Partly in response to this call, a range of promising intervention tools are now ready for furtherinvestigation for prevention of oviposition or biting or suppression of vector populations, whiledevelopments in molecular genetic research have increased the likelihood that novel methods will bedeveloped.Control of immature stagesVector control, either by elimination of breeding sites (‘clean-up’ campaigns) or by larviciding withinsecticides, has led many dengue programmes in the past. Although still widely used and promoted,and successes have been achieved in many contexts, clean-up campaigns are limited by the level ofcompliance by the community and by the fact that often the most productive containers (i.e. thosefrom which most Ae. aegypti emerge) cannot be disposed of or emptied (e.g. drinking-water stores).Larvicidal treatment of such water is possible. Only temephos, permethrin, Bti (see below) andpyriproxyfen are approved by WHO for use in drinking-water . The organophosphate temephos(Abate) has been most widely used and, although it is effective, acceptance levels are often low andcoverage poor. Placing additives with persistent flavour into domestic water, particularly drinking-water is, understandably, often viewed with suspicion and not well accepted. Additionally, regularwater usage or emptying of containers (often encouraged in simultaneous ‘clean-up’ campaigns) canreduce or negate the intended effect. The situation would be improved, first, if attention could belimited to known productive, rather than all potential, breeding sites and secondly, if more acceptablelarvicides were available. A major new multi-country study using pupal/demographic surveys hasdemonstrated that accurate identification of epidemiologically important breeding sites/containers ispossible and should enable targeting of control [24,59]. This is a very important development as,although initial surveys will have to be more intensive, the potential increase in effectiveness gainedby directing control at productive containers is very significant. With this strategy in mind, we havelooked at the potential for a range of new larval control tools.Insect growth regulator (IGR)—pyriproxyfenPyriproxyfen is an insect juvenile-hormone analogue, which is active against many arthropods andwhich has been in use for agricultural pest control for about 15 years. It is extremely effective againstmosquitoes and can prevent the emergence of Ae. aegypti at concentrations as low as 1 ppb or less,while extremely high concentrations do not inhibit oviposition [34,78]. Treated water does not tastetainted. Interestingly, very low doses of pyriproxyfen can also sublethally affect adults by decreasingfecundity or fertility, and the contaminated adult female can transfer effective doses to any breedingsite she subsequently visits [23,34]. New pyriproxyfen formulations can retain efficacy for 6 months, reducing the need for frequent reapplication, and new studies  are indicating its high level ofefficacy. Clearly further trials are now called for. Worryingly, however, this intervention was notaccepted by communities in Mexico , suggesting that suspicion of domestic-water treatments fordengue control remains an obstacle to larval control. Moreover, since pyriproxyfen only preventseclosion, larvae and pupae remain visibly active in breeding sites, conveying the false impression of alack of efficacy and further compromising acceptance. A current trial in Thailand is investigating theseareas further.Predatory copepodsVarious predatory Mesocyclops spp. (Crustacea; Eudecapoda) have been studied for their potential tocontrol mosquito larvae. Two species in particular, M. thermocyclopoides and M. aspericornis, haveproven effective against dengue vectors [9,39,50,52,57,70,73]. However, although copepods cansurvive up to 6 months in containers, they are often lost when water is removed, containers arecleaned or (copepod) food is limited, and reintroduction may be necessary for sustainable control
[14,46,50,73]. However, in certain contexts Mesocyclops can be very effective. A large-scale vector-control programme using copepods and clean-up campaigns in Viet Nam successfully controlleddengue transmission for a number of years and, in this region at least, appeared to be sustainable[39,55,57]. Attempts should now be made to investigate the potential of this method in othercountries where vector-productive containers could sustain copepod populations, and wherecommunities might be willing to accept such a (depending on their experience) novel intervention.Furthermore, the efficacy of this method might be improved by combining it with other compatibletools, such as fish  or other methods (see next section).Bti toxinsThe endotoxin produced by the entomopathogenic bacteria Bacillus thuringiensis var. israelensis (Bti)has high larvicidal activity in mosquitoes, but is non-toxic to important beneficial organisms. Variousformulations (i.e. wettable powders, granules and briquettes) are effective, and newer slow-releaselong-lasting formulations may reduce the need for frequent reapplication. One recent field trial of slow-release long-lasting Bti products in Thailand demonstrated that the treatments could persist for 3months in utility water containers in rural villages (Wiwat et al., personal communication).Combination strategies have often yielded satisfactory results in terms of potency and long-termefficacy [22,60,70,83]. When a combination of M. longisetus and Bti, Bacillus sphaericus, ormethoprene was evaluated , the combined agents were better than either alone. The compatibilityof the commercial formulation Vectobac 12AS with certain chemical insecticides for controlling Aedeslarvae and adults was demonstrated in Malaysia . A mixture of Vectobac 12AS and Actellic 50EC(pirimiphos-methyl) showed promise as a larvicide in the laboratory . These combinations clearlywarrant follow-up. An experiment to integrate Bti with Mesocyclops and mosquito densovirus(AThDNV; see below), conducted in semi-natural conditions, demonstrated that this complexcombination might also be effective (Wiwat et al., personal