Biostimulation Conditions Research Articles

Multi-Omic Profiling of a Newly Isolated Oxy-PAH Degrading Specialist from PAH-Contaminated Soil Reveals Bacterial Mechanisms to Mitigate the Risk Posed by Polar Transformation Products.

Polar biotransformation products have been identified as causative agents for the eventual increase in genotoxicity observed after the bioremediation of PAH-contaminated soils. Their further biodegradation has been described under certain biostimulation conditions; however, the underlying microorganisms and mechanisms remain to be elucidated. 9,10-Anthraquinone (ANTQ), a transformation product from anthracene (ANT), is the most commonly detected oxygenated PAH (oxy-PAH) in contaminated soils. Sand-in-liquid microcosms inoculated with creosote-contaminated soil revealed the existence of a specialized ANTQ degrading community, and Sphingobium sp. AntQ-1 was isolated for its ability to grow on this oxy-PAH. Combining the metabolomic, genomic, and transcriptomic analyses of strain AntQ-1, we comprehensively reconstructed the ANTQ biodegradation pathway. Novel mechanisms for polyaromatic compound degradation were revealed, involving the cleavage of the central ring catalyzed by Baeyer-Villiger monooxygenases (BVMO). Abundance of strain AntQ-1 16S rRNA and its BVMO genes in the sand-in-liquid microcosms correlated with maximum ANTQ biodegradation rates, supporting the environmental relevance of this mechanism. Our results demonstrate the existence of highly specialized microbial communities in contaminated soils responsible for processing oxy-PAHs accumulated by primary degraders. Also, they underscore the key role that BVMO may play as a detoxification mechanism to mitigate the risk posed by oxy-PAH formation during bioremediation of PAH-contaminated soils.

Therapeutic exposure of diabetic blood using 532 nm-wavelength low level laser through electrical impedance spectroscopy (EIS).

Prior information of blood dielectric properties is important and a valuable tool to examine cell morphological structures, functions and mechanisms of their metabolic systems; however, this is under investigated. The study explores the technique of electrical impedance spectroscopy (EIS) measurements to reveal the proper biostimulation conditions for diabetic blood-low level laser therapy. Dielectric spectroscopy of the diabetic patient’s blood was studied using Impedance analyser (Agilent 4294A) within the range of frequency 40 Hz-30 MHz. Blood biostimulations were done using a 532 nm wavelength portable solid-state laser (diode-pumped). The accurate characterization of irradiated diabetic blood impedance and proper observations at power output of 50 mw within exposure duration of 10-15 minutes yield an efficient biostimulation and positively enable rejuvenation and proliferation of the deformed erythrocytes and stimulate healthy blood condition. The irradiated diabetic blood got rejuvenated and had their electrode polarization (α-dispersion region) stretched from frequency of 40 Hz until within a range of 11.7- 11.9 kHz similar to the healthy human blood. This formed appreciable positive effect of the properly stimulated diabetic blood with significant impedance within the cell structures, cytoplasm and plasma of the blood.

Isomer-selective biodegradation of high-molecular-weight azaarenes in PAH-contaminated environmental samples

Polycyclic aromatic nitrogen heterocycles, or azaarenes, normally co-occur with polycyclic aromatic hydrocarbons (PAHs) in contaminated soils. We recently reported that nontarget analysis using high resolution mass spectrometry of samples from four PAH-contaminated sites revealed a previously unrecognized diversity and abundance of azaarene isomers and their methylated derivatives. Here we evaluated their biodegradability by natural microbial communities from each site in aerobic microcosm incubations under biostimulated conditions. The removal of total quantifiable azaarenes ranged from 15 to 85%, and was related to the initial degree of weathering for each sample. While three-ring azaarenes were readily biodegradable, the five-ring congeners were the most recalcitrant. Microbial-mediated removal of four-ring congeners varied for different isomers, which might be attributed to the position of the nitrogen atom that also influences the physicochemical properties of azaarenes and possibly the susceptibility to transformation by relevant microbial enzymes. The presence of methyl groups also influenced azaarene biodegradability, which decreased with increasing degree of methylation. Several oxidation products of azaarenes were detected, including ketones and dioxygenated derivatives of three- and four-ring compounds. Our results indicate the susceptibility of some azaarenes to bioremediation, while suggesting the potential implications for risk from the persistence of less-biodegradable isomers and the formation of oxidized-azaarene derivatives.

