Cavitation Activity Research Articles

Optical analysis of transesterification reaction assisted by ultrasound

Ultrasound can be used during the biodiesel production reaction, aiming to increase the efficiency of the reaction, as well as reduce the catalyst and alcohol content, facilitating the purification process and minimizing environmental impacts in the production of this biofuel. This work evaluated by optical means the influence of the use of ultrasound on the reaction dynamics of transesterification under different catalyst contents. Tests were conducted using a Langevin-type transducer (40 kHz x 20.5 W) to convert 270 mL of soybean oil per batch. Previous tests in the reactor used showed intense cavitation activity resulting in mass loss of up to 27.3% in erosion tests on aluminum foil. Optical transmittance and ester content evaluation analyzes of the samples showed that ultrasound was able to increase the conversion rate and reduce the optical stabilization time (up to 9.9%), suggesting a shorter reaction time, especially for lower ester contents. catalyst.

Magnetic resonance cavitation imaging for the monitoring of ultrasound therapies

Objective.Focused ultrasound (FUS) is a promising non-invasive therapeutic approach that can be used to generate thermal and non-thermal bioeffects. Several non-thermal FUS therapies rely on FUS-induced oscillations of microbubbles (MBs), a phenomenon referred to as cavitation. Cavitation monitoring in real time is essential to ensure both the efficacy and the safety of FUS therapies. This study aims to introduce a new magnetic resonance (MR) method for cavitation monitoring during FUS therapies.Approach.By finely synchronizing the FUS pulse with an accelerated turbo spin-echo MR sequence, the cavitation effect could be quantitatively estimated on the acquired images at 1-Hz refresh rate. The proposed method was assessed in vitro in a water bath. A series of FUS pulses were generated on a silicone tube filled with MBs at different acoustic pressures (0.07-2.07 MPa) and pulse durations (20-2000μs). MR images and passive cavitation detection (PCD) signals were simultaneously acquired for each FUS pulse.Main results.Inertial cavitation was found to induce a quantitatively interpretable signal loss on the MR image. The transition from stable to inertial cavitation was identified on MR cavitation maps with high repeatability. These results were found to be in good agreement with PCD measurements in terms of pressure thresholds between stable and inertial cavitation. MR cavitation imaging was shown to be sensitive to short and even ultrashort FUS pulses, from 2 ms down to 20μs. The presented theoretical model suggests that the signal loss in MR cavitation imaging relies on susceptibility changes related to the diameter of the oscillating MBs.Significance.The proposed MR cavitation imaging method can both locate and characterize cavitation activity. It has therefore the potential to improve the efficacy and safety of FUS therapies, particularly for localized drug delivery applications.

Quantitative evaluation of anti-biofilm cavitation activity seeded from microbubbles or protein cavitation nuclei by passive acoustic mapping

Bacterial biofilms represent a major challenge for effective antibiotic therapy as they confer physical and functional changes that protect bacteria from their surrounding environment. In this work, focused ultrasound in combination with cavitation nuclei was used to disrupt biofilms of Staphylococcus aureus and Pseudomonas aeruginosa, both of which are on the World Health Organization's priority list for new antimicrobial research. 
Approach: Single species biofilms were exposed to ultrasound (0.5 MHz centre frequency, 0.5-1.5 MPa peak rarefactional pressure, 200 cycle pulses, 5 Hz repetition frequency, 30 s duration), in the presence of two different types of cavitation nuclei. Quantitative passive acoustic mapping (PAM) was used to monitor cavitation emissions during treatment using a calibrated linear array. 
Main Results: It was observed that the cumulative energy of acoustic emissions during treatment was positively correlated with biofilm disruption, with differences between bacterial species attributed to differences in biofilm morphology. PCaN provided increased biofilm reduction compared to microbubbles due in large part to their persistence over the duration of ultrasound exposure. There was also good correlation between the spatial distribution of cavitation as characterized by PAM and the extent of biofilm disruption observed with microscopy. 
Significance: Collectively, the results from this work indicate the potential broad applicability of cavitation for eliminating biofilms of priority pathogens and the opportunity presented by Passive Acoustic Mapping for real-time monitoring of antimicrobial processes.

