Larvicidal activity of Stemona collinsiae root extract against Musca domestica and Chrysomya megacephala.

Musca domestica and Chrysomya megacephala, considered synanthropic insects, are medically important flies, as they transmit vector-borne diseases to humans and animals. In Thailand, Stemona (Stemonaceae) plants have been traditionally used as insecticides. This study was designed to determine the larvicidal activity of S. collinsiae root extract against M. domestica and C. megacephala larvae. A 70% ethanol crude extract from S. collinsiae roots was tested against the third-instar larvae of both species using direct and indirect contact methods. The development and mortality rates of the insects were observed, and the LC values were calculated. The extract caused irregular development in both species, shown as segmental puparia that could not emerge as adult flies. The LC50 values of the extract against M. domestica tested by direct and indirect contact methods were 0.0064 ± 0.0005 mg/larva and 0.0165 ± 0.0002 mg/cm2/larva, respectively. In the case of C. megacephala, the LC50 value determined by the indirect contact method was 1.0500 ± 0.0001 mg/cm2/larva. The ethanolic root extract of S. collinsiae was able to kill the larvae of both species after dermal administration. It is of interest to develop S. collinsiae root extract as a natural fly control biopesticide.

Larvicidal Activity of Synthesized Silver Nanoparticles from Curcuma zedoaria Essential Oil against Culex quinquefasciatus.

Culex quinquefasciatus is the major vector of the bancroftian filarial parasite which causes human lymphatic filariasis and St. Louis encephalitis. The simple way to stop the transmission is to control the vector by using synthetic chemicals. However, herbal essential oils have biological properties, such as a larvicidal effect and are ecofriendly to use. In this study, we investigated the larvicidal activity of Curcuma zedoaria essential oil (ZEO) and biosynthesized silver nanoparticles using this essential oil (ZEO-AgNPs). The larvicidal activity against both insecticide-susceptible and -resistant strains of Cx. quinquefasciatus larvae of ZEO were investigated and compared with ZEO-AgNPs. The ZEO-AgNPs showed the utmost toxicity against both strains of Cx. quinquefasciatus. After 24 h of exposure, LC50 and LC99 of ZEO against susceptible strain were 36.32 and 85.11 ppm, respectively. While LC50 and LC99 of ZEO against the resistant strain were 37.29 and 76.79 ppm, respectively. Whereas ZEO-AgNPs offered complete larval mortality within 24 h of exposure, LC50 and LC99 of ZEO-AgNPs against the susceptible strain, were 0.57 and 8.54 ppm, respectively. For the resistant strain, LC50 and LC99 values were 0.64 and 8.88 ppm, respectively. The potency in killing Cx. quinquefasciatus and stability of ZEO-AgNPs have made this product a good candidate for the development of novel natural larvicides.

Courtship activity, copulation & insemination success in a mosquito vector fed a herbal aphrodisiac: Implications for sterile insect technology.

BACKGROUND & OBJECTIVES: In sterile insect technology (SIT), mating competitiveness is a pre-condition for the reduction of target pest populations and a crucial parameter for judging efficacy. Still, current SIT trials are being hindered by decreased effectiveness due to reduced sexual performance of released males. Here, we explored the possible role of a herbal aphrodisiac in boosting the mating activity of Aedes aegypti. METHODS: Males were fed one of two diets in this study: experimental extract of Eurycoma longifolia (MSAs) and sugar only (MSOs). Differences in life span, courtship latency, copulation activity and mating success were examined between the two groups. RESULTS: No deaths occurred among MSA and MSO males. Life span of MSOs was similar to that of MSAs. The courtship latency of MSAs was shorter than that of MSOs (P<0.01). MSAs had greater copulation success than MSOs (P<0.001). In all female treatments, MSAs mated more than MSOs, but the differences in rate were significant only in the highest female density (P<0.05). In MSAs, mating success varied significantly with female density (P<0.01), with the 20-female group (P<0.01) having the lowest rate. Single MSA had better mating success at the two lowest female densities. In MSOs, there were no significant differences in mating success rate between the different female densities. INTERPRETATION & CONCLUSIONS: Our results suggested that the herbal aphrodisiac, E. longifolia, stimulated the sexual activity of Ae. aegypti and may be useful for improving the mating competitiveness of sterile males, thus improving SIT programmes.

