We studied airborne concentrations of fungal spores and the thermal environment in houses with semi-basements surrounded by a natural forest. We examined the relationship between airborne fungi and the thermal environment, surrounding natural environment, structures of houses and use of a dehumidifier. The subject residential area was located in the northern part of Nara city, Nara prefecture, Japan. Six detached houses were included in this study. In residential areas, outdoor airborne concentrations were high during summer and autumn, correlated with humidity. The presence of Basidiomycetes was particularly notable, although the indoor concentration was lower than the outdoor level. In the semi-basement rooms, relative humidity was nearly always >80% when the residence was built; however, both the indoor humidity and fungal concentrations decreased greatly when a dehumidifier was used in this study. High levels of Aspergillus and Basidiomycetes were detected in semi-basements. Basidiomycetes are likely of outdoor origin, whereas Aspergillus might grow indoors. Moreover, the composition of fungal species differed according to room-structure and usage. Due to the health risks associated with high indoor concentrations of fungi, the utilization of the semi-basement or basement space requires adequate ventilation and dehumidification, beginning immediately after construction.
We examined the effectiveness of cocopeat and rice hull powder obtained from agricultural wastes as biocarriers for an oil-degrading bacterial consortium. Scanning electron microscopy revealed colonization and strong attachment of bacterial cells on the surface of both carriers. Results of a 60-day in vitro seawater bioremediation trial showed significant oil reduction and high cultivable bacterial counts in treatments augmented with the carrier-attached bacterial consortia compared to treatments supplemented with the same consortium in free living and encapsulated forms. Significant degradations in both aliphatic and aromatic fractions were obtained in treatments augmented with carrier-immobilized consortia. The developed immobilized cells showed sustained activities and viabilities during storage for six months. Results of this study demonstrated that inexpensive waste materials can be utilized as biocarriers of an oil-degrading consortium and that immobilization on biocarriers can enhance the bioremediation of oil-contaminated seawater.
To analyze the status of the genus Alcaligenes in the agricultural environment, we developed a PCR method for detection of these species from vegetables and farming soil. The selected PCR primers amplified a 107-bp fragment of the 16S rRNA gene in a specific PCR assay with a detection limit of 1.06 pg of pure culture DNA, corresponding to DNA extracted from approximately 23 cells of Alcaligenes faecalis. Meanwhile, PCR primers generated a detectable amount of the amplicon from 2.2×102 CFU/ml cell suspensions from the soil. Analysis of vegetable phylloepiphytic and farming soil microbes showed that bacterial species belonging to the genus Alcaligenes were present in the range from 0.9×100 CFU per gram (or cm2) (Japanese radish: Raphanus sativus var. longipinnatus) to more than 1.1×104 CFU/g (broccoli flowers: Brassica oleracea var. italic), while 2.4×102 to 4.4×103 CFU/g were detected from all soil samples. These results indicated that Alcaligenes species are present in the phytosphere at levels 10-1000 times lower than those in soil. Our approach may be useful for tracking or quantifying species of the genus Alcaligenes in the agricultural environment.
For a microbial ecological analysis, 20 strains of Legionella pneumophila isolated from both unchlorinated Noyu (unattended natural hot spring) samples and chlorinated hot spring bath water samples collected throughout Japan were subjected to a clustering analysis on the basis of a Pulsed-Field Gel Electrophoresis (PFGE) pattern analysis. The PFGE patterns obtained from 19 strains of L. pneumophila after digestion with SfiI were used to divide the strains into two groups (Groups A and B), although the similarity level was very low among the groups. Group A consisted of 8 strains, and all of these strains were isolated from hot spring bath water samples. Group B consisted of 11 strains, and all but two of these strains were isolated from Noyu samples. The chlorine resistance (99.9% CT values) of these isolates was experimentally confirmed, and we attempted to define the relationship between chlorine resistance and the geno-cluster. The average CT value of group A (8 strains from hot spring bath water) was 0.49 mg･min/l and the average of group B (9 strains from Noyu samples) was 0.29 mg･min/l. No remarkable differences in the CT values for the groups were found. A chlorine-sensitive Noyu strain (0.14 mg･min/l) and a chlorine-resistant strain (0.62 mg･min/l) from hot spring bath water were then compared to identify any differences in their lipid composition. There was no notable difference in the ratio of saturated to unsaturated fatty acids between the chlorine-sensitive and chlorine-resistant bacteria. However, the chlorine-sensitive and chlorine-resistant bacteria demonstrated differences in the relative percentages of cell wall and cell membrane fatty acids.
A commercial aflatoxin detection ELISA kit, “RIDASCREEN® FAST Aflatoxin”, was validated with corn samples naturally contaminated with aflatoxin and non-contaminated reference corn samples according to the Japanese Notification Method ShokuAnHatsu 0816-7. The trueness, intra-laboratory repeatability, intermediate precision, limit of detection and limit of quantitation were found to be 91%, 10%, 6.4%, 0.6μg/kg and 2μg/kg, respectively, and the performance of the kit was recognized as complying with all criteria in the Supplement Table of the Notification. These data suggest that this kit is useful as a simplified device to screen out negative corn samples contaminated with less than 4μg/kg.
To clarify the availability of the dielectrophoretic impedance measurement (DEPIM) system as the evaluator for oral care, we evaluated the usefulness of DEPIM system by comparison with the standard plate counting (SPC) method. First, the relationship between the DEPIM results and bacterial concentration measured by SPC was clarified. Next, the measurement of the microorganism number in a mixed suspension was evaluated with DEPIM and SPC. The bacterial counts with DEPIM strongly correlated with those with SPC (r2=0.633-0.997) and this correlation was also shown in the measurement of a mixed bacterial suspension (ranging from 105 to 108 cfu/ml) of two bacterial species. Moreover, the experiments using dissociating enzymes to eliminate the influence of the size of the bacterial aggregates demonstrated that the microbial measurement results with DEPIM are unaffected by bacterial aggregates. This study demonstrated that bacterial counts with DEPIM strongly correlated with those with SPC and were unaffected by bacterial aggregates.
Candida albicans malate dehydrogenase (Mdh1p) has been screened by previous proteome studies as a candidate for a vaccine against candidiasis. In this study, recombinant Mdh1 protein with a His-tag was produced in Escherichia coli and evaluated as an immunogenic protein against candidiasis. Mdh1p was administrated to mice by two methods subcutaneous injection and intranasal administration before challenging them with a lethal dose of C. albicans. After vaccination of Mdh1p, antibody responses were observed. To evaluate the vaccination effect of Mdh1p, survival tests were performed after 35 d. Although all control mice died within 24 d or 25 d, 100% and 80% of mice survived with subcutaneous and intranasal administration, respectively. Therefore, our results indicate that, among C. albicans antigens examined thus far, Mdh1p is currently the most effective antigen for use as a vaccine for C. albicans.
A chemiluminescence system, Milliflex Quantum (MFQ), to detect microcolonies, has been used in the pharmaceutical field. In this study, we investigated aquatic bacteria in hemodialysis solutions sampled from bioburden areas in 4 dialysis faculties. Using MFQ, microcolonies could be detected after a short incubation period. The colony count detected with MFQ after a 48-hour incubation was 92% ± 39%, compared to that after the conventionally used 7-14-day incubation period； in addition, the results also showed a linear correlation. Moreover, MFQ-based analysis allowed the visualization of damaged cells and of the high density due to the excessive amount of bacteria. These results suggested that MFQ had adequate sensitivity to detect microbacteria in dialysis solutions, and it was useful for validating the conditions of conventional culture methods.