Literature overview: Microbial metabolism of high molecular weight polycyclic aromatic hydrocarbons

High molecular weight polycyclic aromatic hydrocarbons (HMW PAHs) increase in hydrophobicity with increases in their molecular weight and ring angularity. Microbial strategies to deal with PAH hydrophobicity include biofilm formation, enzyme induction, and biosurfactants, the effect of which is variable on PAH metabolism depending on the surfactant type and concentration, substrate, and microbial strain(s). Aerobic HMW PAH metabolism proceeds via mineralization, partial degradation, and cometabolic transformations. Generally, bacteria and nonlignolytic fungi metabolize PAHs via initial PAH ring oxidation by dioxygenases to form cis‐dihydrodiols, which are transformed to catechol compounds by dehydrogenases and other mono‐ and dioxygenases to substituted catechol and noncatechol compounds, all ortho‐ or metacleaved and further oxidized to simpler compounds. However, lignolytic fungi form quinones and acids to CO₂. This review discusses the pathways for HMW PAH microbial metabolism. © 2008 Wiley Periodicals, Inc.     

Characterizing In Situ Methane‐Enhanced Biostimulation Potential for 1,4‐Dioxane Biodegradation in Groundwater

This study characterizes the 1,4‐dioxane biodegradation potential for an in situ methane‐enhanced biostimulation field pilot study conducted at Air Force Plant 44, located south of the Tucson International Airport in Arizona. In this study, the use of methane as the primary substrate in aerobic cometabolic biodegradation of 1,4‐dioxane is evaluated using environmental molecular diagnostic tools. The findings are compared to an adjacent pilot study, wherein methane was generated via enhanced reductive dechlorination and where methane monooxygenase and methane‐oxidizing bacteria were also found to be abundant. This article also presents the use of ¹³C and ²H isotopic ratio enrichment, a more recent tool, to support the understanding of 1,4‐dioxane biodegradation in situ. This study is the first of its kind, although alkane gas‐enhanced biodegradation of 1,4‐dioxane has been evaluated extensively in microcosm studies and propane‐enhanced biodegradation of 1,4‐dioxane has been previously studied in the field. ©2016 Wiley Periodicals, Inc.     

Production, Purification and Activity of an Extracellular Depolymerase from Aspergillus fumigatus

A strain of Aspergillus fumigatus, which was observed to rapidly degrade poly-3-hydroxybutyrate (PHB) in a leaf compost, was found to secrete an extracellular hydrolase when grown on PHB as the sole carbon source. Isolation and characterization of the PHB hydrolase (depolymerase) from this fungus revealed that the enzyme had a molecular weight of 57 kDa, an isoelectric point of 7.2, and a PHB hydrolysis activity maxima which occurred at 70°C and pH 8.0. Affinity labeling experiments suggested that this fungal hydrolase is a type of serine esterase. The cyclic trimers of 3-hydroxybutyrate were found to reversibly inhibit the enzymes.     

Abiotic/biotic degradation and mineralization of N‐nitrosodimethylamine in aquifer sediments

The N‐nitrosodimethylamine (NDMA) degradation rate and mineralization rate were measured in two aquifer sediments that received treatments to create oxic, reducing, and sequential reducing/oxic environments. Chemically reduced sediments rapidly abiotically degraded NDMA to nontoxic dimethylamine to parts per trillion levels, then degraded to further products. NDMA was partially mineralized in reduced sediments (6 to 28 percent) at a slow rate (half‐life 3,460 h) by an unknown abiotic/biotic pathway. In contrast, NDMA was mineralized more rapidly (half‐life 342 h) and to a greater extent (30 to 81 percent) in oxic sediments with propane addition, likely by a propane monooxygenase pathway. NDMA mineralization in sequential reduced sediment followed by oxic sediment treatment did result in slightly more rapid mineralization and a greater mineralization extent relative to reduced systems. These increases were minor, so aerobic NDMA mineralization with oxygen and propane addition was the most viable in situ NDMA mineralization strategy. © 2008 Wiley Periodicals, Inc.     

