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Background The Sterile insect technique (SIT) has been successfully used in agricultural pest control, leading to interest in its application for public health, particularly in controlling Aedes mosquitoes in the USA, Italy, Cuba, and Greece. Malaysia has conducted a small-scale SIT pilot trial since 2019 for dengue control. This study evaluates mosquito populations in treated and untreated sites through three objectives: (1) comparing mean larvae per trap (MLT) and dengue cases for Ae. aegypti and Aedes albopictus ; (2) estimating survival rates and wild populations using mark-release-recapture (MRR); and (3) analysing spatial distribution in treated and untreated sites. Methods Ae. aegypti males, irradiated at 55 Gray, were released in three locations: Pangsapuri Kota Laksamana (KT), Malacca (19 months), Pangsapuri Taman Tasik Utama (TTU), Malacca (8 months), and the Customs, Immigration, and Quarantine Complex (CIQ), Johor (7 months). Statistical analyses assessed SIT effectiveness, including T-tests for larval density and ovitrap indices, Mulla’s formula and relative variance (RV) for population reduction, and the Lincoln Index for estimating wild male populations and probability of daily survival. Results Weekly releases of sterile Ae. aegypti males at doses of 1278–7942 males/ha achieved a sterile-to-wild male ratio of 5.85 and a mean daily survival rate of 0.61, leading to significant reductions in larval densities: 76.25% in Kota Laksamana (KT), 96.74% in Taman Tasik Utama (TTU), and 89.00% in CIQ Gelang Patah, thereby supporting dengue control efforts. In KT, the MLT was initially low but increased, although with suppression < 90%, there was a reduce of dengue cases throughout the release period. The MRR’s mean survival rate (± standard deviation) in KT was 0.61 (± 0.08). The spatial clustering of Ae. aegypti was observed in central blocks during the high MLT period. However, larval densities rebounded after releases ceased. Spatial clustering revealed no initial clustering, though clustering patterns emerged over time in KT. Conclusions SIT effectively suppressed Ae . aegypti populations and supported dengue control. Optimizing sterile-to-wild male ratios, spatial distribution, and monitoring strategies is essential for sustainable vector control. These findings provide insights for scaling up SIT field trials, with future efforts focusing on refining release and monitoring strategies to enhance SIT as an effective dengue control tool. Trial registration NMRR-17–2652-39,099 “Field evaluation of Sterile Insect for Aedes aegypti Suppression.”

Dengue is the most important mosquito-borne viral disease. In the absence of specific drugs or vaccines, control focuses on suppressing the principal mosquito vector, Aedes aegypti, yet current methods have not proven adequate to control the disease. New methods are therefore urgently needed, for example genetics-based sterile-male-release methods. However, this requires that lab-reared, modified mosquitoes be able to survive and disperse adequately in the field. Adult male mosquitoes were released into an uninhabited forested area of Pahang, Malaysia. Their survival and dispersal was assessed by use of a network of traps. Two strains were used, an engineered 'genetically sterile' (OX513A) and a wild-type laboratory strain, to give both absolute and relative data about the performance of the modified mosquitoes. The two strains had similar maximum dispersal distances (220 m), but mean distance travelled of the OX513A strain was lower (52 vs. 100 m). Life expectancy was similar (2.0 vs. 2.2 days). Recapture rates were high for both strains, possibly because of the uninhabited nature of the site. After extensive contained studies and regulatory scrutiny, a field release of engineered mosquitoes was safely and successfully conducted in Malaysia. The engineered strain showed similar field longevity to an unmodified counterpart, though in this setting dispersal was reduced relative to the unmodified strain. These data are encouraging for the future testing and implementation of genetic control strategies and will help guide future field use of this and other engineered strains.

