Larvicidal and adulticidal activities of castor oil against the dengue vector, Aedes aegypti

@#Plants contain numerous bioactive compounds that can be used to develop environmentally-safe insect control agents. Castor oil is a vegetable oil derived from the seeds of castor bean and is widely used as an industrial lubricant and medicinal purgative. In search of an alternative natural insecticide, the objective of this study was to evaluate the larvicidal and adulticidal activities of castor oil against the important dengue vector, Ae. aegypti. Larvicidal and adulticidal bioassays were conducted following the World Health Organization methods. Larvicidal activity was observed at castor oil concentrations of 10, 25, 50, 75, and 100 ppm; larval mortality was checked after 48 h of exposure and the lethal concentration (LC) at LC50 and LC90 were 51.38 and 116.26 ppm, respectively. Adulticidal activity was determined by topical application at the concentrations of 1, 5, 10, 15, and 20 µg/ mg on female mosquitoes and the mortality was checked after 24 h of exposure. The effective adulticidal activity was apparent with the LD50 and LD90 values of 6.03 and 25.07 µg/mg against female mosquitoes. The results indicated that castor oil has potential in the practical control of both immature and adult stages of the mosquito vector.

Adulticide efficacy of essential oil from Piper retrofractum Vahl against Aedes aegypti and Culex quinquefasciatus

Essential oil was extracted from the fresh fruit of Piper retrofractum Vahl by water distillation method. The yield of extraction was estimated at 0.125%. Adulticidal activity was tested against 2-5 days-old non-blood-fed female Aedes aegypti and Culex quinquefasciatus by topical application. Six concentrations of essential oil dissolved in acetone were prepared in 5%, 7%, 9%, 11%, 13% and 15%. The solution (0.5 μL) was on the pronotum of mosquito, and mortality was observed after 24 hours of exposure. The LD50 and LD99 of essential oil in acetone against Ae. aegypti (8.86%, 23.21%) and Cx. quinquefasciatus (6.95% and 17.35%,) was measured. This is the first report of adulticidal activities of essential oil from P. retrofractum against mosquito vectors.

Three indigenous Thai medicinal plants for control of Aedes aegypti and Culex quinquefasciatus.

The potential larvicidal activity and insect growth regulator (IGR) properties of three selected indigenous medicinal Thai plants were tested against two species of mosquito with special reference to the late 3rd and early 4th instar larvae (L3 and L4, respectively). In case of larvicidal activity, Thevetia peruviana was the most potent, followed by Pueraria mirifica, and Butea superba was the least effective. In all cases, the late 3rd instar was more susceptible than the early 4th instar larvae, and the 48-hours exposure yielded more potent larvicidal activity than 24-hours exposure. However, at sublethal dosages, both P. mirifica and B. superba showed some dispersed effects interfering with ecdysis. A variety of toxic effects were observed and recorded in eight categories according to the stage of metamorphosis when death occurred. P. mirifica rendered the main deleterious effects in the pupa-adult period in both instar of Ae. aegypti and Cx. quinquefasciatus, whereas B. superba showed highest effect in black-pupa period of the late 3rd instar larval stage. The results were reversed for the early 4th instar larvae of both species of mosquito as the main effect appeared in the pupa-adult category. The overall results indicated that T. peruviana did not show any IGR properties; whereas, P. mirifica and B. superba seemed to exhibit the juvenile hormone type activity which resulted in abnormal death at various stages of development. B. superba was more promising than P. mirifica, and Ae. aegypti was about 2 times more susceptible than Cx. quinquefasciatus. In addition, L3 was always more susceptible than L4 with both mosquito species.

Comparative morphometry, morphology of egg and adult surface topography under light and scanning electron microscopies, and metaphase karyotype among three size-races of Fasciola gigantica in Thailand.

Comparative morphometry of eggs and adults under light microscope, and morphology of adults under scanning electron microscope (SEM) were undertaken in the three size-races (< 25 mm, 25-35 mm, > 35 mm) of Fasciola gigantica (Thailand strain). Morphometric examination revealed intraspecific variation with respects to the dimensions of eggs and adults, whereas surface topography of the three size-race adults under SEM was morphologically similar. The observations on mitotic metaphase chromosomes of spermatogonial cells from testes of the three size-races revealed 2n=20 (diploid type), and no karyotypic difference was observed among them. The meiotic metaphase chromosome was 10 bivalents in primary spermatocyte in diplotene to diakinesis, and many mature spermatozoa were seen in the testicular preparations.

Comparative studies on protein, isoenzyme and DNA restriction endonuclease profiles of Thai Paragonimus metacercariae.

