Toxicity assessment upon augmented biostimulation source to indigenous rhizobium Cupriavidus taiwanensis

This novel-attempt study used chemostat pulse technique (CPT) and transient dynamics of dissolved oxygen (DO) in CSTR to quantify stimulating or inhibitory effects of augmented nutrient sources in the presence of phenol upon Cupriavidus taiwanensis R186. With injected augmented nutrients, phenol degradation performance of R186 was directly dependent on combined toxicity between phenol and the augmented substrate and the biodegradability of phenol. The findings indicated that although phenol was toxic to R186, all augmented nutrient sources still exhibited stimulating effects to bacterial growth of R186 in the presence of phenol. The simulating rankings of augmented nutrients were (1) at 200 mg/L, acetic acid>gluconic acid>yeast extract>glycerol>phenol alone, (2) at 1000 mg/L, gluconic acid>acetic acid>glycerol>yeast extract>phenol alone. This stimulating effect clearly suggested that this combined toxicity was antagonistic. It was also revealed that transient responses of DO seemed to be in parallel with the findings from CPT. It was thus concluded that substrate consumption patterns would play the most significant role in biostimulation to cultures with dual energy sources. This study can help uncover the mysteries of optimal biostimulation for phenol degradation as proposed in previous studies. In addition, this study directly provided a kinetic model to quantify the relative stimulation ranking of augmented nutrients in the presence of phenol for practical bioremediation.     

Optimal exponential feeding strategy for dual-substrate biostimulation of phenol degradation using Cupriavidus taiwanensis

The exponential feeding strategy (EFS) of dual substrates (i.e., phenol and glycerol) was applied to optimize the overall performance of phenol degradation by Cupriavidus taiwanensis R186. Addition of a second substrate (e.g., glycerol) could stimulate the phenol biodegradation efficiency of strain R186. Hence, a feasible EFS was developed for fed-batch phenol biodegradation using the dual-substrate biostimulation technique. The phenol degradation kinetics was well characterized with proposed model and response surface analysis. Our findings quantitatively revealed that glycerol could effectively enhance the phenol degradation performance, as the highest phenol degradation efficiency occurred with the supplementation of 0.8–1.2 g L⁻¹ of glycerol. The optimal dual-substrate EFS was identified via contour analysis and kinetic modeling. With the optimal dual-substrate EFS (i.e., a feeding rate constant (α₁ and α₂) of 0.5 and 0.3, respectively), the shortest time (ca. 13.80 h) for phenol degradation was achieved with a specific growth rate of ca. 0.281 h⁻¹.     

Characteristics of phenol biodegradation in saline solutions by monocultures of Pseudomonas aeruginosa and Pseudomonas pseudomallei

Phenol is a highly toxic and carcinogenic compound and its biodegradation is very important to meet the environmental regulations. Two bacterial strains capable of utilizing phenol as a sole source of carbon were isolated from the wastewater of a pharmaceutical industry. On the basis of morphological and biochemical characteristics these strains were identified as Pseudomonas aeruginosa and Pseudomonas pseudomallei. Both of these strains were very efficient for phenol degradation. P. pseudomallei degraded phenol at a maximum concentration of 1500 mg L(-1) within seven days with a specific growth rate of 0.013 h(-1) and phenol degradation rate of 13.85 mg L(-1)h(-1). Maximum initial concentration of phenol utilized by P. aeruginosa was 2600 mg L(-1) with 0.016 h(-1) specific growth rate and 26.16 mg L(-1)h(-1) phenol degradation rate. Moreover, the effect of various salts i.e., NaCl, KCl, Na(2)SO(4) and K(2)SO(4) on the growth of these strains and phenol degradation rate (at 1000 mg L(-1)) was studied. In the presence of these salts, P. aeruginosa showed up to 1.53 and 1.34 times faster phenol degradation rate and specific growth rate, respectively as compared to P. pseudomallei. In addition, P. aeruginosa exhibited higher chemical oxygen demand (COD) and biochemical oxygen demand (BOD) reduction rates as compared to the strain P. pseudomallei.     

Estimation of the biodegradation rate of 2,3,7,8-tetrachlorodibenzo-p-dioxin by using dioxin-degrading fungus, Pseudallescheria boydii

We are developing a bioreactor system for treating dioxin-contaminated soil or water using the dioxin-degrading fungus, Pseudallescheria boydii (P. boydii). In order to design the bioreactor system, this study estimated the rate at which P. boydii degraded 2,3,7,8-tetrachlorodibenzo-p-dioxin (2,3,7,8-TCDD), which is the most toxic of the dioxins. The experimental results showed that P. boydii degraded 2,3,7,8-TCDD during its logarithmic growth phase, using glucose as a carbon source for growth, and that the growth of P. boydii was not affected by 2,3,7,8-TCDD concentrations usually found at contaminated sites. These results were then used to apply successfully an existing mathematical model to the degradation of 2,3,7,8-TCDD by P. boydii. This allowed an estimation of the rate of degradation of 2,3,7,8-TCDD by P. boydii that can be used in the design of the bioreactor system.     

