To test the efficacy of chemical disinfectants against bacterial biofilms in hemodialysis equipment, a Center for Disease Control and Prevention (CDC)-Biofilm Reactor was used to create biofilms. Methylobacterium radiotolerance was isolated from the hemodialysis fluid and used as the test organism. We examined the efficacy of sodium hypochlorite (NaOCl) in elimination of planktonic cells compared to that in the case of biofilms. Planktonic bacteria were completely eliminated at 50 parts per million (ppm) of NaOCl, which is the lowest concentration for clinical use. The viable cell count in the biofilm reached its minimum value around a logarithmic reduction value (LRV) of 6, when the concentration was raised to 1000 ppm and the reaction time was extended by 1 hour or more. Furthermore, at 200 ppm, the LRV was elevated depending on the time. And the LRV while maintaining static conditions for 6 hours at 200 ppm was similar to that of short time at 1000 ppm. These results suggest that NaOCl has sufficient bactericidal activity even for biofilms at a practical concentration and reaction time, and that the CDC-Biofilm Reactor is an effective tool for finding useful disinfection conditions.

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Sodium percarbonate (SP) is a relatively low-cost and stable solid oxidizer with a small environmental burden. It is often included in cleansers for sanitizing circulating water pipes and as bleaching reagents in laundry, although the bactericidal effect of SP is lower than that of chlorine-based agents. 2-[Bis(carboxymethyl)amino] propanoic acid-chelated copper (MGDA-Cu) was added to increase the effect of SP. The addition of 12 µM MGDA-Cu increased the bactericidal effect of 0.5 wt% SP against Staphylococcus aureus even in the presence of 0.3 wt% BSA, which is an experimental model of organic stain to protect bacteria from SP. MGDA-Cu was effective against Escherichia coli only in the absence of BSA and showed little effect against Bacillus subtilis. It enhanced the effect of SP to decrease the viscosity of sodium alginate, which is one of the major components of biofilms. The effect of MGDA-Cu on sanitization was also evaluated by 16S rRNA amplicon sequencing of the bacterial flora of the biofilm on an experimental model of a circulating water pipe. The structure of the bacterial flora was more influenced by a cleanser containing both MGDA-Cu and SP than a cleanser with only SP, suggesting that MGDA-Cu increases the sanitization effect.

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Dental materials are inevitably contaminated with oral microorganisms. To prevent transmission of infectious diseases, impressions need to be disinfected. In the present study, we examined the disinfection effects on impression materials and biofilm removal by sodium dichloroisocyanurate (SDIC). Exponentially growing Streptococcus mutans, Escherichia coli, Staphylococcus aureus and Candida albicans, and dental plaque bacteria were suspended in phosphate buffered saline (PBS) and exposed for 1, 5 and 10 min to 1 mL of the 10 ppm, 100 ppm, 1,000 ppm, and 10,000 ppm SDIC solutions. The bactericidal effect was evaluated by colony forming units of each microorganisms. Moreover, the effect of SDIC solution on S. mutans biofilm was examined. Bactericidal effects of SDIC solutions on oral bacteria on dental impression surfaces were assessed and the surface quality of dental casts after immersion in SDIC solution for 30 min was observed under a scanning electron microscope. The number of all bacterial strains, including plaque bacteria, were significantly decreased by SDIC solution treatment in a dose-dependent manner. Significant S. mutans biofilm removing activity of SDIC was observed in 1,000 and 10,000 ppm solution. The number of oral bacteria adhering to the surfaces of impressions markedly decreased following 10-min immersion in the 1,000 ppm SDIC solution. The 30-min immersion of dental impression in the 1,000 ppm SDIC solution did not adversely affect the surface roughness of dental casts. The results indicate that SDIC Solution is useful to deactivate oral bacteria on dental impression.

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Bioshell calcium oxide (BiSCaO) is derived from scallop shells and after heat treatment exhibits broad microbicidal activity. BiSCaO Water is a disinfectant prepared by collecting the aqueous layer after adding BiSCaO powder to water, is colorless and transparent, and has a pH of 12.8. We compared the utility of commercially available BiSCaO Water, ethanol, sodium hypochlorite, hypochlorous acid and hydrogen peroxide solutions as sterilization agents to enable the reuse of surgical and N95 face masks. The microbicidal efficacy of each disinfectant was evaluated using pieces of surgical and N95 face masks contaminated with normal bacterial flora. The results suggest that BiSCaO Water has excellent disinfection activity toward contaminated polypropylene masks and has minimal adverse effect on the structure of non-woven masks.

