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Stable isotope evidence for biodegradation of chlorinated ethenes at a fractured bedrock site.

Stable carbon isotope analysis of chlorinated ethenes and ethene was performed at a site contaminated with trichloroethene (TCE), a dense non-aqueous phase liquid (DNAPL). The site is located in fractured bedrock and had variable groundwater hydraulic gradients during the study due to a local excavation project. Previous attempts to biostimulate a pilot treatment area at the site resulted in the production of cis-1,2-dichloroethene (cis-DCE), the first product of reductive dechlorination of TCE. Cis-DCE concentrations accumulated however, and there was no appreciable production of the breakdown products from further reductive dechlorination, vinyl chloride (VC) and ethene (ETH). Consequently, the pilot treatment area was bioaugmented with a culture of KB-1, a natural microbial consortium known to completely reduce TCE to nontoxic ETH. Due to ongoing dissolution of TCE from DNAPL in the fractured bedrock, and to variable hydraulic gradients, concentration profiles of dissolved TCE and its degradation products cis-DCE, VC, and ETH could not convincingly confirm biodegradation of the chlorinated ethenes. Isotopic analysis of cis-DCE and VC, however, demonstrated that biodegradation was occurring in the pilot treatment area. The isotope values of cis-DCE and VC became significantly more enriched in 13C over the last two sampling dates (in one well from -17.6%o to -12.8%o and from -22.5%o to -18.2%o for cis-DCE and VC, respectively). Quantification of the extent of biodegradation in the pilot treatment area using the Rayleigh model indicated that, depending on the well, between 21.3% and 40.7% of the decrease in cis-DCE and between 15.2% and 36.7% of the decrease in VC concentrations can be attributed to the effects of biodegradation during this time period. Within each well, the isotope profile of TCE remained relatively constant due to the continuous input of undegraded TCE due to DNAPL dissolution.

Biodegradation, Environmental↗

Aerobic biodegradation behavior of nonylphenol polyethoxylates and their metabolites in the presence of organic matter.

In this paper, the aerobic biodegradation behavior of nonylphenol polyethoxylates (NPnEOs) with ethoxy (EO) units of specific lengths, which were fractionated using high-performance liquid chromatography(HPLC) with photodiode array detection, was studied in the presence of different types of organic materials. NPnEOs and their related metabolites under a modified OECD 301E biodegradation test were monitored using liquid chromatography/mass spectrometry (LC/MS) and gas chromatography/mass spectrometry (GC/MS). Biodegradation tests in the presence of organic matters, such as methanol, glucose, and yeast extract, showed the formation of the corresponding nonylphenol polyethoxy carboxylates by the oxidation of the terminal alcoholic group. However, aerobic biodegradation tests without organic matter revealed that NP2EO and NP3EO were predominant metabolites of the long-chain-oligomer precursor system which undergo fast and complete shortening. Degradation rates were higher for the long-chain oligomers than for shorter ones. The degradation pathway of NPnEOs was greatly influenced by the presence or absence of organic matter. Organic materials such as those given above apparently play a significant role in the formation of the carboxylated metabolites of NPnEOs.

Aerobiosis↗

Biodegradation of 2,4,6-tribromophenol during transport in fractured chalk.

The effect of physicochemical conditions (residence time, oxygen concentrations, and chalk characteristics) on the biodegradation of 2,4,6-tribromophenol (TBP) during transport was investigated in low-permeability fractured-chalk cores. Long-term (approximately 600 d) biodegradation experiments were conducted in two cores (approximately 21 cm diameter, 31 and 44 cm long, respectively), intersected by a natural fracture. TBP was used as a model contaminant and as the sole carbon source for aerobic microbial activity. Bacterial isolates were recovered and identified by both Biolog identification kit and 16S rDNA sequences from batch enrichment cultures. One of the strains, with 98% similarity (based on the 16S rDNA data) to Achromobacter xylosoxidans, was shown to have the ability to degrade TBP in the presence of chalk. The decrease in TBP concentration along the fracture due to biodegradation was not affected by reducing the residence time from 49 to 8 min. In contrast, adding oxygen to the water at the inlet and increasing the flow rates improved TBP removal. Although the matrix pore-size distribution limits microbial activity to the fracture void, the chalk appears to provide an excellent environment for biodegradation activity. Approximately 90% of TBP removal occurred within 10 cm of the TBP source, indicating that in-situ bioremediation can be used to remove organic contaminants in low-permeability fractured rocks if nutrient-delivery pathways within the aquifer are secured.