Statistical optimization of medium components and growth conditions by response surface methodology to enhance phenol degradation by Pseudomonas putida

In this work, a four-level Box-Behnken factorial design was employed combining with response surface methodology (RSM) to optimize the medium composition for the degradation of phenol by pseudomonas putida (ATCC 31800). A mathematical model was then developed to show the effect of each medium composition and their interactions on the biodegradation of phenol. Response surface method was using four levels like glucose, yeast extract, ammonium sulfate and sodium chloride, which also enabled the identification of significant effects of interactions for the batch studies. The biodegradation of phenol on Pseudomonas putida (ATCC 31800) was determined to be pH-dependent and the maximum degradation capacity of microorganism at 30 degrees C when the phenol concentration was 0.2 g/L and the pH of the solution was 7.0. Second order polynomial regression model was used for analysis of the experiment. Cubic and quadratic terms were incorporated into the regression model through variable selection procedures. The experimental values are in good agreement with predicted values and the correlation coefficient was found to be 0.9980.     

Characterization of phenol degradation by Rhizobium sp. CCNWTB 701 isolated from Astragalus chrysopteru in mining tailing region

To screen high strength phenol degrading bacteria, we selected 108 rhizobial strains isolated from nodules of eight wild legumes species in the mining tailing region of Shaanxi province, northwest of China, and cultivated them in a basal salt (BS) medium supplemented with different phenol concentrations as a sole carbon source. The results showed that some of the strains could use phenol as sole carbon source. In order to study the characteristics of phenol degradation, the strain CCNWTB701 isolated from Astragalus chrysopteru was used as well, due to the fact that it was very efficient in phenol degradation. The phenol degradation was around 99.5 and 78.3%, with an initial concentration of 900 and 1000 mg/l phenol in 62 and 66 h, respectively. Kinetic studies indicated that the strain had a high KS (743.1 microM) and an extremely high KSI (10,469 microM) in Haldane's model. The phylogenetic analysis based on 16S rRNA gene sequences showed that CCNWTB701 belonged to the Rhizobium genus, and it was closely related to Rhizobium mongolense and Rhizobium gallicum.     

Biodegradation of anthracene by Aspergillus fumigatus

An anthracene-degrading strain, identified as Aspergillus fumigatus, showed a favorable ability in degradation of anthracene. The degradation efficiency could be maintained at about 60% after 5d with initial pH of the medium kept between 5 and 7.5, and the optimal temperature of 30 °C. The activity of this strain was not affected significantly by high salinity. Exploration on co-metabolism showed that the highest degradation efficiency was reached at equal concentration of lactose and anthracene. Excessive carbon source would actually hamper the degradation efficiency. Meanwhile, the strain could utilize some aromatic hydrocarbons such as benzene, toluene, phenol etc. as sole source of carbon and energy, indicating its degradation diversity. Experiments on enzymatic degradation indicated that extracellular enzymes secreted by A. fumigatus could metabolize anthracene effectively, in which the lignin peroxidase may be the most important constituent. Analysis of ion chromatography showed that the release of anions of A. fumigatus was not affected by addition of anthracene. GC-MS analysis revealed that the molecular structure of anthracene changed with the action of the microbe, generating a series of intermediate compounds such as phthalic anhydride, anthrone and anthraquinone by ring-cleavage reactions.     

Bioremediation of phenols and polycyclic aromatic hydrocarbons in creosote contaminated soil using ex-situ landtreatment.

Soil from a former creosoting plant containing phenols and polycyclic aromatic hydrocarbons, was remediated using an ex-situ landtreatment process. Total 16 USEPA priority PAH and total phenol were reduced from 290 mg/kg and 40 mg/kg to < 200 mg/kg and 2 mg/kg, respectively. The bioremediation process involved soil mixing, aeration, and slow release fertilizer addition. The indigenous populations of PAH and phenol utilizing populations of microorganisms were shown to increase during the treatment process, indicating that biostimulation was effective. The most extensive degradation was apparent with the 2- and 3-ring PAH, with decreases of 97% and 82%, respectively. The higher molecular weight 3- and 4-ring PAH were degraded at slower rates, with reductions of 45% and 51%, respectively. Six-ring PAH were degraded the least with average reductions of < 35%. The residual concentrations of PAH and total phenol obtained in the study allowed the treated soil to be disposed of as low level contaminated landfill.     

