Biodegradation of hydrocarbons vapors: Comparison of laboratory studies and field investigations in the vadose zone at the emplaced fuel source experiment, Airbase Vaerløse, Denmark

The natural attenuation of volatile organic compounds (VOCs) in the unsaturated zone can only be predicted when information about microbial biodegradation rates and kinetics are known. This study aimed at determining first-order rate coefficients for the aerobic biodegradation of 13 volatile petroleum hydrocarbons which were artificially emplaced as a liquid mixture during a field experiment in an unsaturated sandy soil. Apparent first-order biodegradation rate coefficients were estimated by comparing the spatial evolution of the resulting vapor plumes to an analytical reactive transport model. Two independent reactive numerical model approaches have been used to simulate the diffusive migration of VOC vapors and to estimate degradation rate coefficients. Supplementary laboratory column and microcosm experiments were performed with the sandy soil at room temperature under aerobic conditions. First-order kinetics adequately matched the lab column profiles for most of the compounds. Consistent compound-specific apparent first-order rate coefficients were obtained by the three models and the lab column experiment, except for benzene. Laboratory microcosm experiments lacked of sensitivity for slowly degrading compounds and underestimated degradation rates by up to a factor of 5. Addition of NH3 vapor was shown to increase the degradation rates for some VOCs in the laboratory microcosms. All field models suggested a significantly higher degradation rate for benzene than the rates measured in the lab, suggesting that the field microbial community was superior in developing benzene degrading activity.     

Effect of expandable clays and cometabolism on PAH biodegradability

Pyrene and phenanthrene degradation was examined in both single and binary slurry systems for three different natural soils. It was found that the amount of total expandable clays (smectite and vermiculite) was in a good agreement with the achieved rate and extent of biodegradation. For instance, the intrinsic phenanthrene biodegradation rate was 626 microg/L/day for the soil with the largest expandable clay and 3203 microg/L/day for the soil with the least. Similarly, the smallest total pyrene biodegradation (65%) was found for the soil rich in expandable clays, compared to an 82% pyrene reduction in the soil that had the lowest amount. Mass transfer limitation after compound sorption to the clays was more pronounced for the more hydrophobic pyrene. In the presence of phenanthrene, total pyrene biodegradation increased by 2 to 7% due to cometabolism, while the total phenanthrene biodegradation was only enhanced by 0.5 to 5% in the binary system. This research demonstrated that expandable clays might govern the substrate availability to microorganisms and microbial accessibility to substrates. Therefore, the contribution of organic matter and expandable clays to sorption, desorption and biodegradation should be taken equally into account in order to better understand complex bioremediation issues.     

The lack of microbial degradation of polycyclic aromatic hydrocarbons from coal-rich soils

Analytical techniques used to assess the environmental risk of contamination from polycyclic aromatic hydrocarbons (PAHs) typically consider only abiotic sample parameters. Supercritical fluid extraction and sorption enthalpy experiments previously suggested slow desorption rates for PAH compounds in two coal-contaminated floodplain soils. In this study, the actual PAH availability for aerobic soil microorganisms was tested in two series of soil-slurry experiments. The experimental conditions supported microbial degradation of phenanthrene if it was weakly sorbed onto silica gel. Native coals and coal-derived particles in two soils effectively acted as very strong sorbents and prevented microbial PAH degradation. The long history of PAH exposure and degree of coal contamination apparently had no influence on the capability of the microbial soil community to overcome constraints of PAH availability. Within the context of the experimental conditions and the compounds chosen, our results confirm that coal-bound PAHs are not bioavailable and hence of low environmental concern.     