Soil Microbial Communities Involved in Reductive Dissolution of Arsenic from Arsenate-Laden Minerals with Different Carbon Sources.

The natural microbial communities involved in arsenic (As) extraction under biostimulated conditions are still unclear. In this study, soil slurry was incubated with arsenate [As(V)]-laden Fe(III) or Al (hydr)oxides with lactate or acetate. After 40 d, dissolved As released from As(V)-laden Fe(III) accounted for 54% of the initial solid-phase As in lactate-amended slurries, while much less As was released from acetate-amended slurries. As was released more rapidly from As(V)-laden Al, but the total release was relatively low (45%). High-throughput Illumina sequencing of 16S rRNA genes revealed that dissimilatory metal(loid) reducers such as Desulfitobacterium became predominant in lactate-amended slurries. Moreover, anaerobic fermenters in the Sporomusaceae family were predominant. Interestingly, a Sporomusaceae bacterial strain isolated from the slurry was capable of releasing As from both As(V)-laden (hydr)oxides in the presence of lactate. The strain first released As as As(V) and subsequently reduced it to As(III) in the aqueous phase. These results suggest that lactate is a suitable carbon source for As extraction by natural microbial communities, and that both dissimilatory metal(loid) reducers and certain anaerobic fermenters play significant roles in As extraction. Microbial reductive dissolution of As may be expected to be a cost-effective restoration technique for As-contaminated soils.

Biostimulants application for bacterial metabolic activity promotion and sodium dodecyl sulfate degradation under copper stress

In this study, the metabolic activity (adenosine triphosphate, ATP; electron transfer system, ETS; and dehydrogenase activity, DHA) response of a sodium dodecyl sulfate (SDS) degrading bacterium Pseudomonas sp. SDS-N2 to copper stress conditions were investigated. Results showed that the ATP content, ETS activity, and DHA activity of strain SDS-N2 were significantly correlated with substrate removal efficiency and bacterial growth under copper stress conditions. Based on the metabolic response patterns of strain SDS-N2, biostimulants citric acid, proline as well as FeSO4 were used to promote the metabolic activity of strain SDS-N2 at 0.8 mg L−1 copper stress condition. Plackett–Burman design and analysis proved that citric acid and FeSO4 were significant factors for enhanced SDS removal; and the optimum biostimulation conditions (FeSO4 72 mg L−1 and citric acid 100 mg L−1) for SDS removal were obtained by using steepest ascent experiment and central composite design. Under the optimum biostimulation conditions, ATP, ETS, DHA activity as well as bacterial growth were 14.1, 45.5, 0.5 and 2.3-fold higher than that of the control (without FeSO4 and citric acid addition) after 12.5 h biodegradation, and the substrate removal efficiency was increase by 37.6%.

Assessment of crude oil bioremediation potential of seawater and sediments from the shore of Lebanon in laboratory microcosms

