Background: Uropathogens are group of micro-organisms capable of causing urinary tract infections (UTIs). Treatment of infections due to uropathogens has become difficult because of indiscriminate use of antibiotics and ability to easily acquire resistance mechanisms by these pathogens. The objective of this study was to determine the prevalence and antimicrobial susceptibility profile of bacterial uropathogens implicated in UTI among patients attending the three Mission hospitals in Abakaliki, Ebonyi State of Nigeria.Materials & Methods: This was a descriptive study done from 1st January 2022 to 30th June 2022, in three Mission Hospitals of Nigeria 1-Rural Improvement Mission hospital, Ikwo (Hospital A); 2-Mater Misrecordea Hospital, Afikpo (Hospital B) and 3-Saint Vincent hospital, Ndubia (Hospital C) and all are located in semi-urban areas of the Ebonyi State. Inclusion criteria were urine samples of both out-patient and admit hospital patients. Both genders of all age groups patients were included. The bacterial isolates were detected using standard microbiological methods and re-confirmed using colonial morphology. Antimicrobial susceptibility profile was performed using Kirby Bauer disc diffusion.Results: Total 650 bacterial uropathogens isolates were collected from the laboratory units of the three mission hospitals and among them 529 Gram negative bacterial uropathogens were identified and re-confirmed following standard microbiological methods. Overall prevalence of uropathogens in these mission hospitals were 164 (80%), 222 (86%) and 143 (77%) from Hospital A, B and C respectively. E. coli with 58 (35%), 76 (34%) and 50 (35%) prevalence in hospital A, B and C respectively was the most prevalent and frequently isolated uropathogens. Imipenem and meropenem are the two most potent antibiotics against uropathogens in these Mission hospitals. In contrast, the isolated uropathogens developed 100% resistance to tigecycline, ertapenem, aztreonam and ampicillin/sulbactam.Conclusion: Gram negative uropathogens are predominantly the most cause of UTIs and determination of antimicrobial susceptibility profiles of bacterial uropathogens has become crucial in the treatment of UTIs.