Biodegradation, Environmental↗

Pathway dependent isotopic fractionation during aerobic biodegradation of 1,2-dichloroethane.

1,2-Dichloroethane (1,2-OCA) is a widespread groundwater contaminant known to be biodegradable under aerobic conditions via enzymatic oxidation or hydrolytic dehalogenation reactions. Current literature reports that stable carbon isotope fractionation of 1,2-DCA during aerobic biodegradation is large and reproducible (-27 to -33/1000). In this study, a significant variation in the magnitude of stable carbon isotope fractionation during aerobic biodegradation was observed. Biodegradation in experiments involving microcosms, enrichment cultures, and pure microbial cultures produced a consistent bimodal distribution of enrichment factors (epsilon) with one mean epsilon centered on -3.9 +/- 0.6/1000 and the other on -29.2 +/- 1.9/1000. Reevaluation of epsilon in terms of kinetic isotope effects 12k/13k gave values of 12k/13k = 1.01 and 1.06, which are typical of oxidation and hydrolytic dehalogenation (S(N)2) reactions, respectively. The bimodal distribution is therefore consistent with the microbial degradation of 1,2-DCA by two separate enzymatic pathways. This interpretation is further supported in this study by experiments with pure strains of Xanthobacter autotrophicus GJ10, Ancylobacter aquaticus AD20, and Pseudomonas sp. Strain DCA1 for which the enzymatic degradation pathways are well-known. A small fractionation of -3.0/1000 was measured for 1,2-DCA degradation by Pseudomonas sp. Strain DCA1 (monooxygenase enzyme), while degradation by the hydrolytic dehalogenase enzyme by the other two pure strains was characterized by fractionation of -32.3/1000.

Biodegradation, Environmental↗

Gaseous hexane biodegradation by Fusarium solani in two liquid phase packed-bed and stirred-tank bioreactors.

Biofiltration of hydrophobic volatile pollutants is intrinsically limited by poor transfer of the pollutants from the gaseous to the liquid biotic phase, where biodegradation occurs. This study was conducted to evaluate the potential of silicone oil for enhancing the transport and subsequent biodegradation of hexane by the fungus Fusarium solani in various bioreactor configurations. Silicone oil was first selected among various solvents for its biocompatibility, nonbiodegradability, and good partitioning properties toward hexane. In batch tests, the use of silicone oil improved hexane specific biodegradation by approximately 60%. Subsequent biodegradation experiments were conducted in stirred-tank (1.5 L) and packed-bed (2.5 L) bioreactors fed with a constant gaseous hexane load of 180 g x m(-3)(reactor) x h(-1) and operated for 12 and 40 days, respectively. In the stirred reactors, the maximum hexane elimination capacity (EC) increased from 50 g x m(-3)(reactor) x h(-1) (removal efficiency, RE of 28%) in the control not supplied with silicone oil to 120 g x m(-3)(reactor) x h(-1) in the biphasic system (67% RE). In the packed-bed bioreactors, the maximum EC ranged from 110 (50% RE) to 180 g x m(-3)(reactor) x h(-1) (> 90% RE) in the control and two-liquid-phase systems, respectively. These results represent, to the best of our knowledge, the first reported case of fungi use in a two-liquid-phase bioreactor and the highest hexane removal capacities so far reported in biofilters.

Biodegradation, Environmental↗

Compositional and moisture content effects on the biodegradability of zein/ethylcellulose films.

The effect of moisture content and film composition on biodegradability is the focus of this study. Flexible films were first characterized for the effect on water sorption isotherms of relative humidity, temperature, zein content, and the addition of the plasticizers stearic acid, poly(ethylene glycol), or etoxylated ricine oil. Zein/ethylcellulose (EC) mixture films had a behavior between that for pure zein and EC films, which had the lowest water sorption. For films with plasticizer, the lowest water sorption at 25 degrees C was observed for those with stearic acid. Biodegradability of zein/EC films, evaluated using bacterial cultures selected for their zein proteolytic activity and isolated from a local solid waste landfill and a lagoon, showed no plasticizer effect even though its effect on moisture content was significant. Large differences were observed at different film zein concentration with the highest biodegradability for 100% zein. However, biodegradability did not mimic the water sorption behavior of zein/EC mixture films.

Biodegradation, Environmental↗

Diversity and correlation of specific aromatic hydrocarbon biodegradation capabilities.