Several sites, Z-7L, Z-5 and Z-14, in Sibu district, Sarawak, Malaysia, experienced intense dengue transmission in 2014 that continued into 2015. A pilot study with Bacillus thuringiensis israelensis (Bti) to control Aedes aegypti (L.) and Ae. albopictus (Skuse) was evaluated in Z-7L, a densely populated site of 12 ha. Bti treatments were conducted weekly from epidemiology week (EW) 24/2015 for 4 weeks, followed by fortnight treatments for 2 months, in addition to the routine control activities. Bti was directly introduced into potable containers and the outdoor artificial and natural containers were treated via a wide area spray application method using a backpack mister. Aedes indices significantly reduced during the treatment and post treatment phases, compared to the control site, Z-5 (p<0.05). A 51 fold reduction in the incidence rate per 100,000 population (IR) was observed, with one case in 25 weeks (EW 29-52). In Z-5 and Z-14, control sites, a 6 fold reduction in the IR was observed from EW 29-52. However, almost every week there were dengue cases in Z-14 and until EW 44 in Z-5. In 2016, dengue cases resurfaced in Z-7L from EW 4. Intensive routine control activities were conducted, but the IR continued to escalate. The wide area Bti spray misting of the outdoor containers was then included from EW 27 on fortnight intervals. A 6 fold reduction in IR was observed in the Bti treatment phase (EW 32-52) with no successive weekly cases after EW 37. However, in the control sites, there were dengue cases throughout the year from EW 1-52, particularly in Z-14. We feel that the wide area Bti spray application method is an integral component in the control program, in conjunction with other control measures carried out, to suppress the vector population in outdoor cryptic containers and to interrupt the disease transmission.

Dengue is the most important mosquito-borne viral disease. No specific treatment or vaccine is currently available; traditional vector control methods can rarely achieve adequate control. Recently, the RIDL (Release of Insect carrying Dominant Lethality) approach has been developed, based on the sterile insect technique, in which genetically engineered 'sterile' homozygous RIDL male insects are released to mate wild females; the offspring inherit a copy of the RIDL construct and die. A RIDL strain of the dengue mosquito, Aedes aegypti, OX513A, expresses a fluorescent marker gene for identification (DsRed2) and a protein (tTAV) that causes the offspring to die. We examined whether these proteins could adversely affect predators that may feed on the insect. Aedes aegypti is a peri-domestic mosquito that typically breeds in small, rain-water-filled containers and has no specific predators. Toxorhynchites larvae feed on small aquatic organisms and are easily reared in the laboratory where they can be fed exclusively on mosquito larvae. To evaluate the effect of a predator feeding on a diet of RIDL insects, OX513A Ae. aegypti larvae were fed to two different species of Toxorhynchites (Tx. splendens and Tx. amboinensis) and effects on life table parameters of all life stages were compared to being fed on wild type larvae. No significant negative effect was observed on any life table parameter studied; this outcome and the benign nature of the expressed proteins (tTAV and DsRed2) indicate that Ae. aegypti OX513A RIDL strain is unlikely to have any adverse effects on predators in the environment.

In 2015, fifteen surveillances for larvae of dengue vectors, Aedes aegypti (L.) and Ac. albopictus (Skuse), were conducted in localities, which had dengue outbreaks in 2014 and 2015. The surveillances covered both indoor and outdoor of premises and the surroundings of the locality. All artificial and natural containers holding stagnant water were inspected for the presence of immature larvae. House index (HI) in all 4 localities exceeded the threshold value to implementation of dertgue vector control. Taman Sungai Jelok (TSJ), Selangor, had the highest HI, container index and the highest larval density per surveillance and per hectare. The larval productivity was higher in outdoor containers, irrespective of outdoor of premises or in the general surrounding of the localities. Ae. aegypti was found equally breeding in indoor and outdoor artificial containers. However, Ac. albopictus was the dominant breeder in the outdoor artificial containers, 2.34-fold higher than Ac. aegypti. There is an insignificant Ac. albopictus population in the indoors. Plastic containers, flower pots, vases and tires were key receptacles. A very significant finding was that both vectors were found in a concrete drain holding stagnated clear water in TSJ. As for natural containers, yam, bromeliad plants and a Terminalia cutup pa tree hole (containing both vectors) were key receptacles. These findings will be useful in promoting awareness in the Malaysian Ministry of Health vector control personnel and residents on dengue vectors breeding habitats and the need for their eradication.

