Non-thermal Inhibition of Antibiotic Resistant Bacteria by Oscillating

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Pulsed Electric Fields (OPEF)
1 Browne, M., 1Foley, S., 1Babck, N., 2Olshefsky, S., 3,4Holland, A., and 1Ahern, H.

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1SUNY Adirondack, Queensbury, New York
2Microbiology Department, Montana State University, Bozeman, Montana
3Skidmore College, Saratoga Springs, New York

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4Novobiotronics, Inc., Saratoga Springs, New York

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Background Results Discussion

Figure 1: Comparison of growth rates of Ps. aeruginosa cultures determined by viable cell count.

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Antibiotic-resistant bacteria have emerged as a significant A bacteriostatic effect was observed when
medical problem, prompting research into alternative ------ No OPEF exposure ------ OPEF exposure the bacteria were exposed to OPEF only,

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strategies for destruction. Pseudomonas aeruginosa and with a significant reduction in the growth

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methicillin-resistant Staphylococcus aureus (MRSA) are
two bacterial pathogens with well characterized
rate of the cultures. Addition of an
mechanisms of resistance, including biofilm formation, antibiotic, to which the bacteria initially
showed resistance by disk-diffusion

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efflux pumps, and antibiotic-degrading enzymes. Several
assay, at the time of OPEF exposure was

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studies have shown that exposure to electrical fields
inhibits the growth of planktonic and biofilm bacteria and bacteriocidal, with a 50% (average)
enhances the efficacy of antibiotics. In this study, bacteria reduction in the number of viable cells in

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with demonstrated resistance to specific antibiotics were the cultures. OPEF in a frequency range
exposed to an oscillating, pulsed electric field broadcast of 100 ± 150 kHz reduced the growth rate
by an enclosed gas plasma antenna. Helium gas inside the
of bacteria equivalent to the effect seen

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glass tube was stimulated into a plasma state using an
amplitude modulated customized radio frequency with the antibiotic alone, and when used in
Figure 2: Comparison of growth rates of Ps. aeruginosa cultures determined by viable cell count. conjunction with low concentration of
transmitter. OPEF may be a potential treatment modality

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for infections caused by antibiotic-resistant bacteria. ------ No ampicillin ------ Ampicillin 0.25 mg/ml antibiotic, OPEF was lethal to the bacterial
cells. Because the observed biological

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Method effects are not due to electric field-
induced thermal stress or cavitation (as in
Cultures of Pseudomonas aeruginosa and MRSA other studies) , these results suggests
(Staphylococcus aureus ATCC 43300) were evaluated for

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that OPEF could be applied as a treatment
antibiotic susceptibility and minimal inhibitory modality for infections caused by
concentration using standard techniques. Broth cultures antibiotic resistant bacteria.
of both bacteria were prepared with and without
antibiotic (antibiotic concentration used was 10 times

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less than the MIC). After an initial 2 hour incubation References
period, the cultures were exposed to OPEF at 150 kHz
Figure 3: Comparison of growth rates of methicillin-resistant Staphylococcus aureus (MRSA) Costerton, J.W., Ellis, B., Lam, K., et.al (1994) Mechanism
and 100 kHz, respectively. The bacteria were maintained

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of electrical enhancement of efficacy of antibiotics in
at constant temperature permissive to growth during the determined by viable cell count.
killing biofilm bacteria. Antimicrobial Agents and
exposure period. Growth rates of exposed and non- ------ No cefoxitin ------ Cefoxitin 0.025 mg/ml Chemotherapy 38(12): 2803±2809.

0
exposed cultures were determined over several hours by
Giladi, M., Porat, Y., Blatt, A., et.al. (2008) Microbial growth
spectrophotometric measurement of absorbance and
inhibition by alternating electric fields. Antimicrobial
viable cell counts on plate medium during the growth Agents and Chemotherapy, 52(10): 3517-3522.

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period.
Giladi, M., Porat, Y., Blatt, A., et.al. (2010) Microbial growth
Inhibition by alternating electric fields in mice with
Pseudomonas aeruginosa lung infection. Antimicrobial
Agents and Chemotherapy, 54(8): 3212-3218.

©
Figure 2: Pulsed waveform output
of the plasma antenna at 100Khz,
50% duty cycle.
Dubost, Holland, Bare, Bellossi (2012) Morphological
Transformations of Human Cancer Cells and Microtubules
Caused by Frequency Specific Pulsed Electric Fields
Broadcast by An Enclosed Gas Plasma Antenna,
Proceedings of the 7th International Workshop on
Biological Effects of EMF (Malta)
Acknowledgments: Dr. James Bare, Plasma Sonics Ltd., for assistance with
Figure 1: OPEF exposure of bacterial cultures the device used in these experiments, US patents no. US 8,269,571 B2 and
Plasma Sonics Ltd., James Bare, US patent no. US US5908441.
8,269,571B2 and US 590844.