Mapping of a Stirred Ultrasonic Reactor Operated in Recirculation Mode Using a Cavitation Activity Meter and Validation Using Dye Degradation

Mapping of a Stirred Ultrasonic Reactor Operated in Recirculation Mode Using a Cavitation Activity Meter and Validation Using Dye Degradation

Reconstructing Multiphase Flow Fields with Limited Pressure Observations Based on an Improved Transformer Model

In practical applications, the implementation of active cavitation control can significantly enhance the hydrodynamic performance of underwater vehicles. However, the sparsity of sensor information poses a substantial challenge to acquiring flow field states. Recent advances in deep learning offer reliable solutions to this problem. Specifically, deep learning methods, through the construction of sparse reconstruction models, establish connections between sparse observational information and flow field states with minimal or no prior knowledge. To reconstruct multiphase flow fields from sparse pressure observations on the hydrofoil surface, this work proposes a Transformer-based sparse reconstruction model. Test results on cavitation datasets demonstrate that the model's predictions of cavity contours, cavity lengths, and cavity volumes are highly consistent with actual results during the sheet cavity growth and cavity shedding stages. However, due to the highly unsteady nature of cavitation flow and the lack of far-field information, the model's prediction performance is limited during the cloud cavity aggregation and collapse stages. This model exhibits potential in the sparse reconstruction of multiphase flow fields, providing support for the observation of flow field states and active cavitation control.

Optimizing high-intensity focused ultrasound-induced immunogenic cell-death using passive cavitation mapping as a monitoring tool

Over the past decade, ultrasound (US) has gathered significant attention and research focus in the realm of medical treatments, particularly within the domain of anti-cancer therapies. This growing interest can be attributed to its non-invasive nature, precision in delivery, availability, and safety. While the conventional objective of US-based treatments to treat breast, prostate, and liver cancer is the ablation of target tissues, the introduction of the concept of immunogenic cell death (ICD) has made clear that inducing cell death can take different non-binary pathways through the activation of the patient's anti-tumor immunity. Here, we investigate high-intensity focused ultrasound (HIFU) to induce ICD by unraveling the underlying physical phenomena and resulting biological effects associated with HIFU therapy using an automated and fully controlled experimental setup. Our in-vitro approach enables the treatment of adherent cancer cells (B16F10 and CT26), analysis for ICD hallmarks and allows to monitor and characterize in real time the US-induced cavitation activity through passive cavitation detection (PCD). We demonstrate HIFU-induced cell death, CRT exposure, HMGB1 secretion and antigen release. This approach holds great promise in advancing our understanding of the therapeutic potential of HIFU for anti-cancer strategies.

Pulsed focused ultrasound ablation assisted by a surface modified catheter for thrombolysis: a feasibility study

Interventional procedures for the recanalization of blood vessels to treat deep vein thrombosis carry a high risk of vessel wall injuries or hemorrhaging. Focused ultrasound (FUS) has been used to non-invasively break down blood clots that occlude the vessels in both in vitro and in vivo studies. Previous studies have either used thrombolytic drugs or ultrasound contrast agents (e.g., microbubbles) in combination with FUS. Several studies have applied very high peak-negative-pressures (PNP) during FUS treatment to achieve successful thrombolysis without the use of contrast agents. In the current study, we demonstrated that cavitation activity could be significantly enhanced by placing a nitinol wire, whose surface was roughed by laser etching, in the focal region of a FUS field. We demonstrated in vitro in a mock thrombosis that the thrombolysis efficacy of a 500 kHz FUS transducer was significantly enhanced using a surface-etched nitinol wire as compared to an unetched nitinol wire, whereas FUS-alone at the same pressure level did not result in any thrombolysis. These results suggest that a surface modified nitinol catheter exposed to FUS can result in intense cavitation activities leading to enhanced thrombolysis without the use of additional pharmacological or contrast agents.

2D spatiotemporal passive cavitation imaging and evaluation during ultrasound thrombolysis based on diagnostic ultrasound platform