communication).In Thailand it has been demonstrated that reductions of Ae. aegypti larvae of more than 90% could beachieved for 3-month periods with combined treatment of copepods and Bti, although supplementaryfood for the copepods (community-supplied products such as rice and leaves) was required [14,44]. Apilot community-based intervention using integrated physical and biological control, including acombination of copepods and Bti, in eastern Thailand was successful and showed potential forexpansion into other areas (Kittayapong et al., 2006) .Mosquito densonucleosis virusesDensonucleosis viruses or densoviruses (DNVs) are invertebrate viruses in the genus Brevidensovirusof the family Parvoviridae . The virions of the DNVs contain a 4 kb single-stranded DNA genomewith terminal hairpins packed in a non-enveloped particle [3,7]. Five strains of Aedes densoviruseshave been identified to date [2,5,11,16,36,40]. Current experiments show that the efficiency in vectorcontrol could vary greatly depending on both viral strains and geographic origins of the mosquitovectors (Wiwat et al., personal communication). Application could involve either direct lethal effects ontreated populations, like a biological insecticide, or shortening of adult lifespan, much as describedbelow for Wolbachia. A recent finding has reported improved survival of Ae. aegypti larvae from 15%to 58% after infecting successive generations of mosquitoes with AThDNV, the Thai strain densovirus. Future research to study interactions between the pathogenic viruses and their mosquito hosts,with respect to resistance and mode of action, will generate useful data. Field trials should beencouraged in due course.Vector control of adult mosquitoes
In the past, control directed at adult mosquitoes has been limited to the use of ultra-low-volume (ULV)application of insecticides, usually by vehicle-mounted apparatus. There is controversy regarding theefficacy of this type of control, with a number of studies indicating that its effect is rarely, if ever,significant [65,66,68]. This is likely to be partly the result of a failure of outdoor sprayed insecticide toreach indoor populations of mosquitoes and failures of vertical programmes to deliver at communitylevel. Despite doubts about efficacy, such interventions remain the last resort in combating epidemics.However, new tools open the way to adult mosquito control at community level.Insecticide-treated materialsInsecticide-treated materials (ITMs; typically deployed as insecticide-treated bednets [ITNs]) haveproven highly effective in preventing diseases transmitted by nocturnally-active vectors. Their efficacyin controlling diurnally-active Ae. aegypti is now being evaluated. A cluster-randomized trial in LatinAmerica has demonstrated that insecticide-treated window curtains and/or insecticide-treateddomestic-water container covers can reduce dengue vector densities to low levels and potentially havean impact on dengue transmission . ITMs, particularly curtains, were well accepted and theirefficacy reinforced by the sight of dead insects (cockroaches, houseflies and other pests, as well asmosquitoes) beneath the treated curtains. Importantly, a spill-over effect, whereby the interventionreduced vector populations in neighbouring control clusters, also occurred, such that houses withoutITMs that were located close to treated houses were less likely to have infestations than those furtheraway. Presumably, as with other vectors, the risk of a host-seeking mosquito contacting an ITM atsome point during its frequent visits to houses to blood-feed, means that its life expectancy isreduced; in effect, the age structure of the vector population is altered and few individuals are likely tolive long enough to become infective with dengue. This mass effect will reduce vectors throughout thecommunity [25,29] and is an excellent outcome, given that coverage with any intervention tool isalways less than 100%, sometimes markedly so. New trials are now underway in Venezuela andThailand to examine these interventions further and additional trials in yet more locations and contextsshould be encouraged.Indoor residual treatments are known to kill Ae. aegypti, although such methods have rarely beenused, nor are they recommended today . Indeed, such is the strong endophagic/endophilictendency of this vector [17,64] that the use of insecticidal aerosol cans in the household can alsomarkedly affect dengue transmission . Could other ‘indoor treatments’ also work? The question ofwhether ITNs might also affect Ae. aegypti has also been considered in a pilot study in Haiti (Wiwat etal., personal communication). Although not a fully controlled trial, results indicated that bednetsreduced peridomestic dengue vector breeding (as measured by standard indices), and may havehelped reduce seroconversion rates over 12 months. A community-wide effect was again observed, asdengue-vector breeding also appeared to be reduced in neighbouring control areas. As dengue vectorsbite during the day, how could ITNs exert an effect? Either the presence of the insecticide reducedentry into houses by repelling incoming mosquitoes or, most likely given the community-level effectobserved, the ITNs functioned as ‘residually-treated resting surfaces’ with which contact was maderesulting in killing; an effect that one might expect to be enhanced inside the smaller houses typical inpoorer districts. Effects on Ae. aegypti of ITNs deployed widely for malaria control have not beenrecorded despite the many trials that have been undertaken (presumably because routine sampling fornocturnal malaria vectors would not record Ae. aegypti), although low-level Ae. aegypti biting wasreduced to zero after introduction of ITNs in a village in the Democratic Republic of the Congo .Sustainability of many interventions against all mosquito vectors is a problem in many contexts.However, use of ITNs is invariably higher in urban areas [26,47], precisely where dengue is thegreatest problem. That such an appropriate and widespread intervention might also reduce denguetransmission should be evaluated.
