A 12-mo ecological study of the spatial-temporal distribution of immature stages of Anopheles species was conducted in Sukabumi District, West Java, Indonesia. The study characterized 1,600 sites from a contiguous coastal and hill zone (0-800-m elevation) of which 64% contained Anopheles larvae. Principal component and multiple logistic regression analyses identified ecological parameters associated with presence of nine [Anopheles aconitus Doenitz, Anopheles annularis Van de Wulp, Anopheles barbirostris Van der Wulp, Anopheles flavirostris (Ludlow), Anopheles insulaeflorum (Swellengrebel and Swellengrebel de Graaf), Anopheles kochi Doenitz, Anopheles maculatus Theobald, Anopheles sundaicus (Rodenwaldt), and Anopheles vagus Doenitz] of 15 Anopheles species collected. Combined data for all nine species showed increased Anopheles presence associated with wet season periods and higher elevation habitats exhibiting reduced tree canopy coverage, higher water temperatures, and shallower water depths. Habitat variables measured included topography (elevation), water conditions (temperature, pH, salinity depth, and velocity), habitat characteristics (substrate and canopy cover), density and type of aquatic vegetation coverage (riparian, floating, and emergent), and distance from nearest human habitation. Significant relationships were found for nine species when using all habitats in the analysis. Habitat characteristics for three species were refined. An. aconitus and An. barbirostris were associated with higher elevation rice, Oryza savita L., paddies with relatively shallow water depths, higher water temperatures, higher acidity and salinity concentrations, and a greater average distance from human habitation. An. vagus presence in rice paddies was associated with lower elevation fields, deeper and cooler water, less acidic and saline conditions, and habitats closer to human dwellings. Overall, the distribution of Anopheles species in Sukabumi was found to be nonrandom and predictable on the basis of habitat characteristics.

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Aedes aegypti (L.) is the primary vector of dengue viruses, a group of four serotypic single-stranded RNA viruses. Dengue virus RNA can be readily detected in fresh or dried infected mosquitoes by using reverse transcriptase-polymerase chain reaction (RT-PCR). The current study examined the persistence and limit of dengue virus RNA detection in infected Ae. aegypti killed and exposed to natural ambient tropical conditions of temperature and humidity. Under relatively harsh conditions, dengue RNA retained sufficient integrity to be detected in dried mosquitoes up to 13 wk after exposure to relatively high ambient temperatures (26.3-31.7 degrees C) and relative humidity (49.4-69.9%). These findings confirm that the necessity for testing either fresh or frozen mosquitoes is not a prerequisite when using RT-PCR as the viral detection method, and under particular epidemiological circumstances it allows for a more convenient means of conducting vector-virus surveillance activities where collection methods and logistics may preclude immediate testing or access to a cold chain.

The determination of the presence or absence of malaria sporozoites in wild-caught Anopheles mosquitoes remains an integral component to the understanding of the transmission dynamics in endemic areas. To improve that capability, there has been on-going development of a new device using dipstick immunochromatographic technology for simplifying the testing procedure and reducing the time required to obtain results. As part of a larger multi-center effort, we evaluated the sensitivity and specificity of a prototype malaria sporozoite antigen panel assay (Medical Analysis Systems, Camarillo, CA) against three human Plasmodium species/polymorphs. The wicking (dipstick) assay was compared against a standard parasite antigen capture enzyme-linked immunosorbent assay (ELISA) for the detection of human circumsporozoite protein (CSP) in wild-caught mosquitoes. Over 6,800 Anopheles mosquitoes, representing 20 species collected from malaria endemic areas of Indonesia were tested either individually or in pools of up to 10 mosquitoes each. From 1,442 pooled test strip assays and ELISA formats, nine mosquito pools were found reactive for P.falciparum, P. vivax 210, or P. vivax 247 CSP. There was complete concordance between test strip results and ELISA results. Sensitivity was 100% and given some minor problems with false positives or negatives, specificity (n = 488) was 97%. Most strips judged as false positive produced very weak signals compared with negative control blank strips and paired ELISA-negative samples. The dipstick test proved technically simpler to perform and interpret than the ELISA and results were obtained within 15 min of exposure to mosquito suspension. This qualitative assay appears an attractive alternative to the CSP ELISA for detection of sporozoites in fresh or dried mosquitoes.

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