Urinary Tract Infections (UTIs) are one of the most common infections in the UK and many other parts of the world. The prevalence of the Extended Spectrum ?-Lactamases (ESBLs) producing UTIs, combined with their ability to form a bio film, has significantly risen and is limiting therapeutic options. This study investigated the anti-bio film activity of Manuka honey on two ESBL producing pathogens, Escherichia coli TEM-3 and Klebsiella pneumonia SHV18, commonly found in UTIs. The ESBL production was confirmed by the double disk synergy method used to confirm the ESBL production. The antibacterial activity of Manuka honey was determined using the agar well diffusion method. The Minimum Inhibitory Concentration (MIC) was established using serially diluted honey ranging from 50% to 1.56%. The effect of Manuka honey on the pathogen bio films was analysed using the Tissue Culture Plate method, with an established MIC and under 24h incubation with the honey. The results indicated that K. pneumonia SHV18 is a stronger bio film producer than E. coli TEM 3. 50% (w/v) MIC Manuka honey appears to fully prevent the plank tonic growth of both strains. A significant reduction of 81% of the E. coli TEM3 (p < 0.001) and 52% of the K. pneumonia SHV18 (p = 0.001) bio film biomass was observed. The E. coli bio films were found to be more sensitive to the 50% (w/v) honey dilution than those produced by K. pneumonia. The study indicated the anti-bio film potency of Manuka honey and its potential to become an alternative treatment for the ESBL producing pathogens associated with UTIs.