Anaerobic PAH degradation in soil by a mixed bacterial consortium under denitrifying conditions

Polycyclic aromatic hydrocarbons (PAHs) are one of the main classes of contaminants in the terrestrial environment. Concentrations of biphenyl, fluorene, phenanthrene and pyrene were added to soil samples in order to investigate the anaerobic degradation potential of PAHs under denitrifying conditions. A mixed population of microorganisms obtained from a paddy soil was incubated for 20 days in anaerobic conditions in the presence of soil alone or with nitrate, adding, as electron donors, PAHs and, in some samples, glucose or acetate. At regular time intervals oxidation-reduction potential, PAHs concentration, microbial ATP and nitrate concentration into the solution were measured. Degradation trends for each hydrocarbon are similar under all conditions, indicating that the molecular conformation prevails over other parameters in controlling the degradation. Poor degradation results were obtained when PAHs were the only organic matter available for the inoculum, thus confirming the recalcitrance to degradation of these compounds. Biodegradation was influenced by the addition of other carbon sources. As better degradation results were generally obtained when acetate or glucose were added, the hypothesis of a co-metabolic enhancement of PAH biodegradation seems likely. Thus, anaerobic biodegradation of PAHs studied, biphenyl, fluorene, phenanthrene and pyrene, seems to be possible both through fermentative and respiratory metabolism, provided that low molecular weight co-metabolites and suitable electron acceptors (nitrate) are present.     

Estimating the spatial scale of herbicide and soil interactions by nested sampling, hierarchical analysis of variance and residual maximum likelihood

An unbalanced nested sampling design was used to investigate the spatial scale of soil and herbicide interactions at the field scale. A hierarchical analysis of variance based on residual maximum likelihood (REML) was used to analyse the data and provide a first estimate of the variogram. Soil samples were taken at 108 locations at a range of separating distances in a 9 ha field to explore small and medium scale spatial variation. Soil organic matter content, pH, particle size distribution, microbial biomass and the degradation and sorption of the herbicide, isoproturon, were determined for each soil sample. A large proportion of the spatial variation in isoproturon degradation and sorption occurred at sampling intervals less than 60 m, however, the sampling design did not resolve the variation present at scales greater than this. A sampling interval of 20-25 m should ensure that the main spatial structures are identified for isoproturon degradation rate and sorption without too great a loss of information in this field.     

Solubilization, solution equilibria, and biodegradation of PAH's under thermophilic conditions

Biodegradation rates of PAHs are typically low at mesophilic conditions and it is believed that the kinetics of degradation is controlled by PAH solubility and mass transfer rates. Solubility tests were performed on phenanthrene, fluorene and fluoranthene at 20 degrees C, 40 degrees C and 60 degrees C and, as expected, a significant increase in the equilibrium solubility concentration and of the rate of dissolution of these polycyclic aromatic hydrocarbons (PAHs) was observed with increasing temperature. A first-order model was used to describe the PAH dissolution kinetics and the thermodynamic property changes associated with the dissolution process (enthalpy, entropy and Gibb's free energy of solution) were evaluated. Further, other relevant thermodynamic properties for these PAHs, including the activity coefficients at infinite dilution, Henry's law constants and octanol-water partition coefficients, were calculated in the temperature range 20-60 degrees C. In parallel with the dissolution studies, three thermophilic Geobacilli were isolated from compost that grew on phenanthrene at 60 degrees C and degraded the PAH more rapidly than other reported mesophiles. Our results show that while solubilization rates of PAHs are significantly enhanced at elevated temperatures, the biodegradation of PAHs under thermophilic conditions is likely mass transfer limited due to enhanced degradation rates.     

Persistence of perfluoroalkylated substances in closed bottle tests with municipal sewage sludge

BACKGROUND, AIM, AND SCOPE: Perfluoroalkylated substances (PFAS) are chemicals with completely fluorinated alkyl chains. The specific properties of the F-C bond give PFAS a high stability and make them very useful in a wide range of applications. PFAS also pose a potential risk to the environment and humans because they have been recently characterized as persistent, bioaccumulative, and toxic. The objective of this work is to study the bacterial degradation of PFAS under aerobic and anaerobic conditions in municipal sewage sludge as a contribution toward understanding their environmental fate and behavior. MATERIALS AND METHODS: Bacterial communities from sewage sludge were exposed to a mixture of PFAS under aerobic or anaerobic conditions. Individual PFAS concentrations were determined in the experiment media at different exposure times using liquid chromatography-mass spectrometry analysis after extraction with solid-phase extraction. RESULTS: The PFAS analyses of samples of sludge showed repeatable replicate results, allowing a reliable quantification of the different groups of PFAS analyzed. No conclusive evidence for PFAS degradation was observed under the experimental conditions tested in this work. Reduction in concentrations, however, was observed for some PFAS in sludge under aerobic conditions. DISCUSSION: The largest concentration decrease occurred for the fluorotelomer alcohols (FTOHs), especially for the 8:2 FTOH, which have been described as biodegradable in the literature. However, this concentration decrease could be due to different causes: sorption to glass, septa, or matrix components, as well as bacterial activity. Therefore, it is not certain that biodegradation occurred. CONCLUSIONS: PFAS are very recalcitrant chemicals, especially when fully fluorinated. Although some decreases in concentration have been observed for some PFAS, such as the FTOHs, there is no conclusive evidence for biodegradation. It can be concluded that the PFAS tested in these experiments are non-biodegradable under these experimental conditions. RECOMMENDATIONS AND PERSPECTIVES: Since the presence of PFAS is ubiquitous in the environment and they can be toxic, more research is needed in this field to elucidate which PFAS are susceptible to biodegradation, the conditions required for biodegradation, and the possible routes followed. A possible inhibitory effect of PFAS on bacteria, the threshold concentrations, and conditions of inhibition should also be investigated.     

