Despite historical and contemporary evidence of its effectiveness, larval source management with insecticides remains little used by most malaria control programs worldwide. Here we show that environmentally safe biological larvicides under field conditions can significantly reduce anopheline larval density in fish farming ponds that have became major larval habitats across the Amazon Basin. Importantly, the primary local malaria vector, Anopheles darlingi Root (Diptera: Culicidae), feeds and rests predominantly outdoors, being little affected by interventions such as long-lasting insecticidal bed net distribution and indoor residual spraying. We found >95% reduction in late-instar density up to 7 d after the first application of VectoMax FG or VectoLex CG (both from Valent BioSciences), and up to 21 d after larvicide reapplication in fish ponds (n = 20) situated in the main residual malaria pocket of Brazil, irrespective of the formulation or dosage (10 or 20 kg/ha) used. These results are consistent with a substantial residual effect upon retreatment and support the use of biological larvicides to reduce the density of anopheline larvae in this and similar settings across the Amazon where larval habitats are readily identified and accessible.

Background Larvicides are typically applied to fixed and findable mosquito breeding sites, such as fish farming ponds used in commercial aquaculture, to kill immature forms and thereby reduce the size of adult malaria vector populations. However, there is little evidence suggesting that larviciding may suppress community-wide malaria transmission outside Africa. Here, we tested whether the biological larvicide VectoMax FG applied at monthly intervals to fish farming ponds can reduce malaria incidence in Amazonian Brazil. Methods This study was carried out in Vila Assis Brasil (VAB; population 1700), a peri-urban malaria hotspot in northwestern Brazil with a baseline annual parasite incidence of 553 malaria cases per 1000 inhabitants. The intervention consisted of monthly treatments with 20 kg/ha of VectoMax FG of all water-filled fish ponds in VAB (n ranging between 167 and 170) with a surface area between 20 and 8000 m2, using knapsack power mistblowers. We used single-group interrupted time-series analysis to compare monthly larval density measurements in fish ponds during a 14-month pre-intervention period (September 2017–October 2018), with measurements made during November 2018–October 2019 and shortly after the 12-month intervention (November 2019). We used interrupted time-series analysis with a comparison group to contrast the malaria incidence trends in VAB and nearby nonintervention localities before and during the intervention. Results Average larval densities decreased tenfold in treated fish farming ponds, from 0.467 (95% confidence interval [CI], 0.444–0.490) anopheline larvae per dip pre-intervention (September 2017–October 2018) to 0.046 (95% CI, 0.041–0.051) larvae per dip during (November 2018–October 2019) and shortly after the intervention (November 2019). Average malaria incidence rates decreased by 0.08 (95% CI, 0.04–0.11) cases per 100 person-months (P < 0.0001) during the intervention in VAB and remained nearly unchanged in comparison localities. We estimate that the intervention averted 24.5 (95% CI, 6.2–42.8) malaria cases in VAB between January and December 2019. Conclusions Regular larviciding is associated with a dramatic decrease in larval density and a modest but significant decrease in community-wide malaria incidence. Larviciding may provide a valuable complementary vector control strategy in commercial aquaculture settings across the Amazon. Graphical abstract