Ultrasound histotripsy has shown great potential for replacing surgery in many applications. In this work, a modification of ultrasound histotripsy was used to destroy Escherichia coli (E. coli) biofilms that had been grown on chambered microscope slides. Biofilms are central to the pathogenesis and persistence of nosocomial (hospital-acquired) infections associated with indwelling medical devices. The slides were exposed to 9.1 micros pulses at a pulse repetition frequency of 1000 Hz. The pulses were generated by a 1.1 MHz spherically focused source with a focal length of 6.3 cm and an active diameter of 7 cm. The peak rarefactional pressure for the pulses was varied as 3.1, 4.1, 5.2, 6.2, and 7.6 MPa in addition to a sham where the biofilms were not exposed. The effectiveness of the treatment was assessed by determining the viable number of colony forming units (CFU) remaining in the biofilm. Most of the biofilms treated at the higher exposures of 6.2 and 7.6 MPa had no remaining CFU indicating that the biofilm was completely destroyed. However, the persistence of some CFU for some of the biofioms at the higher exposure settings needs to be resolved prior to implementing the treatment clinically.