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Biofilms, mainly comprised of bacteria, form on materials' surfaces due to bacterial activity. They are generally composed of water, extracellular polymeric substances (polysaccharides, proteins, nucleic acids, and lipids), and bacteria. Some bacteria that form biofilms cause periodontal disease, corrosion of the metal materials that make up drains, and slippage. Inside of a biofilm is an environment conducive to the growth and propagation of bacteria. Problems with biofilms include the inability of disinfectants and antibiotics to act on them. Therefore, we have investigated the potential application of alternating electromagnetic fields for biofilm control. We obtained exciting results using various materials' specimens and frequency conditions. Through these studies, we gradually understood that the combination of the type of bacteria, the kind of material, and the application of an electromagnetic field with various low frequencies (4 kHz-12 kHz) changes the circumstances of the onset of the biofilm suppression effect. In this study, relatively high frequencies (20 and 30 kHz) were applied to biofilms caused by Escherichia coli (E. coli) and Staphylococcus epidermidis ( S. epidermidis ), and quantitative evaluation was performed using staining methods. The sample surfaces were analyzed by Raman spectroscopy using a Laser Raman spectrometer to confirm the presence of biofilms on the surface.

The precise observation of a solid-liquid interface by means of frequency modulation atomic force microscopy (FM-AFM) was performed, demonstrating its applicability to a study on lead acid batteries using an electrochemical test cell for in-liquid FM-AFM embedded with a specialized cantilever holder. The consistency and reproducibility of each surface profile observed via amplitude modulation AFM and FM-AFM were verified properly in a strong acidic electrolyte. In terms of FM-AFM, the ability to observe remarkable changes in the force mapping is the most beneficial, especially near the negative electrode surface. The localization of lignosulfonate (LS) added into the electrolyte as an expander could be visualized since this characteristic force mapping was captured when LS was added to electrolyte.

Biofilms have caused many problems, not only in the industrial fields, but also in our daily lives. Therefore, it is important for us to control them by evaluating them properly. There are many instrumental analytical methods available for evaluating formed biofilm qualitatively. These methods include the use of Raman spectroscopy and various microscopes (optical microscopes, confocal laser microscopes, scanning electron microscopes, transmission electron microscopes, atomic force microscopes, etc.). On the other hand, there are some biological methods, such as staining, gene analyses, etc. From the practical viewpoint, staining methods seem to be the best due to various reasons. Therefore, we focused on the staining method that used a crystal violet solution. In the previous study, we devised an evaluation process for biofilms using a color meter to analyze the various staining situations. However, this method was complicated and expensive for practical engineers. For this experiment, we investigated the process of using regular photos that were quantified without any instruments except for digitized cameras. Digitized cameras were used to compare the results. As a result, we confirmed that the absolute values were different for both cases, respectively. However, the tendency of changes was the same. Therefore, we plan to utilize the changes before and after biofilm formation as indicators for the future.

Biofilms are a result of bacterial activities and are found everywhere. They often form on metal surfaces and on the surfaces of polymeric compounds. Biofilms are sticky and mostly consist of water. They have a strong resistance to antimicrobial agents and can cause serious problems for modern medicine and industry. Biofilms are composed of extracellular polymeric substances (EPS) such as polysaccharides produced from bacterial cells and are dominated by water at the initial stage. In a series of experiments, using Escherichia coli, we developed three types of laboratory biofilm reactors (LBR) to simulate biofilm formation. For the first trial, we used a rotary type of biofilm reactor for stirring. For the next trial, we tried another rotary type of reactor where the circular plate holding specimens was rotated. Finally, a circular laboratory biofilm reactor was used. Biofilms were evaluated by using a crystal violet staining method and by using Raman spectroscopy. Additionally, they were compared to each other from the practical (industrial) viewpoints. The third type was the best to form biofilms in a short period. However, the first and second were better from the viewpoint of "ease of use". All of these have their own advantages and disadvantages, respectively. Therefore, they should be properly selected and used for specific and appropriate purposes in the future.

The sulfuric acid permeation and biofilm formation behaviors of polysiloxane films have been investigated, and simple methods for evaluating the sulfuric acid permeation and biofilm formation behaviors have been proposed in this paper. The polysiloxane films used in these experiments were practically impermeable to the aqueous sulfuric acid solution, and the amount of biofilm formation varied depending on the composition of the films. Further, the amount of sulfuric acid permeation can be estimated by measuring the polarization curves of polysiloxane films with different thicknesses formed on iron electrodes. By measuring the adhesion work of pure water and simulated biofilm droplets on polysiloxane films of different compositions, we can estimate the resistance of biofilm formation on the polysiloxane films.

In this study, we examined the relationship between the effect of a zinc coating on protecting carbon steel against biofilm formation in both air and water environments. SS400 carbon steel coupons were covered with a zinc thermal spray coating or copper thermal spray coating. Coated coupons were exposed to either air or water conditions. Following exposure, the surface conditions of each coupon were observed using optical microscopy, and quantitatively analyzed using an x-ray fluorescence analyzer. Debris on the surface of the coupons was used for biofilm analysis including crystal violet staining for quantification, Raman spectroscopic analysis for qualification, and microbiome analysis. The results showed that the zinc thermal spray coating significantly inhibited iron corrosion as well as biofilm formation in both air and water environments. The copper thermal spray coating, however, accelerated iron corrosion in both air and water environments, but accelerated biofilm formation only in a water environment. microbially-influenced-corrosion-related bacteria were barely detected on any coupons, whereas biofilms were detected on all coupons. To summarize these results, electrochemical corrosion is dominant in an air environment and microbially influenced corrosion is strongly involved in water corrosion. Additionally, biofilm formation plays a crucial rule in carbon steel corrosion in both air and water, even though microbially-influenced-corrosion-related bacteria are barely involved in this corrosion.

A laboratory biofilm reactor (LBR) was modified to a new loop-type closed system in order to evaluate novel stents and catheter materials using 3D optical microscopy and Raman spectroscopy. Two metallic specimens, pure nickel and cupronickel (80% Cu-20% Ni), along with two polymers, silicone and polyurethane, were chosen as examples to ratify the system. Each set of specimens was assigned to the LBR using either tap water or an NB (Nutrient broth based on peptone from animal foods and beef extract mainly)-cultured solution with E-coli formed over 48-72 h. The specimens were then analyzed using Raman Spectroscopy. 3D optical microscopy was employed to corroborate the Raman Spectroscopy results for only the metallic specimens since the inherent roughness of the polymer specimens made such measurements difficult. The findings suggest that the closed loop-type LBR together with Raman spectroscopy analysis is a useful method for evaluating biomaterials as a potential urinary system.