Biodegradation of chlorpyrifos and its hydrolysis product 3,5,6-trichloro-2-pyridinol by Bacillus pumilus strain C2A1

A bacterial strain C2A1 isolated from soil was found highly effective in degrading chlorpyrifos and its first hydrolysis metabolite 3,5,6-trichloro-2-pyridinol (TCP). On the basis of morphology, physiological characteristics, biochemical tests and 16S rRNA sequence analysis, strain C2A1 was identified as Bacillus pumilus. Role of strain C2A1 in the degradation of chlorpyrifos was examined under different culture conditions like pH, inoculum density, presence of added carbon/nutrient sources and pesticide concentration. Chlorpyrifos was utilized by strain C2A1 as the sole source of carbon and energy as well as it was co-metabolized in the presence of glucose, yeast extract and nutrient broth. Maximum pesticide degradation was observed at high pH (8.5) and high inoculum density when chlorpyrifos was used as the sole source and energy. In the presence of other nutrients, chlorpyrifos degradation was enhanced probably due to high growth on easily metabolizable compounds which in turn increased degradation. The strain C2A1 showed 90% degradation of TCP (300 mg L⁻¹) within 8 days of incubation.     

Effects of pH on the chlorination process of phenols in drinking water

Toxic organic compounds detected generally in source water could combine with chlorine and contribute significantly to chlorination disinfection by-products (CDBPs). The effects of pH on species distribution of CDBPs and the kinetics of chlorination were investigated using phenol as a model of ionizable toxic organic compounds in the pH range of 6.0-9.0. It was found that five chlorination products including 2-monochlorophenol (2-MCP), 4-monochlorophenol (4-MCP), 2,6-dichlorophenol (2,6-DCP), 2,4-dichlorophenol (2,4-DCP) and 2,4,6-trichlorophenol (TCP) were produced by successive chlorination substitution. MCP (2-MCP and 4-MCP) were the dominant products and phenol partly remained in acid media, while TCP and DCP (2,6-DCP and 2,4-DCP) were the main components in neutral and alkaline media. A steady equilibrium of phenol and its chlorination products was reached in 20-30 min in acid-, neutral- and slightly alkaline media, and was delayed to 60-180 min in alkaline media. The difference in properties between phenols and phenolates, and those between HOCl and ClO(-) should be considered simultaneously in explaining the effects of pH on the chlorination process with the theory of electrophilic substitution. These results show that pH plays an important regulating role in the species distribution of CDBPs and the kinetics of chlorination for ionizable toxic organic compounds in chlorination.     

In situ degradation of phenol and promotion of plant growth in contaminated environments by a single Pseudomonas aeruginosa strain

For bioremediation of contaminated environments, a bacterial strain, SZH16, was isolated and found to reduce phenol concentration in a selective medium. Using the reaction vessel containing the soil mixed with phenol and bacteria, we found that the single strain degraded efficiently the phenol level in soil samples. The strain was identified as Pseudomonas aeruginosa on the basis of biochemical tests and by comparison of 16S rDNA sequences, and phosphate solubilization and IAA production were not observed in the strain. Simultaneous examination of the role of strain SZH16 in the plant growth and phenol biodegradation was performed. Results showed that inoculation of the single strain in the phenol-spiked soil resulted in corn growth promotion and in situ phenol degradation and the increase in plant biomass correlated with the decrease in phenol content. Colonization experiments showed that the population of the SZH16 strain remained relatively constant. All these findings indicated that the corn growth promotion might be due to reduction in phytotoxicity, a result of phenol biodegradation by the single strain SZH16. Furthermore, the strain was found to stimulate corn growth and reduce phenol concentration simultaneously in phenol-containing water, and even historically contaminated field soils. It is attractive for environment remediation and agronomic applications.     

Modeling of nonylphenol degradation by photo-nanocatalytic process via multivariate approach

Modeling of photocatalytic degradation of nonylphenol (NP), an endocrine disrupter and toxic compound, has been investigated in synthetic aqueous solutions containing ZnO nanoparticles as semiconductor using multivariate approach. In this regard, a full factorial experimental design was performed in order to study the main variables affecting the degradation process as well as their most significant interactions. Initial NP concentrations ([NP](0)) of 0.454-9.08 μM, were treated with UV-vis/ZnO using different pH and nanocatalyst loading rates. Effect of experimental parameters on the NP degradation rate constant was established by the response surface plots. The degradation rate constant decreased with an increase in the initial concentration of NP, while it increased with ZnO loading until a concentration of 0.5 g L(-1). The rate constant increases with increase in pH up to 10, after which a significant decrease is observed. The results showed that most influential factors on NP degradation constant are the [NP](0), pH of reaction media, and ZnO loading rate, and the most significant interaction is [NP]-pH. Finally, two mathematical models have been proposed to estimate NP degradation rate constant (k) on the basis of the significant variables and interactions. Predicted results of models showed good agreement with the experimental data (R(2) = 0.83 and 0.93).     

