Because soy milk is nutrient rich and nearly neutral in pH, it favors the growth of microbial contaminants. To ensure that soy milk meets food-safety standards, it must be pasteurized and have its sterility confirmed. ATP bioluminescence assay has become a widely accepted means of detecting food microorganisms. However, the high background bioluminescence intensity of soy milk has rendered it unsuitable for ATP analysis. Here, we tested the efficacy of an improved pre-treated bioluminescence assay on soy milk. By comparing background bioluminescence intensities obtained by the conventional and improved methods, we demonstrated that our method significantly reduces soy milk background bioluminescence. The dose-response curve of the assay was tested with serial dilutions of Bacillus sp. culture. An extremely strong log-linear relation between the bioluminescence intensity relative light units and colony formation units CFU/ml emerged for the tested strain. The detection limit of the assay was estimated as 5.2×103 CFU/ml from the dose-response curve and an imposed signal limit was three times the background level. The results showed that contaminated samples could be easily detected within 24 h using our improved bioluminescence assay.
A high-level aseptic environment must be maintained in bio-cleanrooms used for the manufacture of sterile products. In the past, formaldehyde gas was most commonly used to sterilize bio-cleanrooms, but due to strict residual limitations there has been a need to develop a less toxic alternative choice. The authors have developed a revolutionary new sterilization system using a high concentration of ozone gas and used this system to sterilize an actual bio-cleanroom. This system integrates the ozone gas generator with the air conditioning system by proper control. The design specifications for the system included an ozone gas concentration of 200ppm or more, relative humidity of 80% or more, and a sterilizing time of 120min. Blow vents and suction ports were placed to ensure a uniform airflow which would extend through the entire room during ozone gas sterilization. Tests regarding long-term material exposure to ozone gas were conducted when the system was introduced to distinguish usable and unusable materials. In an actually constructed cleanroom, simulations were used to predict the evenness of the diffusion of ozone gas concentration and relative humidity during ozone gas sterilization, and measurements of the actual indoor ozone gas concentration, temperature and relative humidity during sterilization revealed that the ozone concentration and relative humidity needed for sterilization had been achieved generally throughout the entire environment. In addition, the CT value (mg/m3 (=ppm) ×min) , derived by multiplying the ozone gas concentration during ozone gas sterilization by the sterilization time, was equal to or greater than the target value of 24×103 (ppm·min) . When the results of sterilization in a cleanroom were confirmed using a biological indicator (BI) , negative results were obtained at all measurement points, demonstrating that sterilization was being performed effectively in the actual factory at which the ozone gas sterilization system had been introduced.
We found that a novel biosurfactant from the cultured broth of red yeast, Rhodotorula mucilaginosa KUGPP-1, originating in the Antarctic, has dispersive power against astaxanthin. The novel biosurfactant was purified from extracts to the ultrafiltration state by acetone precipitation and chromatography on a DEAE-Toyopearl 650M, and gel filtration on a Sephacryl S-400HR. The molecular mass of the novel biosurfactant was estimated to be about 730,000 by gel filtration chromatography. The novel biosurfactant was comprised of sugar and protein in an approximate molar ratio of 9 : 1. The sugars were comprised of mannose, galactose and glucose. The particle size of the astaxanthin (0.13 μ g/ml) micelle was about 410 nm. Astaxanthin was stable to oxidation in the novel biosurfactant micelles. To our knowledge, this is the first report on a glycoprotein type of biosurfactant with astaxanthin-stabilizing ability.
The physicochemical properties and bacterial community in sediments of Lake Shiraishi, a lake with brackish water, were characterized to elucidate the influence of oyster farming and seawater and freshwater inflow. Physicochemical analyses suggested the marine origin of the sediment at the mouth of the lake, while higher organic matter load and the resultant anaerobic, reductive condition of the sediments of the inner part were observed. The bacterial community in the sediments reflects these sediment environments: the bacterial community in the vicinities of oyster farms included sulfate-reducing bacteria (SRB) , although sulfur-oxidizing bacteria (SOB) were found at all the sampling sites. In addition, similarity of the band profiles obtained with 16S ribosomal RNA gene (16S rDNA) -denaturing gradient gel electrophoresis (DGGE) decreased in proportion to the distance from the mouth of the lake to the oyster farms in the inner part. This study was able to characterize the microbial community shift in brackish lake sediments with an oyster aquaculture system through the molecular fingerprinting technique, DGGE, in relation to their physicochemical characteristics.
The effect of an oxygen-releasing compound (ORC) magnesium peroxide (MgO2) on the changes in the bacterial community in organically polluted sediment of aquaculture farms was tested in a microcosm experiment. The sediment, to which fish feed was added, was treated with 1% or 5% MgO2. The addition of fish feed induced a highly reduced environment with low redox potential, high total sulfides, and abundance of sulfate-reducing bacteria (SRB) . Although the sediment remained highly reduced at 1% MgO2, there was a significant reduction of total sulfides, increase of redox potential, and resultant reduction of SRB. The bacterial community clearly changed with the treatments according to denaturing gradient gel electrophoresis (DGGE) analysis of 16S ribosomal RNA gene (16S rDNA) . Aerobes disappeared in the fish feed-added sediment, and some SRB emerged in place of these aerobes. On the other hand, the SRB disappeared in the ORC-amended sediment due to its highly oxic condition. This study revealed the bacterial community in the sediments was affected mainly by the redox potential and resultant sulfides produced by SRB, but total organic carbon and nitrogen were not determinants of the microbial population.
An ecological study of pathogenic vibrios in aquatic environments of Okayama was carried out. The number of Vibrio parahaemolyticus detected in the sea area was comparatively smaler than that found in the survey of about two decades ago. Various reasons for the decrease in the case of food poisoning by V. parahaemolyticus have been suggested but the lower number of the vibrio in aquatic environments may be one explanation. Although the number of V. vulnificus was also not as large, most of the isolates possessed the pathogenic genes, vvp and vvh, suggesting the potential for fatal pathogenicity to patients having underlying diseases. As for V. cholerae, some non-O1/non-O139 serovar isolates were detected in a fresh water area, and many of them had hlyA, the gene for hemolysin which acts as a pathogenic factor in sporadic cases of diarrhea. Thus, the total number of pathogenic vibrios detected was not of concern. However, the marine products of these areas are shipped in wide area and are for general consumption. Therefore, it is necessary to continue to survey pathogenic vibrios in aquatic environments in order to ensure food hygiene.