Production characteristics of lipopeptide antibiotics in biofilm fermentation of Bacillus subtilis

In biofilm fermentation, from the very early moments, surfactin was produced along with the biofilm development in the lipopeptide antibiotic production medium by using Bacillus subtilis. However, almost no iturin A was produced in its first 24 hours of cultivation and the production of iturin A began much later. Volumes of the nutrient medium and available surface area of the biofilm reactors were found to be important with the relative production of these two antibiotics. Production of iturin A was increased from 12 mg to about 50 mg per reactor when the culture size was increased from 5 mL to 20 mL, as the depth of the medium was increased. The production level was saturated thereafter with larger volumes. On the other hand, surfactin production was remained similar, which was about 10 mg per reactor, from all the 5 mL to 80 mL of biofilm culture. Optimized temperature for iturin A and surfactin production was observed at 25 and 37°C, respectively. In the biofilm fermentation, production of surfactin was increased when the incubation temperature was increased within the temperature range of 25 to 37°C, on the other hand, iturin A production was gradually decreased with the increase of the incubation temperature.     

Application of malt residue in submerged fermentation of Bacillus subtilis

Malt residue is a common waste or byproduct from beer industries after brewing and milling of malted barley. In this work, Bacillus subtilis RB14 was used to study the microbial growth and production of secondary metabolites like lipopeptide antibiotic iturin A in the malt residue for its effective recycling. B. subtilis RB14 could grow in submerged fermentation of malt residue and significant growth (10⁹ CFU/mL) was observed without any supplementation. In submerged fermentation iturin A production using malt residue was about 170 mg/L, which was found to be higher than its production in No.3 (Polypepton, glucose, KH₂PO₄, MgSO₄·7H₂O) medium where production was about 120 mg/L. More than 600 mg/L of iturin A production was observed when malt residue was combinedly used with No.3 medium. This production was significantly higher than their summation of their individual production. However, the growth of B. subtilis in combined medium was found to be similar to that of the submerged fermentation in simple malt residue. Therefore, the remarkable enhancement in production of iturin A in supplemented malt residue was attributed to the nutrients supplied from No.3 medium.     

Effects of copper chloride on formation of polychlorinated dibenzo-p-dioxins in model waste incineration

Combustion experiments in a laboratory-scale fluidized-bed reactor were conducted to elucidate the effects of copper chloride as a catalyst on polychlorinated dibenzo-p-dioxins (PCDDs) formation in municipal waste incineration. We used model wastes with and without copper chloride (CuCl₂ · 2H₂O), both of which contained polyvinyl chloride as a chlorine source. Combustion temperature was set to 900 °C, and the amount of air supplied was twice the stoichiometric ratio. The experimental setup was carefully planned to suppress the influences of experimental conditions except the waste composition. Results of these experiments showed that copper chloride in the waste increased the amount of PCDDs formed and made the homologue profile to shift towards the highly chlorinated species. Copper chloride contributes to the PCDDs formation by promoting chlorination, whereby the reaction is important in that organic matter is chlorinated directly by copper compounds. Copper chloride did not exert a great influence on the isomer distribution patterns of PCDDs, while there appeared a significant difference in the case of PCDFs. This points out the difference of the major formation mechanisms between PCDDs and PCDFs. PCDDs are less formed by the catalytic reactions from carbon/polycyclic aromatic hydrocarbons than PCDFs in our experimental conditions.     

Consideration on safety for emerging technology – Case studies of seven service robots

Despite the fact that several manufacturers of service robots launched their innovations into the market, worldwide guidelines or regulations concerning the safety aspects of service robots are not yet available. However the general principles and methodologies of safety of machinery (e.g., ISO/IEC Guide 51, partly ISO14121, ISO12100) could be implemented to a certain degree. The safety of seven service robots as an emerging technology was verified by safety professionals of “NPO – the Safety Engineering Laboratory” a Non Profit Organization. NPO verified the “Critical Hazards” for each service robot mentioned by the respective manufacturers. For those cases, both the “As Low As Reasonably Practicable” principle and the “Reasonable Alternative Design” standard were applied for judging if the risk associated with the Critical Hazards were tolerable or not and if state of the art measures for reducing the risk were applied adequately. These experiences will help to establish guidelines for the safety of service robots as an emerging technology in the future.     

A spatially explicit empirical model of structural development processes in natural forests based on climate and topography

Stand structure develops with stand age. Old-growth forests with well-developed stand structure support many species. However, development rates of stand structure likely vary with climate and topography. We modeled structural development of 4 key stand variables and a composite old-growth index as functions of climatic and topographic covariates. We used a hierarchical Bayesian method for analysis of extensive snap-shot National Forest Inventory (NFI) data in Japan (n = 9244) to account for differences in stand age. Development rates of structural variables and the old-growth index exhibited curvilinear responses to environmental covariates. Flat sites were characterized by high rates of structural development. Approximately 150 years were generally required to attain high values (approximately 0.8) of the old-growth index. However, the predicted age to achieve specific values varied depending on environmental conditions. Spatial predictions highlighted regional variation in potential structural development rates. For example, sometimes there were differences of >100 years among sites, even in the same catchment, in attainment of a medium index value (0.5) after timber harvesting. The NFI data suggested that natural forests, especially old natural forests (>150 years), remain generally on unproductive ridges, steep slopes, or areas with low temperature and deep snow, where many structural variables show slow development rates. We suggest that maintenance and restoration of old natural forests on flat sites should be prioritized for conservation due to the likely rapid development of stand structure, although remaining natural forests on low-productivity sites are still important and should be protected.     

