Improvement in solvent tolerance by exogenous glycerol in Pseudomonas sp. BCNU 106.

Solvent hypertolerant Pseudomonas sp. BCNU 106 still has some underlying growth limitation in solvents. Therefore, efficient mass cultivation methods are needed to pursue its applications in biotechnology. Pseudomonas sp. BCNU 106 was cultured in a medium supplemented with 0·05 mol l-1 glycerol and cell survival was monitored during its cultivation in the presence of 1% (v/v) toluene. Exogenously supplemented glycerol provided more protection against damage caused by toluene stress and conferred higher solvent tolerance of Pseudomonas sp. BCNU 106 to toluene compared to control Pseudomonas sp. BCNU 106 without the supplementation of glycerol. This low-cost mass cultivation method can be used to efficiently apply solvent-tolerant bacteria in biotransformation and biodegradation. SIGNIFICANCE AND IMPACT OF THE STUDY: Protection against toluene and improvement in bacterial cell growth by supplementation of glycerol in the presence of toluene are demonstrated in this study. This result can be used to solve growth-related hindrances of solvent-tolerant bacteria and establish their low-cost mass cultivation, thereby broadening their industrial and environmental applications.

Enhancement of solvent tolerance in Pseudomonas sp. BCNU 106 with trehalose.

UNLABELLED: As the solvent hyper-resistant Pseudomonas sp. BCNU 106 experiences limited growth with solvents, a strategy is therefore needed to allow better growth to broaden its performance in biotechnological applications. Pseudomonas sp. BCNU 106 was cultivated in a medium supplemented with 0·05 mol l(-1) trehalose, and the cell survival was observed during subsequent growth with 1% (v/v) toluene. Exogenously added trehalose was transported into the cells and conferred protection against toluene stress. BCNU 106 grown in the presence of exogenous trehalose showed higher solvent tolerance, it can thus have more potential for biotransformation and biodegradation. SIGNIFICANCE AND IMPACT OF THE STUDY: This study shows that exogenously supplemented trehalose confers protection against toluene stress and enhances the bacterial cell growth in the presence of toluene. This is of importance to the mass cultivation of solvent-tolerant bacteria, where some of the growth-related limitations of solvent-tolerant bacteria can be overcome, and their performance in biotechnological applications for biotransformation and biodegradation broadened.

Microbial community in biofilm on membrane surface of submerged MBR: effect of in-line cleaning chemical agent.

The objective of this study was to investigate the change in microbial community pattern with the effect of cleaning agent using a quinone profile that is used for membrane in-line chemical cleaning in SMBR. The dominant quinone types of biofilm were ubiquinone (UQs)-8, -10, followed by menaquinone (MKs)-8(H4), -7 and UQ-9, but those of suspended microorganisms were UQ-8, UQ-10 followed by MKs-8(H4), -7 and -11. Both UQ and MK contents decreased with increasing NaCIO dosage and it seems that there is more resistance from UQ compared to MK. In addition, COD and DOC concentrations increased with increasing NaClO dosage up to 0.05 g-NaCIO/g-SS. The organic degradation performance of the microbial community in the presence of NaClO was impaired. The present study suggested that larger added amounts of NaClO caused an inhibition of organic degradation and cell lysis.

The stability of nitrite nitrification with strong nitrogenous wastewater: effects of organic concentration and microbial diversity.

A stable achievement of nitritation with strong nitrogenous wastewaters is considered as a difficult task in practice, probably due to the fate of interaction between dominating heterotrophs and nitrifier species. An experimental study was carried out to examine the organic effects in lab-scale biofilm nitritation reactors. The control unit without organic addition showed a stable nitritation performance for more than 220 days of operating period. The nitritation activity gradually failed at the reactors with an organic addition, but the nitritation activity eventually recovered with a prolonged aeration. It was not possible to explain the nitritation recovery with neither free ammonia inhibition concept nor DO competition hypothesis in these cases. The results suggest that the nitritation with organic requires a long start-up period for acclimation. In addition, the results of quinone profile analysis were in agreement with nitritation activity in reactors. The diversity of microbial community in the nitritation reactors could be described by the quinone profiles.

Oxidative treatment characteristics of biotreated textile-dyeing wastewater and chemical agents used in a textile-dyeing process by advanced oxidation process.

The oxidative treatment characteristics of biotreated textile-dyeing wastewater and typical chemicals such as desizing, scouring, dispersing and swelling agents used in the textile-dyeing process by advanced oxidation process were experimentally studied. The refractory organic matters remained in the effluent of biological treatment process without degradation may be suitable for the improvement of biodegradability and mineralized to CO2 by combined ozonation with and without hydrogen peroxide. On the other hand, the refractory chemicals contained in the scouring agent A and swelling agent may not be mineralized and their biodegradability may not be improved by ozonation. However, the BOD/DOC ratio of scouring agent B increased from 0.3 to 0.45 after ozonation. Based on the results described above, advanced treatment process involving the ozonation without and with the addition of hydrogen peroxide, followed by biological treatment was proposed for the treatment of refractory wastewater discharged from the textile-dyeing process.

