Response of the bacterial community to in situ bioremediation of organic-rich sediments

A field trial experiment was carried out to assess the potential of bioremediation for mobilisation of carbon in organic-rich sediments. Both bioaugmentation (bio-fixed microorganisms) and biostimulation (oxygen release compounds--ORC) protocols have been tested and the response of the bacterial community has been described to assess the baseline for bioremediation potential. Multifactorial ANOVA revealed that bioaugmentation protocol had an effect in stimulate mobilisation processes and significantly enhanced extra-cellular enzymatic activity rates. In contrast biostimulation treatment did not have an effect on mobilisation rates but contributed to enhance bacterial efficiency through a maximization of the bacterial production:enzymatic activity ratio. Average calculation of net mobilised carbon showed that 23% increase of mobilised pool was accounted for bioaugmentation in summer. Although biostimulation accounted for a smaller increase in mobilised carbon (<10%), the use of ORC resulted in an increased mineralisation and net carbon loss via respiration. Based on our results, a conceptual model for application of bioremediation to face the problem of sediment eutrophication is discussed.     

Evaluation of bioremediation strategies of a controlled oil release in a wetland

A controlled petroleum release was conducted to evaluate bioremediation in a wetland near Houston, Texas. The 140-day study was conducted using a randomized, complete block design to test three treatments with six replicates per treatment. The three treatment strategies were inorganic nutrients, inorganic nutrients with an alternative electron acceptor, and a no-action oiled control. Samples were analyzed for petroleum chemistry and inorganic nutrients. These results are discussed in the context of our related research involving toxicology and microbiology at the site during the experiment. To evaluate biodegradation, the targeted compounds were normalized to the conservative compound C3017alpha, 21beta-[H]hopane, thus reducing the effects of spatial heterogeneity and physical transport. The two biostimulation treatments demonstrated statistically-higher rates of biodegradation than the oiled no-action control. For the majority of the experiment, target nutrient levels were maintained. Further research may be warranted to optimize these bioremediation strategies as well as evaluating additional treatment strategies for wetlands and other shoreline systems.     

Coupling Interactions between Electrokinetics and Bioremediation for Pyrene Removal from Soil under Polarity Reversal Conditions

This paper investigates the hybrid technology of electrokinetics (EK) coupled with bioremediation (Bio) in the removal of pyrene (PYR) in a soil matrix. Five different treatments were conducted to investigate the coupling interactions between EK and Bio on PYR degradation. A simulated removal curve was obtained by combining the degradation curves in EK‐ and Bio‐only experiments. The results show that the simulated curve fitted well with the actual degradation curve in electro‐bioremediation (EK‐Bio) experiments for the first 30 days of the experiment, while at later stages a discrepancy was found. This discrepancy was caused by adverse effects of low soil pH (3.6) near the anode on bacteria health during EK treatments. With polarity reversal (PR) to control the soil pH, the simulated curve fitted very well (r > 0.99) with the actual degradation curve during the whole treatment period. At the end of the experiment, PYR removal amounted to 63% with EK‐Bio treatments in PR electric fields, which was 1.7 times that of Bio‐only. Moreover, the bacteria counts under electric fields were more than that without EK. The spatial distributions of PYR degradation and bacterial counts were also investigated. The results show that they were both higher nearer the electrodes under PR electric fields.     

Demonstration of Enhanced Bioremediation in a TCE Source Area at Launch Complex 34, Cape Canaveral Air Force Station

