Phage therapy for the treatment for urinary tract infection: Results of in-vitro screenings and in-vivo application using commercially available bacteriophage cocktails Introduction and Objectives Material and MethodsResults In-vitro screening for bacteriophage sensitivity of antibiotic resistant Escherichia coli and Klebsiella pneumoniae strains, isolated from the urine of patients with symptomatic UTI. In-vivo treatment of patients by using traditional phage preparation Pyo-bacteriophage including components active against E. coli, Enterococcus, Proteus, Staphylococcus and Streptococcus Urinary Tract Infections (UTI) are among the most prevalent microbial diseases. Approximately 15% of all community-prescribed antibiotics in the USA are for the UTI at an estimated annual cost of over 1 billion USDs. The use of bacteriophages against bacterial pathogens has gained a renewed interest over the last years, due to a continuing increase in antibiotic resistance worldwide. The goal of our research is to investigate the options of using bacteriophage therapy for treatment of symptomatic UTI and prepare for a double-blind, randomized, placebo- controlled clinical trial In vitro: The results of the initial screening of 61 pure cultures collected between December 2014 and March 2015 showed that 33% of the strains were completely susceptible (S), 23% demonstrated intermediate susceptibility (I) and 42% appeared to be resistant(R). After the first adaptation cycle the collected strains have been screened again for phage susceptibility. The results showed that the number of completely (S) and intermediary susceptible strains increased up to 42% and 32%, correspondingly, while the number of resistant strains decreased from 42% to 26%. (figure 1). In vivo: 170 patients undergoing transurethral surgery (either TURP or laser vaporization/enucleation) have been examined for the presence of uropathogens and phage sensitivity prior to surgery. 107 (62%) of 170 patients have had positive urine culture (≥10 4 colony forming units/mL of uropathogens) and lower urinary tract symptoms like: Urgency, frequency, urge incontinence, etc. The project is funded by the grant from The Swiss National Science Foundation Acknowledgments 265 A. Ujmajuridze 1, G. Jvania 1, N. Chanishvili 2, M. Goderdzishvili 2, W. Sybesma 3, L. Managadze 1, A. Chkhotua 1, T. Kessler 4 (1) National Center of Urology, Dept. of Urology, Tbilisi, Georgia, (2) The Eliava Institute of Bacteriophage, Dept. of Microbiology and Virology, Tbilisi, Georgia, (3) University of Zürich, Institute of Medical Microbiology, Zürich, Switzerland, (4) Balgrist University Hospital, Dept. of Neuro-Urology, Spinal Cord Injury Center & Research, Zürich, Switzerland Sensitivity test has shown that UTI was sensitive to phages in 39.43%, intermediately sensitive in 15.5% and resistant in 45%. Also here adaptation of Pyo-phage was done to increase effectiveness of this preparation, resulting in an increase of sensitivity of phage sensitive urine samples from 28.26% to 39.43% (P=0.0370) (figure 3). Nine patients have been treated so far with bacteriophages: 4 patients with E. coli, 1 patient with P. aeruginosa, 2 patients with Enterococcus, and 2 patients with Streptococcus. The bacteriophage treatment has significantly decreased urine concentration of uropathogens in all patients (P=0.0092). No drug associated side effects have been detected in these patients (figure 4). Pathogens in urine Cultures Patients% E.coli3621,2 Enterococcus3922,9 Pseudomonas74,1 Staphilococcus74,1 Streptococcus1810,6 2 Pathogens or fungus 1810,6 Klebsiela10,6 Nonpathogenous4023,5 Sterile42,4 Total screened170 Infection found in10762,90% p= vs. 1 Figure 3. Influence of phage adaptation on urine culture sensitivity st Adaptation1 2 nd Adaptation2 3 rd Adaptation3 Baseline1 After phagotherapy2 CFU Figure 2. Results of urine culture analyses in 170 patients undergoing surgery for BPO Figure 4. Effect of phage therapy on the number (CFU) of uropathogens Preliminary results of this clinical trial are showing high frequency of UTI in patients undergoing surgery for BPO. Substantial number of these patients will have MDR syndrome precluding the standard antibiotic treatment. Due to the positive results of our study, we will start a multicenter, double-blind, randomized, placebo controlled clinical trial in Q to further assess application of bacteriophages in the treatment of UTI. Conclusion Figure 1. Positive dinamics in efficicncy of Pyo-phage after the first cycle of adaptation to urology pathogens. Of those E.coli was found in 21.2%, Enterococcus in 22.9%, Streptococcus in 10.6%, P. aeruginosa and Staphylococcus in 4.1% cases each. Mixed culture was found in 10.6% of patients. Three percent of patients had sterile urine (figure 2).