In considering ITM-based strategies for dengue control, new tools primarily developed for malariacontrol can also have applications for dengue control. In the case of long-lasting insecticide-treatednetting (LLIN), the netting is loaded with sufficient insecticide during manufacture to avoid the needfor re-impregnation. Both LLIN window curtains and water-container covers are effective (Kroeger etal., 2006). The long-lasting formulation ‘KO-Tab 1-2-3®’, which can be applied in the community toany material, renders it as wash-resistant as LLIN . Treatment of the existing window curtains in ahouse might be possible and should be investigated, although loss in efficacy over time, resulting fromthe degradation of the insecticide by ultraviolet rays in sunlight may be a problem. Renewed interestfrom industry partners that perceive potential markets in this field, particularly for malaria control, islikely to result in new products to meet control needs in the coming years.The mode of action of ITMs should be investigated: do they repel Ae. aegypti (curtains, bednets or jarcovers), attract and kill them (during hostseeking or oviposition) or exert their effect in some ‘passive’or as yet unknown way? Finally, as is always the case with insecticide-based strategies, resistance toexisting insecticides is inevitable and the current global state of insecticide resistance in Ae. aegypti isworrying (see Hemingway, in the present report). This could be resolved by using alternatives toinsecticides, as described in this article, by using different or by rotating insecticides—a real possibility,as some of the ITMs that are proving effective in dengue control can be used with insecticides otherthan the pyrethroids, which, because of their low mammalian toxicity have been first choice in thepast. Eventually, new insecticides will be needed, an approach that is fundamental to the Gates-funded‘Innovative Vector Control Consortium’1 which aims to develop a portfolio of chemical and technologicaltools suitable for vector control .Lethal ovitrapsThe ovitrap or oviposition trap was first recommended by WHO for surveillance of Aedes vectors[87,88], then modified to render it lethal to adults or larvae of Ae. aegypti [12,13,42,65,66,81].Notably, they were used to eradicate Aedes vectors from Singapore International Airport . Theautocidal ovitrap was designed and developed for the control of Aedes vectors in urban areas with ahigh density of Aedes and a high incidence of dengue haemorrhagic fever in Singapore . Inprinciple, ovitraps could kill adult mosquitoes if the ovistrip was treated with insecticide  or destroyprogeny by using fine nylon netting for trapping the larvae . In Brazil, lethal ovitraps withdeltamethrin-treated ovistrips killed 89% of Ae. aegypti adults and produced more than 99% larvalmortality during 1-month field trials . The advantages of lethal ovitraps for controlling Aedesvectors include their simplicity, their specificity for and effectiveness against container breeders likeAe. aegypti and their potential for integration with other chemical or biological control methodologies.Studies are still at an early stage [66,81] and should be encouraged.The future: strategies based on genetic modificationIn the search for novel tools for vector control, genetic modification of Aedes to resist infection withdengue virus is one of the most seductive long-term approaches. Although some methodology hasalready been shown to be effective in the laboratory , the challenges here are technical (andpotentially independent of any kind of community participation in vector-control activities, althoughinitial acceptance of genetically-modified mosquitoes may not always be certain) requiringdevelopment of mosquito lines resistant to or unable to transmit dengue and replacement of thenatural populations with these non-vector lines.1 Innovative Vector Control Consortium: http://www.ivcc.com/
A major obstacle however, has been the slow development of strong driving systems to deliver denguevirus-resistant genes into natural vector populations , although much research has been devotedto identifying candidates such as transposable elements, meiotic drive or endosymbiotic Wolbachiabacteria [8,35,80]. Transposable elements have been shown to be efficient for transformation ofmosquitoes using an external transposase source, but further research is needed to investigate theautonomous transposable elements in which the effector gene is linked to the source of transposase. The use of meiotic drive in genetic control also requires a greater understanding of the molecularmechanism, especially in mosquitoes. One of the proposed gene driving systems which has potential isthe use of maternally inherited Wolbachia endosymbionts . An advantage is the possibility forrepeated spread and invasion of naturally Wolbachia-infected populations via superinfections ofdifferent Wolbachia strains. Much research is still needed.Development of strategies based on genetic modification ofAe. aegyptiOne of the global collaborative Grand Challenges in Global Health (GCGH)-funded projects aims to usegenetic-based strategies to prevent Ae. aegypti from transmitting dengue viruses either by reducingdensities of mosquito populations or by eliminating their ability to transmit dengue viruses (James,personal communication). The main challenges and objectives of this project are summarized in threetopics: first, the effector genes for population replacement and reduction will be optimized; secondly,safe and efficient drive systems for introgressing effector genes into mosquito populations will bedeveloped; and lastly, a field site for genetic control trials will be established in a suitable developingcountry. Different directions are being pursued. Two approaches aim to reduce or eliminate naturalpopulations of Aedes vectors: densoviruses could