Synergistic Repellent and Irritant Effects of a Mixture of ß-Caryophyllene Oxide and Vetiver Oil against Mosquito Vectors.

Repellents play a major role in reducing the risk of mosquito-borne diseases by preventing mosquito bites. The present study evaluated the mosquito-repellent activity of ß-caryophyllene oxide 1% (BCO), vetiver oil 2.5% (VO), and their binary mixtures (BCO + VO (1:1), BCO + VO (2:1), BCO + VO (1:2)) against four laboratory-colonized mosquito species, Aedes aegypti (L.), Aedes albopictus (Skuse), Anopheles minimus Theobald, and Culex quinquefasciatus Say, using an excito-repellency assay system. In general, the compound mixtures produced a much stronger response in the mosquitoes than single compounds, regardless of the test conditions or species. The greatest synergetic effect was achieved with the combination of BCO + VO (1:2) in both contact and noncontact trials with An. minimus (74.07-78.18%) and Cx. quinquefasciatus (55.36-83.64%). Knockdown responses to the binary mixture of BCO + VO were observed for Ae. albopictus, An. minimus, and Cx. quinquefasciatus, in the range of 18.18-33.33%. The synergistic repellent activity of BCO and VO used in this study may support increased opportunities to develop safer alternatives to synthetic repellents for personal protection against mosquitoes.

Behavioral avoidance and biological safety of vetiver oil and its constituents against Aedes aegypti (L.), Aedes albopictus (Skuse) and Culex quinquefasciatus Say.

Numerous plant-based repellents are widely used for personal protection against host-seeking mosquitoes. Vitiveria zizanioides (L.) Nash essential oil and its constituents have demonstrated various mosquito repellent activities. In this study, three chemical actions of vetiver oil and five constituents (terpinen-4-ol, α-terpineol, valencene, vetiverol and vetivone) were characterized against Aedes aegypti, Aedes albopictus and Culex quinquefasciatus by using the high-throughput screening assay system (HITSS). Significant contact escape responses in Ae. aegypti and Ae. albopictus to all test compounds at concentrations between 2.5 and 5% were observed. Spatial repellency responses were also observed in some tested mosquito populations depending upon concentrations. The most significant toxic response on mosquitoes was found at the highest concentration, except for vetivone which had no toxic effect on Ae. aegypti and Ae. albopictus. Results on phototoxic and genotoxic hazard revealed that vetiver oil and their constituents showed no phototoxic potential or any significant genotoxic response. In conclusion, vetiver oil and two constituents, valencene and vetiverol, are potentials as active ingredients for mosquito repellency and present no toxicity.

Evaluation of the Constituents of Vetiver Oil Against Anopheles minimus (Diptera: Culicidae), a Malaria Vector in Thailand.

The development of resistance by mosquitoes to current synthetic compounds has resulted in reduced effectiveness of prevention and control methods worldwide. An alternative nonchemical based control tools are needed to be evaluated particularly plant-derived essential oils. Several components of vetiver oil have been documented as insect repellents. However, detailed knowledge of those components action against insect remains unknown. In this study, behavioral response of Anopheles minimus to four constituents of vetiver oil (valencene, terpinen-4-ol, isolongifolene, vetiverol) was evaluated by using the high-throughput screening assay system. Vetiverol and isolongifolene exhibited strong contact irritancy action at 1.0% (80.2% escaping) and 5.0% (81.7% escaping) concentration, respectively, while moderate action was found in both valencene and terpinen-4-ol at 5.0% (57.6% escaping). Only at 1.0% (0.7 spatial activity index [SAI]) and 5.0% (1.0 SAI) of valencene and 0.5% (0.7 SAI) of isolongifolene showed spatial repellency activity. High mortality (58.9-98.2%) was recorded in all concentration of vetiverol and isolongifolene. Meanwhile, valencene exhibited high mortality only at 5.0%, terpinen-4-ol showed very low toxic action (0-4.3%) in all concentration. These proved that valencene in vetiver oil is the promising constituent that can be developed as an alternative mosquito control mean in efforts to prevent disease transmission.

Evaluation of a Noncontact, Alternative Mosquito Repellent Assay System.

A novel noncontact repellency assay system (NCRAS) was designed and evaluated as a possible alternative method for testing compounds that repel or inhibit mosquitoes from blood feeding. Deet and Aedes aegypti were used in a controlled laboratory setting. Using 2 study designs, a highly significant difference were seen between deet-treated and untreated skin placed behind the protective screens, indicating that deet was detected and was acting as a deterrence to mosquito landing and probing behavior. However, a 2nd study showed significant differences between protected (behind a metal screen barrier) and unprotected (exposed) deet-treated forearms, indicating the screen mesh might restrict the detection of deet and thus influences landing/biting response. These findings indicate the prototype NCRAS shows good promise but requires further evaluation and possible modification in design and testing protocol to achieve more desirable operational attributes in comparison with direct skin-contact repellency mosquito assays.

Plants traditionally used as mosquito repellents and the implication for their use in vector control.

Numerous plants with insect repelling properties are native to the tropics where they are produced for a wide range of medicinal purposes. In Thailand, these native plant species have a history of use for personal protection against biting insects. From our investigation we identified 37 plant species within 14 plant families that showed some mosquito repellent properties. Of these, 9 plant species were characterized using an excito-repellency test system against several Thai mosquito species. Results from these studies revealed that five essential oils extracted from plants demonstrated promising insect repellent activity. These active ingredients show promise for further development into formulations that may serve as alternatives to DEET or possibly be used as natural bio-pesticides to kill mosquitoes.

