Urine is rich in nutrients such as Nitrogen and Phosphorus which are essential for agriculturernand therefore worthy of safe recovery. The aim of this study was to evaluate the effectivenessrnof acidification, storage and pasteurization of source-separated human urine in reducingrncoliform bacteria. Urine samples were collected using acetic acid-treated portable toilets andrnjerrycans from four sites of Addis Ababa namely; Addis Ababa University (AAU) NaturalrnScience campus, Merkato portable toilet, Megenagna public toilet and Gelila Elementaryrnschool. Treatment of urine was conducted using Hydrochloric acid (HCl) heating to 80 ÂºC forrnpasteurization and storage in tight-sealed containers at room temperature. The urine undergoingrnthe three sanitization methods were analyzed for their physico-chemical parameters, coliformrnbacteria and for the stored samples, PCR-based detection of genes resistant to tetracycinern(tetB), sulfonamides (sul2) and quinolones (qnrB) were analyzed. The study showed the pH ofrnfresh urine collected from the four sites to be slightly acidic in the range of pH 5.2 and 5.8,rnwhich increased to a maximum of 10.4 during storage for six months. Total coliforms in freshrnurine collected from the four sites of Addis Ababa ranged between the highest count of 234,000rncfu/ml to 64500 cfu/ml, which decreased steadily during the storage time reaching to nil at thernsixth month of storage. In all the fresh urine samples, fecal coliforms were detected with thernmaximum count of 1500 cfu/100 ml in samples collected from AAU. Storage for one monthrnwas efficient in reducing fecal coliform units to zero in all the samples collected from the fourrnsites. Pasteurization was also efficient in completely removing both total and fecal coliforms.rnAcidification was didnâ€˜t show any reduction in total coliform counts of urine collected from allrnthe four sites. Regarding fecal coliforms, a slight decrease was observed resulting in 20%rnremoval of fecal coliforms in the case of urine collected from Merkato .PCR-based detection ofrntetB resistance gene from the meta-DNA revealed that fresh urine collected from AAU wasrnpositive for the tetB which was absent at the sixth month of storage. Similarly, the fresh urinernsample collected from Merkato was positive for qnrB which persisted in the stored sample for 4rnmonths and finally absent at the sixth month of storage. It was concluded that storage as arnsanitization process is the most effective method in reducing coliform bacteria and some of thernhorizontally transmitted antibiotic resistance genes, with minimal loss of Nitrogen andrnPhosphorus.