With the planned oil and gas exploration activities off the coast of Lebanon, the risk of shoreline contamination with crude oil spills has become a major concern. This study aimed at assessing the crude oil bioremediation potential of the chronically polluted Lebanese shores in light of the continuous discharge of nutrient-rich sewage into the Mediterranean Sea and the long-lasting absence of proper sewage treatment systems. It was anticipated that, with the high pollution levels of the coastline, background concentrations of nutrients would be sufficient to sustain high intrinsic biodegradation rates without human intervention. Biodegradation experiments were conducted using crude oil-spiked beach sediments and seawater under natural attenuation and biostimulation conditions. The experiments were conducted at 18 and 28 °C to account for seasonal variation in temperature, background nutrient levels, and microbial communities. The biodegradability of oil constituents – namely alkanes and polycyclic aromatic hydrocarbons (PAHs), was monitored over a 42-day period using gas chromatography–mass spectrometry (GC–MS). Under biostimulation conditions, significant enhancement in the overall biodegradation rates of alkanes and PAHs was observed in seawater at 18 and 28 °C, while little to no improvement was measured at both temperatures in sediments where background nutrient levels were sufficient to induce near maximum intrinsic biodegradation rates. Under both natural attenuation and biostimulation treatments, the increase in temperature increased the oil biodegradation rates in sediment and seawater microcosms. In both instances, the overall trend in the biodegradation of individual alkanes and PAHs suggested a typical decrease in biodegradation rates with the increase in carbon number/rings and alkyl groups.

Tracing the Biotransformation of Polycyclic Aromatic Hydrocarbons in Contaminated Soil Using Stable Isotope-Assisted Metabolomics.

Biotransformation of organic pollutants may result in the formation of oxidation products more toxic than the parent contaminants. However, to trace and identify those products, and the metabolic pathways involved in their formation, is still challenging within complex environmental samples. We applied stable isotope-assisted metabolomics (SIAM) to PAH-contaminated soil collected from a wood treatment facility. Soil samples were separately spiked with uniformly 13C-labeled fluoranthene, pyrene, or benzo[a]anthracene at a level below that of the native contaminant, and incubated for 1 or 2 weeks under aerobic biostimulated conditions. Combining high-resolution mass spectrometry and automated SIAM workflows, chemical structures of metabolites and metabolic pathways in the soil were proposed. Ring-cleavage products, including previously unreported intermediates such as C11H10O6 and C15H12O5, were detected originating from fluoranthene and benzo[a]anthracene, respectively. Sulfate conjugates of dihydroxy compounds were found as major metabolites of pyrene and benzo[a]anthracene, suggesting the potential role of fungi in their biotransformation in soils. A series of unknown N-containing metabolites were identified from pyrene, but their structural elucidation requires further investigation. Our results suggest that SIAM can be successfully applied to understand the fate of organic pollutants in environmental samples, opening lines of evidence for novel mechanisms of microbial transformation within such complex matrices.

Large‐Scale Experiments in Microbially Induced Calcite Precipitation (MICP): Reactive Transport Model Development and Prediction

AbstractDesign of in situ microbially induced calcite precipitation (MICP) strategies relies on a predictive capability. To date much of the mathematical modeling of MICP has focused on small‐scale experiments and/or one‐dimensional flow in porous media, and successful parameterizations of models in these settings may not pertain to larger scales or to nonuniform, transient flows. Our objective in this article is to report on modeling to test our ability to predict behavior of MICP under controlled conditions in a meter‐scale tank experiment with transient nonuniform transport in a natural soil, using independently determined parameters. Flow in the tank was controlled by three wells, via a complex cycle of injection/withdrawals followed by no‐flow intervals. Different injection solution recipes were used in sequence for transport characterization, biostimulation, cementation, and groundwater rinse phases of the 17 day experiment. Reaction kinetics were calibrated using separate column experiments designed with a similar sequence of phases. This allowed for a parsimonious modeling approach with zero fitting parameters for the tank experiment. These experiments and data were simulated using PHT3‐D, involving transient nonuniform flow, alternating low and high Damköhler reactive transport, and combined equilibrium and kinetically controlled biogeochemical reactions. The assumption that microbes mediating the reaction were exclusively sessile, and with constant activity, in conjunction with the foregoing treatment of the reaction network, provided for efficient and accurate modeling of the entire process leading to nonuniform calcite precipitation. This analysis suggests that under the biostimulation conditions applied here the assumption of steady state sessile biocatalyst suffices to describe the microbially mediated calcite precipitation.