Biodegradation of Aliphatic and Aromatic Hydrocarbons: Specificity among Bacteria Isolated from Refinery Waste Sludge

Indigenous microorganisms, enriched and isolated from refinery waste sludge, were observed to possess a broad range of metabolic activities for mixtures of several classes of substrates of petroleum hydrocarbons, such as monoaromatic and polycyclic aromatic hydrocarbons (PAHs) and n- and branched alkanes. Three of the best-growing bacterial isolates selectively enriched with these compounds were identified by 16S rDNA sequencing as belonging to the genera Enterobacter and Ochrobactrum. Two of them, Enterobacter sp. strain EK3.1 and Ochrobactrum sp. strain EK6 utilise a hydrocarbon mixture of the branched alkane 2,6,10,14-tetramethylpentadecane and the PAHs acenaphthylene and acenaphthene. Enterobacter sp. strain EK4 can grow with a mixture of 2,6,10,14-tetramethylpentadecane, toluene, acenaphthylene and acenaphthene as carbon sources. Nucleic acid fingerprint analysis, by terminal restriction fragment length polymorphism (T-RFLP) of the PCR-amplified 16S rRNA genes, of the autochthonous bacterial community in contaminated soil samples showed complex and different community structures under different treatments of refinery waste sludge in landfarm areas. The characteristic peaks of the T-RFLP profiles of the individual, isolated degrading bacteria Enterobacter spp. and Ochrobactrum sp. were detected in the T-RFLP fingerprint of the bacterial community of the four months old treated landfarm soil, suggesting the enrichment of bacteria belonging to the same operational taxonomic units, as well as their importance in degrading activity.     

Chemical oxidation using stabilized hydrogen peroxide in high temperature,saline groundwater impacted with hydrocarbons and MTBE

In situ chemical oxidation (ISCO) of petroleum hydrocarbons (PHCs) within groundwater is considered a proven approach to addressing PHC‐impacted groundwater in nonsaline environments. One of the most common oxidants used for oxidation of PHCs in groundwater is hydrogen peroxide (H₂O₂). Due to its highly reactive nature, H₂O₂ is often stabilized to aid in increasing its reactivity lifespan. Limited research and application of ISCO has been completed in warm, saline groundwater environments. Furthermore, even fewer studies have been completed in these environments for ISCO using stabilized H₂O₂. In this research, stabilized H₂O₂ was examined to determine its effectiveness in the treatment of PHCs and the additive methyl tert‐butyl ether (MTBE). Three stabilizers (citrate, phytate, silica [SiO₂]) were tested to determine if the stabilizers could enhance and extend the treatment life of H₂O₂ within saline groundwater. To determine the effect of salinity on the three stabilizers, groundwater and aquifer samples were collected from two saline locations that had different salinity (total dissolved solids of about 7,000 mg/L and 18,000 mg/L). Specific target chemicals for treatment were water soluble, mobile components of gasoline including benzene, toluene, ethylbenzene, xylenes, (BTEX) and MTBE. Previous studies using unactivated persulfate indicated that the PHCs within the groundwater could be oxidized, however, only limited oxidation of the MTBE could be affected. The results of the laboratory tests indicated that greater than 95 percent of the target hydrocarbons were removed within 7 days of treatment. Microcosms with citrate‐stabilized H₂O₂ demonstrated a significantly faster and greater decline with most hydrocarbon concentrations reaching < 5 μg/L. The exceptions were ethylbenzene and m‐xylene, which were slightly decreased to about 30 and 20 μg/L, respectively. Initial mean concentrations of the BTEX compounds within the citrate‐stabilized microcosms were 10,554 μg/L, 9,318 μg/L, 6,859 μg/L, and 14,435 μg/L, respectively. The silicate‐stabilized H₂O₂ microcosms showed no significant benefit over the unstabilized control microcosms. The better performance of citrate‐stabilized microcosms was confirmed by increasing δ13C values of remaining hydrocarbons. MTBE declined from > 400 mg/L to < 100 mg/L in all microcosms, again with the best removal (> 90 percent) being measured in the citrate‐stabilized microcosms. Unfortunately, H₂O₂ oxidation in the microcosms also resulted in production of up to 40 mg/L TBA or approximately 10 percent of the MTBE oxidized.     

