Scientific Studies

Institute of Applied Biochemistry, University of Tsukuba, Ibaraki, Japan.

Water Sci Technol. 2002;46(6-7):207-15.
A novel strategy for cyanobacterial bloom control by ultrasonic irradiation.
Lee TJ, Nakano K, Matsumura M.

The application of ultrasonic irradiation to control cyanobacterial blooms was evaluated in actualeutrophic lake water. Ten prototype units of the Ultrasonic Irradiation System (USIS) were installed in the 32 ha Lake Senba, and the water and sediment quality were monitored for 2 years. By incorporating the ultrasonication process with the on-going strategy, particularly flushing with induction water, cyanobacterial blooms can be controlled effectively. In addition, a significant improvement in the conditions of the lake in terms of chlorophyll-a, COD and T-P was attained. Moreover, the feasibility of ultrasonic irradiation and bacterial assisted control of cyanobacterialblooms was also evaluated in laboratory conditions. The destruction of gas vacuoles brought about by ultrasonic irradiation promoted close contact between cyanobacteria and their lysing Myxobacter leading to immediate and accelerated destruction of the cells.

Department of Mechanical Engineering, Tsinghua University, Beijing, PR China.

J Environ Sci Health A Tox Hazard Subst Environ Eng. 2004 Jun;39(6):1435-46.
Cyanobacterial bloom control by ultrasonic irradiation at 20 kHz and 1.7 MHz.
Hao H, Wu M, Chen Y, Tang J, Wu Q.

Ultrasonic irradiations at high frequency of 1.7 MHz and low frequency of 20 kHz were tested to prevent cyanobacteria Spirulina platensis from bloom. The inhibition effectiveness at 1.7 MHz was much greater than that at 20 kHz. The cyanobacteria biomass was reduced by 63% after 5 min ultrasonic irradiation at 1.7 MHz, whereas three days were needed for the tested cyanobacteria to recover its original density. However, longer exposure time did not significantly enhance the inhibition. It was observed after ultrasonic irradiation that the gas vesicles in cells collapsed, which may result in cyanobacterial precipitation and photosynthetic inhibition. The concentration of chlorophyll a (Chla) was reduced and its biosynthesis was delayed in a 4-day continuous culture.The fluorescence spectra at 77K of phycobilisome (PBS) and absorption spectra of intact cells in vivo showed that light energy transfer in PBS was inhibited and phycocyanin (PC) was damaged much more acutely compared with Chla. These results indicated that 5 min ultrasonic  irradiation at 1.7 MHz every third day might be an effective and economic operation mode for practical application.

Environmental Biotechnology Laboratory, Korea Research Institute of Bioscience and Biotechnology, Daejeon 305-333, Korea.

Environ Sci Technol. 2003 Jul 1;37(13):3031-7.
Growth inhibition of Cyanobacteria by ultrasonic radiation: laboratory and enclosure studies.
Ahn CY, Park MH, Joung SH, Kim HS, Jang KY, Oh HM.

The growth of Microcystis aeruginosa UTEX 2388 was repressed by ultrasonic radiation and resulted in an increased chlorophyll a content and cell size, suggesting the inhibition of cell division. However, growth was recovered immediately after the interruption of ultrasonication. In addition to the disruption of gas vesicles, other mechanisms of growth inhibition were also investigated. Although free radicals were produced by ultrasonication and hydrogen peroxide, the resulting lipid peroxidation in the cells was not comparable, indicating minimal damage by the free radicals. Ultrasonic radiation late in the day was found to be most effective in reducing the growth rate of M. aeruginosa, and this timing also corresponded to the phase of daily cell division. In an enclosure experiment, ultrasonic radiation reduced the pH, DO, total nitrogen, and total phosphorus, whereas it increased the water temperature, conductivity, and orthophosphate concentration. The algal cell density and chlorophyll a concentration drastically decreased after 3 d of ultrasonication, plus the cyanobacterial proportion was selectively reduced as compared to other algal species. Accordingly, ultrasonic radiation would appear to have considerable potential as an effective control method for cyanobacterial blooms.

Institute of Applied Biochemistry, University of Tsukuba, Tennodai 1-1-1, Tsukuba, Ibaraki 305-0006, Japan.

Environ Technol. 2001 Apr;22(4):383-90.
Ultrasonic irradiation for blue-green algae bloom control.
Lee TJ, Nakano K, Matsumara M.

A novel application of ultrasonic irradiation for rapid control of blue-green algae (BGA) bloom was investigated. Potassium iodide (KI) experiments demonstrated that frequency and input power are the major factors that affect the ultrasonic irradiation intensity. Short exposure (3 s) to ultrasonic irradiation (120 W input power, 28 kHz) effectively settled naturally growing BGA suspension.Electron microscopy reconfirmed that sedimentation was caused by the disruption and collapse of gas vacuoles after ultrasonic exposure. Moreover, even after 5 min of exposure to ultrasonic irradiation (1200 W input power, 28 kHz) the microcystin concentration in BGA suspensions did not increase. For the same input power (120 W), a lower frequency (28 kHz) was found to be more effective in decreasing the photosynthetic activity of BGA than a higher frequency (100 kHz). The sonicated cells did not proliferate when they were cultured in conditions that simulated the bottom of water bodies (i.e. with limited light (400 lx) or no light and non-aerated or aerated (1 l min-1)). Furthermore, ultrasonic irradiation did not only collapse gas vacuoles and precipitate BGA, but may have also inflicted damage on the photosynthetic system of the BGA.Colloids Surf B Biointerfaces. 2005 Mar 25;41(2-3):197-201. Epub 2005 Jan 25. Influence of ultrasonic field on microcystins produced by bloom-forming algae.

