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Background: The introduction of Wolbachia (wMel strain) into Aedes aegypti mosquitoes reduces their capacity to transmit dengue and other arboviruses. Randomised and non-randomised studies in multiple countries have shown significant reductions in dengue incidence following field releases of wMel-infected Ae. aegypti. We report the public health outcomes from phased, large-scale releases of wMel-Ae. aegypti mosquitoes throughout three contiguous cities in the Aburrá Valley, Colombia. Methodology/Principal findings: Following pilot releases in 2015–2016, staged city-wide wMel-Ae. aegypti deployments were undertaken in the cities of Bello, Medellín and Itagüí (3.3 million people) between October 2016 and April 2022. The impact of the Wolbachia intervention on dengue incidence was evaluated in two parallel studies. A quasi-experimental study using interrupted time series analysis showed notified dengue case incidence was reduced by 95% in Bello and Medellín and 97% in Itagüí, following establishment of wMel at ≥60% prevalence, compared to the pre-intervention period and after adjusting for seasonal trends. A concurrent clinic-based case-control study with a test-negative design was unable to attain the target sample size of 63 enrolled virologically-confirmed dengue (VCD) cases between May 2019 and December 2021, consistent with low dengue incidence throughout the Aburrá Valley following wMel deployments. Nevertheless, VCD incidence was 45% lower (OR 0.55 [95% CI 0.25, 1.17]) and combined VCD/presumptive dengue incidence was 47% lower (OR 0.53 [95% CI 0.30, 0.93]) among participants resident in wMel-treated versus untreated neighbourhoods. Conclusions/Significance: Stable introduction of wMel into local Ae. aegypti populations was associated with a significant and sustained reduction in dengue incidence across three Colombian cities. These results from the largest contiguous Wolbachia releases to-date demonstrate the real-world effectiveness of the method across large urban populations and, alongside previously published results, support the reproducibility of this effectiveness across different ecological settings. Trial registration: NCT03631719. Author summary: Dengue fever is a viral disease transmitted by Aedes aegypti mosquitoes and is an increasing public health concern globally. A novel evidence-based tool for dengue control involves the release of Ae. aegypti mosquitoes infected with a naturally-occurring insect bacteria called Wolbachia (wMel strain). wMel significantly reduces the mosquito's ability to transmit dengue and other viruses and can establish long-term in local Ae. aegypti populations following short-term releases. Previous studies have shown that establishment of wMel in local mosquito populations significantly reduces local dengue incidence. In Colombia, wMel-infected Ae. aegypti were released throughout the cities of Bello, Medellín, and Itagüí in the Aburrá Valley between 2015 and 2022, covering a population of 3.3 million people. The rate of dengue case notifications in the three cities declined by 95–97% after Wolbachia releases, compared to the prior decade, and dengue case numbers since 2020 have been the lowest in twenty years. A case-control study in Medellin in 2019–2021 showed that laboratory-confirmed dengue incidence was significantly lower in neighbourhoods with Wolbachia releases compared to untreated neighbourhoods. These results confirm the real-world effectiveness of the Wolbachia method, and show that it can be implemented at a city-wide scale to protect communities against dengue. [ABSTRACT FROM AUTHOR]

Background: The wMel strain of Wolbachia has been successfully introduced into Aedes aegypti mosquitoes and has been shown to reduce the transmission of dengue and other Aedes-borne viruses. Here we report the entomological results from phased, large-scale releases of Wolbachia infected Ae. aegypti mosquitoes throughout three contiguous cities located in the Aburrá Valley, Colombia. Methodology/principal findings: Local wMel Wolbachia-infected Ae. aegypti mosquitoes were generated and then released in an initial release pilot area in 2015–2016, which resulted in the establishment of Wolbachia in the local mosquito populations. Subsequent large-scale releases, mainly involving vehicle-based releases of adult mosquitoes along publicly accessible roads and streets, were undertaken across 29 comunas throughout Bello, Medellín and Itagüí Colombia between 2017–2022. In 9 comunas these were supplemented by egg releases that were undertaken by staff or community members. By the most recent monitoring, Wolbachia was found to be stable and established at consistent levels in local mosquito populations (>60% prevalence) in the majority (67%) of areas. Conclusion: These results, from the largest contiguous releases of wMel Wolbachia mosquitoes to date, highlight the operational feasibility of implementing the method in large urban settings. Based on results from previous studies, we expect that Wolbachia establishment will be sustained long term. Ongoing monitoring will confirm Wolbachia persistence in local mosquito populations and track its establishment in the remaining areas. Author summary: The introduction of the naturally occurring wMel Wolbachia strain into Aedes aegypti mosquitoes has been shown to reduce the ability of the mosquitoes to transmit dengue and other viruses. Following engagement with communities to gain acceptance and support, a series of large-scale releases of Ae. aegypti mosquitoes that contained wMel Wolbachia, were undertaken across the cities of Bello, Medellín and Itagüí in Colombia. These releases were undertaken under operational conditions with the aim of rapidly scaling the intervention in response to the Zika virus crisis. Mosquito populations were monitored during and after releases to determine the levels of Wolbachia and whether it persisted in the local mosquitoes. Wolbachia was found to be stable and established at consistent levels in local mosquito populations in the majority of areas. On-going monitoring in these areas will determine whether Wolbachia persists and also whether it establishes at a high level in the remaining areas. This intervention forms the basis of an epidemiological study to assess the impact of operational deployment of wMel Wolbachia on the reduction of the incidence of notified dengue cases and virologically-confirmed dengue. [ABSTRACT FROM AUTHOR]