In low-income regions, drinking water is often derived from groundwater sources, which might spread diarrheal disease if they are microbiologically polluted. This study aimed to investigate the occurrence of fecal contamination in 147 improved groundwater sources in Juba, South Sudan and to assess potential contributing risk factors, based on bivariate statistical analysis. Thermotolerant coliforms (TTCs) were detected in 66% of the investigated sources, including 95 boreholes, breaching the health-based recommendations for drinking water. A significant association (p < 0.05) was determined between the presence of TTCs and the depth of cumulative, long-term prior precipitation (both within the previous five days and within the past month). No such link was found to short-term rainfall, the presence of latrines or damages in the borehole apron. However, the risk factor analysis further suggested, to a lesser degree, that the local topography and on-site hygiene were additionally significant. In summary, the analysis indicated that an important contamination mechanism was fecal pollution of the contributing groundwater, which was unlikely due to the presence of latrines; instead, infiltration from contaminated surface water was more probable. The reduction in fecal sources in the environment in Juba is thus recommended, for example, through constructing latrines or designating protection areas near water sources. The study results contribute to the understanding of microbiological contamination of groundwater sources in areas with low incomes and high population densities, tropical climates and weathered basement complex environments, which are common in urban sub-Saharan Africa.