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    Antibiofilm strategies using mixed disinfectants and cubosome nanocarriers to combat Staphylococcus aureus on sandblasted stainless steel disc surfaces
    (2025-09-01)
    Dawan, Jirapat
    ;
    Guo, Yuyuan
    ;
    Lee, Sang Soo
    ;
    Kim, Jin Chul
    Bacterial biofilms are complex communities in which microorganisms are adhered and encapsulated in a self-produced extracellular polymeric material (EPS) matrix. Biofilm acts as a protective barrier, enhancing the resistance of bacteria to environmental stressors, cleaning agents and antimicrobial treatments. Therefore, this study aimed to evaluated the effectiveness of a mixed disinfectant suspension (MIX) against Staphylococcus aureus biofilms on sandblasted stainless steel disc surfaces. A cubic phase containing MIX was prepared, and its anti-biofilm ability was assessed in comparison to free MIX. The susceptibility of S. aureus ATCC 15,564 (SA<sup>ATCC</sup>) and multidrug-resistant S. aureus CCARM 3080 (SA<sup>CCARM</sup>) biofilms to single and mixed disinfectants (MIX) was evaluated. The incorporation of MIX into cubosomes was performed to evaluate its potential enhancement in antibiofilm activity. 1% MIX demonstrated the most effective anti-biofilm activity. Extracellular DNA (eDNA) and polysaccharides were also significantly reduced in biofilms treated with 1% MIX. TEM analysis revealed that 1% MIX effectively disrupted bacterial cell clusters within the biofilms. The incorporation of 1% MIX into cubosomes enhanced its antibiofilm activity, with cubosome-encapsulated MIX exhibiting superior results compared to free MIX. This study demonstrates the effectiveness of a mixed disinfectant suspension in reducing S. aureus biofilms on sandblasted stainless steel disc surfaces. Furthermore, the incorporation of MIX into lipid-based nanocarriers, such as cubosomes, enhanced its antibiofilm efficacy compared to the free disinfectant.
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    Species-specific detection of environmental DNA by endpoint PCR using mismatch base introducing primer design and nested PCR
    (2025-04-01)
    Ishii, Ryosuke
    ;
    Poti, Teeranai
    ;
    Akimitsu, Kazuya
    ;
    Narumi-Kawasaki, Takako
    ;
    Yamada, Yoshihiro
    Environmental DNA (eDNA) has been used effectively for species identification in aquatic macrobiological surveys. To popularize this method, a cost-effective analysis method with high sensitivity is required. In this study, we developed an eDNA detection method that is as sensitive as conventional quantitative real-time PCR (qPCR), which does not require expensive equipment, and with heightened sensitivity that was achieved through mismatch-introducing primer design and nested PCR. The subject of the experiment was Micropterus nigricans (largemouth bass). Standard PCR with mismatch-free primers detected amplicons not only in M. nigricans but also in M. dolomieu and Lateolabrax japonicus. However, the mismatched primers only amplified DNA from M. nigricans and did not show amplicons for the non-target species. Endpoint PCR was less sensitive than qPCR, but it was able to specifically detect the target species in environmental samples. A similar approach was applied to the bitterling fish species, a species to which many other closely related species are known, and Acheilognathus cyanostigma eDNA was specifically detected. This eDNA analysis method is cost-effective because it is comparable in sensitivity to conventional qPCR methods and does not require expensive equipment.