Electrokinetic enhancement of phenanthrene biodegradation in creosote-polluted clay soil

Given the difficulties caused by low-permeable soils in bioremediation, a new electrokinetic technology is proposed, based on laboratory results with phenanthrene, to afford bioremediation of polycyclic aromatic hydrocarbons (PAH) in clay soils. Microbial activity in a clay soil historically polluted with creosote was promoted using a specially designed electrokinetic cell with a permanent anode-to-cathode flow and controlled pH. The rates of phenanthrene losses during treatment were tenfold higher in soil treated with an electric field than in the control cells without current or microbial activity. Results from experiments with Tenax-assisted desorption and mineralization of 14C-labeled phenanthrene indicated that phenanthrene biodegradation was limited by mass-transfer of the chemical. We suggest that the enhancement effect of the applied electric field on phenanthrene biodegradation resulted from mobilization of the PAH and nutrients dissolved in the soil fluids.     

Biodegradation of polycyclic aromatic hydrocarbons by a mixed culture

We investigated the potential biodegradation of polycyclic aromatic hydrocarbons (PAHs) by an aerobic mixed culture utilizing phenanthrene as its carbon source. Following a 3-5 h post-treatment lag phase, complete degradation of 5 mg/l phenanthrene occurred within 28 h (optimal conditions determined as 30 degrees C and pH 7.0). Phenanthrene degradation was enhanced by the individual addition of yeast extract, acetate, glucose or pyruvate. Results show that the higher the phenanthrene concentration, the slower the degradation rate. While the mixed culture was also capable of efficiently degrading pyrene and acenaphthene, it failed to degrade anthracene and fluorene. In samples containing a mixture of the five PAHs, treatment with the aerobic culture increased degradation rates for fluorene and anthracene and decreased degradation rates for acenaphthene, phenanthrene and pyrene. Finally, it was observed that when nonionic surfactants were present at levels above critical micelle concentrations (CMCs), phenanthrene degradation was completely inhibited by the addition of Brij 30 and Brij 35, and delayed by the addition of Triton X100 and Triton N101.     

The use of ozone in the remediation of polycyclic aromatic hydrocarbon contaminated soil

The potential of using ozone for the removal of phenanthrene from several different soils, both alone and in combination with biodegradation using a microbial inoculant (Pseudomonas alcaligenes PA-10), was examined. The greater the water content of the soil the less effective the ozone treatment, with air-dried soils showing the greatest removal of phenanthrene; while soils with higher levels of clay also reduced the effectiveness of the ozone treatments. However, at least a 50% reduction in phenanthrene levels was achieved in air-dried soil after an ozone treatment of 6 h at 20 ppm, with up to 85% removal of phenanthrene achieved in sandy soils. The biodegradation results indicate that P. alcaligenes PA-10 may be useful as an inoculant for the removal of PAHs from contaminated soils. Under the conditions used in our experiments, however, pre-ozonation did not enhance subsequent biodegradation of phenanthrene in the soils. Similar levels of phenanthrene removal occurred in both non-ozonated and ozonated Cruden Bay soil inoculated with P. alcaligenes PA-10. However, the biodegradation of phenanthrene in ozonated Boyndie soil was much slower. This may be due to the release of toxic products in this soil during ozonation.     