A chemox treatment for urea‐ and ammonium‐contaminated groundwater

In a pilot project performed at a fertilizer manufacturing facility, a one‐step chemical oxidation technique successfully treated urea‐ and ammonium‐contaminated groundwater. The oxidation reaction occurred in an 1,100‐gallon batch reactor. The contaminated inflow was buffered by the metered addition of sodium bicarbonate solution and subsequently treated with sodium hypochlorite in an 8:1 weight ratio of Cl₂:N. In an instantaneous reaction, the urea and ammo‐nium‐N were completely oxidized to nitrogen gas that was vented to the atmosphere during mixing. The pH of the reactor discharge was ?6.5. Sodium sulfite was used to reduce residual hypochlorite in the reactor effluent to chloride to provide process water with characteristics suitable for discharge. Oxidation rates were similar with different strengths of hypochlorite; however, a 5 to 6 percent sodium hypochlorite (as Cl₂) solution was the most stable. © 2005 Wiley Periodicals, Inc.     

Comparative degradation between heavy and light crude oil mediated by nitrogen‐induced cell‐surface hydrophobicity

A bacterial strain UKMP‐10M2 isolated from a Malaysian petroleum refinery was able to degrade 84% of heavy Khafji sour crude and 68% of light Tapis sweet crude within seven days. Analysis of gas chromatography‐flame ionization detector chromatograms show the strain UKMP‐10M2 degraded up to 90% pristane and 50% phytane in heavy crude, but significantly lower pristane (50%) and phytane (30%) were degraded from the light crude. A mixture of aliphatic hexadecane and three‐ring phenanthrene better supported the growth of isolate UKMP‐10M2 compared to using phenanthrene alone, suggesting cometabolism influenced how crude oil with different individual hydrocarbon contents affected the degradation. Peptone as the source of nitrogen increases the emulsifying index in UKMP‐10M2 exposed to heavy Khafji sour crude 20% higher than in light Tapis sweet crude. However, BATH assay showed the same nitrogen source increases bacterial cell surface hydrophobicity of UKMP‐10M2 up to 14% higher in light Tapis crude oil compared to heavy Khafji. This study suggest the nitrogen source plays a decisive role in elevating UKMP‐10M2 bacterial cells hydrophobicity, and in correlation with types of crude oil. Phylogenetic tree analysis based on 16S rDNA sequence results identified the strain to be Rhodococcus ruber.     

Study of the VOC emissions from a municipal solid waste storage pilot-scale cell: comparison with biogases from municipal waste landfill site

The emission of volatile organic compounds (VOCs) from municipal solid waste stored in a pilot-scale cell containing 6.4 tonnes of waste (storage facility which is left open during the first period (40 days) and then closed with recirculation of leachates during a second period (100 days)) was followed by dynamic sampling on activated carbon and analysed by GC–MS after solvent extraction. This was done in order to know the VOC emissions before the installation of a methanogenesis process for the entire waste mass. The results, expressed in reference to toluene, were exploited during the whole study on all the analyzable VOCs: alcohols, ketones and esters, alkanes, benzenic and cyclic compounds, chlorinated compounds, terpene, and organic sulphides.The results of this study on the pilot-scale cell are then compared with those concerning three biogases from a municipal waste landfill: biogas (1) coming from waste cells being filled or recently closed, biogas (2) from all the waste storage cells on site, and biogas (3) which is a residual gas from old storage cells without aspiration of the gas. The analysis of the results obtained revealed: (i) a high emission of VOCs, principally alcohols, ketones and esters during the acidogenesis; (ii) a decrease in the alkane content and an increase in the terpene content were observed in the VOCs emitted during the production of methane; (iii) the production of heavier alkanes and an increase in the average number of carbon atoms per molecule of alkane with the progression of the stabilisation/maturation process were also observed.Previous studies have concentrated almost on the analysis of biogases from landfills. Our research aimed at gaining a more complete understanding of the decomposition/degradation of municipal solid waste by measuring the VOCs emitted from the very start of the landfill process i.e. during the acidogenesis and acetogenesis phases.     

