Repeated landmass reformation limits diversification in the widespread littoral zone mosquito Anopheles sundaicus sensu lato in the Indo-Oriental Region.

Southeast Asia harbours abundant biodiversity, hypothesized to have been generated by Pliocene and Pleistocene climatic and environmental change. Vicariance between the island of Borneo, the remaining Indonesian archipelago and mainland Southeast Asia caused by elevated sea levels during interglacial periods has been proposed to lead to diversification in the littoral zone mosquito Anopheles (Cellia) sundaicus (Rodenwaldt) sensu lato. To test this biogeographical hypothesis, we inferred the population history and assessed gene flow of A. sundaicus s.l. sampled from 18 populations across its pan-Asian species range, using sequences from mitochondrial cytochrome c oxidase subunit 1 (CO1), the internal transcribed spacer 2 (ITS2) and the mannose phosphate isomerase (Mpi) gene. A hypothesis of ecological speciation for A. sundaicus involving divergent adaptation to brackish and freshwater larval habitats was also previously proposed, based on a deficiency of heterozygotes for Mpi allozyme alleles in sympatry. This hypothesis was not supported by Mpi sequence data, which exhibited no fixed differences between brackish and freshwater larval habitats. Mpi and CO1 supported the presence of up to eight genetically distinct population groupings. Counter to the hypothesis of three allopatric species, divergence was often no greater between Borneo, Sumatra/Java and the Southeast Asian mainland than it was between genetic groupings within these landmasses. An isolation-with-migration (IM) model indicates recurrent gene flow between the current major landmasses. Such gene flow would have been possible during glacial periods when the current landmasses merged, presenting opportunities for dispersal along expanding and contracting coastlines. Consequently, Pleistocene climatic variation has proved a homogenizing, rather than diversifying, force for A. sundaicus diversity.

A spatial decision support system for guiding focal indoor residual interventions in a malaria elimination zone.

A customized geographical information system (GIS) has been developed to support focal indoor residual spraying (IRS) operations as part of a scaled-up campaign to progressively eliminate malaria in Vanuatu. The aims of the GISbased spatial decision support system (SDSS) were to guide the planning, implementation and assessment of IRS at the household level. Additional aims of this study were to evaluate the user acceptability of a SDSS guiding IRS interventions. IRS was conducted on Tanna Island, Republic of Vanuatu between 26 October and 5 December 2009. Geo-referenced household information provided a baseline within the SDSS. An interactive mapping interface was used to delineate operation areas, extract relevant data to support IRS field teams. In addition, it was used as a monitoring tool to assess overall intervention coverage. Surveys and group discussions were conducted during the operations to ascertain user acceptability. Twenty-one operation areas, comprising a total of 187 settlements and 3,422 households were identified and mapped. A total of 3,230 households and 12,156 household structures were sprayed, covering a population of 13,512 individuals, achieving coverage of 94.4% of the households and 95.7% of the population. Village status maps were produced to visualize the distribution of IRS at the sub-village level. One hundred percent of survey respondents declared the SDSS a useful and effective tool to support IRS. The GIS-based SDSS adopted in Tanna empowered programme managers at the provincial level to implement and asses the IRS intervention with the degree of detail required for malaria elimination. Since completion, SDSS applications have expanded to additional provinces in Vanuatu and the neighbouring Solomon Islands supporting not only specific malaria elimination and control interventions, but also the broader public health sector in general.

Six months of Aedes aegypti control with a novel controlled-release formulation of pyriproxyfen in domestic water storage containers in Cambodia.

A field evaluation of a novel, 5% controlled-release formulation of pyriproxyfen was carried out in 400-500 liter concrete water storage jars, the most common and important larval habitat of the dengue vector, Aedes aegypti, in Cambodia. The formulation consisted of cylindrical resin strands, 3 mm in diameter and 40 mm in length. Pyriproxyfen was applied to 100 jars at a target dose of 0.03-0.04 mg of active ingredient (a.i.) per liter (30-40 ppb) in households in Phum Thmei, a village near the capital city of Phnom Penh, in April 2005. Inhibition of adult emergence (IE) in field populations ofAe. aegypti was measured every 2 weeks for 34 weeks. IE in treated jars exceeded 90% for 20 weeks, and remained above 80% until the end of the study. In 25 untreated jars, failure of pupae to metamorphose into viable adults remained below 4%. No alteration of taste or other undesirable effects of the treatment were reported by householders. This single treatment provided control of Ae. aegypti in water jars for the length of the main dengue transmission season in Cambodia that normally extends from May to November.

Community-based use of the larvivorous fish Poecilia reticulata to control the dengue vector Aedes aegypti in domestic water storage containers in rural Cambodia.

