Background: Urinary tract infections are always treated empirically before the results of bacteriological cultures are obtained. The choice of antibiotics depends upon the causative organism and its expected local antibiotic susceptibility pattern. Objectives: We analyzed the spectrum and resistance patterns of uropathogens against common antimicrobial agents in Ahvaz Abuzar Children's Hospital, a tertiary care pediatric unit in southwest of Iran. Patients and Methods: In this retrospective study, all urine samples of children hospitalized with urinary tract infection (288 patients, aged 1mon -14.5 years) during October 2008 to May 2011 were included in the study. After bacteria were identified by standard methods, antimicrobial susceptibility testing was performed using a panel of antimicrobial agents. Results: The most of patients were girls (n = 226, 78.5%), and the median age was 13 months. The most common pathogens were Escherichia coli (84%), Klebsiella spp. (10.1%), Enterococcus spp. (2.4%), Proteus spp. (1.7%), and Pseudomonas spp. (1.7%). Overall bacterial resistance spectrum was the highest for co-trimoxazole (64.8%), followed by gentamicin (44.6%), amikacin (40.5%), nalidixic acid (37.3%), cefotaxime (28.9%), cefixime (27.5%), ceftriaxone (27.4%), and nitrofurantoin (10.2%). The female:male ratio was 2:1 (67.1% versus 32.9%) in infants aged < 1 year and 8:1 in those aged > 1 year (89.4% vs. 10.6%). Vesicoureteral reflux and abnormal sonography findings were associated with high resistance to cefotaxime (P = 0.017), ceftriaxone (P = 0.004), nitrofurantoin (P = 0.014), and nalidixic acid (P < 0.001). Conclusions: Increasing resistance to third-generation cephalosporins changed our opinion for using them as a single empiric intravenous therapy in hospitalized and very ill patients with acute pyelonephritis; the success will be achieved by concomitant use of an aminoglycoside or using other potent antibiotics.