Integrated approach to PCE‐impacted site characterization,site management,and enhanced bioremediation

Tetrachloroethene (PCE)‐ and trichloroethene (TCE)‐impacted sites pose significant challenges even when site characterization activities indicate that biodegradation has occurred naturally. Although site‐specific, regulatory, and economic factors play roles in the remedy‐selection process, the application of molecular biological tools to the bioremediation field has streamlined the assessment of remedial alternatives and allowed for detailed evaluation of the chosen remedial technology. The case study described here was performed at a PCE‐impacted site at which reductive dechlorination of PCE and TCE had led to accumulation of cis‐dichlorethene (cis‐DCE) with concentrations ranging from approximately 10 to 100 mg/L. Bio‐Trap® samplers and quantitative polymerase chain reaction (qPCR) enumeration of Dehalococcoides spp. were used to evaluate three remedial options: monitored natural attenuation, biostimulation with HRC®, and biostimulation with HRC‐S®. Dehalococcoides populations in HRC‐S‐amended Bio‐Traps deployed in impacted wells were on the order of 10³ to 10⁴ cells/bead but were below detection limits in most unamended and HRC‐amended Bio‐Traps. Thus the in situ Bio‐Trap study identified biostimulation with HRC‐S as the recommended approach, which was further evaluated with a pilot study. After the pilot HRC‐S injection, Dehalococcoides populations increased to 10⁶ to 10⁷ cells/bead, and concentrations of cis‐DCE and vinyl chloride decreased with concurrent ethene production. Based on these results, a full‐scale HRC‐S injection was designed and implemented at the site. As with the pilot study, full‐scale HRC‐S injection promoted growth of Dehalococcoides spp. and stimulated reductive dechlorination of the daughter products cis‐DCE and vinyl chloride. © 2008 Wiley Periodicals, Inc.     

Synthesis of Poly(β-hydroxybutyrate) in Simulated Microgravity: An Investigation of Aeration Profiles in Shake Flask and Bioreactor

The microbial strain Azotobacter vinelandii UWD was grown under conditions of simulated microgravity in the National Aeronautics and Space Administration (NASA) Bioreactor. Bacterial growth in simulated microgravity differed significantly from that observed in conventional shake flask experiments: Cells tended to grow in a cluster-like pattern and polymer production started immediately after exposing them to conditions of simulated microgravity, and no lag time was observed. It was imperative to differentiate between the effects derived from microgravity and those imposed by the altered oxygen supply in the bioreactor. Aeration conditions were studied in both reactor types and a gas supply profile was developed for the bioreactor. This supply profile allowed for similar amounts of dissolved oxygen in the bioreactor and the shake flask in the initial stage of the fermentation and, therefore, for an evaluation of the effects of microgravity on biopolyester-producing bacteria. Since the optical density that is conventionally used as a measure for the cell growth could not be used due to the cluster-like growth pattern of the cells, it was determined that bacterial growth behavior in the bioreactor can be monitored through glucose or oxygen consumption.     

Stable Isotope Probing to Confirm Field‐Scale Co‐Metabolic Biodegradation of 1,4‐Dioxane

1,4‐Dioxane, a common co‐contaminant with chlorinated solvents, is present in groundwater at Site 24 at Vandenberg Air Force Base in California. Historical use of chlorinated solvents resulted in concentrations of 1,4‐dioxane in groundwater up to approximately 2,000 μg/L. Starting in 2013, an in situ propane biosparge system operation demonstrated reductions in 1,4‐dioxane concentrations in groundwater. The work detailed herein extends the efforts of the first field demonstration to a second phase and confirms the biodegradation mechanism via use of stable isotope probing (SIP). After two months of operation, 1,4‐dioxane concentrations decreased approximately 45 to 83 percent at monitoring locations in the test area. The results of the SIP confirmed ¹³C‐enriched 1,4‐dioxane was transformed into dissolved inorganic carbon (suggesting mineralization to carbon dioxide) and incorporated into microbial biomass (likely attributed to metabolic uptake of biotransformation intermediates or of carbon dioxide).  ©2016 Wiley Periodicals, Inc.     

