An advantage of vapor phase hydrogen peroxide (VPHP) is that it can readily react to form reactive free radicals, which perform the sterilization and form water and hydrogen by catalyst. Absorptive hydrophilic materials such as cellulosics hinder penetration due to hydrogen bonding and necessitate the use of hydrophobic materials, i.e., polyethylene or polypropylene, as packaging materials. The 8h TWA (time-weight-average) is 1 ppm. Hydrogen peroxide sterilization is now being used for the sterilization of gloveboxes, freeze dryers, isolators and endoscopes and so on. This paper focuses on the application of VPHP to the sterilization of the endoscopes.

Heterotrophic plate count (HPC) test has been employed to indicate the effectiveness of water treatment processes and the microbiological condition of the distribution system. In Japan, because the majority of HPC bacteria are supposed to be harmless and all tap water should maintain residual chlorine, there are few studies of the virulence of HPC bacteria. In this study, we examined HPC bacteria isolated from finished and tap water for hemolytic activity to determine their virulence potential. 34 of 39 colonies expressing hemolytic activity were identified by partial 16S rDNA sequencing, but some of their percent identity were relatively low. This may have been due to the mismatching of the primer pair with some strains, or these strains may be unidentified new species. A total of 30 of 34 isolates identified have been reported to be opportunistic pathogens or food poisoning bacteria. To control the growth of these opportunistic pathogens among HPC bacteria, appropriate water quality control must always be done and residual chlorine must be maintained in every tap for a safe water supply.

Before the present study, no fungi using anion surfactant as a nutrient had been identified, although some fungi were known to use nonion surfactant. Washing water collected from 63 washing machines was inoculated onto 0.1% LAS (Sodium dodecyl benzenesulfonate) anion surfactant media to identify fungi that can feed on anion-surfactant. Small dark colonies of fungi were found on several of the Petri-dishes from 12 days after inoculation. These were identified as Cladophialophora boppii and Exophiala spinifera using morphological features and rDNA data. A number of the isolates of C. boppii specifically were recognized as using anion surfactant as a nutrient. The growth characteristics of the two fungal species were examined on surfactant media of three kinds. Apart from anion surfactant, the fungi were also able to grow on nonion surfactant and on soap. The application of these fungi for environmental cleansing after detergent pollution is also discussed.

A new type of ultrasonic washer-disinfector-sterilizer, able to clean, disinfect and sterilize most kinds of reusable medical devices, has been developed by using the ultrasonic levitation function with umbrella-shape oscillators and ozone bubbling together with sterilization carried out by silver electrolysis. We have examined the biomedical and physicochemical performance of this instrument. Prokariotic and gram-negative Escherichia coli and eukariotic Saccharomyces cerevisiae were killed by silver electrolysis in 18 min and 1 min, respectively. Prokariotic and gram-positive Geobacillus stearothermophilus and Bacillus atrophaeus, which are most resistant to autoclave and gas sterilization, respectively, were killed by silver electrolysis within 20 min. Prokariotic and gram-negative Pseudomonas aeruginosa was also killed by silver electrolysis in 10 min. The intensity distribution of the ultrasonic levitation waves was homogeneous throughout the tank. The concentration of ozone gas was 2.57 mg/ kg. The concentration of dissolved silver ions was around 0.17 mg/L. The disulfide bond in proteins was confirmed to be destroyed by silver electrolysis.

As part of an epidemiological study on legionellosis, we attempted to isolate Legionella spp. from hot spring water samples, and were able to isolate Legionella micdadei from 3 (5.5%) of 55 samples. All of these isolates were able to grow within Acanthamoeba sp., suggesting that the isolates will be pathogens. We also confirmed that the K-2 strain from hot spring water grew in guinea pig monocytes. Sensitivity tests using 10 drugs showed that the isolates were most sensitive to imipenem, with the MIC90 of 0.032 microg/ml, were least sensitive to minocycline, with the MIC90 of 4 microg/ml, and were not sensitive to low amounts of other drugs.