Acoustic cavitation plays a critical role in various biomedical applications. However, uncontrolled cavitation can lead to undesired damage to healthy tissues. Therefore, real-time monitoring and quantitative evaluation of cavitation dynamics is essential for understanding underlying mechanisms and optimizing ultrasound treatment efficiency and safety. The current research addressed the limitations of traditionally used cavitation detection methods by developing introduced an adaptive time-division multiplexing passive cavitation imaging (PCI) system integrated into a commercial diagnostic ultrasound platform. This new method combined real-time cavitation monitoring with B-mode imaging, allowing for simultaneous visualization of treatment progress and 2D quantitative evaluation of cavitation dosage within targeted area. An improved delay-and-sum (DAS) algorithm, optimized with a minimum variance (MV) beamformer, is utilized to minimize the side lobe effect and improve the axial resolution typically associated with PCI. In additional to visualize and quantitatively assess the cavitation activities generated under varied acoustic pressures and microbubble concentrations, this system was specifically applied to perform 2D cavitation evaluation for ultrasound thrombolysis mediated by different solutions, e.g., saline, nanodiamond (ND) and nitrogen-annealed nanodiamond (N-AND). This research aims to bridge the gap between laboratory-based research systems and real-time spatiotemporal cavitation evaluation demands in practical uses. Results indicate that this improved 2D cavitation monitoring and evaluation system could offer a useful tool for comprehensive evaluating cavitation-mediated effects (e.g., ultrasound thrombolysis), providing valuable insights into in-depth understanding of cavitation mechanisms and optimization of cavitation applications.

Three-dimensional super-resolution passive cavitation mapping in laser lithotripsy.

Kidney stone disease is a major public health issue. By breaking stones with repeated laser irradiation, laser lithotripsy (LL) has become the main treatment for kidney stone disease. Laser-induced cavitation is closely associated with the stone damage in LL. Monitoring the cavitation activities during LL is thus crucial to optimizing the stone damage and maximizing LL efficiency. In this study, we have developed three-dimensional super-resolution passive cavitation mapping (3D-SRPCM), in which the cavitation bubble positions can be localized with an accuracy of 40 μm, which is 1/10th of the acoustic diffraction limit. Moreover, the 3D-SRPCM reconstruction speed has been improved by 300 times by adopting a GPU-based sparse-matrix beamforming approach. Using 3D-SRPCM, we studied LL-induced cavitation activities on BegoStones, both in free space of water and confined space of a kidney phantom. The dose-dependence analysis provided by 3D-SRPCM revealed that accumulated impact pressure on the stone surface has the highest correlation with the stone damage. By providing high-resolution cavitation mapping during LL treatment, we expect that 3D-SRPCM may become a powerful tool to improve the clinical LL efficiency and patient outcome.

Histological characterization of HIFU lesions

Background: High intensity focused ultrasound (HIFU) can destroy tissue by thermal ablation which may be accompanied by acoustic cavitation and/or tissue water boiling, but the biological and histological effects of these treatments have not been fully documented. Here, detailed histological analysis over time using well characterized HIFU exposures in in vivo rat livers is described. Methods: Exposures used invoked either (i) thermal, with acoustic cavitation and/or tissue water boiling or (ii) predominantly thermal damage. Cavitation activity was detected using both active and passive methods. Histological assessment involved hematoxylin and eosin (H&E), picrosirius red and immunohistochemical staining. Results: Distinct concentric damage regions were identified after HIFU exposures. The outermost ring showed a red H&E-stained rim that was characterized by hemorrhage. The adjacent inner band appeared white due to increased extracellular spaces. The morphology of the next zone depended on the exposure. Where there was no tissue acoustic cavitation/water boiling, this was the lesion center, in which heat-fixed cells were seen. Where acoustic cavitation/boiling occurred, a centermost zone with irregular holes up to several hundred microns across was seen. Cleaved caspase-3 and Hsp70 staining in the periphery of both types of HIFU exposures was seen within the outermost ring of hemorrhage, where an inflammatory response was also observed. By day 7, a distinct acellular region in the center of the HIFU lesions had been created. Conclusions: These results identify the morphological effects and elucidate the similarities and differences of HIFU-induced thermal lesions in the presence or absence of acoustic cavitation/tissue water boiling.