The influence of a NAPL on the loss and biodegradation of 14C-phenanthrene residues in two dissimilar soils

This study was carried out to assess the influence of diesel, applied over a log concentration range, on the loss and extractability of phenanthrene (measured as putative 14C-phenanthrene residues) in two different soils. The influence of diesel on the ability of a cyclodextrin based extraction method to predict the microbial bioavailability of 14C-residues was also assessed. An increase in loss of 14C-residues with increasing diesel concentration from 0 to 2000 mg kg-1 was generally observed with time in both soils. It is suggested that this trend is attributable to competitive sorption for soil sorption sites and to a lesser extent to displacement of 14C-residues from soil sorption sites by diesel resulting in greater compound availability and therefore greater loss by degradation via the actions of indigenous microorganisms. However, in the 20000 mg kg-1 diesel treatments of both soils, results indicated a delayed loss. It is suggested that this retarded loss was due to the formation of a discrete NAPL-phase into which 14C-phenanthrene residues partitioned, thereby decreasing their availability and as a consequence their degradation. Furthermore, it is suggested that nutrient limitation may have slowed down degradation rates as diesel concentrations increased. Comparison between cyclodextrin-extractability and microbial mineralisation supported the use of cyclodextrin to assess microbial bioavailability of 14C-residues after 50 d or more ageing up to diesel concentrations of 2000 mg kg-1. However, results suggested that at high diesel concentrations (specifically 20000 mg kg-1) co-extraction of 14C-phenanthrene residues may have occurred as a result of the combined solvation powers of both the cyclodextrin and the diesel. Furthermore, mineralisation of 14C-phenanthrene residues may have been affected by extreme nutrient limitation in this treatment.     

Modeling and interpreting bioavailability of organic contaminant mixtures in subsurface environments

Bioavailability often controls the fate of organic contaminants in surface and subsurface aquatic environments. Bioavailability can be limited by sorption, mass transfer, and intrinsic biodegradation potential and can be further altered by the presence of other compounds. This paper reviews current perspectives on the processes influencing subsurface contaminant bioavailability, how these processes are modeled, and how the relative role of the various processes can be assessed through bioavailability indices. Although these processes are increasingly well understood, the use of sophisticated models and indices often are precluded by an inability to estimate the many parameters that are associated with complex models. Nonetheless, the proper representation of sorption, mass transfer, biodegradation, and co-solute effects can be critical in predicting bio-attenuation. The influence of these processes on contaminant fate is illustrated with numerical simulations for the simultaneous degradation of toluene (growth substrate) and trichloroethylene (nongrowth cometabolite) in hypothetical, aerobic, solid-water systems. The results show how the relative impacts on contaminant fate of the model's various component processes depends upon system conditions, including co-solute concentrations. Slow biodegradation rates increase the inhibition effects of a cometabolite and suppress the rate enhancement effects of a growth substrate. Irrespective of co-solute effects, contaminant fate is less sensitive to biodegradation processes in systems with strong sorption and slow desorption rates. Bioavailability indices can be used to relate these findings and to help identify appropriate modeling simplifications. In general, however, there remains a need to redefine such indices in order that bioavailability concepts can be better incorporated into site characterization, remediation design, and regulatory oversight.     

Combined UV-biological degradation of PAHs

The UV-photolysis of PAHs was tested in silicone oil and tetradecane. In most cases, the degradation of a pollutant provided within a mixture was lower than when provided alone due to competitive effects. With the exception of anthracene, the larger pollutants (4- and 5-rings) were always degraded first, proving that UV-treatment preferentially acts on large PAHs and thereby provides a good complement to microbial degradation. UV-photolysis was also found to be suitable for treatment of soil extract from contaminated soils. The feasibility of UV-biological treatment was demonstrated for the removal of a mixture of phenanthrene and pyrene in silicone oil. UV-irradiation of the silicone oil led to 83% pyrene removal but no phenanthrene photodegradation. Subsequent treatment of the oil in a two-phases partitioning bioreactor (TPPB) system inoculated with Pseudomonas sp. was followed by complete phenanthrene biodegradation but no further pyrene removal. Totally, the combined process allowed 92% removal of the PAH mixture. Further work should focus on characterizing the photoproducts formed and studying the influence of the solvent on the photodegradation process.     