This work investigated the biodegradation capabilities of indigenous microorganisms exposed to different combinations of aromatic hydrocarbons. Considerable diversity was found in the catabolic specificity of 55 strains. Toluene was the most commonly degraded compound, followed by p-xylene, m-xylene and ethylbenzene. Strains capable of degrading o-xylene and benzene, which were the least-frequently-degraded compounds, exhibited broader biodegradation capabilities. Kappa statistics showed a significant correlation between the abilities to degrade toluene and ethylbenzene, p-xylene and m-xylene, and p-xylene and o-xylene. The ability to degrade naphthalene was correlated to the ability to degrade other alkylbenzenes, but not benzene. In addition, the inability to degrade benzene was correlated to the inability to degrade o-xylene. Factorial analysis of variance showed that biodegradation capabilities were generally broader when aromatic hydrocarbons were fed as mixtures than when fed separately. Beneficial substrate interactions included enhanced degradation of benzene, p-xylene, and naphthalene when toluene was present, and enhanced degradation of naphthalene by ethylbenzene. Such heuristic relationships may be useful to predict biodegradation patterns when bacteria are exposed to different aromatic hydrocarbon mixtures.

Bacteria↗

Stoichiometry of the aerobic biodegradation of the organic fraction of municipal solid waste (MSW).

An elemental analysis was applied to describe the composition of the organic fraction of municipal solid waste (MSW). The initial elemental composition was constant at C5H8.5O4N0.2. The changes of the composition during the biodegradation process and the final waste composition were strictly dependent on the process conditions. The decrease in carbon content due to biodegradation increased with temperature at which the experiments were conducted, from 20% at 20 degrees C to about 40% at 37-42 degrees C after 96 hours. It was correlated with the amount of oxygen that was utilised in the investigated processes of aerobic biodegradation of the waste suspension. The amount of oxygen required for biodegradation of organic fraction of MSW was estimated on the basis of stoichiometric equations and increased from 0.92 moles per 1 mole of waste at 20 degrees C to 1.6 moles at 42 degrees C within 96 hours of the experiments.

Aerobiosis↗

Anaerobic biodegradation of linear alkylbenzene sulfonate (LAS) in upflow anaerobic sludge blanket (UASB) reactors.

The anaerobic biodegradation of Linear Alkylbenzene Sulfonate (LAS) was studied in Upflow Anaerobic Sludge Blanket Reactors (UASB). One reactor was fed with easily degradable substrates and commercial LAS solution during a period of 3 months (Reactor 1), meanwhile a second reactor was fed with a commercial LAS solution without co-substrate (Reactor 2) during 4 months. Both reactors were operated with an organic loading rate of 4-5 mg-LAS/l x day and a hydraulic retention time of one day. The LAS biodegradation was determined by full mass balance. LAS was analysed by HPLC in the liquid phase (influent and effluent streams of the reactors) as well as in the solid phase (granular sludge used as biomass). The results indicate a high level of removal (primary biodegradation: 64-85%). Biodegradation was higher in the absence of external co-substrates than in the presence of additional sources of carbon. This indicates that the surfactant can be partially used as carbon and energy source by anaerobic bacteria. Under the operating conditions used, inhibition of the methanogenic activity or any other negative effects on the biomass due to the presence of LAS were not observed. The methanogenic activity remained high and stable throughout the experiment.

Alkanesulfonic Acids↗

Enhanced biodegradation of Casablanca crude oil by a microbial consortium in presence of a rhamnolipid produced by Pseudomonas aeruginosa AT10.

The biodegradation of oil products in the environment is often limited by their low water solubility and dissolution rate. Rhamnolipids produced by Pseudomonas aeruginosa AT10 were investigated for their potential to enhance bioavailability and hence the biodegradation of crude oil by a microbial consortium in liquid medium. The characterization of the rhamnolipids produced by strain AT10 showed the effectiveness of emulsification of complex mixtures. The addition of rhamnolipids accelerates the biodegradation of total petroleum hydrocarbons from 32% to 61% at 10 days of incubation. Nevertheless, the enhancement of biosurfactant addition was more noticeable in the case of the group of isoprenoids from the aliphatic fraction and the alkylated polycyclic aromatic hydrocarbons (PHAS) from the aromatic fraction. The biodegradation of some targeted isoprenoids increased from 16% to 70% and for some alkylated PAHs from 9% to 44%.

Biodegradation, Environmental↗

Modeling intrinsic bioremediation for interpret observable biogeochemical footprints of BTEX biodegradation: the need for fermentation and abiotic chemical processes.

