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Impact of present and future regulations on bioremediation.

Innovative treatment technologies are in increasing demand to clean up the nation's existing environmental contamination. There also are mounting pressures for industry to minimize the production or generation of hazardous pollutants. Bioremediation is a viable, cost-effective treatment option for both field remediation and treatment in enclosed systems. The use of innovative treatment technologies is largely regulatory driven. Over the last two decades, at least a dozen Federal environmental statutes have been enacted and hundreds of regulations implemented to control releases of pollutants into the air, water and on land. These statutes not only have created markets for the use of treatment technologies, they also may regulate some aspect of the application of that technology. Regarding bioremediation, four statutes should be reviewed to determine if compliance is necessary before employing microorganisms in the field or in enclosed systems. This paper summarizes the Federal statutes (i.e., the Toxic Substances Control Act (TSCA); the Resource Conservation and Recovery Act (RCRA); the Comprehensive Environmental Response, Compensation and Liability Act (CERCLA); and the Federal Plant Pest Act (FPPA], and regulations that may impact the bioremediation industry; outlines potential markets for bioremediation that are being driven by regulations; and highlights, within the regulatory framework, promising applications for the bioremediation of hazardous wastes.

Biotechnology↗

Effect of drying on bioremediation bacteria properties.

Bioremediation bacteria with drought-resistance characteristics were selected and compared to a collection of 10 strains selected only for their bioremediation properties. Twenty-six strains were selected from dried diesel-polluted soil, and they exhibit a better level of survival during drying, compared to collection bioremediation strains (two orders of magnitude difference). The lyophilization process does not affect the strains' ability to grow on xenobiotic compound when measured immediately after drying. However, collection bioremediation strains selected only for their bioremediation properties lose up to 80% of their properties when stored at 25 degrees C for 15 d, but the strains selected for their drought resistance lose their properties to a lesser extent during the same period. The maximal growth rate and the rate of xenobiotic degradation of the still-active cells are not affected by the drying process.

Bacteria↗

Bioremediation of hydrocarbon-contaminated polar soils.

Bioremediation is increasingly viewed as an appropriate remediation technology for hydrocarbon-contaminated polar soils. As for all soils, the successful application of bioremediation depends on appropriate biodegradative microbes and environmental conditions in situ. Laboratory studies have confirmed that hydrocarbon-degrading bacteria typically assigned to the genera Rhodococcus, Sphingomonas or Pseudomonas are present in contaminated polar soils. However, as indicated by the persistence of spilled hydrocarbons, environmental conditions in situ are suboptimal for biodegradation in polar soils. Therefore, it is likely that ex situ bioremediation will be the method of choice for ameliorating and controlling the factors limiting microbial activity, i.e. low and fluctuating soil temperatures, low levels of nutrients, and possible alkalinity and low moisture. Care must be taken when adding nutrients to the coarse-textured, low-moisture soils prevalent in continental Antarctica and the high Arctic because excess levels can inhibit hydrocarbon biodegradation by decreasing soil water potentials. Bioremediation experiments conducted on site in the Arctic indicate that land farming and biopiles may be useful approaches for bioremediation of polar soils.

Antarctic Regions↗

Linking genetics, physiology and ecology: an interdisciplinary approach for advancing bioremediation.

Our understanding of microbial catabolic pathways relevant to bioremediation has been shaped by laboratory studies using isolated pollutant-degrading microorganisms. Recent investigations of natural microbial communities have, however, suggested that catabolic populations in the environment are much more diverse than those previously isolated in the laboratory. In addition, most laboratory strains are now thought to constitute minor populations in the environment, sharing only small contributions to bioremediation processes. Currently, attempts to isolate microorganisms that constitute major populations in the environment have been initiated with the aid of molecular ecological techniques. Such studies will provide information more directly relevant to the catabolic reactions occurring in bioremediation processes and are thus expected to help develop new strategies for advancing bioremediation. This article outlines our studies on phenol-degrading bacteria in activated sludge to illustrate a possible scheme of how genetic and physiological information obtained in the laboratory can be applied to advancing bioremediation processes.

Journal Article↗

Bioremediation of 2,4,6-trinitrotoluene contaminated soil in slurry and column reactors.

