>Magister Scientiae - MSc / Informal settlements are often implicated in surface water pollution with faecal matter. In most instances faecal pollution in the associated surface waters persists despite improvements in sewage removal infrastructure. This study evaluates the importance of investigating the water quality of the Plankenbrug River before it reaches Khayamnandi settlement by comparing water quality in spring and in winter upstream (Pre-Khayamnandi) and downstream (Post- Khayamnandi) from the settlement. In this study, faecal indicator bacteria (Escherichia coli and total coliforms) were enumerated using Chromocult agar. E. coli was further characterized with analytical profiling index (API) and haemolysis assays. Both Pre- and Post-Khayamnandi were not significantly different from each other for both total coliforms and E. coli in winter. Pre-Khayamnandi had between 105 and 108 cfu/100 ml for total coliforms while Post-Khayamnandi had total coliform colony count between 106 and 107 cfu/100 ml. E. coli also exhibited a similar pattern with slightly higher counts at Post-Khayamnandi with colony counts from 104 to 107 and 105 to 107 cfu/100 ml. Spring microbial count demonstrated a significant difference to winter counts within each test site (p ≤ 0.01) and across the two sites (p ≤ 0.05). Both total coliforms and E. coli were 102 fold higher at Post-Khayamnandi than at Pre-Khayamnandi in spring. The API assay demonstrated significant difference (p ≤ 0.05) between the two test sites. Pre- Khayamnandi predominantly had two different profiles while Post-Khayamnandi had three. These profiles represented five distinct E. coli biotypes. Sorbitol and sucrose tests within the API assay demonstrated significant differences (p ≤ 0.05) between the two test sites. The prevalence of sorbitol fermenters at Pre-Khayamnandi was 100% while at Post-Khayamnandi it was 73%. Pre-Khayamnandi also demonstrated a significantly higher prevalence of sucrose fermenters than Post-Khayamnandi at 100% and 59% respectively. These differences indicated dissimilar sources of faecal contamination around these sites. Differences in the distributions of sorbitol and sucrose fermenting biotypes demonstrate different toxicity potentials across these two test sites. The haemolysis assay demonstrated that 9% of isolates were haemolytic with reference to both known α- and β-haemolyitic streptococci at Post-Khayamnandi. At Pre-Khayamnandi there was a higher percentage of α- and β-haemolyitic species, 29% and 28%, respectively. Post- Khayamnandi and Pre-Khayamnandi were significantly different from each other with reference to both α- and β-haemolysis (p ≤ 0.05). These haemolytic activities also demonstrate different toxicity potentials across the two sites. In conclusion Khayamnandi contributes to an already heavy faecal load in the Plankenbrug River. Thus remedial measures to maintain high surface water quality of Plankenbrug River should be directed upstream from the Khayamnandi settlement as well as within the settlement equally. This study recommends integration of microbial loads with programs such as the National Microbial Monitoring Program of South Africa to drive prioritization process in directing reclaiming of water quality, inter alia.