Coffee, its roasted form, and their residues cause birth failure and shorten lifespan in dengue vectors.

In dengue mosquitoes, successful embryonic development and long lifespan are key determinants for the persistence of both virus and vector. Therefore, targeting the egg stage and vector lifespan would be expected to have greater impacts than larvicides or adulticides, both strategies that have lost effectiveness due to the development of resistance. Therefore, there is now a pressing need to find novel chemical means of vector control. Coffee contains many chemicals, and its waste, which has become a growing environmental concern, is as rich in toxicants as the green coffee beans; these chemicals do not have a history of resistance in insects, but some are lost in the roasting process. We examined whether exposure to coffee during embryonic development could alter larval eclosion and lifespan of dengue vectors. A series of bioassays with different coffee forms and their residues indicated that larval eclosion responses of Aedes albopictus and Ae. aegypti were appreciably lower when embryonic maturation occurred in environments containing coffee, especially roasted coffee crude extract (RCC). In addition, the lifespan of adults derived from eggs that hatched successfully in a coffee milieu was reduced, but this effect was less pronounced with roasted and green coffee extracts (RCU and GCU, respectively). Taken together, these findings suggested that coffee and its residues have embryocidal activities with impacts that are carried over onto the adult lifespan of dengue vectors. These effects may significantly reduce the vectorial capacity of these insects. Reutilizing coffee waste in vector control may also represent a realistic solution to the issues associated with its pollution.

Insecticidal activity of Stemona collinsiae root extract against Parasarcophaga ruficornis (Diptera: Sarcophagidae).

In Thai indigenous knowledge, Stemona plant has traditionally been used as insecticide on plantations. Recently, S. collinsiae extract was showed to be an anti-feedant and growth inhibitor and to exert strong insecticidal activity. Here, the insecticidal activity of S. collinsiae root extract against Parasarcophaga ruficornis is studied. The larvicidal and pupicidal activities of the ethanolic root extract of S. collinsiae were tested using contact toxicity tests, and adulticidal activity was tested using the topical contact toxicity and sugar bait methods The ethanolic extract at concentration ranging of 0.3-320mg/larva for the direct contact toxicity test and from 0.3 to 3.2mg/cm2/larva for the secondary contact toxicity test showed 3.0-51.0 and 1.0-94.0% corrected mortality, respectively. Against third-instar larvae, the LD50 concentrations of the ethanolic extract were 31.7±0.0mg/larva and 1.4±0.0mg/cm2/larva for direct and secondary contact toxicity tests, respectively. Pupae were not eliminated at all concentrations of the ethanolic extract. Against adult flies, which were killed via oral administration, the LD50 concentration of the ethanolic extract was 0.145±0.070g extract/g glucose. Thus, the ethanolic extract of S. collinsiae was capable of eliminating P. ruficornis in larval and adult stages via topical and ingestion administration, respectively.

Sweet waste extract uptake by a mosquito vector: Survival, biting, fecundity responses, and potential epidemiological significance.

In nature, adult mosquitoes typically utilize nectar as their main energy source, but they can switch to other as yet unidentified sugary fluids. Contemporary lifestyles, with their associated unwillingness to consume leftovers and improper disposal of waste, have resulted in the disposal of huge amounts of waste into the environment. Such refuse often contains unfinished food items, many of which contain sugar and some of which can collect water from rain and generate juices. Despite evidence that mosquitoes can feed on sugar-rich suspensions, semi-liquids, and decaying fruits, which can be abundant in garbage sites, the impacts of sweet waste fluids on dengue vectors are unknown. Here, we investigated the effects of extracts from some familiar sweet home waste items on key components of vectorial capacity of Aedes aegypti. Adult mosquitoes were fed one of five diets in this study: water (WAT); sucrose (SUG); bakery product (remnant of chocolate cake, BAK); dairy product (yogurt, YOG); and fruit (banana (BAN). Differences in survival, response time to host, and egg production were examined between groups. For both males and females, maintenance on BAK extract resulted in marked survival levels that were similar to those seen with SUG. Sweet waste extracts provided better substrates for survival compared to water, but this superiority was mostly seen with BAK. Females maintained on BAK, YOG, and BAN exhibited shorter response times to a host compared to their counterparts maintained on SUG. The levels of egg production were equivalent in waste extract- and SUG-fed females. The findings presented here illustrate the potential of sweet waste-derived fluids to contribute to the vectorial capacity of dengue vectors and suggest the necessity of readdressing the issue of waste disposal, especially that of unfinished sweet foods. Such approaches can be particularly relevant in dengue endemic areas where rainfall is frequent and waste collection infrequent.