The bioremediation of 2,4,6-trinitrotoluene (TNT) contaminated soil was performed on a laboratory scale. To compare bioremediation methods, a soil slurry reactor and a soil column reactor were operated and the effects of supplemental sources were investigated. Optimal conditions for the two bioremediation systems for the removal of TNT were obtained. In the soil slurry reactor, about 60% of the 1000 mg/kg TNT contaminated soil was degraded after 10 d, nearly complete biodegradation (>99%) was achieved within 25 d, and the microorganisms grew and reached a maximum of 9.5 x 10(9) CFU/ml at 15 d. In the soil column reactor, about 50% of the 1000 mg/kg TNT contaminated soil was degraded after 25 d and nearly complete biodegradation (>99%) was achieved within 60 d. Microorganisms grew and reached a maximum of 9.8 x 10(10) CFU/g soil at 40 d. These results should help in determining the best bioremediation method and improving the design and operation of large scale clean up of contaminated sites by bioremediation systems.

Journal Article↗

Cleaning up with genomics: applying molecular biology to bioremediation.

Bioremediation has the potential to restore contaminated environments inexpensively yet effectively, but a lack of information about the factors controlling the growth and metabolism of microorganisms in polluted environments often limits its implementation. However, rapid advances in the understanding of bioremediation are on the horizon. Researchers now have the ability to culture microorganisms that are important in bioremediation and can evaluate their physiology using a combination of genome-enabled experimental and modelling techniques. In addition, new environmental genomic techniques offer the possibility for similar studies on as-yet-uncultured organisms. Combining models that can predict the activity of microorganisms that are involved in bioremediation with existing geochemical and hydrological models should transform bioremediation from a largely empirical practice into a science.

Bacteria↗

Comparison of the potential of coastal materials loaded with bacteria for bioremediating oily sea water in batch culture.

The objective of this paper was to study whether the bioremediation potential of coastal materials for oil-polluted sea water depended on the numbers of hydrocarbon-utilizing bacteria they naturally harbor. Inshore water of the Arabian Gulf was found to contain only about one thousand hydrocarbon-utilizing bacteria per ml. Coastal sand, cyanobacterial mats and epilithic biomass were much richer in these bacteria, with numbers ranging between several thousand fold to several million fold than in the water body. The predominant bacterium in all samples was Acinetobacter calcoaceticus, next in predominance were nocardioforms and Micrococcus sp. Inoculation, in batch cultures, of oily sea water or sea water containing pure hydrocarbons with fresh sea water, coastal sand, cyanobacterial mats or epilithic biomass harboring significantly different numbers of hydrocarbon-utilizing bacteria brought bioremediation effects that depended on fertilizing with KNO3. In the absence of KNO3, the bioremediation effect increased with numbers of hydrocarbon-utilizing bacteria in the inoculum. In the presence of KNO3 similar bioremediation effects were found irrespective of the inoculated materials. The reason may be that bacteria reproduce quickly in closed cultures provided with nitrogen, reaching equal maximum numbers, irrespective of the inoculum size. This information could be useful in constructing technologies for oily sea water bioremediation.

Bacteria↗

Volatile hydrocarbon emissions from a diesel fuel-contaminated soil bioremediation facility.

Concerns have been expressed that emissions of volatile hydrocarbons (HCs) from bioremediation facilities containing soils contaminated with petroleum HCs may negatively impact regional air quality or human health. Little information is available regarding the emission of HCs from bioremediation sites, and few field studies have been performed during which the flux of HCs has been directly measured during bioremediation. To aid in answering questions about the impact of bioremediation facilities on the atmospheric environment, a two-part field study was conducted over summer 1996 at a remote landfarm in northern Ontario where diesel fuel-contaminated soil was undergoing bioremediation. Volatile total hydrocarbon (THC) atmospheric flux measurements were successfully taken over 18 days using a flux gradient micrometeorological technique incorporating a THC detector constructed in-house. Peak THC emissions reached 131 microg C/m2/sec shortly after implementation and tilling of the landfarm soil. The influence of soil temperature and tillage on THC emissions was examined. Off-site inhalation exposure was considered with the aid of an areal source model and results from speciated air samples collected on sorbent tubes and analyzed via gas chromatography/mass spectrometry (GCMS) techniques.

Biodegradation, Environmental↗

Bioremediation potential of terrestrial fuel spills.

