# Effects of Resonant Electromagnetic Fields on Biofilm Formation in _Pseudomonas aeruginosa_
## 4 Discussion
A series of six experiments all showed an inhibitory effect, on average a 27% reduction of biofilm formation. The specific frequency of 3.9 Hz was calculated as the Ion Cyclotron Resonance (ICR) of the potassium ion, K\({}^{+}\), at a specific magnetic field strength. We found that tripling the magnetic field and tuning the ICR frequency did not show the same consistency as observed using the 3.9 Hz frequency. This would be expected if the resonance effect derives purely from a single ion species. However, as mentioned earlier, the effects observed are likely dependent on the protonation of water as well as the biologically relevant ion, in this case K\({}^{+}\). The term protonation of water here means the exposure of the aqueous solution to the ICR frequency of the Zundel Cation, H\({}_{3}\)O\({}^{+}\)(H\({}_{2}\)O). In previous studies on the polymorphism of water, the role of the Zundel Cation, one of the most abundant hydronium hydrates, has been suggested to provide the needed energy for activating other cations. It was observed that the release of Zundel Cations and other hydronium hydrates enhances the conductivity of water, reducing pH and modifying its refraction index, thereby enhancing the small signal of ICR, and allowing ion currents aroused by ICR to overcome the environmental thermal barrier, \(kT\) (where \(k\) is the Boltzmann's constant and \(T\) the absolute temperature). It was also observed that water works as an amplifier in the ELF band when hydronium hydrates are able to enhance such polymorphisms [30; 31]. Such effect of ICR on hydronium hydrates has been confirmed in the present experiment thanks to differential results depending on the presence of the Zundel Cation component in the K\({}^{+}\)-ICR that was administrated to bacteria, in the way described below. As the static magnetic field (10 microT) consistently showed effects on bacterial biofilm, we expect an ICR of K\({}^{+}\) of 3.9 Hz and 4.1 Hz of the Zundel Cation, H\({}_{3}\)O\({}^{+}\)(H\({}_{2}\)O), thus a 0.2 Hz difference. Considering the Half Width at Half Maximum (HWHM), \(\Gamma^{\prime}/2\), may vary depending on many parameters characterizing the resonance, including the Q factor known from radio wave reception theory. It has previously been suggested that the HWHM is in the order of \(10^{-1}\) Hz, thus providing a plausible explanation for activating both K\({}^{+}\) and H\({}_{3}\)O\({}^{+}\). (H\({}_{2}\)O) when using the 10 microT static field [8]. However, when all factors were tripled, the difference in calculated ICR was increased to 0.6 Hz (3 \(\times\) 0.2 Hz) which is in the order of 1 Hz and may reduce the effect of nearby frequencies and the reproducibility of the experiment. Thus, the methodology described here may serve as a starting point for developing novel strategies against the rising global challenge of antimicrobial resistance and impact clinical challenges such as chronic infections in ulcers, catheters, and urinary tracts. Future directions may include the synergistic effects of resonant magnetic fields and antimicrobial compounds, and assay conditions mimicking clinically relevant situations. These may include biofilm formation in flow chambers and behavior of bacteria infecting organoids (small artificial organs). Molecular and biochemical investigations using metabolomics, transcriptomics, and proteomics approaches may also elucidate the biochemical pathways perturbed by resonant magnetic fields.