Udom Chaithong

Larvicidal effect of pepper plants on Aedes aegypti (L.) (Diptera: Culicidae)

ABSTRACT Ethanolic extracts derived from three species of the Piperaceae (pepper) family, Piper longum L., P. ribesoides Wall., and P. sarmentosum Roxb. ex Hunt., were evaluated for efficacy against early 4th instar larvae of Aedes aegypti mosquitoes using larvicidal bioassays. The highest larvicidal efficacy was established from P. longum, followed by P. sarmentosum and P. ribesoides, with LC50 values of 2.23, 4.06, and 8.13 ppm, respectively. Observations of morphological alterations on treated 4th instar larvae revealed that most organs, except anal papillae, had a normal structural appearance that was similar to controls. Under light microscopy, the internal structures of anal papillae in the treated larvae showed shrinkage, while the external features were normal in appearance. Ultrastructural studies, however, clearly demonstrated external destruction, with extensive damage and shrunken cuticle of the anal papillae. The structural deformation of anal papillae probably led to their dysfunction, which may be intrinsically associated with the death of the larvae. This study affords some evidence regarding the action site of the pepper extracts and suggests their potential in developing new types of larvicides used for mosquito control.

Chemical Composition and Larvicidal Activity of Edible Plant-Derived Essential Oils Against the Pyrethroid-Susceptible and -Resistant Strains of Aedes aegypti (Diptera: Culicidae)

ABSTRACT: The chemical compositions and larvicidal potential against mosquito vectors of selected essential oils obtained from five edible plants were investigated in this study. Using a GC/MS, 24, 17, 20, 21, and 12 compounds were determined from essential oils of Citrus hystrix, Citrus reticulata, Zingiber zerumbet, Kaempferia galanga, and Syzygium aromaticum, respectively. The principal constituents found in peel oil of C. hystrix were &bgr;-pinene (22.54%) and d-limonene (22.03%), followed by terpinene-4-ol (17.37%). Compounds in C. reticulata peel oil consisted mostly of d-limonene (62.39%) and y-terpinene (14.06%). The oils obtained from Z. zerumbet rhizome had &agr;-humulene (31.93%) and zerumbone (31.67%) as major components. The most abundant compounds in K. galanga rhizome oil were 2-propeonic acid (35.54%), pentadecane (26.08%), and ethyl-p-methoxycinnamate (25.96%). The main component of S. aromaticum bud oil was eugenol (77.37%), with minor amounts of trans-caryophyllene (13.66%). Assessment of larvicidal efficacy demonstrated that all essential oils were toxic against both pyrethroid-susceptible and resistant Ae. aegypti laboratory strains at LC50, LC95, and LC99 levels. In conclusion, we have documented the promising larvicidal potential of essential oils from edible herbs, which could be considered as a potentially alternative source for developing novel larvicides to be used in controlling vectors of mosquitoborne disease.

Adulticidal activity against Stegomyia aegypti (Diptera: Culicidae) of three Piper spp.

Three Piper species, Piper longum, P. ribesoides and P. sarmentosum, were selected for investigation of adulticidal potential against Stegomyia aegypti, a main vector of dengue and dengue haemorrhagic fever. Successive extraction by maceration with 95% ethanol showed percentage yields of ethanolic extracts, which derived from P. longum, P. ribesoides and P. sarmentosum, of 8.89, 3.21 and 5.30% (w/w), respectively. All Piper extracts illustrated an impressive adulticidal activity when tested against female mosquitoes by topical application. The susceptibility of St. aegypti females to ethanol-extracted Piper was dose dependent and varied among the plant species. The highest adulticidal effect was established from P. sarmentosum, followed by P. ribesoides and P. longum, with LD50 values of 0.14, 0.15 and 0.26 microg/female, respectively. The potential of these Piper species, as possible mosquitocides, established convincing activity for further researches to develop natural substances for combat against adult mosquitoes.