be developed as biological control agents of Ae.aegypti larvae , while release of insects carrying dominant lethal mutations (RIDL) are designed toreduce or eliminate mosquito populations, especially those that are infected with dengue viruses[31,33,84]. Other methods—synthetic transposable elements (TEs) , meiotic drive  andunderdominance —aim to modify vector populations by introgressing genes that eliminate vectorcompetence. These methods are currently at various stages in development and readiness for field-testing.A different approach to is being taken via the development of a novel gene-based sterile insecttechnology (SIT) whereby genetically altered, rather than irradiated, males are released into theenvironment to mate with wild females, thereby reducing population numbers and ultimatelypreventing the transmission of disease . This strategy is very well advanced and likely to lead tofield trials in the very near future (Alphey, personal communication).Development of Aedes vectors with a short lifespanAnother GCGH project aims to modify the age structure of the population of dengue vectors such thatlifespan is reduced and mosquitoes die before reaching the age when they could transmit dengue(O’Neill et al., personal communication). The strategy is based on introducing endosymbioticWolbachia bacteria into wild vector populations, these bacteria being capable of significantly shorteningthe adult mosquito lifespan [49,51]. These bacteria are present naturally in most insects andwidespread in mosquitoes [41,86]. Both single and double strains of Wolbachia, naturally found in Ae.albopictus, have already been successfully transferred into Ae. aegypti [72,92]. Dengue viruses needan extrinsic incubation period of about 12–14 days in the vector before being transmitted intohumans; the ‘Popcorn’ strain of Wolbachia could shorten the life of its insect hosts to about half thisperiod. Theoretical models with realistic parameter estimates suggest that an intervention usingrelease of Popcorn-transinfected vector lines could reduce dengue transmission by 90–100% [10,67].
The primary objective of this project is to generate stable lines of Ae. aegypti and Ae. albopictus thatare infected with the Popcorn Wolbachia. Once established, these Wolbachia-transinfected lines couldbe characterized and tested in experimental cages to determine virulence, transovarial transmission,effect on host age and potential to spread in natural vector populations via cytoplasmic incompatibility.The project is ongoing and, if successful, a field trial in large, confined field cages will be attempted inan endemic country. Importantly, this strategy has the potential to be implemented without the needto release transgenic mosquitoes into the environment.Any studies on post-genomic relationships between mosquito hosts and their endosymbionts willbenefit further development of genetic strategies to control dengue vectors. Application of thepopulation-replacement approach in field situations will require detailed understanding of Wolbachia-transfected laboratory-generated strains compared with the naturally uninfected strains of denguevectors in term of their expressed physiological and behavioural parameters at the post-genomic level.The recently-completed sequencing of the Ae. aegypti genome should facilitate identification of neweffector genes, as well as potential gene-driving mechanisms.2 Future research should include studiesrelated to novel genetic markers to investigate gene flow and genetic diversity among differentgeographic populations, which will be crucial to the success of population replacement strategies. Alsoessential, in due course, will be the fostering of public understanding and support of such tools.Priorities and opportunities for researchA number of major initiatives have brought together scientists from many disciplines. Currentprogrammes include the WHO/TDR/International Development Research Centre (IDRC) ResearchInitiative on Eco-Bio-Social Research on Dengue in Asia, a research initiative aimed at improving theprevention of dengue by understanding its multifarious determinants and how they affect transmission(and therefore influence the likelihood of successful control) at community level. The European Unionis funding two major multidisciplinary studies on dengue, one of which, ‘DENCO’, includes two largecluster-randomized trials of ITMs and pyriproxyfen. The Bill & Melinda Gates Foundation have funded arange of initiatives in vector-borne diseases, three of which at least directly target dengue (seeabove).But many areas still require work. In addition to those directions already mentioned, further studiesare required to confirm the efficacy, acceptance, cost–effectiveness and sustainability of ITMs incontrolled trials; trials of slow-release formulations of Bti and pyriproxyfen; densovirus trials; theefficacy of tools against Ae. albopictus should be investigated wherever this is an important vector; thelong-term effects of interventions on mosquito populations and the long-term effects and benefits tocommunities should also be investigated.Ideal tools should be ‘user-friendly’, requiring little additional work or behaviour change byhouseholders. They should be affordable, safe and effective in reducing vector densities belowthreshold levels estimated by the ‘pupae per person index’; indeed, as might be the case withinsecticide-treated curtains in many societies, they could even be viewed as desirable by householders.The ideal situation that we must strive for is to have a suite of proven effective, safe andenvironmentally friendly tools available for intervention, from which the most appropriate tool orcombination of tools can be selected to suit the specific biological and cultural needs of eachcommunity.2 A special issue of Science will be devoted to the Aedes genome in early 2007.