An improved scheme for passive cavitation imaging in high intensity focused ultrasound

Passive cavitation imaging (PCI) is an effective technique for monitoring high intensity focused ultrasound (HIFU) procedure due to the correlation between cavitation activities and the therapeutic effects. Nevertheless, current PCI techniques are unable to recognize noise signals from non-cavitation situations, and they fail to accurately visualize the highly dynamic cavitation activities due to the lack of a stable baseline reference. In this study, a deep learning-based cavitation detection network (CD-NET) was proposed to determine whether passively received channel signals originated from cavitation activities or were merely noises. Additionally, a cavitation intensity baseline was derived to better display the drastically time-varying cavitation activities. The CD-NET model was trained and validated with data collected from in-vitro and ex-vivo HIFU sonication experiments, and then tested by in-vivo rabbit experiments. Both CD-NET models, trained with data from the in-vitro agar phantoms and ex-vivo bovine liver tissues, successfully distinguished between cavitation and non-cavitation activities from the same study object, achieving accuracies as high as 99.7% and 99.5%, respectively. In addition, the models trained on in-vitro and ex-vivo datasets achieved accuracies of 88.1% and 91.4%, respectively, when detecting cavitation events from unseen in-vivo data. The results demonstrate that the improved PCI scheme, utilizing the CD-NET model and a data-derived cavitation baseline, is resistant to noise from non-cavitation situations and can depict the dynamic characteristics of cavitation more reliably. In conclusion, this study addressed two practical challenges in PCI techniques, allowing for better monitoring of cavitation-related HIFU sonication in clinical applications.

Monitoring holograms for therapeutic ultrasound using passive cavitation beamforming

Acoustic holograms can generate cavitation patterns of complex spatial distribution by shaping and steering the focal spot of therapeutic ultrasound systems. However, when monitoring these systems by passive cavitation detection, off-axis therapeutical targets and the receiver directivity may not be aligned. In this paper, we present passive cavitation beamforming to monitor a therapeutical ultrasound system using holograms targeted to arbitrary locations, in which both therapeutic and passive cavitation monitoring systems use 3D-printed acoustic lenses. The therapeutic system uses an acoustic hologram to focus the ultrasound beam on the target, which is off-axis. Then, a second lens is designed to beamform the cavitation signals which emerge from the therapeutic target, steering the directivity of the passive cavitation detector in the direction of the therapeutic focus and, in addition, compensating for skull aberrations. The system is experimentally tested with an ex-vivo macaque skull and a blood vessel phantom with microbubbles. In addition, results are compared with a standard confocal configuration and an off-axis configuration in the absence of the monitor lens. A parametric study is performed by varying the amplitude of the emitted signal and the impact on the behaviour of the microbubbles is analysed based on the cavitation index values. Results show that monitoring holograms align the passive cavitation detector response with the focal spot of the targeted therapeutic transducer. These holograms encode a fixed beamformer for cavitation signals in reception, increasing the sensitivity of cavitation emission at the target. In this way, cavitation doses can be used to locally monitor the cavitation activity of microbubbles, thus opening a new path to low-cost monitoring of therapeutic ultrasound systems.

Assessment of Cavitation Activity by Cavitation Noise in Different Frequency Ranges

The results of testing the spectral-acoustic cavitometer with measurement modes that differ in the frequency ranges of integration of cavitation noise are presented. For each of the H1–H4 modes, the lower integration frequency fmin is 1, 70, 180 and 300 kHz, respectively. The upper frequency is the same for all modes and is fmax = 10 MHz. Piezoelectric waveguide sensors were used as a cavitation noise receivers. The distributions of cavitation activity in non-uniform ultrasonic fields were recorded: in the volume of the ultrasonic bath (f0 = 35 kHz) and in the field of the submersible emitter (f0 = 21 kHz). The frequency range of integration was varied by changing the lower bound. It has been established that a change in the integration range within a fairly wide range does not have a noticeable qualitative effect on the result of measurements of cavitation activity, only a multiple change in the readings occurs. This conclusion was confirmed using sensors that differ in their spectral characteristics.

Sonochemical oxidation activity in 20-kHz probe-type sonicator systems: The effects of probe positions and vessel sizes

The 20-kHz probe-type sonicator systems were investigated for the enhancement of the cavitational oxidation activity under various geometric conditions including vertical and horizontal probe positions and vessel sizes/volumes as a following study to our previous study. The sonochemical oxidation activity (mass-based I3- ion generation rate) increased significantly for all vessel size conditions as the probe was placed close to the vessel bottom, owing to the expansion of the sonochemical active zone induced by the reflections of ultrasound at the bottom and the reactor wall. A concentric circular active zone is observed at positions close to the bottom. The highest sonochemical activity was obtained at 1 cm (vertical position) in the 20 cm vessels (input power: 50 %). At the vertical positions of 11 cm to 7 cm, no significant difference in the sonochemical activity was observed for all input power conditions (25, 50, and 75 %) because no meaningful reflections occurred. Higher sonochemical activities were obtained at an input power of 75 % owing to the increased power and strong reflection. The highest cavitational yield considering the energy efficiency was obtained at 6 cm (vertical position) for 75 % of all power and geometric conditions. Horizontal probe position tests showed that the asymmetric formation of the sonochemical active zone could significantly enhance the sonochemical activity. The highest activity was obtained at 1 cm (vertical position) and 2.5 cm (horizontal position) in the 20 cm vessel.