General proteins and 14 enzymes from metacercariae of Paragonimus heterotremus, P. siamensis and P. westermani were determined by vertical polyacrylamide gel electrophoresis. The isoenzyme profiles showed considerable interspecific polymorphism for general protein (PT), malate dehydrogenase (MDH), malic enzyme (ME) and tetrazolium oxidase (TO) while those of glucose phosphate isomerase (GPI) and phosphoglucomutase (PGM) showed similarity. The Pt-6 and To-I loci can be used as identification markers for these three species. The preliminary study of the molecular biology of Paragonimus heterotremus P. siamensis and P. westermani was based on analysis of metacercarial genomic DNA with restriction endonuclease Pst I. Agarose gel electrophoresis revealed restriction fragment length differences among the three species studied. The DNA restriction fragments were approximately 4-6 fragments, ranging from 5.35 to 14.67 kb. Among these. P westermani shared two homologous fragments with P. siamensis, ie, 5.35 and 7.22 kh, none with P. heterotremus, while P. heterotremus shared only one with P. siamensis, ie, 8.16 kb. Thus, the DNA restriction fragment length differences can be used to differentiate among these three species.

Isoenzyme study and hybridization of two forms of Anopheles sinensis (Diptera: Culicidae) in northern Thailand.

The screening of ten isoenzymes of two forms of Anopheles sinensis, Form A and B, using electrophoretic gels revealed that Est-5(96) allele was the marker in both 4th larva and adult female of An. sinensis Form B, whereas it was lacking in Form A. Hybridization tests of the two sinensis forms were done by induced copulation. The results of crosses indicated that they were genetically compatible, providing viable progeny and completely synaptic polytene chromosomes.

Differentiation of Anopheles minimus species complex by RAPD-PCR technique.

Amplification of random regions of genomic DNA using 10-base primers in the random-amplified polymorphic DNA polymerase chain reaction (RAPD-PCR) was used to differentiate Anopheles minimus A and An. minimus C. Genomic DNA was extracted from individual mosquitoes of An. minimus A and An. minimus C and amplified in PCR reactions using single primers of arbitrary nucleotide sequence. Fifteen different commercially available primers (Operon oligonucleotides kit M from Operon Technologies, Inc.), six primers were selected on the basis of presence or absence of the bands of A and C. They gave 8 different amplified DNA fragments of these two species of An. minimus. The primers revealed only species A are OPM8 and OPM13 at the 0.8 and 2.15 and 2, while both species A and C were in OPM17 at 0.8 Kb revealed A, at the 0.55 and 1.5 Kb revealed C. OPM 12 gave a 0.5 Kb DNA fragment in A while 0.4 Kb in C. OPM15 showed C at 0.7 Kb. These findings indicated that An. minimus species A and C can be differentiated by RAPD-PCR technique.

Integration of control measures for malaria vectors in endemic areas of Thailand.

Various vector control measures were applied in different endemic areas in two provinces, Saraburi and Chanthaburi, with comparison among different control measures. Application of IGR (insect growth regurator, pyriproxyfen) was introduced at Wat Tam Pra Pothisat, Tab-Kwang District, Saraburi Province. Some integration measures were performed at villages 6 and 8, Patavee, Makham District, Chanthaburi Province. In Tab-Kwang District with low malaria endemicity at the study site predators were not able to be released due to rapid velocity of running water. IGR could effectively control malaria compared to the basin released predators. Another endemic areas villagers 6 and 8, Patavee, Makham, Chanthaburi Province was chosen. Highly endemic multidrug resistant malaria has been prevalent for many years in this area. Integration of Kanda's trapping system, application of IGR, use of both residual spraying and impregnated bed-net methods with etofenprox successfully interrupted malaria infection. The application of these methods as an integrated control system could be adjusted to environmental conditions. The results of this study suggest rapid effective vector control.

Population genetic studies on the Anopheles minimus complex in Thailand.

The population genetic studies on the Anopheles minimus complex revealed that An. minimus is a species complex evident by IK value were less than 0.0534 and 0.4163 for the morphotaxonomy and isozyme studies. There are two sibling species in the An. minimus species complex. The first is the typical minimus and its variations described by Harrison. Esterase-2 alleles 100 and 102 are predominant with a few other alleles of 96 and 104. It is exophagic and zoophilic. It is widely distributed in all provinces where minimus breeds. The second of the two sibling species is An. minimus species C, with 2 pale spots on the humeral dark band and Est-2 allele 98. It is found most frequently in Pu Toei District of Kanchanaburi Province, Thailand. An. minimus probably species B showing M1 + 2 wing vein entirely dark is found in China.