AcknowledgementsThe authors are grateful to the other members of the Scientific Working Group for their discussionsthroughout the meeting. In particular, we thank Dana Focks, Roberto Barrera, Didier Fontenille, LukeAlphey, Chusak Prasittisuk, Axel Kroeger and Mike Nathan.References1. Adelman ZN, et al. Development and applications of transgenesis in the yellow fever mosquito, Aedes aegypti. Mol Biochem Parasitol 121: 1–10.2. Afanasiev B, Carlson J. Densovirinae as gene transfer vehicles. Contrib Microbiol 4: 33–58.3. Bando H, et al. Terminal structure of a densovirus implies a hairpin transfer replication which is similar to the model for AAV. Virology, 179: 57–56.4. Barreau C, et al. An efficient and easy method of infection of mosquito larvae from virus- contaminated cells cultures. J Virol Methods 49: 153–156.5. Barreau C, et al. Pathogenicity of the Aedes albopictus Parvovirus (AaPV), a denso-like virus, for Aedes aegypti mosquitoes. J Invertebr Pathol 68: 299–309.6. Bergoin M, Tijssen P. Molecular biology of Densovirinae. In: Faisst S & Rommelaere J, eds. Parvoviruses: from molecular biology to pathology and therapeutic uses. Contrib Microbiol 4:12– 32.7. Berns KI, et al. Parvoviridae: the viruses and their replication. In: Fields virology, Vol. 2. Philadelphia, Lippincott-Raven Publishers, pp. 2173–2197.8. Braig HR, Yan G. The spread of genetic constructs in natural insect populations. In: Letourneau DK & Burrows BE, eds. Genetically engineered organisms: assessing environmental and human health effects. Boca Raton, Florida, CRC, pp. 251–314.9. Brown MD, et al. Evaluation of Australian Mesocyclops (Cyclopoida: Cyclopidae) for mosquito control. J Med Entomol 28: 618–23.10. Brownstein JS, et al. The potential of virulent Wolbachia to modulate disease transmission by insects. J Invertebr Pathol 84: 24–29.11. Buchatsky LP. Densonucleosis of bloodsucking mosquitoes. Dis Aquat Organ 6: 145–150.12. Chan KL. The eradication of Aedes aegypti at the Singapore Paya Lebar International Airport. In: Chan Yow-Cheong, Chan Kai-Lok & Ho Beng-Chuan, eds. Vector Control in Southeast Asia. Proceedings of the First SEAMEO-TROPMED Workshop, Singapore, 1972, pp. 85–88.13. Chan KL, et al. An autocidal ovitrap for the control and possible eradication of Aedes aegypti. Southeast Asian J Trop Med Public Health, 8: 56–62.14. Chansang U, et al. Combination of Mesocyclops thermocyclopoides and Bacillus thuringiensis var. israelensis: a better approach for the control of Aedes aegypti larvae in water containers. J Vector Ecol 29: 218–226.