A bioremediation treatment that consisted of liming, fertilization, and tilling was evaluated on the laboratory scale for its effectiveness in cleaning up a sand, a loam, and a clay loam contaminated at 50 to 135 mg g of soil by gasoline, jet fuel, heating oil, diesel oil, or bunker C. Experimental variables included incubation temperatures of 17, 27, and 37 degrees C; no treatment; bioremediation treatment; and poisoned evaporation controls. Hydrocarbon residues were determined by quantitative gas chromatography or, in the case of bunker C, by residual weight determination. Four-point depletion curves were obtained for the described experimental variables. In all cases, the disappearance of hydrocarbons was maximal at 27 degrees C and in response to bioremediation treatment. Poisoned evaporation controls underestimated the true biodegradation contribution, but nevertheless, they showed that biodegradation makes only a modest contribution to gasoline disappearance from soil. Bunker C was found to be structurally recalcitrant, with close to 80% persisting after 1 year of incubation. The three medium distillates, jet fuel, heating oil, and diesel oil, increased in persistence in the listed order but responded well to bioremediation treatment under all test conditions. With bioremediation treatment, it should be possible to reduce hydrocarbons to insignificant levels in contaminated soils within one growing season.

Journal Article↗

Field evaluations of marine oil spill bioremediation.

Bioremediation is defined as the act of adding or improving the availability of materials (e.g., nutrients, microorganisms, or oxygen) to contaminated environments to cause an acceleration of natural biodegradative processes. The results of field experiments and trials following actual spill incidents have been reviewed to evaluate the feasibility of this approach as a treatment for oil contamination in the marine environment. The ubiquity of oil-degrading microorganisms in the marine environment is well established, and research has demonstrated the capability of the indigenous microflora to degrade many components of petroleum shortly after exposure. Studies have identified numerous factors which affect the natural biodegradation rates of oil, such as the origin and concentration of oil, the availability of oil-degrading microorganisms, nutrient concentrations, oxygen levels, climatic conditions, and sediment characteristics. Bioremediation strategies based on the application of fertilizers have been shown to stimulate the biodegradation rates of oil in aerobic intertidal sediments such as sand and cobble. The ratio of oil loading to nitrogen concentration within the interstitial water has been identified to be the principal controlling factor influencing the success of this bioremediation strategy. However, the need for the seeding of natural environments with hydrocarbon-degrading bacteria has not been clearly demonstrated under natural environmental conditions. It is suggested that bioremediation should now take its place among the many techniques available for the treatment of oil spills, although there is still a clear need to set operational limits for its use. On the basis of the available evidence, we have proposed preliminary operational guidelines for bioremediation on shoreline environments.

Bacteria↗

Bioprospecting Chromobacterium violaceum for bioremediation: an alternative to environmental lead pollution.

Lead pollution is a major environmental concern, but current decontamination technologies remain limited due to high costs. Therefore, alternative biotechnological processes have been successfully developed and applied due to their reduced cost and lower aggressiveness in the environment. The remarkable adaptive versatility of Chromobacterium violaceum in metal-contaminated environments makes this bacterium a promising candidate for Pb bioremediation. Therefore, the reference strain C. violaceum ATCC 12,472 and the environmental isolate C. violaceum SCV1, the first strain of this species isolated from a Brazilian area with natural Pb occurrence, were evaluated for Pb resistance under different Pb concentrations and exposure times. Pb biosorption was assessed by scanning electron microscopy, while strain-specific protein profiles were characterized using tandem mass spectrometry-based proteomic analysis. The results obtained revealed the potential of C. violaceum to perform lead bioremediation. Scanning electron microscopy analysis confirmed the biosorption of lead by C. violaceum strains. C. violaceum SCV1 was able to remove up to 40% more lead concentration when compared to ATCC 12,472 which suggested adaptation through natural selection process of C. violaceum SCV1. Proteome analysis revealed 1531 proteins, of which several are candidates for lead bioremediation. This is the first study on the resistance proteomics of C. violaceum against lead. The acclimatization of the bacteria linked to the identification of several proteins related to: biosorption; efflux and ionic uptake (bioaccumulation); biomolecule transport; and biomethylation, point out to this organism as a potential lead bioremediation agent.

Chromobacterium↗

Water a major source of endocrine-disrupting chemicals: An overview on the occurrence, implications on human health and bioremediation strategies.