Speciation and quantitation of degradation products of silicones (Silane/Siloxane Diols) by gas chromatography—mass spectrometry and stability of dimethylsilanediol

Gas chromatography-mass spectrometry (GC-MS) methodology was developed to speciate and quantitate several degradation products of polydimethylsiloxane (PDMS) in soil. We have demonstrated that the major degradation product,viz., dimethylsilanediol, can be readily analyzed by GC-MS without derivatization as commonly practiced in analyzing such materials. A mixture of linear siloxane diols (n = 1–5, wheren is the number of Me₂SiO units), and cyclic dimethylsiloxanes (n = 4–6) was resolved by GC-MS. We also found that peak identity of various diols required that GC-MS is done in the chemical ionization (CI) mode, since the electron impact (EI) ionization mode produced similar mass fragmentation patterns for diols and cyclics containing the same number of silicon atoms. For siloxane diols, detection limits ranged from 100 pg (forn = 1) to 1 ng (for n = 5). For cyclics, the detection limit was about 1 pg. Dimethylsilanediol, known to be unstable even in the solid state, was shown by NMR techniques to be stable in aqueous solution at <0.1% concentration. A 100-ppm solution was stable for over a year. Purity check for dimethylsilanediol is best carried out by Si-29 solid-state NMR technique.     

Purification and biochemical characterization of recombinant alcohol dehydrogenase from the psychrophilic bacterium <Emphasis Type="Italic">Pseudomonas</Emphasis><Emphasis Type="Italic">frederiksbergensis</Emphasis>

A gram-negative psychrophilic bacterium, with potential for biodegradation of long-chain n-alkanes was isolated from ice samples collected in Spitzbergen, Denmark. On the basis of results of biochemical and morphological tests and sequence analysis of 16S rRNA, the strain was identified as Pseudomonas frederiksbergensis. In this work, a short-chain NAD⁺-dependent alcohol dehydrogenase (alcDH) (Accession number: AAR13804) from the P. frederiksbergensis was cloned and transformed in E. coli BL21 (3DE) competent cells. The alcDH activity was highest in the crude extract of cells induced with 1.0 mM IPTG. The recombinant alcDH enzyme was purified to 93.4% homogeneity using three consecutive purification steps including ammonium sulphate, Q-Sepharose Fast Flow column and gel filtration chromatography employing Superdex 200 10/30 HR column. Enzyme enrichment and yield levels of 31.4 folds and 25.5%, respectively, were achieved. While the subunit molecular mass of the enzyme was determined on SDS-PAGE to be ~38 kDa, the aggregated native form of the enzyme had a molecular mass of ~238 kDa by gel filtration analysis. Reaction conditions optima for the recombinant alcDH were determined with propan-1-ol as the substrate. While the optimum pH was 9, the optimum temperature was 35 °C. The alcDH enzyme exhibited moderate thermal stability with half-lives of 150 min at 55 °C, 27 min at 65 °C and 8 min at 75 °C. Results for kinetic parameters indicated that the apparent K _m value for alcDH with propan-1-ol as the substrate was found to be 1.42 mM and the V _max value was 0.63 mmol mg⁻¹ min⁻¹. Experimental evidence revealed that the recombinant alcDH exhibited a wide range of substrate specificity, with higher levels of specific activity for aliphatic alcohols as compared to secondary alcohols. Taken together, the present study highlights the potential of alcDH as a member of cold-adapted enzymes in several key biotechnological applications including environmental bioremediation and biotransformations. It is envisaged that, with the ongoing screening of microorganisms and metagenomes, directed evolution approaches and the subsequent overexpression of recombinant proteins, more enzymes will be found that are suitable for bioremediation purposes.     