Biodegradation of petroleum hydrocarbon vapors: laboratory studies on rates and kinetics in unsaturated alluvial sand

Predictions of natural attenuation of volatile organic compounds (VOCs) in the unsaturated zone rely critically on information about microbial biodegradation kinetics. This study aims at determining kinetic rate laws for the aerobic biodegradation of a mixture of 12 volatile petroleum hydrocarbons and methyl tert-butyl ether (MTBE) in unsaturated alluvial sand. Laboratory column and batch experiments were performed at room temperature under aerobic conditions, and a reactive transport model for VOC vapors in soil gas coupled to Monod-type degradation kinetics was used for data interpretation. In the column experiment, an acclimatization of 23 days took place before steady-state diffusive vapor transport through the horizontal column was achieved. Monod kinetic parameters Ks and vmax could be derived from the concentration profiles of toluene, m-xylene, n-octane, and n-hexane, because substrate saturation was approached with these compounds under the experimental conditions. The removal of cyclic alkanes, isooctane, and 1,2,4-trimethylbenzene followed first-order kinetics over the whole concentration range applied. MTBE, n-pentane, and chlorofluorocarbons (CFCs) were not visibly degraded. Batch experiments suggested first-order disappearance rate laws for all VOCs except n-octane, which decreased following zero-order kinetics in live batch experiments. For many compounds including MTBE, disappearance rates in abiotic batch experiments were as high as in live batches indicating sorption. It was concluded that the column approach is preferable for determining biodegradation rate parameters to be used in risk assessment models.     

Atrazine biodegradation modulated by clays and clay/humic acid complexes

The fate of pesticides in the environment is strongly related to the soil sorption processes that control not only their transfer but also their bioavailability. Cationic (Ca-bentonite) and anionic (Layered Double Hydroxide) clays behave towards the ionisable pesticide atrazine (AT) sorption with opposite tendencies: a noticeable sorption capacity for the first whereas the highly hydrophilic LDH showed no interactions with AT. These clays were modified with different humic acid (HA) contents. HA sorbed on the clay surface and increased AT interactions. The sorption effect on AT biodegradation and on its metabolite formation was studied with Pseudomonas sp. ADP. The biodegradation rate was greatly modulated by the material's sorption capacity and was clearly limited by the desorption rate. More surprisingly, it increased dramatically with LDH. Adsorption of bacterial cells on clay particles facilitates the degradation of non-sorbed chemical, and should be considered for predicting pesticide fate in the environment.     

Biodegradability of aged pyrene and phenanthrene in a natural soil

A study was conducted to evaluate the biodegradability of pyrene (PYR) and phenanthrene (PHE) aged in a natural soil. Both the single and binary systems were either biostimulated via a nutrient amendment or bioaugmented via an inoculation of the enriched bacteria and nutrients. Aging resulted in higher concentration of both compounds and smaller bacterial activity in the solution-phase. Surprisingly, the total biodegraded extent was greater in the aged soil system than in the freshly spiked system. As anticipated, biostimulation was not appropriate to attain an effective biodegradation in this study, and bioaugmentation achieved a substantial increase the total biodegradation extent. The above findings were attributed to indigenous Pseudomonas aeruginosa entering a stationary-phase during the 200-day aging and producing rhamnolipid biosurfactants. In addition, a different sampling technique (i.e., after vigorous hand-shaking) revealed a 15 times higher microbial population than the normal sampling from the stagnant solution. Therefore, PAH bioavailability in the aged soils can be underestimated when the microbial activity is determined only from the stagnant solution. Furthermore, cometabolism enhanced PYR degradation when PHE was present as a primary substrate.     