The ability of bioremediation to treat a source area containing trichloroethene (TCE) present as dense nonaqueous phase liquid (DNAPL) was assessed through a laboratory study and a pilot test at Launch Complex 34, Cape Canaveral Air Force Center. The results of microcosm testing indicate that the indigenous microbial community was capable of dechlorinating TCE to ethene if amended with electron donor; however, bioaugmentation with a dechlorinating culture (KB-1; SiREM, Guelph, Ontario, Canada) significantly increased the rate of ethene formation. In microcosms, the activity of the dechlorinating organisms in KB-1 was not inhibited at initial TCE concentrations as high as 2 mM. The initially high TCE concentration in ground water (1.2 mM or 155 mg/L) did not inhibit reductive dechlorination, and at the end of the study, the average concentration of ethene (2.4 mM or 67 mg/L) was in stoichiometric excess of this initial TCE concentration. The production of ethene in stoichiometric excess in comparison to the initial TCE concentration indicates that the bioremediation treatment enhanced the removal of TCE mass (either sorbed to soil or present as DNAPL). Detailed soil sampling indicated that the bioremediation treatment removed greater than 98.5% of the initial TCE mass. Confirmatory ground water samples collected 22 months after the bioremediation treatment indicated that chloroethene concentrations had continued to decline in the absence of further electron donor addition. The results of this study confirm that dechlorination to ethene can proceed at the high TCE concentrations often encountered in source areas and that bioremediation was capable of removing significant TCE mass from the test plot, suggesting that enhanced bioremediation is a potentially viable remediation technology for TCE source areas. Dehalococcoides abundance increased by 2 orders of magnitude following biostimulation and bioaugmentation.     

Gladstone, Australia Field Studies: Weathering and Degradation of Hydrocarbons in Oiled Mangrove and Salt Marsh Sediments With and Without the Application of an Experimental Bioremediation Protocol

This field study was a combined chemical and biological investigation of the relative rates of weathering and biodegradation of oil spilled in sediments and testing the influence of a bioremediation protocol. The aim of the chemistry work presented here was to determine whether the bioremediation protocol affected the rate of penetration, dissipation or long-term retention of a medium range crude oil (Gippsland) and a Bunker C oil stranded in tropical Rhizophora sp. mangrove and Halosarcia sp. salt marsh environments. Permission for the planned oil spills was granted in the Port Authority area of Gladstone, Queensland (Australia). Sediment cores from three replicate plots of each treatment for mangroves and four replicate plots for the salt marsh (oil only and oil plus bioremediation) were analysed for total hydrocarbons (THC) and for individual alkane markers using gas chromatography with flame ionization detection (GC–FID). Sediments were collected at day 2, then 1, 2, 5 or 6 and 12 or 13 months post-spill for mangroves and day 2, 1, 3 and 9 months post-spill for salt marshes. Over this time, hydrocarbons in all of the oil treated plots decreased exponentially. There was no statistical difference in initial oil concentrations, penetration of oil to depth, or in the rates of oil dissipation between untreated oil and bioremediated oil in the mangrove plots. The salt marsh plots treated with the waxy Gippsland oil showed a faster rate of biodegradation of the oil in the bioremediated plots. In this case only, the degradation rate significantly impacted the mass balance of remaining oil. The Bunker C oil contained only minor amounts of highly degradable n-alkanes and bioremediation did not significantly impact its rate of loss in the salt marsh sediments. At the end of each experiment, there were still n-alkanes visible in the gas chromatograms of residual oils. Thus it was concluded that there was unlikely to be any change in the stable internal biomarkers of the oils over this time period. The predominant removal processes in both habitats were evaporation and dissolution, with a lag-phase of 1–2 months before the start of microbial degradation.     

Long‐Term Impacts on Groundwater and Reductive Dechlorination Following Bioremediation in a Highly Characterized Trichloroethene DNAPL Source Area

High‐resolution soil and groundwater monitoring was performed to assess the long‐term impacts of bioremediation using bioaugmentation with a dechlorinating microbial consortium (and sodium lactate as the electron donor) in a well‐characterized trichloroethene (TCE) dense nonaqueous phase liquid (DNAPL) source area. Monitoring was performed up to 3.7 years following active bioremediation using a high‐density monitoring network that included several discrete interval multi‐level sampling wells. Results showed that despite the absence of lactate, lactate fermentation transformation products, or hydrogen, biogeochemical conditions remained favorable for the reductive dechlorination of chlorinated ethenes. In locations where soil data showed that TCE DNAPL sources persisted, local contaminant rebound was observed in groundwater, whereas no rebound or continuous decreases in chlorinated ethenes were observed in locations where DNAPL sources were treated. While ethene levels measured 3.7 years after active treatment suggested relatively low (2 to 30%) dechlorination of the parent TCE and daughter products, carbon stable isotope analysis showed that the extent of complete dechlorination was much greater than indicated by ethene generation and that the estimated first‐order rate constant describing the complete dechlorination of TCE at 3.7 years following active bioremediation was approximately 3.6 y–1. Overall, results of this study suggest that biological processes may persist to treat TCE for years after cessation of active bioremediation, thereby serving as an important component of remedial treatment design and long‐term attenuation.     