Remedial options for chlorinated volatile organics in a partially anaerobic aquifer

A laboratory study was conducted for the selection of appropriate remedial technologies for a partially anaerobic aquifer contaminated with chlorinated volatile organics (VOCs). Evaluation of in situ bioremediation demonstrated that the addition of electron donors to anaerobic microcosms enhanced biological reductive dechlorination of tetrachloroethene (PCE), trichloroethene (TCE), and 1,1,1‐trichloroethane (1,1,1‐TCA) with half‐lives of 20, 22, and 41 days, respectively. Nearly complete reductions of PCE, TCE, 1,1,1‐TCA, and the derivative cis‐dichloroethene were accompanied by a corresponding increase in chloride concentrations. Accumulation of vinyl chloride, ethene, and ethane was not observed; however, elevated levels of ¹⁴CO₂ (from ¹⁴C‐TCE spiked) were recovered, indicating the occurrence of anaerobic oxidation. In contrast, very little degradation of 1,2‐dichloropropane (1,2‐DCP) and 1,1‐dichlorethane (1,1‐DCA) was observed in the anaerobic microcosms, but nutrient addition enhanced their degradation in the aerobic biotic microcosms. The aerobic degradation half‐lives for 1,2‐DCP and 1,1‐DCA were 63 and 56 days, respectively. Evaluation of in situ chemical oxidation (ISCO) demonstrated that chelate‐modified Fenton's reagent was effective in degrading aqueous‐phase PCE, TCE, 1,1,1‐TCA, 1,2‐DCP, etc.; however, this approach had minimal effects on solid‐phase contaminants. The observed oxidant demand was 16 g‐H₂O₂/L‐groundwater. The oxidation reaction rates were not highly sensitive to the molar ratio of H₂O₂:Fe²⁺:citrate. A ratio of 60:1:1 resulted in slightly faster removal of chemicals of concern (COCs) than those of 12:1:1 and 300:1:1. This treatment resulted in increases in dissolved metals (Ca, Cr, Mg, K, and Mn) and a minor increase of vinyl chloride. Treatment with zero‐valent iron (ZVI) resulted in complete dechlorination of PCE, and TCE to ethene and ethane. ZVI treatment reduced 1,1,1‐TCA only to 1,1‐DCA and chloroethane (CA) but had little effect on reducing the levels of 1,2‐DCP, 1,1‐DCA, and CA. The longevity test showed that one gram of 325‐mesh iron powder was exhausted in reaction with > 22 mL of groundwater. The short life of ZVI may be a barrier to implementation. The ZVI surface reaction rates (k_sa) were 1.2 × 10^?2 Lm^?2h^?1, 2 × 10^?3 Lm^?2h^?1, and 1.2 × 10^?3 Lm^?2h^?1 for 1,1,1‐TCA, TCE, and PCE, respectively. Based upon the results of this study, in situ bioremediation appeared to be more suitable than ISCO and ZVI for effectively treating the groundwater contamination at the site. © 2004 Wiley Periodicals, Inc.     

Polyethylene conversion by partial oxidation in supercritical water

This paper gives the results of partial oxidation experiments of polyethylene (PE) in supercritical water (SCW). The experiments were carried out at a reaction temperature of 693K and a reaction time of 30 min using 6 cm³ of a batch-type reactor. The loaded sample weight was 0.3 g and there was 2.52 g water (0.42 g/cm³). The ratio of oxygen atoms to carbon atoms was 0.3. The results show a significant CO formation in O₂–SCW, and the 1-alkene/n-alkane ratio in partial oxidation was higher than that in SCW pyrolysis. These results suggest the possibility of the hydrogenation of hydrocarbon through partial oxidation followed by a water–gas shift reaction. Received: July 19, 2000 / Accepted: September 28, 2000     