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Control of infectious diseases requires switching from usual hygiene such as water wiping and cleaning, to control measures including appropriate and aggressive disinfection using 70% alcohol and/or hypochlorous acid depending on the pathogen. Nevertheless, some nursery schools might not understand or select proper disinfectant methods. A survey was administered by the local government of Ibaraki prefecture, Japan in January 2018 to all 456 nursery schools in Ibaraki prefecture. The surveyed items were the number of nursery teachers, usual manual hygiene among children, usual disinfection for lavatories, diaper changing spaces, tables used by children for meals, and classroom floors when there was no vomiting in the facilities and no outbreak in surrounding area. Moreover, it asked about disinfection procedures if children vomited during a community outbreak of gastroenteritis infection. We defined proper use for usual disinfection of a lavatory or diaper changing space as chlorine-based disinfectant including hypochlorous acid or 70% alcohol according to guidelines. Overall, 403 nursery schools responded to the survey. All nursery schools implemented usual hand hygiene. Association between proper disinfection and the size of nursery schools was not significant. Moreover, association between proper disinfection and nursery schools with nurse presence was not found to be significant.

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In these days, all agar media used for both pharmaceutical and industrial territories were required to meet performance criteria. There were recovery rates of assigned microorganisms as performance criteria in both pharmacopeia and ISO standards. However, in spreading plate method, there is no concrete spreading time even though it is shown only “as quickly as possible” in ISO standards. In this study, we verified the impact of spreading time in spreading plate method for the quality control of SCD (Soybean Casein Digest) agar plate. When 30s, 60s, and 120s of spreading time were compared using Bacillus subtilis ATCC 6633, Escherichia coli ATCC 8739, Pseudomonas aeruginosa ATCC 9027, Staphylococcus aureus ATCC 6538, Candida albicans ATCC 10231, and Aspergillus brasiliensis ATCC 16404, respectively, there is no significant difference in recovery rates of all strains tested between 30s and 60s. However, recovery rates of E. coli and P. aeruginosa were decreased in 120s of spreading time. Our results demonstrated that spreading using plastic rod would be better to complete within 60s in spreading plate method since long spreading time had the impact to recovery rate of certain bacteria.

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We previously identified a new Bacillus sporothermodurans strain, TM-I-3, which produces volatile compounds that show potent inhibitory activity against certain types of fungi, including Aspergillus fumigatus and Cladosporium cladosporioides. Non-contact antifungals derived from this bacterium may provide multidirectional inhibition and may be useful in disease prevention. This study is aimed at identifying the stage of fungal growth that is inhibited by TM-I-3 to elucidate the mechanism of its contact-independent antifungal activity. We evaluated mycelial growth and the gross fungal colony areas after 7 days in each experimental group varying the time in the proximity of TM-I-3 for 24 hours each. The fungal growth inhibition assay showed that TM-I-3 inhibited spore germination: the lag phase in the sigmoid growth curve. The present study demonstrated that TM-I-3 might be an effective fungistatic agent against pathogenic and allergenic fungi.

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A large volume of historical archives was extensively damaged by flood water, following Typhoon Hagibis in Fukushima, Japan, in October 2019. They were rescued from the stricken area within a week, however, the prolonged exposure of paper documents to water caused severe biodegradation by fungal growth. To disinfect fungi, the paper documents were exposed to gamma radiations emitted by a source of Cobalt 60 by the industrial irradiation service. The wet paper documents were mainly contaminated with hydrophilic and cellulolytic fungi, including Trichoderma, Stachybotrys, and Fusarium; no fungi grew after irradiation. These results indicated that the average absorbed dosage from 13.1 kGy to 16.1 kGy were sufficient to disinfect paper documents heavily contaminated with fungi. In the present study, we demonstrated the successful practical use of irradiation in fungi-damaged paper documents using a commercial gamma-irradiation facility.

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Culturable bacteria were isolated from tomato seeds using media selective for the canker pathogen Clavibacter michiganensis subsp. michiganensis. Clustering analysis (>99% identity) revealed the presence of 16 operational taxonomic units (OTUs) among isolates detected on semi-selective media. Three OTUs belonged to the phylum Actinobacteria, including those of Micrococcus and Dermacoccus, and 13 OTUs belonged to the phylum Firmicutes, including Bacillus and related genera. These Gram-positive endophytic bacteria have the potential to provide false-positive results in seed health tests using media considered semi-selective for the cancer pathogen.

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