15. Chavasse DC . Chemical methods for the control of vectors and pests of public health importance. Geneva, World Health Organization (WHO/CTD/WHOPES/97.2).16. Chen S, et al. Genetic, biochemical, and structural characterization of a new densovirus isolated from a chronically infected Aedes albopictus C6/36 cell lines. Virology 318: 123–133.17. Christophers R. Aedes aegypti: the yellow fever mosquito: its life history, bionomics and structure. Cambridge, Cambridge University Press.18. Christophides GK. Transgenic mosquitoes and malaria transmission. Cell Microbiol 7: 325–333.19. Chung YK et al. Evaluation of biological and chemical insecticide mixture against Aedes aegypti larvae and adults by thermal fogging in Singapore. Med Vet Entomol 5: 321–327.20. Coleman PG, Alphey L. Genetic control of vector populations: an imminent prospect. Trop Med Int Health 9: 433–437.21. Davis S et al. Engineered under-dominance allows efficient and economical introgression of traits into pest populations. J Theor Biol 212: 83–98.22. de Andrande CF, Modolo M. Susceptibility of Aedes aegypti larvae to temephos and Bacillus thuringiensis var israelensis in integrated control. Rev Saúde Pública, 25: 184–187.23. Dell Chism B, Apperson CS. Horizontal transfer of the insect growth regulator pyriproxyfen to larval microcosms by gravid Aedes albopictus and Ochlerotatus triseriatus mosquitoes in the laboratory. Med Vet Entomol 17: 211–220.24. Focks D, Alexander N. Multicountry study of Aedes aegypti pupal productivity survey methodology: findings and recommendations. Geneva, World Health Organization (TDR/ IDM/ Den/ 06.1).25. Gimnig JE, et al. Effect of permethrin-treated bed nets on the spatial distribution of malaria vectors in Western Kenya. Am J Trop Med Hyg 68(Suppl. 4): 115–120.26. Gimnig JE, et al. Laboratory wash resistance of long-lasting insecticidal nets. Trop Med Int Health 10: 1022–1029.27. Gubler DJ, Clark GG. Community-based integrated control of Aedes aegypti: a brief overview of current programmes. Am J Trop Med Hyg 50(Suppl.): 50–60.28. Harrington LC, et al. Dispersal of the dengue vector Aedes aegypti within and between rural communities, Part 1. Am J Trop Med Hyg 72: 209–220.29. Hawley WA, et al. Community-wide effects of permethrin-treated bed nets on child mortality and malaria morbidity in Western Kenya. Am J Trop Med Hyg 68(Suppl. 4): 121–127.30. Hemingway J, et al.The Innovative Vector Control Consortium: improved control of mosquito-borne diseases. Trends Parasitol 22: 308–312.31. Heinrich JC, Scott MJ. A repressible female-specific lethal genetic system for making transgenic insect strains suitable for a sterile-release program. Proc Natl Acad Sci U S A 97: 8229–8232.
32. Honorio NA, et al. Dispersal of Aedes aegypti and Aedes albopictus (Diptera: Culicidae) in an urban endemic dengue area in the State of Rio de Janeiro, Brazil. Memorias do Instituto Oswaldo Cruz, 98: 191–198.33. Horn C, Wimmer EA. A versatile vector set for animal transgenesis. Dev Genes Evol 210: 630–637.34. Itoh T. Utilisation of bloodfed females of Aedes aegypti as a vehicle for the transfer of the insect growth-regulator Pyriproxyfen to larval habitats. Trop Med 36: 243–248.35. James AA. Gene drive systems in mosquitoes: Rules of the road. Trends Parasitol 21: 64–67.36. Jousset FX, et al. A parvo-like virus persistently infecting a C6/36 clone of Aedes albopictus mosquito cell line and pathogenic for Aedes aegypti larvae. Virus Res 29: 99–114.37. Karch S, et al. Impact of deltamethrin-impregnated bednets on biting rates of mosquitoes in Zaire. J Am Mosq Control Assoc 11: 191–194.38. Kay BH, et al. Laboratory evaluation of Brazilian Mesocyclops (Copepoda: Cyclopidae) for mosquito control. J Med Entomol 6: 9–15.39. Kay BH et al. Control of Aedes vectors of dengue in three provinces of Vietnam by use of Mesocyclops (Copepoda) and community-based methods validated by entomologic, clinical, and serological surveillance. Am J Trop Med Hyg 66: 40–48.40. Kittayapong P, et al. A mosquito densovirus infecting Aedes aegypti and Aedes albopictus from Thailand. Am J Trop Med Hyg 61: 612–617.41. Kittayapong P, et al. (2000). The distribution and diversity of Wolbachia infections in Southeast Asian mosquitoes. J Med Entomol 37: 340–345.42. Kittayapong P, et al. Community participation and appropriate technologies for dengue vector control at transmission foci. J Am Mosq Control Assoc 22: 538–546.43. Kokoza V, et al. Engineering blood meal-activated systemic immunity in the yellow fever mosquito Aedes aegypti. Proc Natl Acad Sci U S A 97: 10895–10898.44. Kosiyachinda, P et al. Enhancement of the efficacy of a combination of Mesocyclops aspericornis and Bacillus thuringiensis var. israelensis by community-based products in controlling Aedes aegypti larvae in Thailand. Am J Trop Med Hyg 69: 206–212.45. Kroeger A, et al. Effective dengue vector control with curtains and water container covers treated with insecticide in Mexico and Venezuela: cluster randomised trials. BMJ, 332: 1247–1250.46. Lardeux FJR. Biological control of Culicidae with the copepod Mesocyclops aspericornis and larvivorous fish (Poeciliidae) in a village of French Polynesia. J Med Entomol 6: 9–15.47. Lindblade KA, et al. Evaluation of long-lasting insecticidal nets after 2 years of household use. Trop Med Int Health, 10: 1141–1150.48. Lok CK, et al. An autocidal ovitrap for the control and possible eradication of Aedes aegypti. Southeast Asian J Trop Med Public Health 8: 56–62.