Isolation and identification of novel high strength phenol degrading bacterial strains from phenol-formaldehyde resin manufacturing industrial wastewater

Phenols are toxic to all types of organisms. Two bacterial strains capable of utilizing phenol as a sole carbon source were isolated from the phenol bearing industrial wastewater. Based on the biochemical test results the organisms were identified as Pseudomonas cepacia and Bacillus brevis. The organisms were very efficient in phenol degradation, the lag phase increased with increase in phenol concentration. The well-acclimatized cultures of P. cepacia and B. brevis degraded 2500 and 1750 mg l(-1) of phenol in 144 h, respectively. The organisms degrade phenol even in the presence of toxicants like thiocyanate, sulphide and cyanide. The organisms can be effectively used for treating high strength phenol containing thiocyanate, sulphide and cyanide. The P. cepacia degrades phenol with a faster rate than B. brevis. P. cepacia can be used effectively for treating high strength phenolic wastewater.     

Effect of key parameters on the removal of formaldehyde and methanol in gas-phase biotrickling filters

The effect of some important operation parameters, as pH, pollutant load and composition of the nutrient media, on the biodegradation of a mixture of formaldehyde and methanol in a gas-phase biotrickling filter was studied. pH proved to affect the degradation of both compounds at moderately acidic values. Replacing ammonium with nitrate as nitrogen source in the liquid solution led to a slight decrease in performance, though this difference was not really significant. A slight decrease in the elimination rate was also observed when reducing the N-NO(3)(-) concentration to 60% of its original value. No interactions between the two pollutants were found under our working conditions.     

Biodegradation of phenol at high initial concentration by Alcaligenes faecalis

Strain Alcaligenes faecalis was isolated and identified as a member of the genus Alcaligenes by using BIOLOG and 16S rDNA sequence analysis. The phenol biodegradation tests showed that the phenol-degrading potential of A. faecalis related greatly to the different physiological phases of inoculum. The maximum phenol degradation occurred at the late phase of the exponential growth stages, where 1600 mg L(-1) phenol was completely degraded within 76 h. A. faecalis secreted and accumulated a vast quantity of phenol hydroxylase in this physiological phase, which ensured that the cells could quickly utilize phenol as a sole carbon and energy source. In addition, the kinetic behavior of A. faecalis in batch cultures was also investigated over a wide range of initial phenol concentrations (0-1600 mg L(-1)) by using Haldane model. It was clear that the Haldane kinetic model adequately described the dynamic behavior of the phenol biodegradation by the strain of A. faecalis.     

Performance analysis of a continuous bioreactor for ethanol and acetate synthesis from syngas via <Emphasis Type="Italic">Clostridium ljungdahlii</Emphasis> using exergy concept

In this paper, the exergetic performance of a continuous bioreactor for ethanol and acetate synthesis from syngas via a strictly anaerobic autotrophic bacterium Clostridium ljungdahlii was carried out for the first time. The fermentation process was evaluated using both conventional exergy and eco-exergy principles for measuring the productivity and renewability of the process at various liquid media flow rates. The microorganisms successfully upgraded the syngas into invaluable ethanol and acetate through the Wood–Ljungdahl pathway. The exergy efficiency was found to be in the range of 6.5–77.5 and 6.8–77.5 % during the fermentation using conventional exergy and eco-exergy concepts, respectively. The subtle differences observed in the exergetic parameters using the two exergetic concepts were ascribed to the slow growth rate of the microorganisms. Nevertheless, the eco-exergy concept would strongly be recommended for commercial bioreactor containing living organisms due to the inclusion of the information carried by microorganisms in the exergetic calculation. A desired liquid media flow rate of 0.55 mL/min was found according to a newly defined thermodynamic indictor namely exergetic productivity index. More specifically, the maximum exergetic productivity index of the fermentation process was found to be 8.0 using both approaches when the rate of inflow liquid was adjusted at the optimal value. The results of this study revealed that process yield alone cannot be a reliable performance metric for decision making on the productivity of various biofuel production pathways. Finally, the proposed exergetic framework could assist engineers and researchers to link biochemical and physical knowledge more robustly and to quantify and elucidate the general purpose of productivity and renewability.     