Endocrine disrupting chemicals (EDCs) are toxic compounds that occur naturally or are the output of anthropogenic activities that negatively impact both humans and wildlife. A number of diseases are associated with these disruptors, including reproductive disorders, cardiovascular disorders, kidney disease, neurological disorders, autoimmune disorders, and cancer. Due to their integral role in pharmaceuticals and cosmetics, packaging companies, agro-industries, pesticides, and plasticizers, the scientific awareness on natural and artificial EDCs are increasing. As these xenobiotic compounds tend to bioaccumulate in body tissues and may also persist longer in the environment, the concentrations of these organic compounds may increase far from their original point of concentrations. Water remains as the major sources of how humans and animals are exposed to EDCs. However, these toxic compounds cannot be completely biodegraded nor bioremediated from the aqueous medium with conventional treatment strategies thereby requiring much more efficient strategies to combat EDC contamination. Recently, genetically engineered microorganism, genome editing, and the knowledge of protein and metabolic engineering has revolutionized the field of bioremediation thereby helping to breakdown EDCs effectively. This review shed lights on understanding the importance of aquatic mediums as a source of EDCs exposure. Furthermore, the review sheds light on the consequences of these EDCs on human health as well as highlights the importance of different remediation and bioremediation approaches. Particular attention is paid to the recent trends and perspectives in order to attain sustainable approaches to the bioremediation of EDCs. Additionally, rigorous restrictions to preclude the discharge of estrogenic chemicals into the environment should be followed in efforts to combat EDC pollution.

Animals↗

Effects of humic substances and soya lecithin on the aerobic bioremediation of a soil historically contaminated by polycyclic aromatic hydrocarbons (PAHs).

The high hydrophobicity of polycyclic aromatic hydrocarbons (PAHs) strongly reduces their bioavailability in aged contaminated soils, thus limiting their bioremediation. The biodegradation of PAHs in soils can be enhanced by employing surface-active agents. However, chemical surfactants are often recalcitrant and exert toxic effects in the amended soils. The effects of two biogenic materials as pollutant-mobilizing agents on the aerobic bioremediation of an aged-contaminated soil were investigated here. A soil historically contaminated by about 13 g kg(-1) of a large variety of PAHs, was amended with soya lecithin (SL) or humic substances (HS) at 1.5% w/w and incubated in aerobic solid-phase and slurry-phase reactors for 150 days. A slow and only partial biodegradation of low-molecular weight PAHs, along with a moderate depletion of the initial soil ecotoxicity, was observed in the control reactors. The overall removal of PAHs in the presence of SL or HS was faster and more extensive and accompanied by a larger soil detoxification, especially under slurry-phase conditions. The SL and HS could be metabolized by soil aerobic microorganisms and enhanced the occurrence of both soil PAHs and indigenous aerobic PAH-degrading bacteria in the reactor water phase. These results indicate that SL and HS are biodegradable and efficiently enhance PAH bioavailability in soil. These natural surfactants significantly intensified the aerobic bioremediation of a historically PAH-contaminated soil under treatment conditions similar to those commonly employed in large-scale soil bioremediation.

Biodegradation, Environmental↗

Ecotoxicity monitoring of hydrocarbon-contaminated soil during bioremediation: a case study.

The ecotoxicity of hydrocarbon-contaminated soil originating from a brownfield site was evaluated during a 17-month biodegradation pilot test. The initial concentration of total petroleum hydrocarbons (TPHs) in the soil was 6380 microg/g dry weight. An amount of 200 kg soil was inoculated with 1.5 L of the bacterial preparation GEM-100 containing Pseudomonas sp. and Acinetobacter sp. strains (5.3 x 10(10) CFU.mL(-1)) adapted to diesel fuel. The concentration of TPHs in the soil decreased by 65.5% after bioremediation. Different organisms such as the bacterium Vibrio fischeri, terrestrial plants Sinapis alba, Lactuca sativa, and Hordeum vulgare, the water plant Lemna minor, the earthworm Eisenia fetida, and the crustacean Heterocypris incongruens were used for ecotoxicity evaluation. The highest toxicity was detected in the first period of bioremediation. However, certain toxic effects were detectable during the whole bioremediation process. The contact tests with plants, earthworms, and crustaceans were the most sensitive of all of the bioassays. Therefore, the contact tests performed directly on soil samples were shown to be a better tool for ecotoxicity evaluation of hydrocarbon-contaminated soil than the tests performed on soil elutriates. The ecotoxicity measured by the responses of the tests did not always correlate with the decrease in TPH concentrations in the soil during bioremediation.

Aliivibrio fischeri↗

Petroleum hydrocarbon bioremediation: sampling and analytical techniques, in situ treatments and commercial microorganisms currently used.

The sampling and analytical methods, along with available microorganisms, used for in situ hydrocarbon bioremediation are reviewed. Each treatment method is briefly described and its advantages and limitations pertaining to potential applications are evaluated. Bioremediation provides cost-effective, contaminant- and substrate-specific treatments equally successful in reducing the concentrations of single compounds or mixtures of biodegradable materials. In situ treatments rarely yield undesirable byproducts, but precautions and preliminary baseline tests are always recommended. Sampling methods should adhere to good laboratory and field practices and usually do not require highly trained personnel. Analytical methods vary in sensitivity, cost, duration of sample analysis and personnel training required. Voucher specimens of bacterial strains used in bioremediation exist in various repositories (e.g. ATCC, DSM, etc.) or are commercially available, and are usually covered by patent rights. Each one of these strains may yield spectacular results in vitro for specific target compounds. However, the overall success of such strains in treating a wide range of contaminants in situ remains limited. The reintroduction of indigenous microorganisms isolated from the contaminated site after culturing seems to be a highly effective bioremediation method, especially when microorganism growth is supplemented by oxygen and fertilizers.