49. McGraw EA, et al. Wolbachia density and virulence attenuation following transfer into a novel host. Proc Natl Acad Sci U S A 99: 2918–2923.50. Marten GG, et al. Use of cyclopoid copepoda for mosquito control. Hydrobiology, 293: 491–496.51. Min K, Benzer S. Wolbachia, normally a symbiont of Drosophila, can be virulent, causing degeneration and early death. Proc Natl Acad Sci U S A 94: 10792–10796.52. Mittal PK, et al. Laboratory evaluation of the biocontrol potential of Mesocyclops thermocyclopoides (Copepoda: Cyclopidae) against mosquito larvae. Southeast Asian J Trop Med Public Health, 28: 857–861.53. Mori A, et al. Reinvestigation of an endogeneous meiotic drive system in the mosquito, Aedes aegypti (Diptera: Culicidae) J Med Entomol 41: 1027–1033.54. Muir LE, Kay BH. Aedes aegypti survival and dispersal estimated by mark-release-recapture in northern Australia. Am J Trop Med Hyg 58: 277–82.55. Nam VS, et al. Eradication of Aedes aegypti from a village in Vietnam, using copepods and community participation. Am J Trop Med Hyg 59: 657–660.56. Nam VS, et al. National progress in dengue vector control in Vietnam: survey for Mesocyclops (Copepoda), Micronecta (Corixidae), and fish as biological control agents. Am J Trop Med Hyg 62: 5–10.57. Nam VS, et al. Elimination of dengue by community programs using Mesocyclops (Copepoda) against Aedes aegypti in central Vietnam. Am J Trop Med Hyg 72: 67–73.58. Nathan BM, Knudsen AB. Aedes aegypti infestation characteristics in several Caribbean countries and implications for integrated community-based control. J Am Mosq Control Assoc 7: 400–404.59. Nathan MB, et al. Pupal/demographic surveys to inform dengue-vector control. Ann Trop Med Parasitol 100(Suppl.): S1–S3.60. Neri-Barbosa JF, et al. Use of bactomos briquets (Bti formulation) combined with the backswimmer Notonectar irrorata (Hemiptera: Notonectidae) for control of mosquito larvae. J Am Mosq Control Assoc 13: 87–89.61. Osaka K, et al. Control of dengue fever with active surveillance and the use of insecticidal aerosol cans. Southeast Asian J Trop Med Public Health, 30: 484–48862. PAHO. Dengue and dengue haemorrhagic fever in the Americas: guidelines for prevention and control. Washington DC, Pan American Health Organization (Scientific Publication).63. Parks W, Lloyd L. Planning social mobilization and communication for dengue fever prevention and control. Geneva, World Health Organization (WHO/CDS/WMC/2004.2.).64. Perich MJ, et al. Comparison of ground and aerial ultra-low volume applications of malathion against Aedes aegypti in Santo Domingo, Dominican Republic. J Am Mosq Control Assoc 6: 1–6.65. Perich MJ, et al. Behavior of resting Ae. aegypti and its relation to ULV efficiency in Panama Journal of Medical Entomology, 35: 541–546.