Repellent properties of celery, Apium graveolens L., compared with commercial repellents, against mosquitoes under laboratory and field conditions

In our search for new bioactive products against mosquito vectors, we reported the slightly larvicidal and adulticidal potency, but remarkable repellency of Apium graveolens both in laboratory and field conditions. Repellency of the ethanolic preparation of hexane‐extracted A. graveolens was, therefore, investigated and compared with those of 15 commercial mosquito repellents including the most widely used, DEET. Hexane‐extracted A. graveolens showed a significant degree of repellency in a dose‐dependent manner with vanillin added. Ethanolic A. graveolens formulations (10–25% with and without vanillin) provided 2–5 h protection against female Aedes aegypti. Repellency that derived from the most effective repellent, 25% of hexane‐extracted A. graveolens with the addition of 5% vanillin, was comparable to the value obtained from 25% of DEET with 5% vanillin added. Moreover, commercial repellents, except formulations of DEET, showed lower repellency than that of A. graveolens extract. When applied on human skin under field conditions, the hexane‐extracted A. graveolens plus 5% vanillin showed a strong repellent action against a wide range of mosquito species belonging to various genera. It had a protective effect against Aedes gardnerii, Aedes lineatopennis, Anopheles barbirostris, Armigeres subalbatus, Culex tritaeniorhynchus, Culex gelidus, Culex vishnui group and Mansonia uniformis. The hexane‐extracted A. graveolens did not cause a burning sensation or dermal irritation when applied to human skin. No adverse effects were observed on the skin or other parts of the human volunteers’ body during 6 months of the study period or in the following 3 months, after which time observations ceased. Therefore, A. graveolens can be a potential candidate for use in the development of commercial repellents that may be an alternative to conventional synthetic chemicals, particularly in community vector control applications.

Mosquito repellency of the seeds of celery (Apium graveolens L.).

Abstract When the mosquito repellencies of four fractions of Apium graveolens seeds (one hexane, two dichloromethane and one methanolic) were investigated in the laboratory, all four were found to offer human volunteers some protection against female, adult Aedes aegypti. The hexane fraction, however, was found to exhibit the highest repellency in the laboratory, with median effective doses (ED50) and ED95 of 0.41 and 2.93 mg/cm2 skin, respectively. Only this fraction, which was also found to provide protection against mosquito bites for 3.5 h when applied, in the laboratory, at a concentration of 250 mg/ml, was then investigated for its repellency in the field and its stability. In storage, it was found to retain its repellency for at least 2 months, although significant reductions in its repellency were observed (in terms of shortened complete-protection times) after 3 months, whatever the temperature of storage (-20°C, 4°C, or room temperature). When applied to the skin of volunteers under field conditions, the hexane fraction showed strong repellent activity against a wide range of mosquito species belonging to various genera (Ae. gardnerii, Ae. lineatopennis, Armigeres subalbatus, Culex tritaeniorhynchus, Cx. vishnui group, Cx. quinquefasciatus and Mansonia uniformis). It appeared not to cause dermal irritation or any other adverse effect, either during 6 months of use or in the following 3 months of follow-up. Mosquito repellents based on extracts of Ap. graveolens seeds could be developed commercially, as an effective personal-protection measure against mosquito bites and the diseases caused by mosquito-borne pathogens.

Susceptibility of eight species members in the Anopheles hyrcanus group to nocturnally subperiodic Brugia malayi