Aerobic degradation of nitrobenzene by immobilization of Rhodotorula mucilaginosa in polyurethane foam

Rhodotorula mucilaginosa Z1 capable of degrading nitrobenzene was immobilized in polyurethane foam. The nitrobenzene-degrading capacity of immobilized cells was compared to free cells in batches in shaken culture. Effects of pH and temperature on the nitrobenzene degradation showed that polyurethane-immobilized Z1 had higher tolerances toward acid, alkali, and heat than those of free cells. Kinetic studies revealed that higher concentrations of nitrobenzene were better tolerated and more quickly degraded by polyurethane-immobilized Z1 than by free cells. Moreover, the ability of polyurethane-immobilized Z1 to resist nitrobenzene shock load was enhanced. Experiments on the nitrobenzene degradation in different concentrations of NaCl and in the presence of phenol or aniline demonstrated that polyurethane-immobilized Z1 exhibited higher tolerance toward salinity and toxic chemicals than those of free cells. Immobilization therefore could be a promising method for treating nitrobenzene industrial wastewater. This is the first report on the degradation of nitrobenzene by a polyurethane-immobilized yeast strain.     

Isolation and characterization of a nitrobenzene degrading yeast strain from activated sludge

Strain Z1 was isolated from nitrobenzene-contaminated sludge. Strain Z1 was able to utilize nitrobenzene as a sole source of carbon, nitrogen, and energy under aerobic condition. Based on the morphology, physiological biochemical characteristics, and 26S rDNA D1/D2 domain sequence, strain Z1 was identified as Rhodotorula mucilaginosa. Strain Z1 mineralized up to 450mg L(-1) nitrobenzene. Kinetics of nitrobenzene degradation was described using the Andrews equation. The kinetic parameters were as follows: q(max)=1.50h(-1), K(s)=31.31mg L(-1), and K(i)=101.34mg L(-1). Strain Z1 had a high-salinity tolerance. It degraded nitrobenzene effectively in 5% NaCl (quality concentration). Even in the presence of aniline or phenol, strain Z1 degraded nitrobenzene efficiently. Strain Z1 therefore could be an excellent candidate for the bio-treatment of nitrobenzene industrial wastewaters.     

Degradation of pyridine by one Rhodococcus strain in the presence of chromium (VI) or phenol

A Rhodococcus strain, Chr-9, which has the ability to degrade pyridine and phenol and reduce chromium (VI) (Cr (VI)) was isolated. The strain could grow with pyridine as the sole carbon and nitrogen source, and its pyridine-degradation capability was enhanced by 100 mg l(-1) phenol; however, the degradation of pyridine was inhibited when the phenol concentration was greater than 400 mg l(-1). The hydroxylation of pyridine suggested that the stimulation and inhibition of phenol to the pyridine degradation may be attributed to competition of phenol and pyridine for the hydroxylase gene. Strain Chr-9 was also able to reduce Cr (VI) when glucose and LB was used as the carbon source; however, the Cr (VI) reduction did not occur when pyridine was the sole carbon and energy source. In addition, strain Chr-9 could reduce Cr (VI) and simultaneously degrade pyridine in the presence of glucose. To the best of our knowledge, strain Chr-9 is the first Rhodococcus strain reported to degrade pyridine in the presence of Cr (VI), and the first strain with the pyridine degradation being stimulated by low concentrations of phenol.     

Profiling of organic acids during fermentation by ultraperformance liquid chromatography-tandem mass spectrometry

A method has been developed for rapid quantification of organic acids using ultraperformance liquid chromatography/electrospray-tandem mass spectrometry (UPLC/ESI-MS-MS) to monitor the metabolism of 10 organic acids during microbial fermentation. Because comprehensive chromatographic separation is not required, analysis time is less than traditional ion chromatography assays, with complete organic acid analyses by UPLC/ESI-MS-MS being achieved in less than 3 min. Quantification is accomplished using nine isotopically labeled organic acids as internal standards. Intrasample precisions for organic acid measurements in fermentation supernatants using this method average 8.9% (RSD). Calibration curves are linear over the range of 0.06-100 microg/mL, and detection limits are estimated at 0.06-1 microg/mL. This method has the potential to demonstrate correlation of organic acid consumption and production by microorganisms with observed growth profiles, novel media formulations, and cellular growth events. Data visualization software has been used to profile organic acid levels during fermentation and correlate these profiles to nutrient supplementation protocols employed during microbial production. The potential use of this capability in computational modeling and simulation of microbial metabolism to accelerate the bioprocess development cycle is recognized.