66. Perich M, et al. Field evaluation of a lethal ovitrap against dengue vectors in Brazil. Med Vet Entomol 17: 205–210.67. Rasgon JL, et al. Wolbachia-induced mortality as a mechanism to modulate pathogen transmission by vector arthropods. J Med Entomol 40: 125–13268. Reiter P, Gubler D. Surveillance and control of urban dengue vectors. In: Gubler DJ & Kuno H, eds. Dengue and dengue hemorrhagic fever. Wallingford UK, CABI Publishing, pp. 425–462.69. Reiter P, et al. Short report: dispersal of Aedes aegypti in an urban area after blood feeding as demonstrated by rubidium-marked eggs. Am J Trop Med Hyg 52: 177–179.70. Riviere F, et al. Mesocyclops aspericornis (Copepoda) and Bacillus thuringiensis var. israelensis for the biological control of Aedes and Culex vectors (Diptera: Culicidae) breeding in crab holes, tree holes and artificial containers. J Med Entomol 24: 425–430.71. Roekring S, et al. Challenging successive mosquito generations with a densonucleosis virus yields progressive survival improvement but persistent, innocuous infections. Dev Comp Immunol 30: 878–892.72. Ruang-areerate T, Kittayapong P. Wolbachia transinfection in Aedes aegypti: a potential gene driver of dengue vectors. Proc Natl Acad Sci U S A 103: 12534–12539.73. Schaper S (1999). Evaluation of Costa Rican copepods (Crustacea: Eudecapoda) for larval Aedes aegypti control with special reference to Mesocyclops thermocyclopoides. J Am Mosq Control Assoc 15: 510–519.74. Scientific Working Group on Dengue. Meeting Report, Scientific Working Group on Dengue, 3–5 April 2000, Geneva, Switzerland. Geneva, World Health Organization (http://www.who.int/tdr/publications/publications/pdf/dengue-swg.pdf).75. Scott TW, et al. The ecology of genetically modified mosquitoes. Science, 298: 117–119.76. Seleena P, et al. Compatibility of Bacillus thuringiensis serovar. israelensis and chemical insecticides for the control of Aedes mosquito. J Vector Ecol 24: 216–223.77. Seng CM, et al. Inhibition of adult emergence of Aedes aegypti in simulated domestic water- storage containers by using a controlled-release formulation of Pyriproxyfen. J Am Mosq Control Assoc 22: 152–154.78. Sihuincha M, et al. Potential use of pyriproxyfen for control of Aedes aegypti (Diptera: Culicidae) in Iquitos, Peru. J Med Entomol 42: 620–630.79. Sinkins SP, Godfrey HC. Use of Wolbachia to drive nuclear transgenes through insect populations. Proc R Soc Lond B Biol Sci 271: 1421–1426.80. Sinkins SP, Gould F. Gene drive systems for insect disease vectors. Nat Rev Genet 7: 427–435.81. Sithiprasasna R, et al. Field evaluation of a lethal ovitrap for the control of Aedes aegypti (Diptera: Culicidae) in Thailand. J Med Entomol 40: 455–462.82. Slosek J. Aedes aegypti mosquitoes in the Americas: a review of their interaction with the human population. Soc Sci Med 23: 249–257.
83. Tietze NS, et al. Integrated management of waste tire mosquitoes utilizing Mesocyclops longisetus (Copepoda: Cyclopidae), of Bacillus thuringiensis serovar. israelensis, Bacillus sphaericus and methoprene. J Am Mosq Control Assoc 10: 363–373.84. Thomas DD, et al. Insect population control using a dominant, repressible, lethal genetic system. Science 287: 2474–2476.85. UNPP. World population prospects: the 2004 revision and world urbanization prospects: the 2004 revision. United Nations Population Division of the Department of Economic and Social Affairs of the United Nations Secretariat (http://esa.un.org/unpp, accessed 28 August 2006).86. Werren JH, et al. Distribution of Wolbachia among neotropical arthropods. Proc R Soc Lond B Biol Sci 262: 197–204.87. WHO (1972). An international system for the surveillance of vectors. Wkly Epidemiol Rec 47: 73– 80.88. WHO (1975). Manual on practical entomology in malaria. Part II. Methods and techniques. WHO Offset Publication, Geneva, World Health Organization.89. WHO (2000). Strengthening implementation of the global strategy for dengue fever/dengue haemorrhagic fever prevention and control: report of the informal consultation, 18–20 October 1999, WHO HQ, Geneva. Geneva, World Health Organization (WHO/CDS(DEN)/IC/2000.1)90. WHO (2006). Pesticides and their application for the control of vectors and pests of public health importance. 6th ed. Geneva, World Health Organization (WHO/CDS/NTD/WHOPES/GCDPP/2006.1).91. Winch P, et al. Beliefs about the prevention of dengue and other febrile illnesses in Mérida, Mexico. J Trop Med Hyg 94: 377–387.92. Xi Z, et al. Wolbachia establishment and invasion in an Aedes aegypti laboratory population. Science 310: 326–328.93. Yates A, et al. Evaluation of KO-Tab 1-2-3®: a wash-resistant ‘dip-it-yourself’ insecticide formulation for long-lasting treatment of mosquito nets. Malar J 4: 5294. Zeichner BC, Perich MJ. Laboratory testing of a lethal ovitrap for Aedes aegypti. Med Vet Entomol 13: 234–238.