Background: Long-lasting insecticidal nets (LLINs) are one of the main vector control strategies recommended by the World Health Organization for the control and elimination of malaria. This study aimed to evaluate the use and retention of LLINs during the 5 years of implementing an integrated control strategy in a malaria-endemic area in the Brazilian Amazon. Methods: This intervention study was conducted in localities of the municipality of Barcelos, Amazonas, from 2008 to 2014. Four rural localities situated along the Padauiri River were the object of this study. Two localities (Bacabal-rio Aracá and the São Sebastião district) were used as controls. LLINs were distributed to all residents of the Padauiri River; assessments were made regarding their use and retention via a semistructured questionnaire, a household register, and direct observation during 5 years. Results: Overall, 208 individuals participated in the study. In the baseline pilot study (2008), 9.9% of the subjects in the intervention group had slept with mosquito nets the previous night compared with 37.8% of the subjects in the control group. In 2010, this percentage was 43.2% in the intervention group and 50.9% in the control group. Therefore, 1 year after the implementation of the strategy, although there was an increase in the use of mosquito nets in both groups, this increase was signifcantly higher in the intervention group. This increase in LLINs use did not persist after 5 years of intervention. The households’ evaluation in 2014 showed that 80% of the houses in the intervention group owned at least one LLIN compared with 66% in the control group (p=0.11); 76% of households in the intervention group owned sufcient LLINs for all family members compared with 50% in the control group (p=0.007). Conclusions: High ownership and retention of the LLINs was observed in the intervention group. At 1 year after the distribution of these LLINs, there was a signifcant increase in their use that was not maintained over the long term.