Research of Acoustic Cavitation: Results, Practical Implementation and Development Prospects

The most significant results of research performed in the Belarusian State University University of Informatics and Radioelectronics, aimed at creating methods and instruments for monitoring cavitation and controlling its activity, are presented. To achieve this goal, it was necessary to clarify the mechanisms and patterns of new and little-studied phenomena: ultrasonic capillary effect, sonoluminescence, cavitation noise and its spectral components and evaluate the possibilities of their use for hardware implementation as cavitation indicators. New cavitation phenomena have been discovered: reverse ultrasonic capillary effect, generation of constant EMF under the influence of alternating pressure of ultrasonic frequency; acoustic clearing of the cavitation area with pulsed modulation of the ultrasonic field, the effect of long-term memory of the cavitation properties of the liquid. Previously unknown patterns have been established: it is shown that the cavitation region goes through four stages of development with increasing ultrasound intensity; a theorem on the maximum cavitation activity was formulated; the spectral characteristics of cavitation noise were established, correlating with the intensity of sonoluminescence. It has been shown that pulsed modulation of the ultrasonic field makes it possible to control the dynamics of the development of the cavitation region and the activity of cavitation. Based on the data obtained, new methods and instruments have been developed to study cavitation and control its activity. In a number of parameters, the created equipment surpasses the best world samples, what is confirmed by deliveries to the Russia, EU countries, China, USA and South Korea.

Experimental and numerical comparison of multiple passive beamformers for separating intra- and extra-canal cavitation activity during transvertebral spinal cord therapy

Distinguishing intra-canal cavitation activity during transvertebral focused ultrasound sonication of the spinal cord is a challenge due to the strong prefocal cavitation emissions in the spinalis musculature overwhelming emissions originating in the canal. To achieve mapping of all cavitation sources simultaneously, two methods for enhancing detection sensitivity are investigated: (1) multiple dynamic ranges within a reconstructed volume and (2) utilizing alternative beamformers to delay-and-sum (DAS) during map reconstruction. The performance of DAS beamforming is compared to a delay-multiply and-sum beamformer (DMAS), with and without a paired multiplicative compounding method (pDMAS). Experiments and simulations were performed on a 128-element, dual-aperture transvertebral array, through stacks of ex vivo human vertebra. Numerically and experimentally obtained point spread functions were compared in 3D, producing voxel-wise cross correlation values of 0.84, 0.89, 0.97 (N = 1) for the beamformers listed above, respectively, in water. Experimental, transvertebral localizations of canal sources in isolation produced localization error of 2.8 ± 1.2, 2.9 ± 1.4, 2.7 ± 1.3 mm for a single vertebral target (N = 30 sonications), respectively. A large numerical data set investigating the prefocal cavitation problem (N > 160) is presented and compared with (N = 9) experimental data demonstrating enhancement of intracanal sensitivity in cavitation maps.

Characterization and impact of dissolved gases in a high-frequency flow reactor

Addressing the growing demands for sustainable green technology in chemical processes, we aim to develop a novel high-frequency ultrasonic flow reactor that utilizes converting ultrasound waves, resulting in intense localized acoustic pressure fields. Our hypothesis that our design of converging waves will establish a uniform spatial distribution of cavitation activity with high energy density, thereby augmenting the rate of radical formation and significantly enhancing overall reactor performance. To validate the hypothesis, KI and terephthalic acid dosimetry were performed. A comprehensive calorimetric study was employed for acoustic power, yielding valuable insight into the energy dynamics of the system. Furthermore, our research systematically assesses the impact of dissolved gases and varying flow rates on reactor performance. Notably, we observe that dissolved gases in the reaction medium exert a substantial influence on reactor efficiency. Additionally, an increased flow rate beyond the optimum decelerates hydroxyl radical generation due to a lower residence time. The detailed characterization, facilitated by dosimetry techniques, unveils the potential of the sonochemical reactor for industrial applications.