In Situ Bioremediation of a Gasoline‐Contaminated Vadose Zone: Implications from Direct Observations

In Situ Bioremediation of a Gasoline‐Contaminated Vadose Zone: Implications from Direct Observations

Potential of wheat bran to promote indigenous microbial enhanced oil recovery

Microbial enhanced oil recovery (MEOR) is an emerging oil extraction technology that utilizes microorganisms to facilitate recovery of crude oil in depleted petroleum reservoirs. In the present study, effects of wheat bran utilization were investigated on stimulation of indigenous MEOR. Biostimulation conditions were optimized with the response surface methodology. The co-application of wheat bran with KNO3 and NH4H2PO4 significantly promoted indigenous MEOR (IMEOR) and exhibited sequential aerobic (O-), facultative (An-) and anaerobic (A0-) metabolic stages. The surface tension of fermented broth decreased by approximately 35%, and the crude oil was highly emulsified. Microbial community structure varied largely among and in different IMEOR metabolic stages. Pseudomonas sp., Citrobacter sp., and uncultured Burkholderia sp. dominated the O-, An- and early A0-stages. Bacillus sp., Achromobacter sp., Rhizobiales sp., Alcaligenes sp. and Clostridium sp. dominated the later A0-stage. This study illustrated occurrences of microbial community succession driven by wheat bran stimulation and its industrial potential.

Evaluation of Biostimulation and Bioaugmentation To Stimulate Hexahydro-1,3,5-trinitro-1,3,5,-triazine Degradation in an Aerobic Groundwater Aquifer.

Hexahydro-1,3,5-trinitro-1,3,5,-triazine (RDX) is a toxic and mobile groundwater contaminant common to military sites. This study compared in situ RDX degradation rates following bioaugmentation with Gordonia sp. strain KTR9 (henceforth KTR9) to rates under biostimulation conditions in an RDX-contaminated aquifer in Umatilla, OR. Bioaugmentation was achieved by injecting site groundwater (6000 L) amended with KTR9 cells (10(8) cells mL(-1)) and low carbon substrate concentrations (<1 mM fructose) into site wells. Biostimulation (no added cells) was performed by injecting groundwater amended with low (<1 mM fructose) or high (>15 mM fructose) carbon substrate concentrations in an effort to stimulate aerobic or anaerobic microbial activity, respectively. Single-well push-pull tests were conducted to measure RDX degradation rates for each treatment. Average rate coefficients were 1.2 day(-1) for bioaugmentation and 0.7 day(-1) for high carbon biostimulation; rate coefficients for low carbon biostimulation were not significantly different from zero (p values ≥0.060). Our results suggest that bioaugmentation with KTR9 is a feasible strategy for in situ biodegradation of RDX and, at this site, is capable of achieving RDX concentration reductions comparable to those obtained by high carbon biostimulation while requiring ~97% less fructose. Bioaugmentation has potential to minimize substrate quantities and associated costs, as well as secondary groundwater quality impacts associated with anaerobic biostimulation processes (e.g., hydrogen sulfide, methane production) during full-scale RDX remediation.

Laboratory studies of rice bran as a carbon source to stimulate indigenous microorganisms in oil reservoirs

There is a great interest in developing cost-efficient nutrients to stimulate microorganisms in indigenous microbial enhanced oil recovery (IMEOR) processes. In the present study, the potential of rice bran as a carbon source for promoting IMEOR was investigated on a laboratory scale. The co-applications of rice bran, K2HPO4 and urea under optimized bio-stimulation conditions significantly increased the production of gases, acids and emulsifiers. The structure and diversity of microbial community greatly changed during the IMEOR process, in which Clostridium sp., Acidobacteria sp., Bacillus sp., and Pseudomonas sp. were dominant. Pressurization, acidification and emulsification due to microbial activities and interactions markedly improved the IMEOR processes. This study indicated that rice bran is a potential carbon source for IMEOR.

Combination of aquifer thermal energy storage and enhanced bioremediation: resilience of reductive dechlorination to redox changes.