The responses of Aspergillus oryzae to the stimuli from near-UV irradiation were investigated by using a black light fluorescent lamp in the presence or absence of TiO2 particles. Light irradiation at an intensity of 6 W/m2 strongly inhibited the growth of germinated pellets of A. oryzae. This growth inhibition was weakened by TiO2 particles (0.05 g/l), especially in the initial growth phase, in which the expression level of catalase gene (catB) was approximately three times higher than that in the absence of TiO2 particles. However, the initial induction of catB expression by H2O2 pretreatment did not restore the growth under the black light irradiation. The weakening of growth inhibition is thought to result from alternative physiological responses of A. oryzae against stimulus by photo-excited TiO2.

The colonial growth of Bacillus subtilis vegetative and spore cells was evaluated by determining the time course of the growth heat changes (growth thermogram) with a microbial calorimeter. The actual heat evolution curve (f (t) curve) obtained from the thermogram was in good agreement with the viable cell number changes. From the logarithmic f (t) curve, the exponential growth phase was estimated to be the period of between 31 and 35 h after inoculation and the growth rate constant (mu') was 0.36 h(-1). The f (t) curve was also obtained from the thermogram of the spore cells. The starting time of growth of the spore cells was 9 h later than that of the vegetative cells. This period corresponded to the germination period. The growth profile after germination was almost similar to that of the vegetative cells. The suppressive activities of sucrose monopalmitate (SMP) against the vegetative and spore cells were evaluated by the f (t) curves. Our results show that SMP inhibited metabolic inductions during the lag phase of the vegetative cells and the germination of the spore cells. Microbial calorimetry is a method suitable for non-destructive growth measurements of microbial colonies on the surface of a solid medium.

Particles of ZnO coated with carbon (ZnOCC) were prepared and evaluated for their antibacterial activity. ZnO powder and poly(vinyl alcohol) (PVA) (polymerization degree: 2,000-95,000) were mixed at a mass ratio (ZnO/PVA) of 1, and then heated at 500-650 degree C for 3 h under argon gas with a flow rate of 50ml/min. Carbon deposited on the ZnOCC surface was amorphous as revealed by X-ray diffraction studies. The ZnOCC particles maintained their shape in water, even under agitation. The antibacterial activity of ZnOCC powder against Staphylococcus aureus was evaluated quantitatively by measuring the change in the electrical conductivity of the growth medium caused by bacterial metabolism (conductimetric assay). The conductivity curves obtained were analyzed using the growth inhibition kinetic model proposed by Takahashi for calorimetric evaluation, allowing the estimation of the antibacterial efficacy and kinetic parameters of ZnOCC. In a previous study, when ZnO was immobilized on materials, such as activated carbon, the amount of ZnO immobilized was approximately 10-50%, and the antibacterial activity markedly decreased compared to that of the original ZnO. On the other hand, the ZnOCC particles prepared in this study contained approximately 95% ZnO and possessed antibacterial activity similar to that of pure ZnO. The carbon-coating treatment could maintain the antibacterial efficacy of the ZnO and may be useful in the develop-ment of multifunctional antimicrobial materials.

We investigated characteristics of the corrosion of stainless steel specimens by bacteria and the effects of using antimicrobial coating on the surface for inhibiting corrosion. Bacillus sp. 2-A and Staphylococcus sp. 2-1 cells adhered tightly to a stainless steel SUS304 specimen, formed a microcolony or biofilm, and had highly corrosive activities. Microbially influenced corrosion (MC) was observed under or around adhering cells. However, dead cells were markedly less active than viable cells not only in corroding the specimen but also in adhering to its surface. The culture supernatant was not able to induce the corrosion of SUS304 effectively. A protamine coating on the specimen killed bacterial cells only on its surface, interfered with cell adhesion, and inhibited MC. From these results, adhesion of viable cells to the surface of a SUS304 specimen led to the outbreak of MC. Protamine was also found to be an effective substance tested for protecting the specimen from both cell adhesion and surface MC. We suggest that a protamine coating can be applied as a convenient and inexpensive corrosion prevention method.

Following the former version of the database on the thermal death of microorganisms, ThermoKill Database R9100, we constructed an expanded version, ThermoKill Database R8100. This database includes the thermal death data together with as many as 70 items of experimental conditions described in research papers which appeared in 23 academic journals published worldwide from 1981 to 2000. The novel version contains 7,454 records as data sets, 12,261 D values, and 2,038 z values. Some statistical data based on the data input in the database are presented. The significance of this novel database in practical use is then discussed.