The intrinsic bioremediation of BTEX must be documented by the stoichiometric consumption and production of several other compounds, called 'footprints' of the biodegradation reaction. Although footprints of BTEX biodegradation are easy to identify from reaction stoichiometry, they can be confounded by the stepwise nature of the biodegradation reactions and by several abiotic chemical reactions that also produce or consume the footprints. In order to track the footprints for BTEX biodegradation, the following reactions need to be considered explicitly: (1) fermentation and methanogenesis as separate processes, (2) precipitation and dissolution of calcite, (3) precipitation and dissolution of amorphous iron monosulfide (FeS), (4) conversion of FeS into the thermodynamically stable pyrite (FeS2) with loss of sulfide and abiotic formation of H2, and (5) reductive dissolution of solid iron(III) by oxidation of sulfide. We critically review the research that underlies why these mechanisms must be included and how to describe them quantitatively. A companion manuscript develops and applies a mathematical model that includes these reactions.

Biodegradation, Environmental↗

In vitro enzymatic biodegradation of amino acid based poly(ester amide)s biomaterials.

A systematic in vitro biodegradation study of regular poly(ester amide)s (PEAs) composed of naturally occurring hydrophobic alpha-amino acids, fatty diols and dicarboxylic acids was carried out in the presence of hydrolases like trypsin, alpha-chymotrypsin, and lipase. An automatic potentiometric titration method was used to examine the biodegradation property of the PEAs. Spontaneous immobilization (absorption) of the enzymes onto the PEAs films surfaces was observed. The surface immobilized enzyme not only accelerated the erosion of the PEAs but also was able to catalyze the hydrolysis of both low-molecular-weight (ATEE) and high-molecular-weight (protein) external substrates. It was found that the enzyme surface absorption process is reversible by nature. A kinetic method for a quantitative determination of the enzyme desorbed from the film surface was developed. The enzymes could also be impregnated into the PEAs to make them "self-destructive" at a target rate. A comparison of the PEAs' in vitro biodegradation data with polylactide (PDLLA) showed that PEAs exhibited a far more tendency toward enzyme catalyzed biodegradation than PDLLA.

Amides↗

Biodegradation of refractory hydrocarbon biomarkers from petroleum under laboratory conditions.

Biomarkers are of great value in petroleum exploration because they provide essential information about the geological history of oils and source rocks. Steranes are of particular importance as they can be related to naturally occurring precursors. These compounds generally experience intense biodegradation, however, which alters their original distribution and obscures the information that they carry regarding oil maturity and source material. In an attempt to identify the microorganisms responsible for this degradation, we have investigated the capacity of 73 aerobic bacteria to degrade steranes present in Rozel Point (Utah) oil. Seven Gram-positive strains, belonging to a limited number of genera, were found to be active. Using Nocardia sp. SEBR 16, which caused the most extensive alteration, we have determined biodegradation rates for several isomers of steranes and methylsteranes. The preference for alteration of different isomers reflects that observed in natural environments, suggesting that the degradation intermediates could be used as indicators of the extent of the biodegradation in an oil. In addition, the microorganisms used here might be effective in biodegrading oil spills.

Arthrobacter↗

Volatile hydrocarbon biodegradation by a mixed-bacterial culture during growth on crude oil.

Volatile hydrocarbon biodegradation by a mixed-bacterial culture during growth on Bow River crude oil was investigated using solid phase microextraction (SPME). Inoculum treatments were examined in relation to C(5)-C(11) hydrocarbon degradation. Up to 1600 mg/l biomass (dry weight) was tested without achieving significant volatile hydrocarbon partitioning and affecting analysis. Inoculum age rather than concentration had the most profound impact on biodegradation. When late log phase crude oil-grown inocula were used, C(5)-C(11) biodegradation reached 55-60%; methylcyclohexane and other branched compounds eluting before n-C(8) were recalcitrant. Increasing the late log inoculum concentration from 0.63 to 63 mg/l resulted in a twofold increase in degradation rate without improving the substrate range. Methylcyclohexane recalcitrance was correlated with reduced levels of hydrocarbon-degrading bacteria and volatile hydrocarbon evaporation from the inoculum flasks. A decreased lag phase prior to degradation was observed when using early stationary phase cultures as inocula and most compounds up to C(11), including methylcyclohexane, were biodegraded.

Bacteria↗

Biodegradation studies of polyaromatic hydrocarbons in aqueous media.

