Background: Urinary tract infection (UTI) is among the most common reasons for patients to seek health assistance that is commonly encountered in office practices. This is also a leading cause of Gram negative sepsis in hospitalized patients. Objectives: This study was carried out in order to isolate, characterize and identify the pathogens associated with UTI in two teaching hospitals at Osun state, Nigeria and to determine their antimicrobial susceptibility patterns. Methods: This was a prospective observational study involving standard microbiological procedures for analysing urine samples of inpatient and outpatient cases of UTI. Identification of these pathogens was performed using Microbact/API identification system. Results: Out of the 300 urine samples cultured, 88 (29.3%) yielded significant growth of urinary pathogens while 212 (70.7%) yielded either insignificant growth or no growth of any urinary pathogen. Escherichia coli 19 (21.6%) were the commonest pathogen isolated followed by Klebsiella pneumonia 14 (15.9%), Staphylococcus aureus 12(13.6%), Candida albicans 12(13.6%), Pseudomonas aerugenosa 9 (10.2%), Klebsiella oxytoca 8 (9.1%), Staphylococcus saprophyticus 6 (6.8%), Serratia rubidaea 3 (3.4%), Enterobacter agglomerans 2 (2.3%), Acinetobacter iwoffii 1 (1.1%), Acinetobacter baumannii 1 (1.1%), Providencia retgerri 1 (1.1%). The susceptibility of Gram negative bacteria (GNB) were mainly toward parenteral antibiotic rather than oral one, while most of the common antibiotic showed a resistant pattern. UTI was more prevalent among patients within hospital setting 71(80.7%) than out-patients 17(19.3%). Conclusion: This study justifies the necessity to treat patients with UTI based on antimicrobial susceptibility test result in order to prevent evolution of resistant pathogens. Since UTI has large impact on the socio-economy and emergence of bacterial resistance, periodic surveillance of antibiotic susceptibility is strongly recommended. RÉSUMÉ Contexte: L'infection des voies urinaires (IVU) est parmi les raisons les plus courantes pour les patients à demander de l'assistance médicale qui est couramment rencontré dans les clientèles privés. C'est aussi la principale cause de septicémie de bactéries à Gram négatif chez les patients hospitalisés. Objectifs: Cette étude a été réalisée afin d'isoler, caractériser et identifier les agents pathogènes associés aux infections urinaires dans les deux centres hospitaliers universitaires de l'Etat d'Osun au Nigéria et pour déterminer leurs profils de sensibilité aux antimicrobiens. Méthodes: C'est une étude prospective observationnelle impliquant des procédures microbiologiques standard pour analyser des échantillons d'urine de patients hospitalisés et ambulatoires de cas d'infection des voies urinaires. L'identification de cesagents pathogènes a été réalisée en utilisant le système d'identification Microbact /API. Résultats: Sur les 300 échantillons de culture urinaire, 88 (29,3%) ont eu une croissance importante de pathogènes urinaires tandis que 212 (70,7%) ...