Department of Mechanical Engineering, Tsinghua University, Beijing 100084, PR China.

Ma B, Chen Y, Hao H, Wu M, Wang B, Lv H, Zhang G.

Under the background of algae removal and growth inhibition by ultrasonic irradiation, the effects of ultrasonic irradiation on removal of Microcystis, the concentration variation of microcystins (MC) produced by Microcystis in Microcystis suspension, and sonochemical degradation of microcystins in water, were studied in the paper. The results showed that ultrasonic irradiation could efficiently inhibit the growth of Microcystis, and ultrasonic irradiation shorter than 5min would not introduce the increase of microcystins dissolved in Microcystis suspension simultaneity. Also, microcystins dissolved in Microcystis suspension would not increase as ultrasonic power increasing. Further research showed that microcystins were effectively degraded in ultrasonic fields. After20min ultrasonic irradiation at 150kHz and 30W, the removal rate of microcystins reached 70%.

Research Institute for Advanced Science and Technology, Osaka Prefecture University, 1-2 Gakuen-cho, Sakai, Osaka 599-8570, Japan.

2004 Apr;11(2):57-60.
Inactivation of Escherichia coli by ultrasonic irradiation.
Furuta M, Yamaguchi M, Tsukamoto T, Yim B, Stavarache CE, Hasiba K, Maeda Y.


Ultrasonic inactivation of Escherichia coli XL1-Blue has been investigated by high-intensity ultrasonic waves from horn type sonicator (27.5 kHz) utilizing the “squeeze-film effect”. The amplitude of the vibration face contacting the sample solution was used as an indication of the ultrasonic power intensity. The inactivation of the E. coli cells by ultrasonic irradiation shows pseudo first-order behavior. The inactivation rate constant gradually increased with increasing amplitude of the vibration face and showed rapid increase above 3 microm (p-p). In contrast, the H2O2 formation was not observed below 3 microm (p-p), indicating that the ultrasonic shock wave might be more important than indirect effect of OH radicals formed by ultrasonic cavitation in this system. The optimal thickness of the squeeze film was determined as 2 mm for the E. coli inactivation. More than 99% of E. coli cells was inactivated within 180-s sonication at the amplitude of 3 microm (p-p) and 2 mm of the thickness of the squeeze film.

Department of Pharmacy, University of Manchester, England, UK.

J Basic Microbiol. 1996;36(1):3-11.
The effect of ultrasound on Escherichia coli viability.
Allison DG, D’Emanuele A, Eginton P, Williams AR.

The effect of continuous-wave ultrasound on the viability of Escherichia coli HB101 was assessed using a 20 kHz ultrasonic processor. A standardised cell suspension of fixed concentration was used to investigate the influence of different physical and environmental conditions on ultrasound susceptibility. Cell viability decreased exponentially with time at different intensities of ultrasound. Increasing intensity caused a decrease in decimal reduction times. Loss of cell viability occurred primarily from the mechanical effects of ultrasound rather than free radical damage. E. coli susceptibility was also shown to vary with growth conditions, whereby cells cultivated either on agar or harvested from the stationary phase of liquid culture were significantly more susceptible to ultrasound than an equivalent population obtained from the exponential phase of liquid growth. The implication of these results is discussed in relation to the use of ultrasound as a novel means of bacterial transformation.

Graduate School of Engineering, Osaka Prefecture University, 1-1 Gakuen-cho, Sakai, Osaka 599-8531, Japan.

2004 Apr;11(2):61-5.
Inactivation of Saccharomyces cerevisiae by ultrasonic irradiation.
Tsukamoto I, Yim B, Stavarache CE, Furuta M, Hashiba K, Maeda Y.

We have investigated the inactivation of Saccharomyces cerevisiae (yeast cells) by ultrasonic irradiation. The amplitude on the vibration face contacting the sample solution was used as an indication of the ultrasonic power intensity. The effects of the amplitude on the vibration face and the initial cell numbers on the sonolytic inactivation of yeast cells have been investigated using a horn-type sonicator (27.5 kHz). The inactivation of the yeast cells by ultrasonic irradiation shows pseudo first-order behavior. The inactivation rate constant varied from 0.0007 to 0.145 s(-1) when the amplitude on the vibration face was in the range of 1-7 microm(p-p). The change in the inactivation rate constant as a function of the amplitude on the vibration face was similar to that of the OH radical formation rate under the same conditions. The threshold of this sonicator was 3 microm(p-p) with the amplitude on the vibration face. The initial cell numbers (from 10(2) to 10(5) mL(-1)) had an influence on the inactivation of the yeast cells by ultrasonic irradiation. The inactivation rate constants varied from 0.023 to 6.4 x 10(-3) s(-1), and the inactivation by ultrasonic irradiation was fastest at the lowest initial cell numbers. In a squeeze-film-type sonicator (26.6 kHz), 90% inactivation of the yeast cells was achieved by ultrasonic irradiation for 60 min.