Effect of crude extract of Solanum xanthocarpum against snails and mosquito larvae.

The ethanolic crude extract from Solanum xanthocarpum was investigated for its molluscicidal activity against Biomphalaria glabrata, the snail vector of Schistosoma mansoni, and Indoplanorbis exustus, the snail vector of intestinal echinostomiasis and Schistosoma spindale, together with the larvicidal activity against the larvae of Aedes aegypti, mosquito vector of dengue hemorrhagic fever and Culex quinquefasciatus, the mosquito vector of urban bancroftian filariasis. The bioassays were carried out following the methods recommended by the World Health Organization. For molluscicidal activity, the LC50 against Bi. glabrata and I. exustus were reported at 163.85 and 198.00 mg/l while the LC90 were 219.33 and 236.80 mg/l, respectively. Regarding mosquito larvicidal activity, the LC50 against the larvae of Ae. aegypti and Cx. quinquefasciatus were 788.10 and 573.20 mg/l, while the LC90 were 1288.91 and 1066.93 mg/l, respectively. These results suggest a preparation of ingredients from this plant may be used as a biological larvicide for these vectors in the field.

Biochemical detection of pyrethroid resistance mechanism in Aedes aegypti in Ratchaburi province, Thailand.

The emergence of insecticide resistance in mosquito vectors was an important issue to be considered as one of factors influencing the success of vector control. The early detection of resistance could help the health personnel to plan and select appropriate alternative control measures or insecticide for effective control. Therefore biochemical assay of enzymes in mosquito was conducted to detect the emergence of insecticide resistance and to define the machanisms involved in pyrethroid resistance. Adults of Aedes aegypti from two localtities in Ratchaburi province were subjected to permethrin and deltamethrin selection in laboratory. After three generations of selection, permethrin-selected and deltamethrin-selected strains were established. Their LT 50 increased to 7.46 and 1.18 folds in the F3 strains that were selected with permethrin and deltamethrin respectively. The enzymes of these mosquitoes were assayed biochemically to study the mechanisms of resistance. The results revealed significant increase of esterase activity and monooxygenase levels in both strains when compared with labolatory susceptible strain. Glutathione-S-transferase activity was found to increase in permethrin-selected strain but not in deltamethrin-selected strain. This suggested that not only esterase and monooxygenase but also glutathione-S-transferase were associated with permethrin resistance in Ae. aegypti. The exposing of permethrin-selected and deltamethrin-selected mosquitoes to diagnostic concentration of permethrin (0.75%) and deltamethrin (0.05%) indicated no cross resistance for permethrin to deltamethrin while slight cross resistance from deltamethrin to permethrin was evident. It seemed that glutathione S-tranferase was not associated with cross resistance since its activity in deltamethrin-selected strain remained unchanged as compared with that of laboratory susceptible strain.

Surveillance of commensal rat and shrew populations in the Bangkok area with references to flea index as the risk indicator of plague.

Commensal rats and shrews were trapped from 47 fresh food markets in Bangkok during the two study periods in the same markets: 21st June to 28th December 1999 and 1st March to 31st May 2000. Trapping was performed using wire live traps on three consecutive nights in each period. The trapped animals were identified for taxonomic species and flea infestation. Fleas were collected, identified and counted. Four species of rodents: Rattus norvegicus, Rattus rattus, Rattus exulans and Mus musculus, and one species of shrew: Suncus murinus were trapped in comparable numbers during the two study periods. Among the 1177 animals trapped, 84.3 per cent were R. norvegicus. Regarding sex prevalence, a higher number of female animals were trapped compared to males. Almost all the fleas collected were Xenopsylla cheopis, and there were very few Ctenocephalidesfelis-felis. Flea index based on the number of X. cheopis was 0.65 for all over Bangkok. Based on the geographical area of Bangkok, the inner area had the highest rodent population and the highest flea index of 0.86. Therefore, the inner region should be the priority for sanitation improvement.

Potential development of temephos resistance in Aedes aegypti related to its mechanism and susceptibility to dengue virus.

The addition of temephos to water containers as a larvicide against Aedes aegypti was commonly used as a part of DHF control programs. The widespread, or long-term, application of insecticides can lead to the development of mosquito resistance to the insecticides through selection pressure. This presents a problem for disease control. Therefore, this study was conducted in the laboratory to observe the potential development of resistance to temephos and the mechanism involved in Ae. aegypti, and to study the significance for dengue infection. The larvae were selected in consecutive generations. The level of resistance to temephos was detected by WHO assay technique. After 19 generations of selection, a low level of resistance was found. The resistance ratio at LC50 was 4.64 when compared with the non-selected group. The assay for major enzyme-based resistance mechanisms was done in a microtiter plate to detect elevated non-specific esterases, monooxygenase, and insensitive acetylcholinesterase in the temephos-selected and non-selected groups. It revealed a significant increase in esterase activity when compared with the non-selected group. There was no elevation of monooxygenase or insensitive acetylcholinesterase activities. However, when an esterase inhibitor (S, S, S-tributyl phosphorotrithioate, or DEF) was added to temephos and the susceptibility in the selected group was studied, the resistance ratio was reduced from 16.92 to 3.57 when compared with a standard susceptible strain (Bora Bora). This indicates that the esterases play an important role in temephos resistance. Dengue-2 virus susceptibility was studied by oral feeding to females of the temephos-selected (S19) and the non-selected groups. The dissemination rates, when the titer of virus in the blood meal was 7.30 MID50/ml, were 11.11% and 9.38% for the selected and non-selected groups, respectively. When the titer of virus in the blood meal was 8.15 MID50/ml, the dissemination rates increased to 24.24% and 33.33%, respectively. A statistical difference in viral susceptibility was not found between the two groups. This suggested that the low level of temephos resistance might not affect oral susceptibility. However, this needs further study.

Temephos resistance in two forms of Aedes aegypti and its significance for the resistance mechanism.

Aedes aegypti, at the larval stage, has been subjected to the temephos selection in laboratory. The level of temephos resistance was detected in a microplate by biochemical assay using WHO bioassay technique. The major enzyme-based resistance mechanisms involved in temephos resistance include elevated nonspecific esterase, oxidase and insensitive acetylcholinesterase. After 19 generations of temephos selection, the selected group showed resistance ratios of 4.64 and 16.92, when compared with a non-selected group and the WHO susceptible strain, respectively. The two seperated forms, type form and the pale form of Ae. aegypti showed low levels of resistance to temephos after 19 generations of selection, with resistance ratios of 4.82 and 4.07 for the type form and the pale form, respectively; when compared with the non-selected strain, 17.58 and 14.84, when compared with the WHO susceptible strain. This showed that the type form could develop higher level resistance than the pale form. The esterase inhibitor (S,S,S-tributyl phosphorotrithioate, DEF) or synergist implicated detoxifying esterase in all the temephos selected groups and the presence of elevated esterase were confirmed by biochemical assay. There were significant differences in elevated esterase activity between the temephos selected groups and the non-selected group. However no significant difference between the type form and the pale form was found. Besides the elevated esterase, there was no change in monooxygenase activity and no evidence of insensitive acetylcholinesterease for all temephos selected groups. These results suggest that temephos resistance could be developed in Ae. aegypti under selection pressure and that the main mechanism is based only on esterase detoxification.