Introduction: Patients with renal failure undergoing hemodialysis face an increased risk of urinary tract infections due to impaired immunity and altered physiology. Distinguishing asymptomatic bacteriuria from clinically significant infection is vital to combat rising antimicrobial resistance. Objectives: This study aimed to characterize the prevalence and microbial profiles of bacteriuria in hemodialysis dependent patients. Materials and Methods: A retrospective study was conducted at a tertiary care center in Gujarat, India. Data was collected from electronic medical record of patients along with urine culture finding and other clinical details to study further for the duration of April 2021 and March 2025. Clinically significant isolates were reviewed while excluding duplicate isolates from same patients. Study was approved by institutional ethics committee. Results: Out of total 17755 various culture samples received from dialysis-patients, 3022 urine cultures were received and from those total 772 urine cultures reported with bacterial growth during the studied duration. The cohort had a mean age of 55.1 years with a female predominance (58.7%). Gram-negative bacteria (≈80%) dominated, primarily Escherichia coli (57.1%) and Klebsiella pneumoniae (13.9%). High resistance was observed against cephalosporins and fluoroquinolones. Enterococcus faecium exhibited significant resistance to vancomycin (20.8%). Prior antibiotic exposure was high at 76.9%. Resistance was more frequently associated with patients having no fixed dialysis schedule and those receiving antibiotics within 24 hours of enrolment. Conclusion: Hemodialysis patients frequently harbor multidrug-resistant pathogens. The high prevalence of ESBL producing and MDR strains necessitates robust antibiotic stewardship and reliance on local antibiograms to guide therapy and minimize unnecessary treatment of asymptomatic cases.