Change of microbial community structure by respiratory quinone profile at intermittently aerated membrane bioreactor.

In the present study, the microbial community structure in an intermittently aerated submerged membrane bioreactor treating domestic wastewater was observed using the respiratory quinone profiles. The effects of different time interval for the aerobic and anoxic period on microbial community structure were examined with 60/90 min (Step 1) and 90/60 min (Step 2) as anoxic/aerobic periods. There was an observable slight difference in microbial community structure between Step 1 and Step 2 in the submerged membrane bioreactor. The dominant quinone types for Step 1 at both anoxic and aerobic conditions were UQ-8 followed by UQ-10 and MK-6, but those for Step 2 were UQ-8, MK-6 and MK-10(H4). The microbial diversity of Step 1 and Step 2 based on the composition of all quinones was 10.6-11.7 and 13.3-13.0 for anoxic and aerobic conditions, respectively. The present results suggest that the introduction of intermittent aeration into the submerged membrane bioreactor has little influence on the bacterial community structure.

Investigation of biological and fouling characteristics of submerged membrane bioreactor process for wastewater treatment by model sensitivity analysis.

In this study, a mathematical model for the submerged membrane bioreactor (SMBR) was developed. The activated sludge model No. 1 (ASM1) was modified to be suitable for describing the characteristics of the SMBR, and the resistance-in-series model was integrated into the ASM1 to describe membrane fouling. Using the newly developed model, the biological and fouling characteristics of the submerged membrane bioreactor process for wastewater treatment was investigated by sensitivity analysis. The sensitivity of effluent COD and nitrogen, TSS in the reactor and membrane flux with respect to each parameter (K(h), mu(H), K(S), K(NHH), K(NOH), b(H), Y(H), mu(A), K(NHA), b(A), Y(A), K(m) and alpha) was investigated by model simulation. As a result, the most important factors affecting membrane fouling were hydrolysis rate constant (K(h)) and cross-flow effect coefficient (K(m)). Heterotrophic yield coefficient (Y(H)) had a great influence on effluent quality. Effluent quality was also somewhat sensitive to K(h). Peculiar operating conditions of the SMBR such as long solids retention time (SRT), absolute retention of solids by membrane and high biomass concentration in bioreactor could explain these model simulation results. The model developed in this study would be very helpful to optimize operating conditions as well as design parameters for a SMBR system.

Microbial community structure of membrane fouling film in an intermittently and continuously aerated submerged membrane bioreactor treating domestic wastewater.

There was an observable difference in microbial community structure between suspended microorganisms and membrane biofouling film in intermittently and continuously aerated SMBRs. The dominant quinone type of membrane biofouling film in an intermittently aerated SMBR was ubiquinone (UQs)-8, -10 followed by menaquinone (MKs)-8(H4) and -8(H2). But that of the continuously aerated SMBR was UQs-10, -8 followed by MKs-6 and -8(H4). The experimental results also showed that the conditions of an intermittently aerated SMBR may contribute to biofouling by Pseudomonas, Moraxella, Vibrio (quinone type UQ-8), Staphylococcus warneri (quinone type MK-7), Micrococcus sp. (quinone type MK-8(H2)) and Nocardia sp. (quinone type MK-8(H4)), but biofouling in a continuously aerated SMBR may be due to Paracoccus sp. (quinone type: UQ-10) and Flavobacterium species (quinone type: MK-6). The microbial diversities in the intermittently aerated SMBR were 10.9 and 9.4 for biofouling film and suspended microorganisms, respectively. For the continuously aerated SMBR, the results were 10.4 and 10.5 for biofouling film and suspended microorganisms, respectively.

Toward the bioremediation of dioxin-polluted soil: structural and functional analyses of in situ microbial populations by quinone profiling and culture-dependent methods.

In order to obtain basic information toward the bioremediation of dioxin-polluted soil, microbial communities in farmland soils polluted with high concentrations of polychlorinated dibenzo-p-dioxins (PCDDs) and polychlorinated dibenzofurans (PCDFs) were studied by quinone profiling as well as conventional microbiological methods. The concentration of PCDD/Fs in the polluted soils ranged from 36 to 4,980 pg toxicity equivalent quality (TEQ) g(-1) dry weight of soil. There was an inverse relationship between the levels of PCDD/Fs and microbial biomass as measured by direct cell counting and quinone profiling. The most abundant quinone type detected was either MK-6 or Q-10. In addition, MK-8, MK-8(H2), and MK-9(H8) were detected in significant amounts. Numerical analysis of quinone profiles showed that the heavily polluted soils (> or = 1,430 pg TEQ g(-1)) contained different community structures from lightly polluted soils (< or = 56 pg TEQ g(-1)). Cultivation of the microbial populations in the heavily polluted soils with dibenzofuran or 2-chlorodibenzofuran resulted in enrichment of Q-10-containing bacteria. When the heavily polluted soil was incubated in static bottles with autoclaved compost as an organic nutrient additive, the concentrations of PCDD/Fs in the soil were decreased by 22% after 3 months of incubation. These results indicate that dioxin pollution exerted a significant effect on microbial populations in soil in terms of quantity, quality, and activity. The in situ microbial populations in the dioxin-polluted soil were suggested to have a potential for the transformation of PCDD/Fs and oxidative degradation of the lower chlorinated ones thus produced.

