Introduction: Urinary tract infections (UTIs) are among the most common bacterial infections, posing a growing public health challenge due to the prevalence of multidrug-resistant bacteria capable of biofilm formation, particularly strains that produce β-lactamase enzymes and spectrum clocks. This has led to increased infection recurrence rates, treatment failure, and reduced effectiveness of many conventional antibiotics. This study aimed to identify the bacterial characteristics of UTI pathogens, their biofilm formation capabilities, and resistance patterns, and to evaluate the antibacterial and anti-biofilm activity of Saussurea costus root extract. Materials and Methods: Ninety urine samples were collected from patients with symptoms of urinary tract infections between June and October 2025 from four hospitals in Baghdad: Ibn Al-Baladi Hospital, Al-Kindi Hospital, Al-Shaheed Al-Sadr Hospital, and Ibn Al-Qaf Hospital. Bacterial isolates were identified using conventional microbiological methods and the VITEK 2 automated system. The disk diffusion method was used for antibiotic susceptibility testing. Biofilm formation was assessed using microplate assays and Congo red staining. An alcoholic extract of Saussurea costus root was prepared and then chemically analyzed using high-performance liquid chromatography (HPLC). Its antibacterial activity was evaluated by determining the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) against selected multidrug-resistant isolates. Results: Out of 90 urine samples, 62 bacterial isolates were obtained, with Escherichia coli being the most common isolate (44/62, 71.0%), followed by Staphylococcus (18/62, 29.0%). Antibiotic susceptibility testing showed high resistance rates in E. coli isolates to ampicillin (71.4%) and tetracycline (64.3%). The test also showed 100% sensitivity to levofloxacin and imipenem. All Staphylococcus aureus isolates showed complete resistance to oxacillin and cefoxitin, confirming the presence of methicillin-resistant Staphylococcus aureus (MRSA). However, they showed 100% sensitivity to linezolid, vancomycin, tigecycline, and teicoplanin. All multidrug-resistant isolates demonstrated a high capacity (100%) for biofilm formation using both the microtiter plate and Congo red agar methods. The flavonoid fraction of the Saussurea Costus root extract showed significant antibacterial activity, with MIC values ranging from 31.2-15.6 µg/ml against Staphylococcus aureus and 75-125 µg/ml against E. coli. MIC/MBC ratio analysis indicated a bacteriostatic effect on Staphylococcus aureus and a bactericidal effect on E. coli. Conclusion: This study highlights the important role of multidrug resistance, as the results confirm the promising therapeutic potential of costus root extract as an adjunct treatment against multidrug-resistant urinary tract pathogens, and emphasize the need to develop alternative or complementary strategies to improve the optimal use of antibiotics.