Bacteria↗

Plant aided bioremediation in the vadose zone: model development and applications.

Phytoremediation has the potential to enhance clean up of land contaminated by various pollutants. A mathematical model that includes a two-fluid phase flow model of water flow as well as a two-region soil model of contaminant reactions was developed and applied to various bioremediation scenarios in the unsaturated zone, especially to plant-aided bioremediation. To investigate model behavior and determine the main parameters and mechanisms that affect bioremediation in unplanted and planted soils, numerical simulations of theoretical scenarios were conducted before applying the model to field data. It is observed from the results that parameters affecting the contaminant concentration in the water phase, such as aqueous solubility, the octanol-water partition coefficient, and organic carbon content of the soil controlled the contaminant fate in the vadose zone. Simulation using the developed model also characterized the fate and transport of the contaminants both in planted and unplanted soils satisfactorily for field applications. Although phytoremediation has the potential for remediation of contaminated soils, results from both modeling and field studies suggested that plants may not always enhance the remediation efficiency when the soil already has a high microbial concentration, when the contaminant bioavailability is low, or when the overall reaction is mass transfer-limited. Therefore, other steps to increase contaminant bioavailability are needed in phytoremediation applications; natural purification mechanisms such as aging, volatilization, and natural bioremediation should be considered to maximize the plant effect and minimize the cost.

Biodegradation, Environmental↗

Application of a new electrolyte circulation method for the ex situ electrokinetic bioremediation of a laboratory-prepared pentadecane contaminated kaolinite.

Ex situ electrokinetic (EK) bioremediation of a laboratory-prepared pentadecane-contaminated kaolinite was carried out. Extraneous bacteria and ionic nutrients were continuously supplied to the soil specimen by a new electrolyte circulation method, which controlled electrical pH change of electrolyte solution to keep bacterial activity. During the EK bioremediation the anode region showed the highest colony forming unit (CFU) due to electrical attraction between anode and bacteria. Simultaneous increases of CFU and uniform pentadecane removal in most soil regions demonstrated that electro-osmosis as well as electrophoresis affected the bacterial transport in soil. At 3.13 mA/cm2, increase in soil temperature to above 45 degrees C inhibited bacterial activity, which caused the decrease of removal efficiency. The removal amount of pentadecane increased with initial pentadecane concentration at the same current densities (0.63 and 1.88 mA/cm2) because of the increased amount of weakly bound pentadecane onto the soil surface. The highest removal efficiency (77.6%) was obtained at 0.63 mA/cm2 for 1000 mg/kg pentadecane after 14 days. Consequently, the present methods of EK bioremediation demonstrated superiority over the conventional bioremediation, which had inherent demerits of slow degradation and low removal efficiency.

Alkanes↗

Application of microbial enumeration technique to evaluate the occurrence of natural bioremediation.

Natural bioremediation is believed to be the major processes that account for both containment of the petroleum-hydrocarbon plume and reduction of the contaminant concentrations. In this study, the feasibility of applying the microbial enumeration technique was assessed for natural biodegradation evaluation at three selected gasoline spill sites. At each site, two monitor wells were installed along the groundwater flow, and one multilevel sampler (MLS) was installed to delineate the vertical distribution of the contaminant plume. Two continuous soil cores were collected at each site to evaluate the horizontal distribution of the microbial activity. Soil samples were used for microbial enumeration, grain-size distribution analysis, and sediment extraction. In situ tracer study was performed using 1,2,4-trimethylbenzene (1,2,4-TMB) as the tracer to study the efficiency of contaminant biodegradation. Investigation results demonstrate the agreement between microbial enumeration and other evaluation techniques for natural bioremediation (e.g., geochemical indicator analysis, tracer study). Results suggest that the microbial enumeration is useful in assessing the occurrence, efficiency, and status of the bioremediation. Because this technique provides insight into factors controlling contaminant biodegradation, it could be performed as the supplemental method for natural bioremediation evaluation at petroleum-hydrocarbon spill sites.

Benzene Derivatives↗