Objective: To detect bacteriuria or funguria by urine culture taken from the renal pelvis directly before Anderson-Hynes pyeloplasty. Methods: 290 patients who underwent Anderson-Hynes pyeloplasty for ureteropelvic junction obstruction (UPJO) were included in a retrospective analysis. Urine was obtained directly before the renal pelvis was opened, and was carried to the laboratory for bacterial culture. Clinical features were analyzed to evaluate risk factors for bacteriuria or funguria by comparing patients whose urine yielded positive cultures to those whose urine cultures were negative for bacteria or yeast. Results: Eighteen patients (6.2%) had positive urine cultures, including six cultures positive for Escherichia coli (E. coli), four for Pseudomonas aeruginosa, three for klebsiella pneumoniae, one for maltophilia monad, one for Enterococcus faecium, one for Candida albicans, one for Candida parapsilosis, and one for yeast not otherwise specified. Bacteriuria or funguria was significantly correlated with four clinical features: fever, urinary urgency, and history of nephrostomy or pyeloplasty. Conclusions: Bacteriuria or funguria was less common in children with UPJO, and the majority of organisms were identified as Escherichia coli, Pseudomonas aeruginosa, or Klebsiella pneumoniae. Prophylactic antibacterial agents were probably necessary in those patients who had signs of urinary tract infection (UTI), or history of nephrostomy or pyeloplasty.