Analytical precision and repeatability of respiratory quinones for quantitative study of microbial community structure in environmental samples.

Microbial community structure is one of the important factors controlling the pollutant-degrading capacity of ecosystems. The analysis of microbial quinones has gained increased recognition as a simple and useful approach for studying microbial structure in environmental samples. The analytical precision of quinone characterization using high performance liquid chromatograph (HPLC) with a UV-detector was studied in this study. Activated sludge was used as a typical mixed culture. The coefficient of variation of quinone content was lower than 6%, and that of microbial diversity calculated from the composition of quinones was as low as 3%. Statistical analyses on the analytical precision of quinones demonstrated that the critical value of dissimilarity between two quinone profiles of activated sludge, which is used to make a judgement whether the two quinone profiles are different or not, is 0.1 for the analytical method used in this study. The values of minimum biomass required for quinone analysis to have a reliable analytical result of microbial quinones were 2 mg-dry-cell for activated sludge.

Analysis of material and energy flow in sewage treatment facilities in Japan.

Energy consumption in sewage treatment facilities in Japan has increased due to increasing tap water consumption. To reduce the resource/energy consumption in sewage treatment facilities, measures such as the selection of optimum treatment processes and operating conditions should be considered. The objective of this study is to gather information necessary for the determination of optimum sewage treatment processes and optimum operating conditions. The energy consumption and material flow in sewage treatment facilities in Japan are analyzed using statistical data. In 1994, reuse rate of treated sewage outside the treatment facilities in Japan was 18% of the amount of domestic treated water. In this regard, reuse of water outside facilities should be encouraged. Average electric power consumption per unit volume of wastewater in sewage treatment facilities varies widely from facility to facility and closely correlates with the facility scale. For example, the smaller the facility scale, the larger the electric power consumption. Treatment volume of sewage in smaller facilities is much less than their capacity. 3.7 million t year-1 of dehydration cake is incinerated and 0.1 million t year-1 of it is converted by composting. The recycle rate of the cake was low. Developing a new sludge treatment process other than incineration is necessary.

Effects of temperature on biodegradation characteristics of organic pollutants and microbial community in a solid phase aerobic bioreactor treating high strength organic wastewater.

The BOD removal rate and microbial community structure in a solid phase aerobic bioreactor using polyvinyl alcohol gel particles as packing material for the treatment of high strength organic wastewater were investigated at various temperatures. The BOD removal rate in the bioreactor increased when the temperature increased from 20 degrees C to 30 degrees C, 40 degrees C, and 50 degrees C, but it decreased when the temperature increased from 50 degrees C to 60 degrees C. Higher temperature enhanced the endogenous respiration of microbes in the bioreactor. The microbial community structure in the bioreactor was analyzed with quinone profile. The experimental results showed that the microbial community structure in the bioreactor was significantly affected by temperature. The dominant quinone of the microbes inhabiting the bioreactor was ubiquinone-8 at 30 degrees C, but that at 50 degrees C and 60 degrees C was menaquinone-7. It was estimated that the thermophilic Bacillus having menaquinone-7 dominated in the bioreactor at higher temperature. The microbial diversity in the bioreactor varied with temperature.

Characterization of microbial community in an activated sludge process treating domestic wastewater using quinone profiles.

The dynamics of microbial community structure of activated sludges in a small-scale domestic wastewater treatment process were examined using a novel approach of quinone profiles. The composition and content of quinones in the activated sludges were analyzed monthly over a period of one year. More than 4 types of ubiquinones and 12 types of menaquinones were observed in the activated sludges, with the dominant quinones being ubiquinone (UQ)-8, menaquinone (MK)-7, followed by UQ-10, MK-8 and MK-6. The total quinone contents in the activated sludges varied from 0.93 to 2.68 mumol per gram of particle organic carbon. The molar ratio of ubiquinones to menaquinones (UK/MK) changed from 0.38 to 0.98, indicating that anaerobic bacteria dominated the microbial community of the activated sludges examined. The ratio of UQ/MK varied similar to that of dissolved oxygen in the bulk. The microbial diversity of the activated sludges calculated from the quinone compositions was 13.4-16.8. The diversity of menaquinones was much higher than that of ubiquinones, and increased slightly with increasing temperature. The microorganisms containing menaquinones appear to be sensitive to the change in temperature than those containing ubiquinones.