Bacteria and organic matter dynamics during a bioremediation treatment of organic-rich harbour sediments

We studied the dynamics of bacteria and organic matter in the Ancient Port of Genoa (Italy) during a bioremediation treatment of sediment (during summer-autumn 1998) in an area characterised by continuous sewage discharge. A strong increase in total benthic bacterial density (TBN) was recorded at the end of the study, from 14 x 10(8) to 58-172 x 10(8) cell g(-1) in different parts of the treated area. The TBN increase was linked to organic matter depletion, from more than 40 to less than 20 mg x g(-1). In order to highlight the main ecological mechanisms involved in bioremediation, a laboratory experiment based on both water and sediment from the basin studied was carried out. We observed an increase in TBN during the first 20 days and a decrease in sediment organic matter (up to about 20%). Increases of organic matter (about 2-fold) and TBN (from 21 to 33 x 10(9) cell l(-1)) occurred in the overlying water, suggesting a strong association between the sediments and water column processes. Hydrolytic activities, which double in the sediment and increase up to a 300-fold in the water, are consistent with the decrease in sediment organic matter and with the water fraction dynamics.     

Assessing TCE Source Bioremediation by Geostatistical Analysis of a Flux Fence

Mass discharge across transect planes is increasingly used as a metric for performance assessment of in situ groundwater remediation systems. Mass discharge estimates using concentrations measured in multilevel transects are often made by assuming a uniform flow field, and uncertainty contributions from spatial concentration and flow field variability are often overlooked. We extend our recently developed geostatistical approach to estimate mass discharge using transect data of concentration and hydraulic conductivity, so accounting for the spatial variability of both datasets. The magnitude and uncertainty of mass discharge were quantified by conditional simulation. An important benefit of the approach is that uncertainty is quantified as an integral part of the mass discharge estimate. We use this approach for performance assessment of a bioremediation experiment of a trichloroethene (TCE) source zone. Analyses of dissolved parent and daughter compounds demonstrated that the engineered bioremediation has elevated the degradation rate of TCE, resulting in a two‐thirds reduction in the TCE mass discharge from the source zone. The biologically enhanced dissolution of TCE was not significant (～5%), and was less than expected. However, the discharges of the daughter products cis‐1,2, dichloroethene (cDCE) and vinyl chloride (VC) increased, probably because of the rapid transformation of TCE from the source zone to the measurement transect. This suggests that enhancing the biodegradation of cDCE and VC will be crucial to successful engineered bioremediation of TCE source zones.     

Response of estuarine meio- and macrofauna to in situ bioremediation of oil-contaminated sediment

Numerous studies have demonstrated the efficacy of bioremediation for enhancing oil removal but the ecological effect on shoreline biota is unclear. Therefore, a field experiment was designed at an intertidal sandflat in SW England to assess the effects of nutrient addition to oiled sediments on meio- and macrofauna for a period of up to 45 weeks. Natural assemblages were exposed to different types of experimental treatments (no oil, oil alone, oil treated with slow-release fertiliser or liquid fertiliser). Bioremediation stimulated the microbial population and increased oil biodegradation. This, however, did not result in faster recolonisation rates of fertilised versus non-fertilised oiled sediments. Mild effects of oil and bioremediation treatments on benthic fauna were observed, including short-term shifts in dominance patterns. Decreased abundance of dominant species in the oiled compared to unoiled sediments resulted in significantly higher evenness of benthic assemblages within the first 11 weeks of the experiment.     

Bioremediation of Chlorinated Pesticides in Field‐Contaminated Soils and Suitability of Tenax Solid‐Phase Extraction as a Predictor of Its Effectiveness