Continued outbreaks of dengue in endemic areas, unabated despite use of conventional vector control methods, necessitate development of new control tools as existing dengue mosquito control technologies are effective only to a limited extent. An insect growth regulator (IGR)-treated auto-dissemination trap was developed against Aedes spp, in which a female mosquito ovipositing in the treated trap is contaminated with an IGR [0.004% pyriproxyfen (PPF)] and transfers the IGR to other containers as the mosquito continues to oviposit. Four PPF auto-dissemination traps were placed on each floor of a 3-block condominium complex in Sri Subang, Selangor, Malaysia for 44 weeks (February to December 2014). Traps were replenished with PPF solution biweekly. Dengue epidemiological monitoring was also conducted by national health authorities. Oviposition in auto-dissemination traps increased over the study period, indicating an attraction for gravid female Aedes spp. No single live larva was observed in any auto-dissemination trap, indicating complete larval mortality induced by PPF. Following introduction of eight additional treated traps on every floor from week 16 onwards, a reduction in ovitrap index from 90 to 33% by week 20 was observed. Correspondingly, number of reported dengue cases was reduced from 53 in 2013 to 13 cases in 2014 (p-value = 0.006). Although Aedes spp populations fluctuated over the course of the study period, the results suggest auto-dissemination traps as a promising dengue control tool. Future research should be directed to determine the optimal PPF concentration and number of PPF treated auto-dissemination traps required to be deployed for ensuring maximum control of dengue.

Dengue is the most important mosquito-borne viral disease. In the absence of specific drugs or vaccines, control focuses on suppressing the principal mosquito vector, Aedes aegypti, yet current methods have not proven adequate to control the disease. New methods are therefore urgently needed, for example genetics-based sterile-male-release methods. However, this requires that lab-reared, modified mosquitoes be able to survive and disperse adequately in the field. Adult male mosquitoes were released into an uninhabited forested area of Pahang, Malaysia. Their survival and dispersal was assessed by use of a network of traps. Two strains were used, an engineered 'genetically sterile' (OX513A) and a wild-type laboratory strain, to give both absolute and relative data about the performance of the modified mosquitoes. The two strains had similar maximum dispersal distances (220 m), but mean distance travelled of the OX513A strain was lower (52 vs. 100 m). Life expectancy was similar (2.0 vs. 2.2 days). Recapture rates were high for both strains, possibly because of the uninhabited nature of the site. After extensive contained studies and regulatory scrutiny, a field release of engineered mosquitoes was safely and successfully conducted in Malaysia. The engineered strain showed similar field longevity to an unmodified counterpart, though in this setting dispersal was reduced relative to the unmodified strain. These data are encouraging for the future testing and implementation of genetic control strategies and will help guide future field use of this and other engineered strains.

Dengue is the most important mosquito-borne viral disease. In the absence of specific drugs or vaccines, control focuses on suppressing the principal mosquito vector, Aedes aegypti, yet current methods have not proven adequate to control the disease. New methods are therefore urgently needed, for example genetics-based sterile-male-release methods. However, this requires that lab-reared, modified mosquitoes be able to survive and disperse adequately in the field. Adult male mosquitoes were released into an uninhabited forested area of Pahang, Malaysia. Their survival and dispersal was assessed by use of a network of traps. Two strains were used, an engineered 'genetically sterile' (OX513A) and a wild-type laboratory strain, to give both absolute and relative data about the performance of the modified mosquitoes. The two strains had similar maximum dispersal distances (220 m), but mean distance travelled of the OX513A strain was lower (52 vs. 100 m). Life expectancy was similar (2.0 vs. 2.2 days). Recapture rates were high for both strains, possibly because of the uninhabited nature of the site. After extensive contained studies and regulatory scrutiny, a field release of engineered mosquitoes was safely and successfully conducted in Malaysia. The engineered strain showed similar field longevity to an unmodified counterpart, though in this setting dispersal was reduced relative to the unmodified strain. These data are encouraging for the future testing and implementation of genetic control strategies and will help guide future field use of this and other engineered strains.