The effects of plant essential oils on escape response and mortality rate of Aedes aegypti and Anopheles minimus.

The High Throughput Screening System (HITSS) has been applied in insecticide behavioral response studies with various mosquito species. In general, chemical or natural compounds can produce a range of insect responses: contact irritancy, spatial repellency, knock-down, and toxicity. This study characterized these actions in essential oils derived from citronella, hairy basil, catnip, and vetiver in comparison to DEET and picaridin against Aedes aegypti and Anopheles minimus mosquito populations. Results indicated the two mosquito species exhibited significantly different (P<0.05) contact irritant escape responses between treatment and control for all tested compound concentrations, except with the minimum dose of picaridin (P>0.05) against Ae. aegypti. Spatial repellency responses were elicited in both mosquito species when exposed to all compounds, but the strength of the repellent response was dependent on compound and concentration. Data show that higher test concentrations had greatest toxic effects on both mosquito populations, but vetiver had no toxic effect on Ae. aegypti and picaridin did not elicit toxicity in either Ae. aegypti or An. minimus at any test concentration. Ultimately, this study demonstrates the ability of the HITSS assay to guide selection of effective plant essential oils for repelling, irritating, and killing mosquitoes.

Comparison of Field and Laboratory-Based Tests for Behavioral Response of Aedes aegypti (Diptera: Culicidae) to Repellents.

The repellent and irritant effects of three essential oils-clove, hairy basil, and sweet basil-were compared using an excito-repellency test system against an insecticide-resistant strain of Aedes aegypti (L.) females from Pu Teuy, Kanchanaburi Province. DEET was used as the comparison standard compound. Tests were conducted under field and controlled laboratory conditions. The most marked repellent effect (spatial noncontact assay) among the three test essential oils was exhibited by sweet basil, Ocimum basilicum L. (53.8% escaped mosquitoes in 30-min exposure period) under laboratory conditions while hairy basil, Ocimum americanum L. and clove, Syzygium aromaticum (L.) Merill et. L.M. Perry from laboratory tests and sweet basil from field tests were the least effective as repellents (0-14%). In contrast, the contact assays measuring combined irritancy (excitation) and repellency effects found the best contact irritant response to hairy basil and DEET in field tests, whereas all others in laboratory and field were relatively ineffective in stimulating mosquitoes to move out the test chambers (0-5.5%). All three essential oils demonstrated significant differences in behavioral responses between field and laboratory conditions, whereas there was no significant difference in contact and noncontact assays for DEET between the two test conditions (P > 0.05).

Behavioral responses of Aedes aegypti, Aedes albopictus, Culex quinquefasciatus, and Anopheles minimus against various synthetic and natural repellent compounds.

The behavioral responses of colony populations of Aedes aegypti, Aedes albopictus, Culex quinquefasciatus, and Anopheles minimus to four essential oils (citronella, hairy basil, catnip, and vetiver), two standard repellents (DEET and picaridin), and two synthetic pyrethroids (deltamethrin and permethrin) were conducted in the laboratory using an excito-repellency test system. Results revealed that Cx. quinquefasciatus and An. minimus exhibited much stronger behavioral responses to all test compounds (65-98% escape for contact, 21.4-94.4% escape for non-contact) compared to Ae. aegypti (3.7-72.2% escape (contact), 0-31.7% (non-contact)) and Ae. albopictus (3.5-94.4% escape (contact), 11.2-63.7% (non-contact)). In brief, essential oil from vetiver elicited the greatest irritant responses in Cx. quinquefasciatus (96.6%) and An. minimus (96.5%) compared to the other compounds tested. The synthetic pyrethroids caused a stronger contact irritant response (65-97.8% escape) than non-contact repellents (0-50.8% escape for non-contact) across all four mosquito species. Picaridin had the least effect on all mosquito species. Findings from the current study continue to support the screening of essential oils from various plant sources for protective properties against field mosquitoes.

Comparative Behavioral Responses of Pyrethroid-Susceptible and -Resistant Aedes aegypti (Diptera: Culicidae) Populations to Citronella and Eucalyptus Oils.

The objective of this study was to compare the behavioral responses (contact irritancy and noncontact spatial repellency) between susceptible and resistant populations of Aedes aegypti (L.) (=Stegomyia aegypti) to essential oils, citronella, and eucalyptus, Eucalyptus globulus, extracts, using an excito-repellency test system. N, N-diethyl-meta-toluamide (DEET) was used as the standard reference repellent. Mosquitoes included two long-standing insecticide susceptible colonies (U.S. Department of Agriculture and Bora Bora) and two pyrethroid-resistant populations recently obtained from Phetchabun and Kanchanaburi provinces in Thailand. Both DEET and citronella produced a much stronger excitation ("irritancy") and more rapid flight escape response in both pyrethroid-resistant populations compared with the laboratory populations. Noncontact repellency was also greater in the two resistant populations. Eucalyptus oil was found to be the least effective compound tested. Differences in responses between long-established pyrethroid-susceptible colonies and newly established and naturally resistant colonies were clearly demonstrated. These findings also demonstrate the need for further comparisons using natural pyrethroid-susceptible populations for elucidation of factors that might contribute to different patterns of escape behavior.