Urban heat islands are characterized by high land surface temperature, low humidity, and poor vegetation, and considered to favor the transmission of the mosquito-borne dengue fever that is transmitted by the Aedes aegypti mosquito. We analyzed the recorded dengue incidence in Sao Paulo city, Brazil, in 2010-2011, in terms of multiple environmental and socioeconomic variables. Geographical information systems, thermal remote sensing images, and census data were used to classify city areas according to land surface temperature, vegetation cover, population density, socioeconomic status, and housing standards. Of the 7415 dengue cases, a majority (93.1%) mapped to areas with land surface temperature >28°C. The dengue incidence rate (cases per 100,000 inhabitants) was low (3.2 cases) in high vegetation cover areas, but high (72.3 cases) in low vegetation cover areas where the land surface temperature was 29±2°C. Interestingly, a multiple cluster analysis phenogram showed more dengue cases clustered in areas of land surface temperature >32°C, than in areas characterized as low socioeconomic zones, high population density areas, or slum-like areas. In laboratory experiments, A. aegypti mosquito larval development, blood feeding, and oviposition associated positively with temperatures of 28-32°C, indicating these temperatures to be favorable for dengue transmission. Thus, among all the variables studied, dengue incidence was most affected by the temperature.

The signal sequence trap method was used to isolate cDNAs corresponding to proteins containing secretory leader peptides and whose genes are expressed specifically in the salivary glands of the malaria vector Anopheles gambiae . Fifteen unique cDNA fragments, ranging in size from 150 to 550 bp, were isolated and sequenced in a first round of immunoscreening in COS-7 cells. All but one of the cDNAs contained putative signal sequences at their 5′ ends, suggesting that they were likely to encode secreted or transmembrane proteins. Expression analysis by reverse transcription–PCR showed that at least six cDNA fragments were expressed specifically in the salivary glands. Fragments showing a high degree of similarity to D7 and apyrase, two salivary gland-specific genes previously found in Aedes aegypti , were identified. Of interest, three different D7-related cDNAs that are likely to represent a new gene family were found in An. gambiae . Moreover, three salivary gland-specific cDNA fragments that do not show similarity to known proteins in the databases were identified, and the corresponding full length cDNAs were cloned and sequenced. RNA in situ hybridization to whole female salivary glands showed patterns of expression that overlap only in part those observed in the culicine mosquito A. aegypti .

Dengue is considered to be the most important mosquito-borne viral disease in the world. The Aedes aegypti mosquito, its vector, is highly anthropophilic and is very well adapted to urban environments. Although several vaccine candidates are in advanced stages of development no licensed dengue vaccine is yet available. As a result, controlling the spread of dengue still requires that mosquitoes be targeted directly. We review the current methods of dengue vector control focusing on recent technical advances. We first examine the history of Brazil’s National Dengue Control Plan in effect since 2002, and we describe its establishment and operation. With the persistent recurrence of dengue epidemics, current strategies should be reassessed to bring to the forefront a discussion of the possible implementation of new technologies in Brazil’s mosquito control program.

When considering a mosquito release programme, one of the first issues to be addressed is how to eliminate/separate the females. The greatest number of options might eventually be available for those who can use transgenic mosquitoes, but the inherent characteristics of the target species may also provide possibilities for interim measures until more efficient methods can be developed. Differences in intrinsic size, in behaviour and in development rate between females and males are often available and useful for sexing. Efficient species-specific systems for eliminating females at the embryo stage have been developed, but most have since been discarded due to lack of use. Ideal systems specifically kill female embryos using some treatment that can be manipulated during production. Such killing systems are far more efficient than using intrinsic sexual differences, but they systems require selectable genetic markers and sex-linkage created by rare random chromosomal rearrangements. While intrinsic sexual differences should not be considered as long-term candidates for the development of robust and efficient sexing approaches, in the absence of these, the accessibility and integration of less efficient systems can provide a stop-gap measure that allows rapid start up with a minimum of investment. The International Atomic Energy Agency is funding over a 5 year period (2013-2018) a new Coordinated Research Project on "Exploring Genetic, Molecular, Mechanical and Behavioural Methods of Sex Separation in Mosquitoes" to network researchers and to address the critical need of genetic sexing strains for the implementation of the sterile insect technique (using radiation-sterilised or transgenic male mosquitoes) and for insect incompatibility technique programmes against disease-transmitting mosquitoes.

New techniques and methods are being sought to try to win the battle against mosquitoes. Recent advances in molecular techniques have led to the development of new and innovative methods of mosquito control based around the Sterile Insect Technique (SIT)^1-3. A control method known as RIDL (Release of Insects carrying a Dominant Lethal)⁴, is based around SIT, but uses genetic methods to remove the need for radiation-sterilization^5-8. A RIDL strain of Ae. aegypti was successfully tested in the field in Grand Cayman^9,10; further field use is planned or in progress in other countries around the world. Mass rearing of insects has been established in several insect species and to levels of billions a week. However, in mosquitoes, rearing has generally been performed on a much smaller scale, with most large scale rearing being performed in the 1970s and 80s. For a RIDL program it is desirable to release as few females as possible as they bite and transmit disease. In a mass rearing program there are several stages to produce the males to be released: egg production, rearing eggs until pupation, and then sorting males from females before release. These males are then used for a RIDL control program, released as either pupae or adults^11,12. To suppress a mosquito population using RIDL a large number of high quality male adults need to be reared^13,14. The following describes the methods for the mass rearing of OX513A, a RIDL strain of Ae. aegypti ⁸, for release and covers the techniques required for the production of eggs and mass rearing RIDL males for a control program.