Bioremediation is intensively studied today as a treatment method for soil contaminated with chlorinated pesticides, chemicals counted among persistent organic pollutants. In the presented work, results of desorption kinetics study using consecutive Tenax TA solid phase extraction (SPE) were tested as predictors of 3‐wk anaerobic soil bioremediation effectiveness for chlorinated pesticides γ‐HCH, DDT, and methoxychlor. Field‐contaminated samples were used in these experiments, and conditions of bioremediation tests were based on previous research. Amounts of pesticides removed during bioremediation (43–98% of initial concentrations) were in most cases much larger (average ratio 1.37) than rapidly desorbing fractions estimated in SPE using two‐compartment model of desorption kinetics. The scatter of results was also considerable (standard deviation 0.45). However, there was a statistically significant correlation between amounts removed and rapidly desorbing fractions (R² = 0.64), indicating a relationship between degradability and desorbability. Nonetheless, determination of rapidly desorbing fractions was considered rather a poor indicator of soil bioremediation efficiency for chlorinated pesticides. The total amounts of pesticides desorbed by Tenax in 72 h performed better in this respect (R² = 0.73, fraction removed/desorbed = 1.10 ± 0.20, average ± standard deviation). Disappearance of DDT during bioremediation was accompanied by DDD formation but this was considerably lower than results expected from stoichiometry.     

Integration of Microalgae-Based Wastewater Bioremediation–Biorefinery Process to Promote Circular Bioeconomy and Sustainability: A Review

Bioremediation of wastewater using microalgae is inexpensive, energy efficient, and effective in pollutant reduction as compared to conventional wastewater treatment technologies. Wastewater is a huge resource of minerals, nutrients, bioenergy, and valuable organic compounds and can be used for cultivation of microalgae. The microalgal biomass can be further used as biorefinery feedstock to produce biofuels and commercially important high-value products. The potential of microalgae toward bioremediation and biorefinery applications presents the avenues for integrating the two processes to support circular bioeconomy and sustainability. This review presents a holistic view of integration of bioremediation and biorefinery processes using microalgae for deriving multiple benefits like pollutant removal, resource recovery, biofuel production, and generation of high-value commercial products. The current status of high-throughput microalgal screening technologies is also discussed since the selection of suitable microalgal strains is crucial for the application. The review further summarizes various processes involved in bioremediation and biorefinery systems such as cultivation, bioremediation, harvesting, and downstream processing. Recent trends in microalgal strain improvement for bioremediation and biorefinery applications through genetic engineering, bioinformatics, omics technologies, and genome editing tools are highlighted, while addressing the risks, biosafety issues, and regulatory affairs associated with genetically modified algae.     

Isolation and characterization of two crude oil-degrading yeast strains, Yarrowia lipolytica PG-20 and PG-32, from the Persian Gulf

Among six crude oil-degrading yeasts that were isolated from an oil-polluted area in the Persian Gulf, two yeast strains showed high degradation activity of aliphatic hydrocarbons. From an analysis of 18S rRNA sequences and biochemical characteristics, these strains were identified as Yarrowia lipolytica strains PG-20 and PG-32. Gas Chromatography (GC) analysis of the crude oil remaining in the culture medium after 1 week at 30°C showed that the strains PG-20 and PG-32 degraded 68% and 58% of crude oil, respectively. The optimal growth condition and biodegradation of hydrocarbons was in ONR medium with an acidic pH (pH5). These two strains may degrade aliphatic hydrocarbons more efficiently than aromatic hydrocarbons, although strain PG-20 had better degradation than strain PG-32. The two Y. lipolytica strains reduce surface tension when cultured on hydrocarbon substrates (1% v/v). These strains showed a cell surface hydrophobicity higher than 70%. These results suggested that Y. lipolytica strains PG-20 and PG-32 have high crude oil degrading activity due to their high emulsifying activity and cell hydrophobicity. In conclusion, these yeast strains can be useful for the bioremediation process in the Persian Gulf and decreasing oil pollution in this marine ecosystem.     

Bacterial Production Stimulated Across the Zone of Influence of a Ground Water Circulation Well in a BTEX-Contaminated Aquifer

