This study investigated the prevalence, distribution, and genotypes of adenoviruses (AdVs) and noroviruses (NoVs) in the Puzi River and surrounding areas in Taiwan. The viruses in the water samples were isolated using the membrane filtration method and the viral nucleic acids were extracted. The RNA of NoVs was reverse-transcribed into complementary DNA using reverse transcriptase-polymerase chain reaction. AdVs and NoVs were detected using nested PCR. Genotyping and phylogenetic analyses were performed to identify the various viral genotypes in the water samples. Human adenovirus (HAdVs) and porcine adenovirus (PAdVs) were the predominant genotypes in the water samples. The prevalence of F species HAdVs serotype 41 (79.2%) and C species PAdVs serotype 5 (18.1%) was higher than that of other serotypes. Among NoVs, genogroup GII was more prevalent than GI. In particular, GII.4 (21.2%) and GII.17 (18.2%) were the predominant genotypes, which was consistent with the clinical findings. The prevalence of both AdVs and NoVs was higher in the winter than spring, summer and autumn seasons. AdVs and NoVs detection results were statistically analyzed by investigating their association with water quality indicators. The results revealed that the presence of AdVs was significantly correlated with the heterotrophic bacterial count, total coliform Escherichia coli, turbidity, salinity, and dissolved oxygen. Meanwhile, the presence of NoVs was only significantly correlated with temperature, pH, and dissolved oxygen. Microbial pollution sources may include urban runoff and discharge of water from livestock farms situated near the river and tributaries within this region of Taiwan. Future studies should include comparisons of the presence of AdVs and NoVs in these known pollution sources and water quality monitoring of these watersheds, as this will allow potential identification of pollution sources. Additionally, remediation strategies must be developed to minimize viral contamination in the river ecosystem.