Rotavirus specie A (RVA) infection is the leading cause of severe acute diarrhea among young children worldwide. To reduce this major RVA health impact, the Rotarix ® vaccine (GlaxoSmithKline, Rixensart, Belgium) was introduced in the Brazilian Expanded Immunization Program in March 2006 and became available to the entire birth cohort. The aim of this study was to evaluate the spread of RVA in the environment after the introduction of Rotarix ® in Brazil. For this purpose, a Wastewater Treatment Plant (WTP) in Rio de Janeiro was monitored for one year to detect, characterize and discriminate RVA genotypes and identify possible circulation of vaccine strains. Using TaqMan ® quantitative PCR (qPCR), RVA was detected in 100% (mean viral loads from 2.40 × 10 5 to 1.16 × 10 7 genome copies (GC)/L) of sewage influent samples and 71% (mean viral loads from 1.35 × 10 3 to 1.64 × 10 5 GC/L) of sewage effluent samples. The most prevalent RVA genotypes were P4, P6 and G2, based on VP4 and VP7 classification. Direct nucleotide sequencing (NSP4 fragment) and restriction enzyme digestion (NSP3) analysis did not detect RVA vaccine-like strains from the sewage samples. These data on RVA detection, quantification and molecular characterization highlight the importance of environmental monitoring as a tool to study RVA epidemiology in the surrounding human population and may be useful on ongoing vaccine monitoring programs, since sewage may be a good screening option for a rapid and economical overview of the circulating genotypes. Display omitted ► Rotavirus A was detected in sewage samples during a year. ► Rotavirus temporal distribution and genotypes were examined. ► Two methodologies were applied to discriminate between rotavirus wild and vaccine strains.