Benzene, toluene, ethylbenzene, and xylene (BTEX) hydrocarbons are typically the most abundant carbon source for bacteria in gasoline-contaminated ground water. In situ bioremediation strategies often involve stimulating bacterial heterotrophic production in an attempt to increase carbon demand of the assemblage. This may, in turn, stimulate biodegradation of contaminant hydrocarbons. In this study, ground water circulation wells (GCWs) were used as an in situ treatment for a fuel-contaminated aquifer to stimulate bacterial production, purportedly by increasing oxygen transfer to the subsurface, circulating limiting nutrients, enhancing bioavailability of hydrocarbons, or by removing metabolically inhibitory volatile organics. Bacterial production, as measured by rates of bacterial protein synthesis, was stimulated across the zone of influence (ZOI) of a series of GCWs. Productivity increased from ∼10² to >10⁵ ng C/L hour across the ZOI, suggesting that treatment stimulated overall biodegradation of carbon sources present in the ground water. However, even if BTEX carbon met all bacterial carbon demand, biodegradation would account for <4.3% of the total estimated BTEX removed from the ground water. Although bacterial productivity measurements alone cannot prove the effectiveness of in situ bioremediation, they can estimate the maximum amount of contaminant that may be biodegraded by a treatment system.     

Experimental study on reduction of U (Ⅵ) by an anaerobic bacterium, Shewanella putrefaciens: Application to sandstone-hosted interlayer oxidation-zone type uranium deposits, China

An experimental study on reduction of U (Ⅵ) by anaerobic bacteria, Shewane//a putrefaciens, is first reported here in China. The experimental conditions were: 35℃ and pH =7.0-7.4, corresponding to a physicochemical environments in which the sandstone-hosted interlayer oxidation-zone type uranium deposit formed in Northwest China's Xinjiang. Bacteria adopted in the present experiment, Shewanella putrefaciens, occur extensively in natural environment. Our study shows that nano-crystal precipitates of uraninite quickly occurred on the surface of the cells within one week. It was found that the pitchblende was characterized by a random arrangement of uraninite nanocrystals (2-4 nm) in it, significantly different from natural pitchblende in which uraninite nanocrystals are arranged in order. Finally, a possible mechanism of uranium biomineralization by microorganisms in the deposits is discussed. Our investigation may supply a technical train of thoughts for bioremediation of nuclear-contaminated water environments and for underground dissolving extraction of the sandstone-hosted uranium ores.     

Estimating Reaction Rate Coefficients Within a Travel‐Time Modeling Framework

A generalized, efficient, and practical approach based on the travel‐time modeling framework is developed to estimate in situ reaction rate coefficients for groundwater remediation in heterogeneous aquifers. The required information for this approach can be obtained by conducting tracer tests with injection of a mixture of conservative and reactive tracers and measurements of both breakthrough curves (BTCs). The conservative BTC is used to infer the travel‐time distribution from the injection point to the observation point. For advection‐dominant reactive transport with well‐mixed reactive species and a constant travel‐time distribution, the reactive BTC is obtained by integrating the solutions to advective‐reactive transport over the entire travel‐time distribution, and then is used in optimization to determine the in situ reaction rate coefficients. By directly working on the conservative and reactive BTCs, this approach avoids costly aquifer characterization and improves the estimation for transport in heterogeneous aquifers which may not be sufficiently described by traditional mechanistic transport models with constant transport parameters. Simplified schemes are proposed for reactive transport with zero‐, first‐, nth‐order, and Michaelis‐Menten reactions. The proposed approach is validated by a reactive transport case in a two‐dimensional synthetic heterogeneous aquifer and a field‐scale bioremediation experiment conducted at Oak Ridge, Tennessee. The field application indicates that ethanol degradation for U(VI)‐bioremediation is better approximated by zero‐order reaction kinetics than first‐order reaction kinetics.     

Oxygen and Ethene Biostimulation for a Persistent Dilute Vinyl Chloride Plume

Contamination of groundwater with chlorinated ethenes is common and represents a threat to drinking water sources. Standard anaerobic bioremediation methods for the highly chlorinated ethenes PCE and TCE are not always effective in promoting complete degradation. In these cases, the target contaminants are degraded to the daughter products DCE and/or vinyl chloride. This creates an additional health risk, as vinyl chloride is even more toxic and carcinogenic than its precursors. New treatment modalities are needed to deal with this widespread environmental problem. We describe successful bioremediation of a large, migrating, dilute vinyl chloride plume in Massachusetts with an aerobic biostimulation treatment approach utilizing both oxygen and ethene. Initial microcosm studies showed that adding ethene under aerobic conditions stimulated the rapid degradation of VC in site groundwater. Deployment of a full‐scale treatment system resulted in plume migration cutoff and nearly complete elimination of above‐standard VC concentrations.     