Optimizing remediation strategies for a former dry‐cleaner site

Residual tetrachloroethene (PCE) contamination at the former Springvilla Dry Cleaners site in Springfield, Oregon, posed a potential risk through the vapor intrusion, direct contact, and off‐site beneficial groundwater uses. The Oregon Department of Environmental Quality utilized the State Dry Cleaner Program funds to help mitigate the risks posed by residual contamination. After delineation activities were complete, the source‐area soils were excavated and treated on‐site with ex situ vapor extraction to reduce disposal costs. Residual source‐area contamination was then chemically oxidized using sodium permanganate. Dissolved‐phase contamination was subsequently addressed with in situ enhanced reductive dechlorination (ERD). ERD achieved treatment goals across more than 4 million gallons of aquifer impacted with PCE concentrations up to 7,800 micrograms per liter prior to remedial activities. The ERD remedy introduced electron donors and nutrient amendments through groundwater recirculation and slug injection across two aquifers over the course of 24 months. Adaptive and mass‐targeted strategies reduced total remedy costs to approximately $18 per ton within the treatment areas. © 2010 Wiley Periodicals, Inc.     

In‐well sediment incubators to evaluate microbial community stability and dynamics following bioimmobilization of uranium

An in‐well sediment incubator (ISI) was developed to investigate the stability and dynamics of sediment‐associated microbial communities to prevailing subsurface oxidizing or reducing conditions. Herein we describe the use of these devices at the Old Rifle Uranium Mill Tailings Remedial Action (UMTRA) site. During a seven‐month period in which oxidized Rifle Aquifer background sediment (RABS) were deployed in previously biostimulated wells under iron‐reducing conditions, cell densities of known iron‐reducing bacteria, including Geobacteraceae, increased significantly, showing the microbial community response to local subsurface conditions. Phospholipid fatty acid (PLFA) profiles of RABS following in situ deployment were strikingly similar to those of adjacent sediment cores, suggesting ISI results could be extrapolated to the native material of the test plots. Results for ISI deployment with laboratory‐reduced sediments showed only slight changes in community composition and pointed toward the ability of the ISI to monitor microbial community stability and response to subsurface conditions. © 2009 Wiley Periodicals, Inc.     

Compositional Limitations in Poly–3–Hydroxyalkanoates Produced by Pseudomonas oleovorans

It is well known that Pseudomonas oleovorans can utilize sodium octanoate for both cell growth and the synthesis of polyhydroxyalkanoates (PHAs), but it can utilize sodium butyrate only for limited cell growth and not for the polyester formation when this substrate is the sole carbon source. Therefore, these two substrates were evaluated as cofeeds for the possible incorporation of 3-hydroxybutyryl groups in the resulting PHA. When sodium butyrate and sodium octanoate were fed to P. oleovorans as cosubstrates in various proportions, the resultant cell density and polymer content were proportional to the amount of sodium octanoate in the feed. The PHA extracted from cells grown in all combinations of these cosubstrates had similar unit compositions of approximately 8 mole % 3-hydroxyhexanoate, 91 mole % 3-hydroxyoctanoate and 1 mole % 3-hydroxydecanoate. 3-Hydroxybutyrate units were not detected in any of the PHAs isolated, indicating that these units could not be incorporated in the copolymer synthesized by P. oleovorans either because the cell did not synthesize that monomer or, if it did, the PHA synthase could not copolymerize it with the longer chain monomers.     

Evaluation of white‐rot fungi for the remediation of creosote‐contaminated soil

Application of fungal‐based bioaugmentation was evaluated for the remediation of creosote‐contaminated soil at a wood‐preserving site in West Virginia. Soil at the site contained creosote‐range polycyclic aromatic hydrocarbons (PAHs) at concentrations in some areas that exceed industrial risk‐based levels. Two white‐rot fungi (Pleurotus ostreatus and Irpex lacteus) were evaluated for remediation effectiveness in a two‐month bench‐scale treatability test. Both fungi produced similar results, with up to 67.3 percent degradation of total PAHs in 56 days. Pilot‐scale testing was performed at the site using Pleurotus ostreatus grown on two local substrate mixtures. During the 276‐day field trial, total PAHs were degraded by up to 93.2 percent, with all individual PAHs except one achieving industrial risk‐based concentrations. It was recommended that fungal‐based remediation be applied to all contaminated soil at the site. © 2002 Wiley Periodicals, Inc.