Aim We evaluated the periphyton on artificial substrate in the treated sewage effluent, effluent patch, inside and after the macrophyte stand in a shallow hypereutrophic reservoir. Specifically, we investigated the relationship between N and P contents and algal biomass in the periphyton with N and P availability, focusing on nutrient retention. Methods Periphyton sampling was performed at the effluent inlet, effluent path, inside, and two sites after macrophyte stand. Periphyton sampling was performed after 30 days of colonization. Abiotic variables were determined in the sewage effluent and in the reservoir water. Results Biomass and N and P contents in the periphyton were significantly different among sampling sites. The highest nutrient concentrations were found in the sampling sites with effluent. The highest periphyton chlorophyll-a were found inside and after the macrophyte stand, while N and P contents were the highest in the effluent inlet and effluent. Conclusions In conclusion, N and P contents in the periphyton were associated with N and P availability, evidencing the ability of nutrient retention of the community. Our findings suggest that periphyton on artificial substrate can as a potential tool for removing N and P from the effluent from the sewage treatment system, contributing to minimizing the nutrient load discharged in a shallow reservoir.