BackgroundFilariasis, caused by Brugia malayi, is a public health problem in Thailand. Currently, at least two locations in southern Thailand are reported to be active endemic areas. Two and four Mansonia species are primary and secondary vectors, respectively, of the nocturnally subperiodic race, whereas, Coquillettidia crassipes is a vector of the diurnally subperiodic race. Although several Anopheles species have been incriminated extensively as natural and/or suspected vectors of B. malayi, little is known about vector competence between indigenous Anopheles and this filaria in Thailand. Thus, the susceptibility levels of eight species members in the Thai An. hyrcanus group to nocturnally subperiodic B. malayi are presented herein, and the two main refractory factors that affect them in different degrees of susceptibility have been elucidated.MethodsAedes togoi (a control vector), An. argyropus, An. crawfordi, An. nigerrimus, An. nitidus, An. paraliae, An. peditaeniatus, An. pursati and An. sinensis were allowed to feed artificially on blood containing B. malayi microfilariae, and dissected 14 days after feeding. To determine factors that take effect at different susceptibility levels, stain-smeared blood meals were taken from the midguts of Ae. togoi, An. peditaeniatus, An. crawfordi, An. paraliae, An. sinensis and An. nitidus immediately after feeding, and their dissected-thoraxes 4 days post blood-feedings were examined consecutively for microfilariae and L1 larvae.ResultsThe susceptibility rates of Ae. togoi, An. peditaeniatus, An. crawfordi, An. nigerrimus, An. argyropus, An. pursati, An. sinensis, An. paraliae and An. nitidus to B. malayi were 70–95%, 70–100%, 80–85%, 50–65%, 60%, 60%, 10%, 5%, and 0%, respectively. These susceptibility rates related clearly to the degrees of normal larval development in thoracic muscles, i.e., Ae. togoi, An. peditaeniatus, An. crawfordi, An. paraliae, An. sinensis and An. nitidus yielded normal L1 larvae of 93.15%, 96.34%, 97.33%, 23.60%, 15.38% and 0%, respectively.ConclusionsAn. peditaeniatus, An. crawfordi, An. nigerrimus, An. argyropus and An. pursati were high potential vectors. An. paraliae and An. sinensis were low potential vectors, while An. nitidus was a refractory vector. Two refractory mechanisms; direct toxicity and/or melanotic encapsulation against filarial larval were involved in the refractoriness of development in the thoracic muscles of the mosquito.

Fine structure of the alimentary canal of the larval blow fly Chrysomya megacephala (Diptera: Calliphoridae)

Morphology of the alimentary canal of the mature third instar larva of the blow fly, Chrysomya megacephala (F.), was examined using light, scanning, and transmission electron microscopy. Salivary structures consist of a single median deferent duct that bifurcates into efferent ducts connected to paired, tubular salivary glands comprised of closely packed conical-shaped epithelial cells with large nuclei. The crop occurs as a large, swollen diverticulum of the digestive tube and is lined internally with convoluted cuticle (epicuticle and endocuticle). The esophagus is a simple, straight tube internally lined with cuticle and externally encompassed by muscle fibers. The cardia is a bulb-like structure composed of anterior foregut tissue and posterior midgut tissue from which the peritrophic membrane (PM) is produced. The midgut begins within the cardia which is flanked posteriorly by four tubular gastric caeca that are lined internally with four to five layers of cuboidal epithelial cells bearing microvilli. Midgut tissue is lined with simple cuboidal epithelium whose cells are filled with numerous secretory granules and possessed long microvilli facing the lumen. A peritrophic membrane is contained within the midgut lumen. The larval hindgut consists of the pylorus, Malpighian tubules, ileum, colon, rectum, and anus, posteriorly. The pylorus is characterized by a single layer of epithelial cells encircled by a muscular layer and the presence of PM within the lumen. Malpighian tubules each diverge into two tubular structures totalling four long tubules of long chained cuboidal cells bearing microvilli internally. The wall of the ileum is comprised primarily of a monolayer of cuboidal epithelial cells with large oval nuclei and more intense muscular fibers surrounding the periphery. A cuticular layer surrounds the lumen containing the PM. This inner cuticle consists of a thin epicuticle that is electron-dense; whereas, the endocuticle is much thicker but less electron-dense. Myo-epithelial cells are dense in the anal region, where the PM persists.

