Background: Increasing population has led to increased water demand in major towns across the world. This study focused on Kitui Town in Kenya where about 80% of the inhabitants depend on Masinga-Kitui water supply system. There are limited alternative water sources including groundwater that are not reliable due high contamination with coliforms. The aim of this study was to determine bacteriological quality of Masinga-Kitui water and establish the potential sources of contaminants.Methods: A total of 90 water samples were collected for analysis whereby, 72 were from the households’ water storage facilities selected through stratified purposive sampling technique. Additional 18 samples were representatively collected from the main distribution network/including two community water points located in Kitui Town (Kalundu and Bondeni). Water quality analysis was performed at Kenya Water Institute water laboratory in Nairobi.Results: All the analyzed water samples were grossly contaminated with E. coli and total Coliforms which exceeded the acceptable WHO/KEBS limit of zero (0) CFU/100 ml of water sample. Majority of the Kitui Town residents associated poor water quality with poor sanitation, poor waste management, poorly maintained sewage and waste waters, contamination of water at the main distribution tank at Kwa-Ngindu, poor water storage at the household and unhygienic practices. About 34% of Kitui residents were at the risk of contracting waterborne diseases due to consumption of contaminated water.Conclusions: Regular monitoring of water quality, surveillance of Masinga-Kitui water infrastructure, and enhanced awareness programme should be adopted to encourage Kitui residents to treat drinking water.