Ultrasound imaging and characterization of biofilms based on wavelet de-noised radiofrequency data

The ability to non-invasively image and characterize bacterial biofilms in children during nasopharyngeal colonization with potential otopathogens and during acute otitis media would represent a significant advance. We sought to determine if quantitative high-frequency ultrasound techniques could be used to achieve that goal. Systematic time studies of bacterial biofilm formation were performed on three preparations of an isolated Haemophilus influenzae (NTHi) strain, a Streptococcus pneumoniae (Sp) strain and a combination of H. influenzae and S. pneumoniae (NTHi + Sp) in an in vitro environment. The process of characterization included conditioning of the acquired radiofrequency data obtained with a 15-MHz focused, piston transducer by using a seven-level wavelet decomposition scheme to de-noise the individual A-lines acquired. All subsequent spectral parameter estimations were done on the wavelet de-noised radiofrequency data. Various spectral parameters-peak frequency shift, bandwidth reduction and integrated backscatter coefficient-were recorded. These parameters were successfully used to map the progression of the biofilms in time and to differentiate between single- and multiple-species biofilms. Results were compared with those for confocal microscopy and theoretical evaluation of form factor. We conclude that high-frequency ultrasound may prove a useful modality to detect and characterize bacterial biofilms in humans as they form on tissues and plastic materials.