In a previous screening program for mosquitocides from local edible plants in Thailand, essential oils (EOs) of Cyperus rotundus, Alpinia galanga and Cinnamomum verum, were found to possess promising adulticidal activity against Aedes aegypti. With the aim of reducing usage of conventional insecticides and improving the management of resistant mosquito populations, this study was designed to determine the potential synergism in the adulticidal efficacy of EOs on permethrin toxicity against Ae. aegypti, both pyrethroid-resistant and -susceptible strains. EOs extracted from rhizomes of C. rotundus and A. galanga as well as C. verum barks were evaluated for chemical compositions and adulticidal activity against Muang Chiang Mai-susceptible (MCM-S) and Pang Mai Dang-resistant (PMD-R) strains of Ae. aegypti. Adulticidal bioassays of EO-permethrin mixtures for synergistic activity were also performed on these Ae. aegypti strains. Chemical characterization by the GC-MS analytical technique demonstrated that 48 compounds were identified from the EOs of C. rotundus, A. galanga and C. verum, representing 80.22%, 86.75% and 97.24%, respectively, of all compositions. Cyperene (14.04%), β-bisabolene (18.27%) and cinnamaldehyde (64.66%) were the main constituents of C. rotundus, A. galanga and C. verum oils, respectively. In adulticidal bioassays, EOs of C. rotundus, A. galanga and C. verum were effective in killing Ae. aegypti, both MCM-S and PMD-R strains, with LD50 values of 10.05 and 9.57 μg/mg female, 7.97 and 7.94 μg/mg female, and 3.30 and 3.22 μg/mg female, respectively. The adulticidal efficacy against MCM-S and PMD-R Ae. aegypti of these EOs was close to that of piperonyl butoxide (PBO, LD50 values = 6.30 and 4.79 μg/mg female, respectively) but less pronounced than that of permethrin (LD50 values = 0.44 and 3.70 ng/mg female, respectively). Nevertheless, combination-based bioassays discovered the accomplished synergism of EOs together with permethrin. Significant synergistic effects with permethrin against both the strains of Ae. aegypti were recorded in the EOs of C. rotundus and A. galanga. Addition of C. rotundus and A. galanga oils decreased the LD50 values of permethrin against MCM-S dramatically from 0.44 to 0.07 and 0.11 ng/mg female, respectively, with synergism ratio (SR) values of 6.28 and 4.00, respectively. Furthermore, EOs of C. rotundus and A. galanga also reduced the LD50 values of permethrin against PMD-R drastically from 3.70 to 0.42 and 0.003 ng/mg female, respectively, with SR values of 8.81 and 1233.33, respectively. The synergy of enhanced adulticidal toxicity recorded from EO-permethrin combinations against both strains of Ae. aegypti presents a promising role of EOs as a synergist for improving mosquitocidal efficacy, particularly in situations where conventional compounds are ineffective or inappropriate.

In ongoing screening research for edible plants, Petroselinum crispum essential oil was considered as a potential bioinsecticide with proven antimosquito activity against both the pyrethroid susceptible and resistant strains of Aedes aegypti. Due to the comparative mosquitocidal efficacy on these mosquitoes, this plant essential oil is promoted as an attractive candidate for further study in monitoring resistance of mosquito vectors. Therefore, the aim of this study was to evaluate the impact of P. crispum essential oil on the biochemical characteristics of the target mosquito larvae of Ae. aegypti, by determining quantitative changes of key enzymes responsible for xenobiotic detoxification, including glutathione-S-transferases (GSTs), α- and β-esterases (α-/β-ESTs), acetylcholinesterase (AChE), acid and alkaline phosphatases (ACP and ALP) and mixed-function oxidases (MFO). Three populations of Ae. aegypti, comprising the pyrethroid susceptible Muang Chiang Mai-susceptible (MCM-S) strain and the pyrethroid resistant Pang Mai Dang-resistant (PMD-R) and Upakut-resistant (UPK-R) strains, were used as test organisms. Biochemical study of Ae. aegypti larvae prior to treatment with P. crispum essential oil revealed that apart from AChE, the baseline activity of most defensive enzymes, such as GSTs, α-/β-ESTs, ACP, ALP and MFO, in resistant UPK-R or PMD-R, was higher than that determined in susceptible MCM-S. However, after 24-h exposure to P. crispum essential oil, the pyrethroid susceptible and resistant Ae. aegypti showed similarity in biochemical features, with alterations of enzyme activity in the treated larvae, as compared to the controls. An increase in the activity levels of GSTs, α-/β-ESTs, ACP and ALP was recorded in all strains of P. crispum oil-treated Ae. aegypti larvae, whereas MFO and AChE activity in these mosquitoes was decreased. The recognizable larvicidal capability on pyrethroid resistant Ae. aegypti, and the inhibitory effect on AChE and MFO, emphasized the potential of P. crispum essential oil as an attractive alternative application for management of mosquito resistance in current and future control programs.

