AC Electromagnetic Field Controls the Biofilms on the Glass Surface by Escherichia coli & Staphylococcus epidermidis Inhibition Effect.

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.

Quantitative Analyses of Biofilm by Using Crystal Violet Staining and Optical Reflection.

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.

Proposal for Some Affordable Laboratory Biofilm Reactors and Their Critical Evaluations from Practical Viewpoints.

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.

Impedance Characteristics of Monolayer and Bilayer Graphene Films with Biofilm Formation and Growth.

Biofilms are the result of bacterial activity. When the number of bacteria (attached to materials' surfaces) reaches a certain threshold value, then the bacteria simultaneously excrete organic polymers (EPS: extracellular polymeric substances). These sticky polymers encase and protect the bacteria. They are called biofilms and contain about 80% water. Other components of biofilm include polymeric carbon compounds such as polysaccharides and bacteria. It is well-known that biofilms cause various medical and hygiene problems. Therefore, it is important to have a sensor that can detect biofilms to solve such problems. Graphene is a single-atom-thick sheet in which carbon atoms are connected in a hexagonal shape like a honeycomb. Carbon compounds generally bond easily to graphene. Therefore, it is highly possible that graphene could serve as a sensor to monitor biofilm formation and growth. In our previous study, monolayer graphene was prepared on a glass substrate by the chemical vapor deposition (CVD) method. Its biofilm forming ability was compared with that of graphite. As a result, the CVD graphene film had the higher sensitivity for biofilm formation. However, the monolayer graphene has a mechanical disadvantage when used as a biofilm sensor. Therefore, for this new research project, we prepared bilayer graphene with high mechanical strength by using the CVD process on copper substrates. For these specimens, we measured the capacitance component of the specimens' impedance. In addition, we have included a discussion about the possibility of applying them as future sensors for monitoring biofilm formation and growth.

Research Activities in Materials Science and Engineering with Academic-Industrial Alliances during the COVID-19 Pandemic.

During the COVID 19 pandemic, the importance of global academia-industrial alliances has increased. It is hoped that the alliances will help us to solve the current problems caused by the pandemic. In this paper, we introduce the application of IT tools and communication skills utilized in a special educational project for an academia-industrial collaboration. Some concrete examples from 2020 are provided from the viewpoint of the national alliance project in Japan. A discussion is included that describes the plans available to increase and strengthen the national project in the future.

Technologies for teaching during a pandemic.

The coronavirus pandemic has had a major impact on education. As a result of this crisis, instructors world-wide had to quickly adapt and shift to remote teaching. This paper presents and describes technologies (such as Microsoft Teams, Zoom, Virtual Worlds, etc.) to promote active student learning during a pandemic. It also provides general information about the coronavirus and the learning environment that it created.

Rotation Disk Process to Assess the Influence of Metals and Voltage on the Growth of Biofilm.

Biofilms consist of not only bacteria but also extracellular polymer substrates (EPS). They are groups of microorganisms that adhere to each other on a surface, especially as a result of exposure to water and bacteria. They can pose health risks to humans as they grow in hospital settings that include medical supplies and devices. In a previous study, the researchers discovered that bacteria/biofilm grew well on wetted external latex, male catheters. These results concerned the investigators and encouraged them to find ways for prohibiting the growth of bacteria/biofilm on the male catheters (which are made of natural rubber). They carried out a new study to assess the influence of metals and voltage for the growth of bacteria on these latex samples. For this purpose, a unique Rotation Disk Reactor was used to accelerate biofilm formation on external male catheter samples. This setup included a dip tank containing water and a rotating wheel with the attached latex samples (some of which had single electrodes while others had paired electrodes with applied voltage). The process allowed the samples to become wetted and also exposed them to microorganisms in the ambient air during each revolution of the wheel. The results (as viewed from SEM images) showed that when compared to the control sample, the presence of metals (brass, stainless steel, and silver) was generally effective in preventing bacterial growth. Also the use of voltage (9.5 volt battery) essentially eliminated the appearance of rod shaped bacteria in some of the samples. It can be concluded that the presence of metals significantly reduced bacterial growth on latex and the application of voltage was able to essentially eliminate bacteria, providing appropriate electrode combinations were used.