Bottle and biochemical assays on temephos resistance in Aedes aegypti in Thailand.

The bottle bioassay measuring the time-mortality rate is a simplified procedure for detecting insecticide resistance. It can be used with a biochemical microplate assay to identify the mechanism involved. This integrated approach was used to detect temephos resistance in Aedes aegypti from Nonthaburi (lowest use) and Roi Et (highest use). Ae. aegypti BKK1 laboratory strain was used as the susceptible reference strain. The appropriate concentration of insecticide for bottle bioassay was determined empirically for Ae. aegypti BKK1 strain and found to be in the range of 800-1,050 microg/bottle. The time-mortality rate at 800 microg/bottle was 170 +/- 8.66 minutes, significantly different from the time-mortality rates in the 850, 900, 950, and 1,050 microg/bottle (p = 0.008) concentrations, which were 135 +/- 15.00, 140 +/- 8.66, 135 +/- 15.00, and 125 +/- 8.66 minutes, respectively. The cut-off concentration selected for resistance detection was 850 microg/bottle. The time-mortality rate for the Roi Et strain was 382 +/- 26.41 minutes, significantly higher than the Nonthaburi (150 +/- 25.10 minutes) and BKK1 strains (145 +/- 20.49 minutes) (p < 0.001). The temephos resistance ratio (RR100) for the Ae. aegypti Roi Et strain was 2.64-fold higher at lethal time (LT100) than for the reference Ae. aegypti BKK1 strain. The mean optical density (OD) value from the biochemical microplate assay for the non-specific esterase of the Roi Et strain was higher than the mean OD for the non-specific esterase of both the Nonthaburi and BKK1 strains. Insensitive acetylcholinesterase was not found to be responsible for the resistance in the field-collected mosquitos. This study suggests that esterase detoxification is the primary cause of resistance in the Ae. aegypti population from Roi Et. Both the bottle bioassay and the biochemical microplate assay were proven to be promising tools for initial detection and field surveillance for temephos resistance.

The geographic information system as an epidemiological tool in the surveillance of dengue virus-infected Aedes mosquitos.

A Geographic Information System (GIS) was used as analysis tool to study the spatial distribution of dengue virus-infected Aedes mosquitos in Thailand. Global Positioning System (GPS) instruments were used to map villages involved in dengue epidemiological studies in Ratchaburi Province, Thailand. Differentially processed GPS data, with a spatial resolution of approximately 1 meter, were incorporated into a GIS for analysis and mapping. Databases associated with a village GIS included village number, Aedes aegypti populations, and test results. Epidemiological surveillance for dengue infection through the detection of the dengue virus type(s) infecting Aedes mosquitos during epidemic periods constitutes a reliable sentinel system for dengue outbreaks. Various techniques were applied including: enzyme linked immunosorbent assay (ELISA), indirect immunofluorescent assay (IFA), and reverse transcriptase-polymerase chain reaction (RT-PCR) assay for the virologic surveillance of the type-specific detection of dengue viruses in artificially infected and in field-caught adult Aedes mosquitos. In laboratory experiments, all assays showed sufficient sensitively to detect one virus infected mosquito and the rapid RT-PCR clearly showed serotype-specificity with very high detection sensitivity. In the field study conducted from April to September 2000, female adult Aedes mosquitos were collected from selected dengue-sensitive areas in Chom Bung district, Ratchaburi Province and assayed by ELISA, IFA and RT-PCR with 18.3% (44/240), 28.98% (20/69) and 15% (3/20) positive for dengue virus, respectively. Geographic distribution of the virus-infected Aedes mosquitos and household locations were demonstrated by the GPS and the GIS. The development of disease mapping data coupled with RT-PCR laboratory-based surveillance of dengue virus infection can successfully serve as epidemiologic tools in an early warning system for dengue hemorrhagic fever (DHF) epidemics.

Relationship between male hydrocele and infection prevalences in clustered communities with uncertain transmission of Wuchereria bancrofti on the Thailand-Myanmar border.

A cross-sectional community-based study was conducted in three clustered communities, belonging to a single small village in Mae Chan subdistrict, Umphang district, Tak Province, close to the Thailand-Myanmar border, where regular night blood survey have been discontinued since 1997 and no epidemiological study had been conducted. In order to understand prevalences of distribution of male hydrocele and infection in clinically diagnostic and epidemiologic implications in uncertain transmission of Wuchereria bancrofti, we analyzed the relationship between male hydrocele and community infection prevalences in 219 (90.5% coverage) subjects aged > or =1 year old, including 54.8% migratory and 45.2% local Karen inhabitants. Migratory inhabitants tended to have high prevalence of antigenemia (p < 0.05) and hydrocele. Overall rates of 23.7% antigenemia, 3.7% microfilaremia, and 4.6% male hydrocele were observed. Male hydrocele prevalence was significantly correlated (r = 0.348, p < 0.0001) with antigenemia prevalence, but not with microfilaremia prevalence (r = 0.065, p = 0.493). However, high antigenemia prevalence in local inhabitants was evident, particularly antigenemia prevalence in children suggesting that transmission in the village may have occurred in recent years.