Mosquitoes are hematophagous insects that transmit parasites and pathogens with devastating effects on humans, particularly in subtropical regions. Different mosquito species display various behaviors, breeding sites, and geographic distribution; however, they can be difficult to distinguish in the field due to morphological similarities between species and damage caused during trapping and transportation. Vector control methods for controlling mosquito-borne disease epidemics require an understanding of which vector species are present in the area as well as the epidemiological patterns of disease transmission. Although molecular techniques can accurately distinguish between mosquito species, they are costly and laborious, making them unsuitable for extensive use in the field. Thus, alternative techniques are required. Geometric morphometrics (GM) is a rapid and inexpensive technique that can be used to analyze the size, shape, and shape variation of individuals based on a range of traits. Here, we used GM to analyze the wings of 1,040 female mosquitoes from 12 different species in Thailand. The right wing of each specimen was removed, imaged microscopically, and digitized using 17 landmarks. Wing shape variation among genera and species was analyzed using canonical variate analysis (CVA), while discriminant function analysis was used to cross-validate classification reliability based on Mahalanobis distances. Phenetic relationships were constructed to illustrate the discrimination patterns for genera and species. CVA of the morphological variation among Aedes, Anopheles, Armigeres, Culex, and Mansonia mosquito genera revealed five clusters. In particular, we demonstrated a high percentage of correctly-distinguished samples among Aedes (97.48%), Armigeres (96.15%), Culex (90.07%), and Mansonia (91.67%), but not Anopheles (64.54%). Together, these findings suggest that wing landmark-based GM analysis is an efficient method for identifying mosquito species, particularly among the Aedes, Armigeres, Culex, and Mansonia genera.

For personal protection against mosquito bites, user-friendly natural repellents, particularly from plant origin, are considered as a potential alternative to applications currently based on synthetics such as DEET, the standard chemical repellent. This study was carried out in Thailand to evaluate the repellency of Ligusticum sinense hexane extract (LHE) against laboratory Anopheles minimus and Aedes aegypti, the primary vectors of malaria and dengue fever, respectively. Repellent testing of 25% LHE against the two target mosquitoes; An. minimus and Ae. aegypti, was performed and compared to the standard repellent, DEET, with the assistance of six human volunteers of either sex under laboratory conditions. The physical and biological stability of LHE also was determined after keeping it in conditions that varied in temperature and storage time. Finally, LHE was analysed chemically using the qualitative GC/MS technique in order to demonstrate a profile of chemical constituents. Ethanol preparations of LHE, with and without 5% vanillin, demonstrated a remarkably effective performance when compared to DEET in repelling both An. minimus and Ae. aegypti. While 25% LHE alone provided median complete-protection times against An. minimus and Ae. aegypti of 11.5 (9.0–14.0) hours and 6.5 (5.5–9.5) hours, respectively, the addition of 5% vanillin increased those times to 12.5 (9.0–16.0) hours and 11.0 (7.0–13.5) hours, respectively. Correspondingly, vanillin added to 25% DEET also extended the protection times from 11.5 (10.5–15.0) hours to 14.25 (11.0–18.0) hours and 8.0 (5.0–9.5) hours to 8.75 (7.5–11.0) hours against An. minimus and Ae. aegypti, respectively. No local skin reaction such as rash, swelling or irritation was observed during the study period. Although LHE samples kept at ambient temperature (21–35°C), and 45°C for 1, 2 and 3 months, demonstrated similar physical characteristics, such as similar viscosity and a pleasant odour, to those that were fresh and stored at 4°C, their colour changed from light- to dark-brown. Interestingly, repellency against Ae. aegypti of stored LHE was presented for a period of at least 3 months, with insignificantly varied efficacy. Chemical analysis revealed that the main components of LHE were 3-N-butylphthalide (31.46%), 2, 5-dimethylpyridine (21.94%) and linoleic acid (16.41%), constituting 69.81% of all the extract composition. LHE with proven repellent efficacy, no side effects on the skin, and a rather stable state when kept in varied conditions is considered to be a potential candidate for developing a new natural alternative to DEET, or an additional weapon for integrated vector control when used together with other chemicals/measures.