PAH dissipation in a contaminated river sediment under oxic and anoxic conditions

A batch experiment was conducted to compare PAH degradation in a polluted river sediment under aerobic and anaerobic conditions, and to investigate whether input of fresh organic material (cellulose) could enhance such degradation. All measurements were checked against abiotic control treatments to exclude artifacts of sample preparation and non-biological processes like aging. Three- and four-ring PAHs could be degraded by the indigenous microbial community under aerobic conditions, but anaerobic metabolism based on iron and sulphate reduction was not coupled with PAH degradation of even the simplest 3-ring compounds like phenanthrene. Cellulose addition stimulated both aerobic and anaerobic respiration, but had no effect on PAH dissipation. We conclude that natural attenuation of PAHs in polluted river sediments under anaerobic conditions is exceedingly slow. Dredging and biodegradation on land under aerobic conditions would be required to safely remediate and restore polluted sites.     

The equilibria of bisolute sorption on soil

The goal of this study was to investigate the effects of both concentration levels and loading sequence or contamination history of each pollutant on the equilibrium sorption of mixed organic pollutants on soils. We measured binary sorption equilibria for a soil using ten concentration levels for both phenanthrene and naphthalene. Both solutes were either simultaneously loaded or sequentially loaded (i.e., the second sorbate was loaded after the sorption of the first sorbate had attained equilibrium) on soil. The results showed different competitive sorption equilibria between phenanthrene and naphthalene. In the presence of phenanthrene and regardless of loading sequence, naphthalene exhibited consistently lower sorption capacities and the ideal adsorbed solution theory (IAST) slightly underestimates the naphthalene sorption equilibria. Conversely, the sorption equilibria of phenanthrene in the presence of naphthalene depended upon the loading sequence of the two sorbates on the soil. Little competition from naphthalene on the sorption equilibria of phenanthrene was observed when phenanthrene was loaded either simultaneously with or sequentially after naphthalene, but appreciable competition from naphthalene was observed when the soil had been pre-contaminated with phenanthrene. IAST slightly underestimates the phenanthrene sorption equilibria observed in the latter system, but it cannot estimate the phenanthrene sorption equilibria in the former two systems. We proposed that adsorption on internal surfaces of ink-bottle shaped pores within relatively flexible sorbent matrix may have caused the competitive sorption phenomena observed in this study. The study suggests that contamination history may have strong influence on the equilibrium sorption of organic pollutant mixtures.     

Biodegradation of phenanthrene in river sediment

The aerobic biodegradation potential of phenanthrene (a polycyclic aromatic hydrocarbon [PAH]) in river sediment was investigated in the laboratory. Biodegradation rate constants (k1) and half-lives (t1/2) for phenanthrene (5 microg/g) in sediment samples collected at five sites along the Keelung River in densely populated northern Taiwan ranged from 0.12 to 1.13 l/day and 0.61 to 5.78 day, respectively. Higher biodegradation rate constants were noted in the absence of sediment. Two of the sediment samples were capable of biodegrading phenanthrene at initial concentrations 5-100 microg/g; lower biodegradation rates occurred at higher concentrations. Optimal biodegradation conditions were determined as 30 degreesC and pH 7.0. Biodegradation was not significantly influenced by the addition of such carbon sources as acetate, pyruvate, and yeast extract, but was significantly influenced by the addition of ammonium, sulfate, and phosphate. Results show that anthracene, fluorene, and pyrene biodegradation was enhanced by the presence of phenanthrene, but that phenanthrene treatment did not induce benzo[a]pyrene biodegradation during a 12-day incubation period.     

Phenanthrene sorption/desorption sequences provide new insight to explain high sorption coefficients in field studies

The sorption coefficients obtained in field investigation vary greatly from laboratory sorption experiments. The possible reasons were discussed in literature. Observing the commonly reported desorption hysteresis, this study proposed that the unclear sorption history of the field study could also result in the diverse sorption coefficients. This study conducted a comparative study regarding phenanthrene sorption/desorption behavior in low-concentration multi-time sorption process and the commonly applied high-concentration one-time sorption process. The sorption coefficients determined during the desorption process were much higher than those at sorption process. Thus, the prediction of sorption coefficient should be related with sorption history. Desorption hysteresis was increased with increased equilibration time and decreased solid-phase concentration. In addition, although the apparent contact time between sorbate and sorbent was shorter for low-concentration multi-time sorption, the desorption hysteresis was much stronger, which consequently result in higher sorption coefficients in comparison to high-concentration one-time sorption. Pore swelling or diffusion-controlled sorption kinetics could not explain this phenomenon. This study calls for research attention on sorption history, especially for field investigations.