was lower than 120 min. Conclusions: It can be concluded that oils of lemon grass and combination from eucalyptus-sweet basil are the most effective in repellent activity.
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Insect repellent activity of medicinal plant oils against aedes aegypti (LINN.), anopheles minimus (Theobald) and culex quinquefasciatus say based on protection time and biting rate
(2010-01-01)
Phasomkusolsil, Siriporn
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Soonwera, Mayura
This study investigated insect bite protection and length of the protection with 30 repellents which were divided into 3 categories: plant oil, essential oil and essential oil with ethyl alcohol, tested against three mosquito species, Aedes aegypti, Anopheles minimus and Culex quinquefasciatus, under laboratory conditions. The plant oil group was comprised of Phlai (Zingiber cassumunar) and Sweet basil (Ocimum basilicum). Both substances were effective as repellents and feeding deterrents against An. minimus (205 minutes protection time and a biting rate of 0.9%), Cx. quinquefasciatus (165 minutes protection time and 0.9% biting rate) and Ae. aegypti (90 minutes protection time and 0.8% biting rate). Essential oil from citronella grass (Cymbopogon nardus) exhibited protection against biting from all 3 mosquito species: for An. minimus, Cx. quinquefasciatus and Ae. aegypti, the results were 130 minutes and 0.9%, 140 minutes and 0.8%, and 115 minutes and 0.8%, respectively. The period of protection time against Ae. aegypti for all repellent candidates tested was lower than the Thai Industrial Standards Institute (TISI) determined time of greater than 2 hours.

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