Chemical Constituents and Combined Larvicidal Effects of Selected Essential Oils against Anopheles cracens (Diptera: Culicidae)

A preliminary study on larvicidal activity against laboratory-colonized Anopheles cracens mosquitos revealed that five of ten plant oils at concentration of 100 ppm showed 95–100% larval mortality. The essential oils of five plants, including Piper sarmentosum, Foeniculum vulgare, Curcuma longa, Myristica fragrans, and Zanthoxylum piperitum, were then selected for chemical analysis, dose-response larvicidal experiments, and combination-based bioassays. Chemical compositions analyzed by gas chromatography coupled to mass spectrometry demonstrated that the main component in the oil derived from P. sarmentosum, F. vulgare, C. longa, M. fragrans, and Z. piperitum was croweacin (71.01%), anethole (63.00%), ar-turmerone (30.19%), safrole (46.60%), and 1,8-cineole (21.27%), respectively. For larvicidal bioassay, all five essential oils exerted promising efficacy in a dose-dependent manner and different performances on A. cracens after 24 hours of exposure. The strongest larvicidal potential was established from P. sarmentosum, followed by F. vulgare, C. longa, M. fragrans, and Z. piperitum, with LC50 values of 16.03, 32.77, 33.61, 40.00, and 63.17 ppm, respectively. Binary mixtures between P. sarmentosum, the most effective oil, and the others at the highest ratio were proved to be highly efficacious with a cotoxicity coefficient value greater than 100, indicating synergistic activity. Results of mixed formulations of different essential oils generating synergistic effects may prove helpful in developing effective, economical, and ecofriendly larvicides, as favorable alternatives for mosquito management.

Field evaluation of G10, a celery ( Apium graveolens )-based topical repellent, against mosquitoes (Diptera: Culicidae) in Chiang Mai province, northern Thailand

The potential of G10, a celery (Apium graveolens)-based topical product, as a repellent against natural mosquito populations was evaluated in comparison to commercial (Insect Block 28) and standard (25% DEET) repellents in Chiang Mai province, northern Thailand. These repellent products afforded encouragingly excellent personal protection against a broad range of mosquito species belonging to various genera, including Aedes, Anopheles, Armigeres, Culex, and Mansonia. No mosquito bite was observed on the volunteers treated with G10 and Insect Block 28 throughout the field study, whereas two species, i.e., six A. barbirostris and two A. subalbatus, came to bite or land on 25% DEET-treated volunteers. Thus, it can be concluded that while G10 and Insect Block 28 exhibited similarly powerful repellent activities with complete (100%) protection, 25% DEET was effective in minimizing bites with 99.68% protection. G10 formula was also studied for physical properties and biological stability after being kept under two conditions; a heating and cooling cycle, and varying temperature and time storage. Most samples of stored G10 not only demonstrated a similarity in appearance and physical properties, but also provided comparable repellency to that of the fresh preparation. These findings encourage commercial development of G10 formula as an alternative to conventional synthetic repellents.

Fine structure of the reproductive system of Chrysomya megacephala (Diptera: Calliphoridae): the external sexual organ

The fine structure of the reproductive system of Chrysomya megacephala (Fabricius), a blow fly species of medical importance, was studied using scanning electron microscopy (SEM) to contribute detailed morphological information on the external sexual organs or genitalia of this species. Analysis of the male genitalia or hypopygium revealed that the epandrium is a broad organ resembling a crescent shape, while the ejaculatory apodeme and aedeagal apodeme show similarity in their lengths. The cercus is significantly longer than the surstylus with its apical end being more or less rounded. Dense patches of long bristles, morphologically similar to the sensilla chaetica and sensilla trichodea, were found along the lower half of the cercus. The surstylus has a stout triangular shape, and the proximal half is greatly endowed with sensilla thought to be sensilla chaetica and sensilla trichodea. The aedeagus per se is prominent and has a clavate shape formed by the base theca and elongated phallus. The vesica of the phallus is a smooth bilobed structure, which curves inward. The juxta and juxta process are armed with many rows of strong spines resembling thorns in their appearance. The harpe is slender, recurved anteriorly, and distally pointed like a sickle. Regarding the female genitalia or ovipositor, SEM observation of the supraanal plate revealed it to be sclerotized and more or less triangular-shaped, lying between a pair of short cerci. Several types of sensilla were found on the supraanal plate, subanal plate and cercus, including the sensilla trichodea, sensilla basiconica, sensilla placodea, and probably, sensilla styloconica. The possible functions of sensilla distributed in particular regions of the fly genitalia are discussed.