The aim of this study was to evaluate the efficiency of a passive point-of-use treatment system, namely, a polyvinyl (alcohol) (PVA) nanofiber membrane/activated carbon column, for the treatment of harvested rainwater. The efficiency of SMI-Q10 [quaternized poly (styrene-co-maleimide)] nanofiber membrane disks placed in a filtration assembly for the treatment of surface water (Plankenburg River, Western Cape, South Africa) and harvested rainwater was also assessed. Two rainwater harvesting tanks were installed at the Welgevallen Experimental farm, Stellenbosch, South Africa, with the filtration system intermittently attached to the tanks for collection of rainwater samples throughout the study period. Parameters used to monitor the filtration systems included heterotrophic bacteria, Escherichia coli, and total coliform enumeration and the presence/absence of adenovirus. When compared to drinking water guidelines, the results indicated that 3 L of potable water could be produced by the synthesized PVA nanofiber membrane/activated carbon column. However, PCR assays indicated that adenovirus and numerous bacteria such as Klebsiella spp., Legionella spp., Pseudomonas spp., and Yersinia spp. were not effectively removed by the filtration system utilized. Additionally, the SMI-Q10 nanofiber membrane disks did not remove viruses from the river or tank water samples as bovine adenovirus 3 strain, simian adenovirus, and human adenovirus A strain were detected in all water samples analyzed. Thus, while the microfiltration system was efficient in reducing the level of indicator organisms to within drinking water standards, further optimization of the electrospun filtration membranes is required as molecular analysis revealed that numerous opportunistic bacterial pathogens and viruses persisted after filtration.