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Mosquito-transmitted viruses such as dengue are a global and growing public health challenge. Without widely available vaccines, mosquito control is the primary tool for fighting the spread of these viruses. New mosquito control technologies are needed to complement existing methods, given current challenges with scalability, acceptability, and effectiveness. A field trial was conducted in collaboration with the Communities Organized to Prevent Arboviruses project in Ponce, Puerto Rico, to measure entomological and epidemiological effects of reducing Aedes aegypti populations using Wolbachia incompatible insect technique. We packed and shipped Wolbachia-males from California and released them into 19 treatment clusters from September 2020 to December 2020. Preliminary evaluation revealed sub-optimal Wolbachia-male densities and impact on the wild-type population. In 2021, we shifted to a phased release strategy starting in four clusters, reducing the mosquito population by 49% (CI 29–63%). We describe the investigation into male quality and other factors that may have limited the impact of Wolbachia-male releases. Laboratory assays showed a small but significant impact of packing and shipping on male fitness. However, mark-release-recapture assessments suggest that male daily survival rates in the field may have been significantly impacted. We compared induced-sterility levels and suppression of the wild population and found patterns consistent with mosquito population compensation in response to our intervention. Analysis of epidemiological impact was not possible due to very low viral transmission rates during the intervention period. Our entomological impact data provide evidence that Wolbachia incompatible-male releases reduced Ae. aegypti populations, although efficacy will be maximized when releases are part of an integrated control program. With improvement of shipping vessels and shipped male fitness, packing and shipping male mosquitoes could provide a key solution for expanding access to this technology. Our project underscores the challenges involved in large and complex field effectiveness assessments of novel vector control methods. Author summary: Mosquito control is a key component of strategies to reduce the increasing occurrence of mosquito-borne diseases like dengue. New effective technologies are needed to complement existing methods. In the Wolbachia-based incompatible insect technique, male mosquitoes with Wolbachia are released repeatedly to mate with wild female mosquitoes; the resulting eggs don't hatch, hence reducing the mosquito population over time. We assessed the effectiveness of this technique using mosquitoes reared and packed in California and shipped and released in 19 community clusters in Ponce, Puerto Rico. Analysis of the impact of the releases in decreasing dengue occurrence was not possible, given the low transmission rate during the project period. Preliminary analysis showed sub-optimal mosquito population reduction; we then focused the releases in 4 clusters, reaching up to 49% (CI 29–63%) population reduction. Our extensive laboratory investigations suggest that the packing and shipping process had significant effects on mosquito fitness and daily survival rates, which may have limited the impact of the releases and prevented reaching a higher population reduction. Our data confirms that this technique is effective in reducing mosquito populations in Ponce, PR, but efficacy can increase when releases are part of an integrated control strategy. [ABSTRACT FROM AUTHOR]