Yih-Shen Hwang

Isolation and identification of mosquito repellents inArtemisia vulgaris

The mugwortArtemisia vulgaris L. (Compositae: Anthemideae) contains insect repellents which can be released from the plant tissues by combustion. Work was carried out to isolate and identify the repellent compounds. The dried, pulverized whole plants were steam-distilled to give a repellent essential oil which was fractionated by column chromatography. Active fractions were analyzed by capillary GC and by combined GC-MS. A number of compounds, mainly monoterpenoids, were identified. When tested as repellents against the yellow fever mosquitoAedes aegypti L. (Diptera: Culicidae), (±)-linalool, (±)-camphor, (+)-camphor, (−)-camphor, isoborneol, (−)-borneol, terpinen-4-ol, and isobornyl acetate were active at 0.14 mg/cm2 or higher. Nonanone-3, (α+β)-thujone, and bornyl acetate were active at 0.28 mg/cm2 or higher. β-Pinene, myrcene, α-terpinene, (+)− limonene, and cineole were active at 1.4 mg/cm2. Of the repellent compounds identified, terpinen-4-ol was the most active and was as effective as dimethyl phthalate.

Attractancy and species specificity of 6-acetoxy-5-hexadecanolide, a mosquito oviposition attractant pheromone.

Four stereoisomers of 6-acetoxy-5-hexadecanolide, a mosquito oviposition attractant pheromone, were bioassayed for their activity on mosquitoes. Only (−)-(5R,6S) isomer was active in attractingCulex quinquefasciatus Say females for oviposition at dosages of 0.5 μg/100 ml water and above with the floating-cap method. The activity of this isomer increased 50-fold when it was applied directly to the water surface. The other three isomers, (+)-(5S,6R), (+)-(5R,6R), and (−)-(5s, 6s), were not active. The active isomer was ovipositionally attractive not only toC. quinquefasciatus but also toC. tarsalis Coquillett; however, it was 100 times more active in the former than in the latter species.Aedes aegypti (L.) andAnopheles quadrimaculatus Say were not attracted to the pheromone, thus suggesting that the pheromone is genus-specific.

Allelochemics produced by the hydrophyteMyriophyllum spicatum affecting mosquitoes and midges

An extract of the hydrophyteMyriophyllum spicatum L. was found to be toxic to first- and fourth-instar larvae of the mosquitoes,Culex quinquefasciatus Say,Culex tarsalis Coquillett,Culiseta incidens (Thomson),Aedes aegypti L., and chironomid midges in the laboratory. When first-stage larvae were exposed to the extract, the lowest concentration (6.4 mg extract/100 ml H2O) produced 86,60, and 48% mortality inC. incidens, C. quinquefasciatus, andA. aegypti, respectively. Higher concentrations caused 100% mortality in both first and fourth instars (63.75 mg/100 ml H2O or more). In general, the biocidal activity of the extract was found to be similar when first- and fourth-stage larvae were exposed. Some mortality in the successive pupal and adult stages was observed when fourth-stage larvae were exposed to the extract, but mortality occurred only in the various larval instars when first instars were exposed. Among the chironomids,Tanytarsus spp. was more tolerant to the extract thanChironomus spp. In addition to the biocidal activity against immature mosquitoes, the extract was found to show a unique activity acting as an attractant to both sexes of adultC. quinquefasciatus andA. aegypti. The response elicited inA. aegypti appeared to be somewhat higher than that inC. quinquefasciatus. In general, the percentage of males responding to water treated with the extract was higher than that of the females. The extract at higher concentrations seemed to show some repellency at the outset to both species, but induced positive responses later on.

Biocidal activity of algal toxins against immature mosquitoes

Extracts of two species of green algae, filamentousRhizoclonium hieroglyphicum Kütz and a phytoplankton,Chlorella ellipsoidea Gerneck, obtained with solvents in the laboratory were assayed againstAedes aegypti L.,Culex quinquefasciatus Say, andCuliseta incidens (Thomson). On extraction with petroleum ether, groundR. hieroglyphicum yielded an active crude extract which was chromatographed on a neutral alumina column and eluted consecutively with petroleum ether, benzene, and methanol. All three eluted fractions were found to induce significant mortality in test mosquito species. The benzene-eluted fraction was the least toxic. The methanol-eluted fraction was the most toxic to all species and exhibited juvenile hormone-like activity; it also caused morphogenetic changes in emerging adults. All three fractions delayed the rate of development of mosquito larvae by 2–5 days. Three supernatants ofC. ellipsoidea obtained on different occasions were tested against first instars ofC. quinquefasciatus. After the confirmation of their activity, all supernatants were extracted with diethyl ether, combined, and assayed against first- and fourth-stage larvae of the three mosquito species. The first-stage larvae ofC. quinquefasciatus andC. incidens were approximately three times more suspectible than those ofA. aegypti. However, fourth-stage larvae of the former two species were about twice as susceptible to the extract as those of the latter species. Dead first-stage larvae of all the species had a shrunken appearance. In general,C. ellipsoidea extract was quicker acting than that ofR. hieroglyphicum.