Experimental study on reduction of U (VI) by an anaerobic bacterium, <Emphasis Type="Italic">Shewanella putrefaciens</Emphasis>: Application to sandstone-hosted interlayer oxidation-zone type uranium deposits,China

An experimental study on reduction of U (VI) by anaerobic bacteria, Shewanella putrefaciens, is first reported here in China. The experimental conditions were: 35°C and pH= 7.0-7.4, corresponding to a physicochemical environments in which the sandstone-hosted interlayer oxidation-zone type uranium deposit formed in Northwest China’s Xinjiang. Bacteria adopted in the present experiment, Shewanella putrefaciens, occur extensively in natural environment. Our study shows that nano-crystal precipitates of uraninite quickly occurred on the surface of the cells within one week. It was found that the pitchblende was characterized by a random arrangement of uraninite nanocrystals (2-4 nm) in it, significantly different from natural pitchblende in which uraninite nanocrystals are arranged in order. Finally, a possible mechanism of uranium biomineralization by microorganisms in the deposits is discussed. Our investigation may supply a technical train of thoughts for bioremediation of nuclear-contaminated water environments and for underground dissolving extraction of the sandstone-hosted uranium ores.     

Analytical approximation to characterize the performance of in situ aquifer bioremediation

The performance of in situ bioremediation to remove organic contaminants from contaminated aquifers depends on the physical and biochemical parameters. We characterize the performance by the contaminant removal rate and the region where biodegradation occurs, the biologically active zone (BAZ). The numerical fronts obtained by one-dimensional in situ bioremediation modeling reveal a traveling wave behavior: fronts of microbial mass, organic contaminant and electron acceptor move with a constant velocity and constant front shape through the domain. Hence, only one front shape and a linear relation between the front position and time is found for each of the three compounds. We derive analytical approximations for the traveling wave front shape and front position that agree perfectly with the traveling wave behavior resulting from the bioremediation model. Using these analytical approximations, we determine the contaminant removal rate and the BAZ. Furthermore, we assess the influence of the physical and biochemical parameters on the performance of the in situ bioremediation technique.     

Biocurtain Design Using Reactive Transport Models

Bioremediation is an attractive alternative to traditional remediation methods for a variety of ground water contaminants. However, widespread implementation of bioremediation is currently limited by the complexity of the dynamic chemical and biological processes that need to be understood and incorporated into the design approach. Reactive transport models provide a powerful tool to simulate these complex interactions and, thus, can be used to improve and guide the design of bioremediation systems. We present a remediation design approach for intermittently stimulated biodegradation using multicomponent reactive transport models, parameterized using a series of nondimensional Damkohler numbers. Designs were based on either (1) a target aqueous phase concentration at the exit of the treatment system, or (2) the total contaminant mass fraction removed from a region of interest. The equation set used to develop this design approach is specific to the case of intermittent electron donor addition to drive cometabolic transformations. We illustrate the design procedure for a biocurtain that removes carbon tetrachloride. Our results for this case indicate that intermittent injection is significantly more efficient than strategies based on continuous pumping. Example design parameters include the length of the biologically active zone (i.e., biocurtain), the effective rate of degradation in this zone, and the interval between electron donor injection cycles. The presented dimensionless parametric approach can be used to design bench-scale column studies and should be helpful for scale-up to field-scale remediation systems.     

Dispersant Use and a Bioremediation Strategy as Alternate Means of Reducing Impacts of Large Oil Spills on Mangroves: The Gladstone Field Trials

Over a three-year period (1995–1998), we studied short-term effects of dispersant use and a bioremediation strategy in two consecutive field trials in sub-tropical Australian mangroves. In each case, weathered oil was applied, and a large spill simulated, in mature Rhizophora stylosa trees around 4–9 m tall. In the first trial, we used Gippsland light crude oil with or without dispersant, Corexit 9527. In the second, a bioremediation strategy followed application of Gippsland oil or Bunker C fuel oil. Bioremediation involved forced aeration with supplemental application of nutrients. Dispersant use had an overall positive benefit shown as reduced tree mortality. By contrast, there was no apparent reduction in mortality of trees with bioremediation. However, one year after oiling, leaf densities of surviving trees were greater in bioremediation plots than in controls, and less in oil-only plots. These and other results have been incorporated into spill response management strategies in Australia.