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BIOREMEDIATION

What is Bioremediation ?
Contamination caused by pollution may occur due to : i. ii. iii. iv. the draining of liquid wastes on open ground, dumping wastes; sludges on unused plots, accidental leakage of storage tanks and underground pipelines or spillage from tankers on land or in the sea.

Natural biodegradation in such circumstances may be slow or nil for any of the following reasons i. ii. iii. iv. v. (i) bioavailability (ii) physical, chemical factors unfavourable (pH, temperature, moisture, redox potential, electron acceptor) iii) recalcitrant nature of substance due to basic chemistry of pollutant (iv) xenobiotic nature (v) lack of useful microflora.

'Bioremediation' encompasses biological methods for clean-up of contaminated soil and groundwater and even marine environment (in case of oil spills). It is also referred to as 'Biorestoration'. 'Bioremediation' means giving nature a helping hand. 'Bioremediation' involves establishing the conditions in contaminated environment so that appropriate micro-organisms flourish and carry out the metabolic activities to detoxify the contaminants. During bioremediation micro-organisms may use the contaminants (hazardous chemicals) as nutrients or energy source or it may be degraded by cometabolism. 'Establishing suitable conditions for bioremediation' may mean adding nutrients to promote the growth of particular organisms, adding terminal electron acceptor (O2 or NO3), adjusting moisture conditions or raising the temperature etc. The basic concept is to provide critical environmental requirements which may be adverse in particular site. Unavailability of critical environmental factors for microbial activity may be responsible for the persistence of the otherwise degradable substances in many places. Bioremediation is expected to degrade toxic hazardous organic contaminants from the site to environmentally safe levels. Bioremediation has a vast potential to treat soil and groundwater contaminated by a variety of hazardous chemicals, including refractory organics, oils, benzene, styrene, vinyl chloride, pentachlorophenols, polyaromatic hydrocarbons (PAHs), toluene, xylene, phenols etc.

Advantages of Bioremediation (1) Bioremediation harnesses natural biogeochemical processes. (2) Cost-effective alternative. (3) Toxic chemicals are destroyed or removed from environment and not merely separated. (4) Low capital expenditure (5) Less energy required when compared with other technologies. (6) Less manual supervision.

Constraints of Bioremediation 1.
The process of bioremediation is slow. Time required is in day to months.

e.e. petroleum refining. For other sites at which receptors are at risk. leather tanning. Types of Bioremediation Bioremediation operations can be 'on-site' or 'off-site'. y y Soil Banking (composting) This is an improvement further. Bioremediation may be required of surface soils and sludges or subsurface soil. The mixture is carefully cultivated.5.3. sewage sludge. kerosene contaminated soil etc. The actual treatment zone may be several feet deep.5. Landfarming is suggested for oil sludges as a low cost disposal method. aeration). For sites at which contaminants of concern pose no significant risk to sensitive receptors such as water supply wells and surface water bodies. y y Landfarming The simplest biotechnological solution in use today is the landfarming. 3. chemical degradation also may occur along with biodegradation.7. Wastes are stabilized to less complex materials and water contents and mass decrease. solids i. A stronger scientific base is required for rational designing of process and success. oil sludges from oil refinery operations are treated with micro-organisms after addition of lime. food processing.2. nitrogen-phosphorus fertilizers and carrying out tillage (i. Hydrolysis. Design and engineering of the system based on treatability results. Bioremediation is the naturally occurring process by which microorganisms transform environmental contaminants to innocuous end products. Wastes from brewing. Volatilization. Innovative biotreatment technologies should be responsive to public concern.5 triazine (RDX) and octahydro 1. paper and pulp production. tatraazocine (HMX) are the explosives which are significantly transformed by composting process. For in situ bioremediation site must have soil with high permeability. Least attention required during the process is the advantage of these methods while longer time duration is the disadvantage. physiology and genetic expression and site engineering.3. Land farming involves degradation in upper 6-12 inches of contaminated soil. leaching. The technique requires more land space and more time. in situ or ex situ. sometimes mixed in with a layer of normal soil on high-density plastic liners. soils contaminated with petroleum products are treated by composting. ex situ treatment may be appropriate. tetranitro 1. 5. Trinitrotoluene (TNT). It does not remove all quantities of contaminants. Tilting is done periodically for aeration and mixing. (Often biotreatment will be combined with other technologies).land farming and soil banking or (2) Liquid-solid slurry process. food processing. Wastes are excavated and treated in one metre high heaps. . in situ treatment may be an appropriate remediation strategy. Runoff leachate is collected and may be recycled on heap or treated separately in a bioreactor. Treatability or feasibility studies to determine the best technology to be used. Here contaminated soil is spread in thin layers on a plot and is supplemented by nutrients and micro-organisms and proper pH. concentration of the contaminants and other interfering problems. photolysis. Heavy metals are not removed. water run-off is prevented from the area under treatment. It has a drainage and leachate collection system. Nutrients and microbes may be added to the mix. Considerations required before bioremediation y y y y y y y y Understanding what regulations govern the problem and what action needs to occur.5 trinitro 1. encouraging the microbes within the soil to degrade the waste. water.The land farming and soil banking processes are used to treat the wastes from coal gassification/liquification. antibiotic fermentation wastes. municipal wastewater and sludge.3.3. y y Ex situ Technologies Processes for surface soils and sludges are either (1) Solid phase processes (land treatment) . Substantial gaps exist in the understanding of microbial ecology. hexahydro 1. Characterization of the site. Here oil. Composting is done generally for organic biodegradable wastes. including the type and volume of the matrix to be treated. 4.