To meet the demand for sustainable energy, aquifer thermal energy storage (ATES) is widely used in the subsurface in urban areas. However, contamination of groundwater, especially with chlorinated volatile organic compounds (CVOCs), is often being encountered. This is commonly seen as an impediment to ATES implementation, although more recently, combining ATES and enhanced bioremediation of CVOCs has been proposed. Issues to be addressed are the high water flow velocities and potential periodic redox fluctuation that accompany ATES. A column study was performed, at a high water flow velocity of 2 m/h, simulating possible changes in subsurface redox conditions due to ATES operation by serial additions of lactate and nitrate. The impacts of redox changes on reductive dechlorination as well as the microbial response of Dehalococcoides (DHC) were evaluated. The results showed that, upon lactate addition, reductive dechlorination proceeded well and complete dechlorination from cis-DCE to ethene was achieved. Upon subsequent nitrate addition, reductive dechlorination immediately ceased. Disruption of microorganisms’ retention was also immediate and possibly detached DHC which preferred attaching to the soil matrix under biostimulation conditions. Initially, recovery of dechlorination was possible but required bioaugmentation and nutrient amendment in addition to lactate dosing. Repeated interruption of dechlorination and DHC activity by nitrate dosing appeared to be less easily reversible requiring more efforts for regenerating dechlorination. Overall, our results indicate that the microbial resilience of DHC in biosimulated ATES conditions is sensitive to redox fluctuations. Hence, combining ATES with bioremediation requires dedicated operation and monitoring on the aquifer geochemical conditions.Electronic supplementary materialThe online version of this article (doi:10.1007/s00253-015-7241-6) contains supplementary material, which is available to authorized users.

Biostimulation of Indigenous Microbial Communities for Anaerobic Transformation of Pentachlorophenol in Paddy Soils of Southern China

This study explored biostimulation mechanisms with an electron donor and a shuttle for accelerating pentachlorophenol (PCP) transformation in iron-rich soils. The results indicated that indigenous microbial communities are important for PCP transformation in soils. Biostimulation of indigenous microbial communities by the addition of lactate and anthraquinone-2,6-disulfonate (AQDS) led to the enhanced rates of PCP dechlorination by the dechlorinating- and iron-reducing bacteria in soils. The electrochemical studies using cyclic voltammograms and microbial current measurements confirmed the high reduction potential and the large amount of electrons generated under biostimulation conditions, which were responsible for the higher rates of PCP transformation. After biostimulation treatments by the additions of lactate and/or AQDS during PCP dechlorination processes, microbial community analysis by the terminal restriction fragment length polymorphism (T-RFLP) method showed the abundance terminal restricted fragments (T-RFs), an indicator of bacterial abundance, which represents the dechlorinating- and iron-reducing bacteria, suggesting their critical roles in PCP dechlorination in soils.

Biodegradation of ethylene dibromide (1,2-dibromoethane [EDB]) in microcosms simulating in situ and biostimulated conditions

Although 1,2-dibromoethane (EDB) is a common groundwater contaminant, there is the lack of knowledge surrounding EDB biodegradation, especially under aerobic conditions. We have performed an extensive microcosm study to investigate the biodegradation of EDB under simulated in situ and biostimulated conditions. The materials for soil microcosms were collected from an EDB-contaminated aquifer at the Massachusetts Military Reservation in Cape Cod, MA. This EDB plume has persisted for nearly 40 years in both aerobic and anaerobic EDB zones of the aquifer. Microcosms were constructed under environmentally relevant conditions (field EDB and DO concentrations; incubated at 12°C). The results showed that natural attenuation occurred under anaerobic conditions but not under aerobic conditions, explaining why aerobic EDB contamination is so persistent. EDB degradation rates were greater under biostimulated conditions for both the aerobic and anaerobic microcosms. Particularly for aerobic biostimulation, methane-amended microcosms degraded EDB, on average, at a first order rate eight times faster than unamended microcosms. The best performing replicate achieved an EDB degradation rate of 7.0 yr−1 (half-life (t1/2)=0.10 yr). Residual methane concentrations and the emergence of methanotrophic bacteria, measured by culture independent bacterial analysis, provided strong indications that EDB degradation in aerobic methane-amended microcosms occurred via cometabolic degradation. These results indicate the potential for enhanced natural attenuation of EDB and that methane could be considered co-substrate for EDB bioremediation for the EDB-contaminated groundwater in aerobic zone.

