Wei-June Chen

Chemical compositions and larvicidal activities of leaf essential oils from two eucalyptus species.

In the current study, the mosquito larvicidal activity of leaf essential oils and their constituents from two eucalyptus species (Eucalyptus camaldulensis and Eucalyptus urophylla) against two mosquito species, Aedes aegypti and Aedes albopictus, was investigated. In addition, the chemical compositions of the leaf essential oils were analyzed using gas chromatography-mass spectrometry. Results from the larvicidal tests revealed that essential oil from the leaves of E. camaldulensis had an excellent inhibitory effect against both A. aegypti and A. albopictus larvae. The 12 pure constituents extracted from the two eucalyptus leaf essential oils were also tested individually against two mosquito larvae. Among the six effective constituents, alpha-terpinene exhibits the best larvicidal effect against both A. aegypti and A. albopictus larvae. Results of this study show that the leaf essential oil of E. camaldulensis and its effective constituents might be considered as a potent source for the production of fine natural larvicides.

Bioactivity of selected plant essential oils against the yellow fever mosquito Aedes aegypti larvae.

The bioactivity of 14 essential oils from five plants has been studied using the brine shrimp lethality test and the Aedes aegypti larvicidal assay. All essential oils screened had LC50 values smaller than 200 microg/ml, showing significant lethality against brine shrimp. In addition, nine of the 14 essential oils tested showed toxicity against the fourth-instar A. aegypti larvae in 24 h (LC50<100 microg/ml). Of these, the leaf and bark essential oils of Cryptomeria japonica demonstrated high larvicidal activity, the most active being the leaf essential oil of C. japonica, with a LC50=37.6 microg/ml (LC90=71.9 microg/ml), followed by the bark essential oil of C. japonica also showing high activity against A. aegypti larvae, with a LC50=48.1 microg/ml (LC90=130.3 microg/ml). The results obtained from this study suggest that the leaf and bark essential oils of C. japonica are promising as larvicides against A. aegypti larvae and could be useful in the search for new natural larvicidal compounds.

Variations in insecticidal activity and chemical compositions of leaf essential oils from Cryptomeria japonica at different ages

The larvicidal effects of the essential oils extracted from the leaves of Cryptomeria japonica at different ages (58, 42, and 26 years old) against 2 mosquito species, Aedes aegypti and Aedes albopictus, were studied. The analysis of major constituents of these essential oils was also investigated. Results obtained from the larvicidal tests, using essential oil from the leaves of 58-year-old C. japonica was found to be most effective against both A. aegypti and A. albopictus larvae, indicating tree age has significant influence on mosquito larvicidal activity. In addition, the eleven pure constituents from C. japonica leaf essential oil were also tested individually against the two mosquito larvae. Among them, alpha-terpinene, gamma-terpinene, p-cymene, 3-carene, terpinolene, and beta-myrcene shows strong larvicidal effect against the two mosquito larvae. Among these pure constituents, 3-carene exhibits the best larvicidal effect against A. aegypti and terpinolene shows an excellent inhibitory action against A. albopictus larvae. The results of this study show that the leaf essential oil and its effective constituents might be considered as a potent source for the production of fine natural larvicides.

Insecticidal activities of leaf essential oils from Cinnamomum osmophloeum against three mosquito species.

The larvicidal activities of leaf essential oils and their constituents from six chemotypes of indigenous cinnamon (Cinnamomum osmophloeum Kaneh.) trees were evaluated against three mosquito species. Results of larvicidal tests demonstrated that the leaf essential oils of cinnamaldehyde type and cinnamaldehyde/cinnamyl acetate type had an excellent inhibitory effect against Aedes albopictus larvae, and their LC(50) values in 24h were 40.8 microg/ml (LC(90)=81.7 microg/ml) and 46.5 microg/ml (LC(90)=83.3 microg/ml), respectively. Results of the 24-h mosquito larvicidal assays also showed that the effective constituents in leaf essential oils were trans-cinnamaldehyde and benzaldehyde and that the LC(50) values of these constituents against A. albopictus larvae were below 50 mug/ml. In addition, cinnamaldehyde type leaf essential oil and trans-cinnamaldehyde have also exhibited great larvicidal performance against Culex quinquefasciatus and Armigeres subalbatus larvae. Comparisons of mosquito larvicidal activity of trans-cinnamaldehyde congeners revealed that alpha-methyl cinnamaldehyde, benzaldehyde, and trans-cinnamaldehyde exhibited strong mosquito larvicidal activity.

