Enteric viruses are present in aquatic environments due to contamination by raw sewage, even in the absence of fecal coliforms, which are considered to be significant indicators when it comes to microbial water quality assessment. This study investigated the presence of group A rotavirus (RVA) in surface water from a river basin in Minas Gerais, Brazil, assessing the influence of the urbanization, the rainfall, and the microbiological and physico-chemical parameters regarding water quality. Forty-eight surface water samples collected in urbanized and non-urbanized areas, both in dry and rainy periods, were obtained throughout the study. The viral particles were concentrated by adsorption-elution in a negatively charged membrane. Fecal coliforms, as well as physico-chemical water parameters were determined at each point in all collections. The RVA was detected in 62.5 % (30/48) of the water samples through a real-time PCR assay. All the sequenced RVA strains belonged to genotype I1. The RVA was detected in 50.0 % (11/22) of the water samples regarded as being acceptable by current microbiological standards. The presence of the RVA and the viral load were influenced by the collection area (p < 0.05). It was also observed a significant association between the RVA and detecting the turbidity of water (p < 0.05). The collected data showed a high level of contamination in this watershed by the discharge of raw sewage, highlighting the need for urgent measures to improve water quality, ensuring the safe use of this matrix.