Characterization of the bottom sediments contaminated with polychlorinated biphenyls: Evaluation of ecotoxicity and biodegradability

At the locality of the former producer of PCBs Chemko Strážske in East Slovakia, a large amount of PCBs (the commercial mixture DELOR 103, an equivalent of AROCLOR 1242) is still persisting in sediments and negatively influences health of the population. The objective of this work was to provide a study of ecotoxicity and genotoxicity of PCBs in contaminated sediments. Toxicity of the PCB-contaminated sediments sampled from Zemplínska šírava and Strážsky canal (surroundings of the former producer of PCBs) was determined applying a standard aquatic plant toxicity test using Lemna minor. The endpoints for the test were frond numbers and frond areas. The sediment sampled from Zemplínska šírava was more toxic to L. minor than the one sampled from Strážsky canal. The results on genotoxicity showed that both sediments were not mutagenic toward the standard strains of the Ames test, Salmonella typhimurium TA98 and TA100. This work deals also with biodegradation of PCBs in two samples of the above mentioned contaminated sediments: a) in the natural sediments by autochthonous microbial consortium and b) in the bioaugmented sediments inoculated by allochthonous bacterial strains, two bacterial isolates from long-term PCB-contaminated soil Pseudomonas stutzeri and Alcaligenes xylosoxidans. Both approaches were applied under the biostimulation conditions, with addition of glucose or biphenyl as co-substrates, as well. The highest PCB degradation was observed in the bioaugmented sediment inoculated with bacterial strain P. stutzeri. Addition of biphenyl, as the co-substrate and the inducer, positively affected degradation of PCBs. The bphA1 gene, encoding enzyme biphenyldioxygenase, responsible for the start of PCB degradation, was identified in genome of P. stutzeri, a potential PCB-degrader isolated from long-term PCB-contaminated soil, but not in genome of A. xylosoxidans.

Microbial community dynamics in uranium contaminated subsurface sediments under biostimulated conditions with high nitrate and nickel pressure

The subsurface at the Oak Ridge Field Research Center represents an extreme and diverse geochemical environment that places different stresses on the endogenous microbial communities, including low pH, elevated nitrate concentrations, and the occurrence of heavy metals and radionuclides, including hexavalent uranium [U(VI)]. The in situ immobilization of U(VI) in the aquifer can be achieved through microbial reduction to relatively insoluble U(IV). However, a high redox potential due to the presence of nitrate and the toxicity of heavy metals will impede this process. Our aim is to test biostimulation of the endogenous microbial communities to improve nitrate reduction and subsequent U(VI) reduction under conditions of elevated heavy metals. Column experiments were used to test the possibility of using biostimulation via the addition of ethanol as a carbon source to improve nitrate reduction in the presence of elevated aqueous nickel. We subsequently analyzed the composition of the microbial communities that became established and their potential for U(VI) reduction and its in situ immobilization. Phylogenetic analysis revealed that the microbial population changed from heavy metal sensitive members of the actinobacteria, alpha- and gamma-proteobacteria to a community dominated by heavy metal resistant (nickel, cadmium, zinc, and cobalt resistant), nitrate reducing beta- and gamma-proteobacteria, and sulfate reducing Clostridiaceae. Coincidentally, synchrotron X-ray absorption spectroscopy analyses indicated that the resulting redox conditions favored U(VI) reduction transformation to insoluble U(IV) species associated with soil minerals and biomass. This study shows that the necessary genetic information to adapt to the implemented nickel stress resides in the endogenous microbial population present at the Oak Ridge FRC site, which changed from a community generally found under oligotrophic conditions to a community able to withstand the stress imposed by heavy metals, while efficiently reducing nitrate as electron donor. Once nitrate was reduced efficient reduction and in situ immobilization of uranium was observed. This study provides evidence that stimulating the metabolism of the endogenous bacterial population at the Oak Ridge FRC site by adding ethanol, a suitable carbon source, results in efficient nitrate reduction under conditions of elevated nickel, and a decrease of the redox potential such that sulfate and iron reducing bacteria are able to thrive and create conditions favorable for the reduction and in situ immobilization of uranium. Since we have found that the remediation potential resides within the endogenous microbial community, we believe it will be feasible to conduct field tests. Biostimulation of endogenous bacteria provides an efficient tool for the successful in situ remediation of mixed-waste sites, particularly those co-contaminated with heavy metals, nitrate and radionuclides, as found in the United States and other countries as environmental legacies of the nuclear age.