A community-based study of the distribution of larvivorous fish, Poecilia reticulata (common name: guppy), in water storage containers for dengue control was undertaken in 14 villages and approximately 1,000 households in Cambodia. Community volunteers reared guppies and distributed them in water jars and tanks in households for which they were responsible. A nearby control area received no intervention. One year after project commencement, 56.9% of eligible containers contained guppies and there was a 79.0% reduction in Aedes infestation in the intervention community compared with the control. Smaller or discarded containers unsuitable for guppy distribution in the intervention area also had 51% less infestation than those in the control area, suggesting a "community-wide" protective effect. In addition, there was less infestation in villages with higher rates of fish uptake, suggesting that the presence of fish was responsible for a reduction in Aedes infestation. This applied vector control model was well-accepted, effective, efficient, and shows promise as a sustainable community-based, non-insecticidal intervention for dengue vector control in large domestic water storage containers in rural Cambodia and elsewhere.

The effect of long-lasting insecticidal water container covers on field populations of Aedes aegypti (L.) mosquitoes in Cambodia.

Dengue in Cambodia is mainly transmitted by Aedes aegypti (L.) mosquitoes that primarily breed in large, concrete jars (> or =200 liters) used for the storage of water for domestic use. Following a preliminary risk assessment, long-lasting insecticidal netting (LN) treated with deltamethrin was incorporated into the design of the covers for these jars. Their effect on immature and adult female populations of Ae. aegypti in six villages in a peri-urban area of Cambodia were compared with populations in six nearby control villages before and for 22 weeks after distribution of the jar covers. There were significantly fewer pupae per house in intervention villages than in control villages (6.6 and 31.9, respectively, p<0.01). Fewer pupae were recovered from intervention houses than from control houses at every post-intervention assessment. Two weeks after the intervention, the average number of indoor resting female Ae. aegypti per house in the intervention villages had declined approximately three-fold, whereas in the controls there was only a slight reduction (16%). The magnitude of the difference between the two areas diminished over time, which contact bioassays confirmed was likely due to a gradual reduction of insecticidal effect of the jar covers. In the study area, insecticide-treated covers for large concrete water storage jars were efficacious for controlling Ae. aegypti in the protected water jars and with a demonstrable effect on adult densities and survival. Further studies of this targeted container strategy in Cambodia, and elsewhere, are recommended. However, improvements in technology that would extend the duration of insecticidal effectiveness of LN materials may be needed for the development of cost-effective public health applications.

Inhibition of adult emergence of Aedes aegypti in simulated domestic water-storage containers by using a controlled-release formulation of pyriproxyfen.

Concrete domestic water-storage jars are a common larval habitat of the dengue vector Aedes aegypti in countries of Southeast Asia. The efficacy of a novel controlled-release formulation of the insect growth regulator pyriproxyfen, designed to inhibit adult emergence for 6 months (the approximate duration of the main dengue transmission season in many endemic countries) was tested in Cambodia against a local strain of Ae. aegypti in 200-liter jars. The resin-based formulation contained 4.8% active ingredient (AI). At target dosages of 18, 27, and 36 ppb of AI, inhibition of adult emergence remained above 95% for at least 2 months. After 3 months at 18 ppb AI, the residual efficacy was significantly lower than for the higher dosages (P < 0.05). At the higher dosages, inhibition of adult emergence was > or = 87% for 6 months. At a dosage of 27 ppb AI, monthly removal and replacement of two thirds of the water did not reduce efficacy (P > 0.05). Potential operational advantages and challenges of using the formulation are discussed. Larger-scale efficacy studies are recommended in community settings.

Molecular evidence of speciation between island and continental populations of Anopheles (Cellia) sundaicus (Diptera: Culicidae), a principal malaria vector taxon in Southeast Asia.

Anopheles sundaicus s.l. is a principal malaria vector taxon on islands and along the coastal areas of Southeast Asia. It has a wide geographical distribution and exhibits a high level of ecological and behavioral variability. Study of this taxon is crucial for understanding its biology and implementing effectise vector control measures. We compared populations of An. sundaicus from Vietnam, Thailand, and Malaysian Borneo by using two mitochondrial DNA markers: cytochrome oxidase I and cytochrome b. Genetic divergence, geographic separation, and cladistic analysis of relationships revealed the presence of two cryptic species: Anopheles sundaicus s.s. on Malaysian Borneo and An. sundaicus species A in coastal areas of Thailand and Vietnam. A polymerase chain reaction (PCR) assay was developed to easily identify these two species throughout their geographic distributions. The assay was based on sequence characterized amplified region derived from random amplified polymorphic DNA. This PCR identification method needs to be validated and adapted for the recognition of other possible species in the Sundaicus Complex.