Magnesium and magnesium alloy dissolution by high intensity focused ultrasound: erosion/cavitation vs. Wave propagation

The dissolution of metals, influenced by mechanical and chemical factors, plays a crucial role in various applications. Ultrasonic irradiation has been explored for its ability to enhance dissolution rates and modify surface characteristics. In this study, we investigate the dissolution of magnesium (Mg) and magnesium alloys under high-intensity focused ultrasound (HIFU) conditions with frequency sweeping (wobbling). Our findings reveal distinct effects of cavitation and acoustic streaming on the dissolution process. For pure magnesium, ultrasonic treatment significantly increases dissolution rates compared to silent conditions. Negative frequency sweeps result in the highest dissolution rates, linked to increased cavitation activity, while positive sweeps reduce dissolution rates but maintain acoustic streaming effects. The removal of surface oxides is accelerated in all sonication conditions. Macro- and micro-roughness patterns on the surface correspond to the wobbling frequency range, with wavelengths matching the average ultrasonic frequency. However, dissolution is not uniform across the sample, and preferential attack occurs at the focal point during negative frequency sweeps. In contrast, magnesium alloys exhibit lower dissolution rates than pure Mg. The alloy's mechanical properties make it less susceptible to cavitation erosion but more sensitive to acoustic streaming-induced dissolution. Grain boundaries are preferentially attacked, revealing differences between ductile pure Mg and the harder, more cavitation-resistant, alloy. This study highlights the complex interplay between cavitation and acoustic streaming in the dissolution of magnesium and its alloys under HIFU conditions, shedding light on the limits and potential applications of this technique, particularly in microstructure analysis.

Passive Acoustic Mapping for Convex Arrays With the Helical Wave Spectrum Method.

Passive acoustic mapping (PAM) has emerged as a valuable imaging modality for monitoring the cavitation activity in focused ultrasound therapies. When it comes to imaging in the human abdomen, convex arrays are preferred due to their large acoustic window. However, existing PAM methods for convex arrays rely on the computationally expensive delay-and-sum (DAS) operation limiting the image reconstruction speed when the field-of-view (FOV) is large. In this work, we propose an efficient and frequency-selective PAM method for convex arrays. This method is based on projecting the helical wave spectrum (HWS) between cylindrical surfaces in the imaging field. Both the in silico and in vitro experiments showed that the HWS method has comparable image quality and similar acoustic cavitation source localization accuracy as the DAS-based methods. Compared to the frequency-domain and time-domain DAS methods, the time-complexity of the HWS method is reduced by one order and two orders of magnitude, respectively. A parallel implementation of the HWS method realized millisecond-level image reconstruction speed. We also show that the HWS method is inherently capable of mapping microbubble (MB) cavitation activity of different status, i.e., no cavitation, stable cavitation, or inertial cavitation. After compensating for the lens effects of the convex array, we further combined PAM formed by the HWS method and B-mode imaging as a real-time dual-mode imaging approach to map the anatomical location where MBs cavitate in a liver phantom experiment. This method may find use in applications where convex arrays are required for cavitation activity monitoring in real time.

Histotripsy-induced bactericidal activity correlates to size of cavitation cloud in vitro.

Large abscesses are walled-off collections of pus and bacteria that often do not respond to antibiotic therapy. Standard of care involves percutaneous placement of indwelling catheter(s) for drainage, a long and uncomfortable process with high rehospitalization rates. The long-term goal of this work is to develop therapeutic ultrasound approaches to eradicate bacteria within abscesses as a noninvasive therapeutic alternative. Inertial cavitation induced by short pulses of focused ultrasound (histotripsy) is known to generate lethal mechanical damage in bacteria. Prior studies with Escherichia coli (E. coli) in suspension demonstrated that bactericidal effects increase with increasing peak negative amplitude, treatment time and duty cycle. The current study investigated correlates of bactericidal activity with histotripsy cavitation cloud size. Histotripsy was applied to E. coli suspensions in 10-mL sample vials at 810 kHz, 1.2 MHz, or 3.25 MHz for 40 minutes. The cavitation activity in the sample vials was separately observed with high-speed photography. The cavitation cloud area was quantified from those images. A linear relationship was observed between bacterial inactivation and cavitation cloud size (R2 = 0.96), regardless of the acoustic parameters (specifically frequency, pulse duration and power) used to produce the cloud.