Environmental quality standards for surface waters have been significantly expanded through recent amendments to German regulations. Limit values are only established for applicable regulations if the water is indicated for certain uses, e.g., abstraction of irrigation water. Nevertheless, surface water bodies are often used for hygiene-sensitive purposes. In the course of climate change, stronger precipitation events will occur, which may lead to more frequent loading and discharge of combined sewer overflow (CSO) into surface water bodies. Retention soil filters (RSFs) are attracting attention as an extensive treatment technology for CSO and additional wastewater treatment. This study examined large-scale RSFs for CSO treatment, as well as the effectiveness of RSFs as a fourth purification stage. A RSF test facility was established at a municipal wastewater treatment plant (WWTP), consisting three semi-technical RSFs that were fed exclusively with treated water from the WWTP. The reduction of microorganisms mostly occurred within the first centimeters of the RSFs. For most hygienic-microbiological parameters, a 1–2 log unit reduction could be detected in addition to the reduction within the WWTP. Antibiotic-resistant bacteria were reduced to the same extent. Investigation of the large-scale RSFs showed that a flow rate reduced by half corresponded to better reduction performances.