Ligusticum sinense Oliv. cv. is a species of Umbelliferae (Apiaceae), a large plant family in the order Apiales. In this study, L. sinense hexane extract nanoemulsion gel (LHE-NEG) was investigated for mosquito repellency and compared to the standard chemical, N,N-diethyl-3-methylbenzamide (DEET), with the goal of developing a natural alternative to synthetic repellents in protecting against mosquito vectors. The results demonstrated that LHE-NEG afforded remarkable repellency against Aedes aegypti, Anopheles minimus, and Culex quinquefasciatus, with median protection times (MPTs) of 5.5 (4.5–6.0), 11.5 (8.5–12.5), and 11.25 (8.5–12.5) h, respectively, which was comparable to those of DEET-nanoemulsion gel (DEET-NEG: 8.5 (7.0–9.0), 12.0 (10.0–12.5), and 12.5 (10.0–13.5) h, respectively). Evaluation of skin irritation in 30 human volunteers revealed no potential irritant from LHE-NEG. The physical and biological stability of LHE-NEG were determined after being kept under heating/cooling cycle conditions. The stored samples of LHE-NEG exhibited some changes in appearance and differing degrees of repellency between those kept for 3 and 6 heating/cooling cycles, thus providing slightly shorter MPTs of 4.25 (4.0–4.5) and 3.25 (2.5–3.5) h, respectively, when compared to those of 5.0 (4.5–6.0) h in fresh preparation. These findings encourage commercially developed LHE-based products as an alternative to conventional synthetic repellents in preventing mosquito bites and helping to interrupt mosquito-borne disease transmission.

Dengue fever has been a significant disease in Thailand for a long time, ranking it as one of the major health problems in the country. Management of the adult stage of mosquito vectors is approached by applying various synthetic chemicals such as adulticides, attractants, deterrents, and repellents. In Thailand, mosquito control and personal protection from mosquito bites are currently the most important measures for preventing and controlling mosquito-borne diseases. Although there are various control strategies for dengue disease, participation from the local community plays a vital role in the success of disease control. At present, a lot of local people have seen the value of local indigenous knowledge and used this to improve their life. The local community in the southern part of Thailand has used mosquito repellent from local knowledge for a long time. The problem regarding mosquito repellent made from local indigenous knowledge is that it has not yet been tested to determine its effectiveness. Therefore, this research aims to assess the effectiveness of mosquito repellent from local learning from Nakhon Si Thammarat provinces in Thailand. From the survey, out of 23 districts, six mosquito repellents were found in 3 communities, including Nabon, Muang, and Thasala. The repellent efficacy against the laboratory strain of Aedes aegypti by using the human-bait technique of the WHO (1996) standard method, with slight modifications. Approximately 0.1 ml of each test sample was applied evenly onto a 30 cm² test site on one forearm of each human volunteer. Exposure experiments continued at 30 min intervals until at least two bites occurred in a 3-min period, or when a first bite was followed by a confirming bite (second bite) in the subsequent observation period. Each test was duplicated on different days for the two human volunteers. The result shows that three mosquito repellents made from local indigenous knowledge that have protection that lasts for more than 2 h are Ban Ko Sa Child Development Center's citronella spray (Nabon district, Kaew Saen subdistrict), Khun Lang's citronella spray, and Khun Lang's citronella ointment (Muang district, Pak Phun subdistrict). The result of this research was reported back to the local community to re-evaluate their self-reliance on their protection against mosquito biting.