Mass spectrometric study of secondary organic aerosol formed from the photo-oxidation of aromatic hydrocarbons

From measurements by an Aerodyne Aerosol Mass Spectrometer (AMS), secondary organic aerosol (SOA) formed in laboratory chambers is believed to be less oxidized than well-oxidized ambient organic aerosol (OA). However, the mass spectrum of SOA formed from the photo-oxidation of aromatic hydrocarbons has not been sufficiently studied by using AMS though these reactions are potential sources of urban SOA. In this study, we studied SOA formed from the photo-oxidation of seven aromatic hydrocarbons by using Time-of-Flight AMS. Strong mass signals from SOA were found at m/z 43 (m43) and 44 (m44) in all the experiments. The m44 to total organic aerosol mass ratio (m44/OA) increased with irradiation time. For example, the m44/OA ratio increased from 10.6% to 13.3% during irradiation for 11 h in an experiment with toluene. The average m44/OA ratios were determined to be 5.8–17.1% for all the experiments. The m44/OA decreased and the m43/OA increased with increasing number of alkyl substituents of precursor aromatic hydrocarbons. This is because low-reactive ketones are preferentially produced rather than aldehydes with increasing number of alkyl substituents. The m44/OA ratios of the benzene and monoalkylbenzene oxidation were 12.2–17.1% and were close to those of well-oxidized ambient OA. These findings are consistent with the hypothesis that the photo-oxidation of aromatic hydrocarbons is a potential source of urban SOA. In addition to oxygenated organic compounds, organic nitrogen oxides were also shown to be present in SOA by high-resolution mass spectra.     

Tritium concentrations in the atmospheric environment at Rokkasho, Japan before the final testing of the spent nuclear fuel reprocessing plant

This study aimed at obtaining background tritium concentrations in precipitation and air at Rokkasho where the first commercial spent nuclear fuel reprocessing plant in Japan has been under construction. Tritium concentration in monthly precipitation during fiscal years 2001-2005 had a seasonal variation pattern which was high in spring and low in summer. The tritium concentration was higher than that observed at Chiba City as a whole. The seasonal peak concentration at Rokkasho was generally higher than that at Chiba City, while the baseline concentrations of both were similar. The reason for the difference may be the effect of air mass from the Asian continent which is considered to have high tritium concentration. Atmospheric tritium was operationally separated into HTO, HT and hydrocarbon (CH₃T) fractions, and the samples collected every 3 d-14 d during fiscal year 2005 were analyzed for these fractions. The HTO concentration as radioactivity in water correlated well with that in the precipitation samples. The HT concentration was the highest among the chemical forms analyzed, followed by the HTO and CH₃T concentrations. The HT and CH₃T concentrations did not have clear seasonal variation patterns. The HT concentration followed the decline previously reported by Mason and Östlund with an apparent half-life of 4.8 y. The apparent and environmental half-lives of CH₃T were estimated as 9.2 y and 36.5 y, respectively, by combining the present data with literature data. The Intergovernmental Panel on Climate Change used the atmospheric lifetime of 12 y for CH₄ to estimate global warming in its 2007 report. The longer environmental half-life of CH₃T suggested its supply from other sources than past nuclear weapon testing in the atmosphere.     

Real-time reverse transcription PCR analysis of trichloroethylene-regulated toluene dioxygenase expression in Pseudomonas putida F1

Toluene dioxygenase (tod) is a multicomponent enzyme system in Pseudomonas putida F1. Tod can mediate the degradation of Trichloroethylene (TCE), a widespread pollutant. In this study, we try to explore the TCE-regulated tod expression by using real-time qRT-PCR. The minimal culture media were supplemented with glucose, toluene, or a mixture of glucose/toluene respectively as carbon and energy sources. The TCE was injected into each medium after a 12-hour incubation period. The TCE injection severely affected bacterial growth when cultured with toluene or toluene/glucose mixtures. The cell density dropped 61 % for bacteria growing in toluene and 36 % for bacteria in the glucose/toluene mixture after TCE injection, but the TCE treatment had little effect on bacteria supplied with glucose alone. The decrease in cell number was caused by the cytotoxicity of the TCE metabolized by tod. The results from the real-time qRT-PCR revealed that TCE was capable of inducing tod expression in a toluene-dependent manner and that the tod expression level increased 50 times in toluene and 3 times in the toluene/glucose mixture after 6 hours of TCE treatment. Furthermore, validation of the rpoD gene as a reference gene for P. putida F1 was performed in this study, providing a valuable foundation for future studies to use real-time qRT-PCR in the analysis of the P. putida F1 strain.     

Optimizing nutrient supply in a rotatory-switching biofilter for toluene vapor treatment

The influence of nutrient conditions on the degradation of toluene vapor in a rotatory-switching biofilter (RSB) was investigated. The biofilter consists of four segments connected in series, each with a packing layer made of polyvinyl formal. The influent airstreams including toluene vapors were passed through segments 1-3 as up-flow with a toluene concentration of 0.9-1.2 g m(-3) and with an empty-bed retention time of 26-52 sec. Nutrient solutions were fed to all packed segments once a day by means of immersion. The nutrient solution was used repeatedly and replenished by the addition of (NH4)2SO4. The result at 155 days showed nitrogen depletion was particularly obvious and the lack of nitrogen affected toluene removal. By adding 161 g of nitrogen solution per volumetric cubic meter of reactor, toluene removal efficiency was immediately increased to greater than 99%. With long-term biofilter operation, 21%-32% of ammonium was utilized for nitrification because of the growth of nitrifying bacteria such as Nitrosomonas sp. Based on the carbon-nitrogen balance, the daily nitrogen demand for toluene removal was estimated 2.1 g day(-1) at a toluene load of 70 g m(-3) hr(-1).