Background: Over the last two decades, there has been remarkable progress in malaria control in sub-Saharan Africa, due mainly to the massive deployment of long-lasting insecticidal nets and indoor residual spraying. Despite these gains, it is clear that in many situations, additional interventions are needed to further reduce malaria transmission. Larval source management (LSM) is a potential supplementary measure that could be used to control malaria. However, prior to its roll-out, knowledge on ecology of larval aquatic habitats is required. Methods: Aquatic habitats colonized by Anopheles vectors were characterised at three sites of low, medium and high malaria transmission in Uganda from October 2011 to June 2016. Larval surveys were conducted along transects in each site and aquatic habitats described according to type and size. Anopheles mosquito larvae and pupae from the described habitats were sampled using standard dipping methods to determine larval densities. Larvae were identified as anopheline or culicine and counted. Pupae were not identified further. Binary logistic regression analysis was used to identify factors associated with the presence of Anopheles larvae in each site. Results: A total of 1,205 larval aquatic habitats were surveyed and yielded a total of 17,028 anopheline larvae, 26,958 culicine larvae and 1,189 pupae. Peaks in larval abundance occurred in all sites in March-May and August-October coinciding with the rainy seasons. Anopheles larvae were found in 52.4 % (n = 251) of aquatic habitats in Tororo, a site of high transmission, 41.9 % (n = 536) of habitats in Kanungu, a site with moderate malaria transmission, and 15.8 % (n=418) in Jinja, a site with low malaria transmission. The odds of finding Anopheles larvae was highest in rice fields compared to pools in both Tororo (odds ratio, OR = 4.21, 95% CI 1.22-14.56, p = 0.02) and Kanungu (OR= 2.14, 95% CI 1.12-4.07, p =0.02). In Kanungu, Anopheles larvae were less likely to be found in open drains compared to pools (OR = 0.15, 95% CI 0.03-0.72, p = 0.02) and river fringe (OR = 0.19, 95% CI 0.07-0.52, p = 0.001). Conclusions: These findings show that Anopheles larvae were common in areas of high and moderate transmission but were rare in areas of low transmission. Although Anopheles larvae were found in all types of water bodies, they were most common in rice fields and less common in open drains and on river fringes. Methods are needed to reduce the aquatic stages of anopheline mosquitoes in human-made habitats, particularly rice fields.