Biofilm growth kinetics on hydrocarbon in a porous medium under biostimulation conditions

AbstractThe formation of biofilm on hydrocarbon in a porous medium under biostimulation conditions was investigated in order to a better understanding of microbial development during bioremediation of hydrocarbon‐contaminated soils. Pseudomonas alkanolytica ATCC 21034 was studied as it grows embedded in a biofilm on dodecane surface. First, an experimental protocol for determination of biofilm growth kinetics on hydrocarbon in a porous medium was proposed. Six identical columns packed with sand were run simultaneously. The growth kinetics was obtained by measuring the biofilm dry weight of each column at different times. The reproducibility of the experimental protocol was validated using statistical tests. Then biofilm growth kinetics was established in the porous medium for two specific surfaces of dodecane (large or small droplets). A linear growth was observed due to transport limitation of both dodecane and dissolved oxygen in the biofilm. So the degradation of dodecane was uncompleted. Moreover, difficulty to bring dissolved oxygen in the deeper part of the porous medium was pointed out. All these phenomena could explain some failures encountered during biostimulation processes. © 2007 American Institute of Chemical Engineers Environ Prog, 2007

石油汚染土壌におけるアルカン分解菌の同定

In this study, we conducted a weathered crude oil (w. oil)-degrading experiment with five different soil samples in order to investigate the biodegradation of alkanes. Two soil samples, soil one (I) and five (V), showed the excellent ability to biodegradade both normal alkanes (C10-C36 alkanes) and branched alkanes (pristane, phytane). We isolated several alkane-degrading bacteria from these soil samples. Isolated bacteria were classified into 3 groups based on their 16S rDNA sequences. The changes in population of these alkane-degrading bacteria were analyzed quantitatively by polymerase chain reaction (PCR) using specific primer sets. As a result, the concentration of group 1 bacteria (Rhodococcus spp.) that were able to degrade both normal and branched alkanes markedly increased due to enrichment with w.oil in soil one (I) and five (V). These results indicated that the group 1 bacteria that belong to the Rhodococcus genus played an important role in degrading alkanes in petroleum-contaminated soil under biostimulated conditions.

Bioluminescence Activity and Degradation Kinetics of Genetically Engineered Strain P. putida mt-2 RB 1401 in Soil System with m-Xylene

Strain RB1401 contains a plasmid with a fusion between the upper pathway promoter of the TOL plasmid xyl genes and a lux gene cassette. Bioluminescence activity appeared in the early period of biodegradation of m-xylene, and was observed at all tested conditions of temperature (15, 27, 35°C) and pH (5, 7, 9), showing in the range of 300 ∼ 6100 RLU. Biokinetics of strain RB1401 in the presence of m-xylene were as follows: Y 0.56 mg/mg, μmax 2.67 day-1, and Ks 19.25 mg/L. A ratio of C:N(KNO3):P(KH2PO4) = 100:8:1.5 and 0.01% (w/v) of H2O2 were chosen for optimum biostimulation conditions of RB1401 with m-xylene in soil system. These results demonstrate the capability of such a genetically engineered bacteria for the development of biomonitoring and bioremediation in soil environments contaminated with specific volatile organic compounds.