ObjectiveTo delineate the epidemiology and antimicrobial resistance (AMR) trends of pathogens causing urinary tract infections (UTIs) during (June 2019 – June 2020) and after (March – July 2023) the implementation of the National Action Plan on AMR 2017-2022 (NAP-AMR) in Mwanza, Tanzania. MethodsThis cross-sectional study was conducted among 2097 patients with clinical symptoms of UTIs during (n=1144) and after (n=953) NAP-AMR. Quantitative urine culture was done to isolate significant bacteria causing UTI, which were then identified to species level and tested for antimicrobial susceptibility. Tabulations, descriptive, and logistic regression analyses were used to analyze categorical and continuous variables, as well as the association between outcome and independent variables. Statistical significance was defined as p ≤ 0.05 at a 95% confidence interval (95%CI). ResultsThe overall prevalence of culture positive UTIs was 22.8% (479/2097, [95%CI: 21.1% - 24.7%]) with no significant difference between the study periods (21.8% (249/1144; [95%CI: 19.5% - 24.3%]) vs. 24.1% (230/953; [95%CI: 21.5% - 26.9%]), p=0.274). We observed a significant increase in resistance to ciprofloxacin (32.0% vs. 45.8%, p=0.0481) and 3rd generation cephalosporins (marked by extended-spectrum beta-lactamase-producing Enterobacterales (ESBL-PE), 38.7% vs. 56.9%, p=0.0307). There were increased odds of ESBL-PE UTIs among patients in higher-tier hospitals than in lower-tier hospitals (29.1% to 49.1%, OR [95%CI]: 3.09 [1.39 – 6.88], p=0.006). ConclusionThere was a significant increase in bacterial resistance to ciprofloxacin and 3rd generation cephalosporins, as well as ESBL-PE. These results emphasize the critical need to enhance AMR surveillance, improve IPC measures, and strengthen antimicrobial stewardship programs.