DNA barcoding and wing morphometrics to distinguish three Aedes vectors in Thailand.

Aedes aegypti (Diptera: Culicidae) (L.), Ae. albopictus (Skuse), and Ae. scutellaris (Walker) are important mosquito vectors of dengue and chikungunya viruses. They are morphologically similar and sympatric in some parts of their distribution; therefore, there is a risk of incorrect morphological identification. Any confusion could have a negative impact on epidemiological studies or control strategies. Therefore, we explored two modern tools to supplement current morphological identification: DNA barcoding and geometric morphometric analyses. Field larvae were reared to adults and carefully classified based on morphological traits. The genetic analysis was based on the 658bp each of 30COI sequences. Some Culex spp., Mansonia bonneae, were included as outgroups, and inclusion of a few other Aedes spp. facilitated phylogenetic inference of the relationship between Ae. albopictus and Ae. scutellaris. The two species were separated by an average interspecific divergence of 0.123 (0.119-0.127). Morphometric examination included landmark- (392 specimens) and outline-based (317 specimens) techniques. The shape of the wing showed different discriminating power based on sex and digitizing technique. This is the first time that Ae. scutellaris and Ae. albopictus have been compared using these two techniques. We confirm that these morphologically close species are valid, and that geometric morphometrics can considerably increase the reliability of morphological identification.

Efficacy of three insecticides against Anopheles dirus and Anopheles minimus, the major malaria vectors, in Kanchanaburi Province, Thailand.

We conducted this study to determine the insecticide susceptibility of two malaria vectors, Anopheles dirus and Anopheles minimus from Kanchanaburi Province, Thailand. The mosquitoes were collected and reared under laboratory conditions. The test was carried out on unfed F-1 female mosquitoes using a standard WHO testing protocol. The LD50 and LD90 of deltamethrin in both species were tested for by exposing the mosquitoes to various doses of deltamethrin for 1 hour. The lethal time was also tested among mosquitoes by exposing them to deltamethrin (0.05%), permethrin (0.75%) and malathion (5%), for different exposure times, ranging from 0.5 to 15 minutes. Percent knockdown at 60 minutes and mortality at 24 hours were calculated. The resistance ratio (RR) was determined based on the LD50 and LT50 values. LD50 of deltamethrin against An. dirus and An. minimus were 0.00077% and 0.00066%, respectively. LT50 values for deltamethrin (0.05%), permethrin (0.75%) and malathion (5%) against An. dirus and An. minimus were 1.20, 3.16 and 10.07 minutes and 0.48, 1.92 and 5.94 minutes, respectively. The study revealed slightly increased tolerance by both mosquito species, compared with laboratory susceptible strains, based on LD50 values. The two anopheline species had the same patterns of response to the three insecticides, based on LT50 values, although the LT50 values were slightly higher in the An. dirus population. Both An. dirus and An. minimus were fully susceptible to all the insecticides tested, with 100% mortality at 24 hours post-exposure. Deltamethrin was the most effective insecticide, followed by permethrin and malathion.

Identification of ricinoleic acid as an inhibitor of Ca2+ signal-mediated cell-cycle regulation in budding yeast.

Free fatty acids exhibit diverse biological effects such as the regulation of immune responses in humans and animals. To investigate the biological effect of fatty acids in the model eukaryotic organism yeast, we examined the activity of various fatty acids in a yeast-based drug-screening system designed to detect the small-molecule compounds that inhibit Ca(2+)-signal-mediated cell-cycle regulation. Among the fatty acids examined, ricinoleic acid markedly alleviated the deleterious physiological effects induced by the compelled activation of Ca(2+) signaling by external CaCl(2), such as the polarized bud growth and the growth arrest in the G(2) phase. In accordance with the physiological consequences induced by ricinoleic acid, it diminished the Ca(2+)-induced phosphorylation of Cdc28p at Tyr-19, concomitant with the decrease in the Ca(2+)-stimulated expression levels of Cln2p and Swe1p.