Insecticidal activities of leaf and twig essential oils from Clausena excavata against Aedes aegypti and Aedes albopictus larvae.

BACKGROUND The current study investigates, for the first time, the mosquito larvicidal activities of leaf and twig essential oils from Clausena excavata Burm. f. and their individual constituents against Aedes aegypti L. and Aedes albopictus Skuse larvae. The yields of essential oils obtained from hydrodistillation were compared, and their constituents were determined by GC-MS analyses. RESULTS The LC(50) values of leaf and twig essential oils against fourth-instar larvae of Ae. aegypti and Ae. albopictus were 37.1-40.1 microg mL(-1) and 41.1-41.2 microg mL(-1) respectively. This study demonstrated that C. excavata leaf and twig essential oils possess mosquito larvicidal activity, inhibiting the growth of mosquito larvae for both species at a low concentration. In addition, results of larvicidal assays showed that the effective constituents in leaf and twig essential oils were limonene, gamma-terpinene, terpinolene, beta-myrcene, 3-carene and p-cymene. The LC(50) values of these constituents against both mosquito larvae were below 50 microg mL(-1). Among these effective constituents, limonene had the best mosquito larvicidal activity, with LC(50) of 19.4 microg mL(-1) and 15.0 microg mL(-1) against Ae. aegypti and Ae. albopictus larvae respectively. CONCLUSION The findings suggested that the essential oils from Clausena excavata leaf and twig and their effective constituents may be explored as a potential natural larvicide.

Potential Role of Armigeres subalbatus (Diptera: Culicidae) in the Transmission of Japanese Encephalitis Virus in the Absence of Rice Culture on Liu-Chiu Islet, Taiwan

Abstract Mosquitoes known to be involved in the transmission of Japanese encephalitis virus (JE) on Taiwan typically develop in rice fields. However, recent serological evidence indicated that JE virus was being transmitted on Liu-Chiu, a rice-free islet. To identify the mosquito vector in this unusual epidemiological situation, 4 mosquito species commonly found in Liu-Chiu were evaluated for their vector competence for a strain of JE (CH1392) virus isolated from central Taiwan. Armigeres subalbatus (Coquillett) was the most susceptible species, indicating its status as a potential vector. In addition, an isolate of JE (T1P1) virus from Ar. subalbatus collected on Liu-Chiu readily infected the salivary glands of orally infected Ar. subalbatus originating from Liu-Chiu. The infection rate reached 79% (11/14) after a 20-d period of extrinsic incubation at 28°C. We conclude that JE likely was transmitted between vertebrate hosts by Ar. subalbatus in this rice-free islet and that this species should be considered as a potential vector in similar ecological conditions.

Parallel infection of Japanese encephalitis virus and Wolbachia within cells of mosquito salivary glands.

Abstract The endosymbiont Wolbachia usually causes cytoplasmic incompatibility in dipteran hosts, including mosquitoes. However, some important arbovirus-transmitting mosquitoes such as Aedes aegypti (L.) are not heritably infected by Wolbachia. In Wolbachia-harboring mosquito Armigeres subalbatus Coquillett, colocalization of Wolbachia and inoculated Japanese encephalitis virus (family Flaviviridae, genus Flavivirus, JEV) in salivary gland (SG) cells was shown by electron microscopy. The infection rate of JEV in SGs, detected with either immunofluorescent antibody test or reverse transcription-polymerase chain reaction, did not show significant differences between Wolbachia-infected and -free colonies. It is suggested that Wolbachia did not mediate resistance of SG cells to superinfection by JEV, although both microorgamisms coexist in the same niche, i.e., the same SG cell. Therefore, a SG escape barrier may not be elevated due to Wolbachia infection, which presumably has no deleterious effects on vector competence in Wolbachia-harboring mosquitoes.