Sixteen bacterial strains isolated from an activated sludge and Mycobacterium ssp. PYR-1 were tested for their ability to degrade polyaromatic hydrocarbons (PAHs). The bacterial strains Pasteurella ssp. (B-2) and Mycobacterium ssp. PYR-1 (AM) showed a high biodegradation potential of three- and four-ring PAHs. Bacterial strain AM was able to degrade up to 80% of three and four-ring PAHs (phenanthrene, fluoranthene and pyrene) within the first month of incubation, while the bacterial strain B-2 achieved the same biodegradation in 2 months. The metabolic pathway of PAH degradation was studied using fluoranthene and the bacterial strain AM. Ninety per cent of fluoranthene was biodegraded within the first 9 d of incubation when applied as a single substrate. Retention factor values from thin-layer chromatography studies, gas chromatography with mass selective detection and tandem mass spectrometry identified 9-fluorenone-1-carboxylic acid as one of the stable metabolic products and from this a fluoranthene biodegradation pathway is proposed.

Biodegradation, Environmental↗

Kinetic studies of biodegradation of insoluble compounds by continuous determination of oxygen consumption.

Continuous determination of oxygen consumption by electrolytic respirometry has been experimented as a means to study the biodegradation kinetics of scarcely soluble environmental pollutants. The substrates used were the polycyclic aromatic hydrocarbons (PAH), naphthalene, phenanthrene, anthracene, fluoranthene and pyrene. The definition of an appropriate mode of PAH supply, either as crystals or more generally as a solution in a water non-miscible solvent, was found essential for yielding reproducible biodegradation kinetics. In these conditions, for all compounds tested, oxygen determination was found suitable for quantitative evaluation of PAH biodegradation and formation of biomass and soluble metabolites. The study of biodegradation kinetics with this methodology showed that a first phase of exponential growth could be characterized in most cases, followed by a phase of limited growth. Possible mechanisms involved in insoluble substrate uptake are discussed. During exponential growth, the bacteria utilized (although not necessarily exclusively) the PAH solubilized in the aqueous medium.

Biodegradation, Environmental↗

Biodegradation of aniline, anthracene, chlornitrophen, fenitrothion and linear alkylbenzene sulphonate in pond water.

Biodegradation of five chemicals (aniline, anthracene, chlornitrophen (CNP), fenitrothion (FNT) and linear alkylbenzene sulphonate (LAS)) by aquatic bacteria in three different types of ponds was determined according to the cultivation method developed by this group. The degradability toward these chemicals was varied among the ponds, except for LAS which was decomposed well in all samples. Higher degradability towards the two agrochemicals, CNT and FNT, was found in the pond surrounded by paddy fields, whereas aniline and anthracene were decomposed more rapidly in the pond located in the industrial area. Water from the pond in the botanical garden, with the least exposure to any chemicals, exhibited the lowest degradation toward all chemicals tested. There was no significant seasonal variation in the biodegradation of chemicals in these ponds. It was deduced that biodegradability toward certain chemicals could be a result of acclimatization of the microbial community by chemical contamination present and past, suggesting the possible use of biodegradation profiles as an indicator for chemical pollution in the aquatic environment.

Biodegradation, Environmental↗

Mechanisms for enhanced biodegradation of petroleum hydrocarbons by a microbe-colonized gas-liquid foam.

AIMS: A microbe-colonized gas-liquid foam formulation has been previously shown to provide enhanced biodegradation capabilities in soil microcosms. The present study considers the reservoir properties of this foam and how this affects hydrocarbon degradation rates. METHODS AND RESULTS: Oxygen solubility in protein hydrolysate solutions draining from aerated and oxygenated foams was measured. The suitability of oxygenated foam to enhance the degradation of n-hexadecane in soil microcosms was assessed. Sorption of bacterial isolates at the gas-liquid interface was also investigated using a range of microscopy techniques. CONCLUSIONS: Oxygenated bioactive foam enhanced biodegradation rates by improving oxygen availability and transfer. Biodegradation of n-hexadecane was also stimulated by the protein hydrolysate used and by the inclusion of known bacterial hydrocarbon-degrading bacteria. The interaction of bacteria with the gas-liquid interface was shown to be a significant factor governing the drainage of the bacteria from the bioactive foam. SIGNIFICANCE AND IMPACT OF THE STUDY: Protein hydrolysate-based bioactive foam may be a suitable treatment technology to enhance the biodegradation of petroleum hydrocarbons in soil.

Alkanes↗