Molecular (sub) grouping of endosymbiont Wolbachia infection among mosquitoes of Taiwan.

Abstract Wolbachia are maternally inherited bacteria that infect a wide range of arthropods as well as filarial worms. The infection usually results in reproductive distortions of the host, primarily cytoplasmic incompatibility, parthenogenesis, and feminization. This study showed that Wolbachia infection (15/29; 51.72%) was prevalent among field-caught mosquitoes in Taiwan. Three mosquito species were identified as having Wolbachia A infection, eight species as having Wolbachia B, and four other species were dually infected by both groups. Each Wolbachia isolate from different mosquitoes was further divided into a specific subgroup. However, there were still some isolates that did not belong to any known subgroup, suggesting that more subgroups remain to be identified. Investigation of tissue tropism in either Aedes albopictus (Skuse) or Armigeres subalbatus (Coquillett) revealed that Wolbachia were extensively distributed within the host, although the ovary was most susceptible to infection. This report provides preliminary features of molecular relationships among Wolbachia groups of mosquitoes from Taiwan.

Intestinal Expression of H+ V‐ATPase in the Mosquito Aedes albopictus is Tightly Associated with Gregarine Infection

ABSTRACT. Vacuolar ATPase (V‐ATPase) is a family of ATP‐dependent proton pumps expressed on the plasma membrane and endomembranes of eukaryotic cells. Acidification of intracellular compartments, such as lysosomes, endosomes, and parasitophorous vacuoles, mediated by V‐ATPase is essential for the entry by many enveloped viruses and invasion into or escape from host cells by intracellular parasites. In mosquito larvae, V‐ATPase plays a role in regulating alkalization of the anterior midgut. We extracted RNA from larval tissues of Aedes albopictus, cloned the full‐length sequence of mRNA of V‐ATPase subunit A, which contains a poly‐A tail and 2,971 nucleotides, and expressed the protein. The fusion protein was then used to produce rabbit polyclonal antibodies, which were used as a tool to detect V‐ATPase in the midgut and Malpighian tubules of mosquito larvae. A parasitophorous vacuole was formed in the midgut in response to invasion by Ascogregarina taiwanensis, confining the trophozoite(s). Acidification was demonstrated within the vacuole using acridine orange staining. It is concluded that gregarine sporozoites are released by ingested oocysts in the V‐ATPase‐energized high‐pH environment. The released sporozoites then invade and develop in epithelial cells of the posterior midgut. Acidification of the parasitophorous vacuoles may be mediated by V‐ATPase and may facilitate exocytosis of the vacuole confining the trophozoites from the infected epithelial cells for further extracellular development.

Repellency of essential oils of Cryptomeria japonica (Pinaceae) against adults of the mosquitoes Aedes aegypti and Aedes albopictus (Diptera:Culicidae).

The purpose of this study was to investigate the repellent activities of essential oils from Cryptomeria japonica (sugi) against adults of mosquitoes Aedes aegypti and Aedes albopictus . Comparison of essential oils from four different plant parts of C. japonica revealed that essential oil from its leaf exhibited the best repellent activity against mosquitoes. To understand the relationship between volatile organic compounds and repellent activity, the solid-phase microextraction (SPME) method was employed to analyze volatile organic compounds of leaf essential oil. The SPME fiber was coated with divinylbenzene/carboxen/polydimethylsiloxane (DVB/CAR/PDMS). The major volatile organic compounds in the cage were 3-carene, alpha-terpinene, limonene, gamma-terpinene, and terpinolene at 0 min. Results demonstrated that (-)-terpinen-4-ol was the major volatile organic compound adsorbed by SPME fiber during repellent assays. Furthermore, the repellent activities of six compounds against adults of the mosquitoes were evaluated, and the results revealed that (-)-terpinen-4-ol exhibited the best repellent activity against A. aegypti and A. albopictus.