organic or inorganic nutrients. dispersants are added to hasten the degradation process. straw. carbon sources such as manure. sediments. Bioslurry process has been demonstrated with a removal rate over 99% for explosive chemicals and a high degree of mineralization. in which simple fuel components like n-alkanes are biodegraded in first step. bioslurry is an option. toluene. The process is similar to the conventional activated sludge process. is required. It may stop or slow the movement of radionuclides in the groundwater. ranging from small (1-5m ) portable units to large plants built specifically for polluted site. and acceptance of the technology by people y . The contaminated materials are mixed into a slurry to allow contact between the micro-organisms and the contaminants.y y Windrow composting Windrow Composting was successfully demonstrated to be effective in treating explosivescontaminated soils in US. For sites requiring greater process control. For groundwater which is contaminated with volatile organic compounds 'Pump and Treat' method is largely used. cables prevent or when groundwater is to be treated). Materials most often treated by this method are wood-preserving and oil refinery wastes containing phenol. alfalfa. Because conditions are optimised for the micro-organisms. windrows are turned periodically using a compost turner. The clean-up of soils and sub-soils is totally dependent on permeabilities and dispersed status of pollutant. similar to composted soils. Windrow composting and Bioslurry process are found to have just half the cost of treatment when compared with incineration process. In situ treatment of groundwater is faster and safer with worker exposure to chemical contaminants being minimum. or where the cost of importing compost amendments is prohibitive. But in situ treatment of groundwater will be more cost-effective. slurry processes are faster than many other biological processes. This helps for proper optimisation of conditions for different organisms. Oxygen is supplied with spargers or floating aerators. Soil leachates or groundwater is pumped to the surface. while more recalcitrant contaminants with multi-ring PAHs are bioattacked in next step. The entire area then acts like a bioreactor. this method is desirable. surfactants. 3 y y In situ Technologies Non-disruptive treatment of contaminated prime site (when excavation is not possible or advisable) is also possible with in situ treatment. groundwater may be extracted and supplemented with organisms and nutrients and re-introducedor nutrient solution is added along with useful micro-organisms by injections into the subsurface through wells and infiltration system. windrow oxygen level. appropriately treated or disposed off. and clays that act as sinks. To facilitate the microbial growth. When contaminated soil is inaccessible (i. The rector may be mobile above ground or lined in situ lagoons or pits. Windrow composting mixes the soil with compost in long piles known as windrows. In. The treated slurry is suitable for direct land application. treated under controlled conditions in the bioreactor and cleaned water is returned to the ground. Neutrlizing agents. Single batch reactor or sequence batch reactor is used. benzene. In situ bioremediation provides a potentially significant benefit for volatile organic compounds (VOCs) and other contaminants that are held up in adsorptive soils or less permeable silts. A slurry biocascade has been developed.e. The liquid phase is discharged in environmentally safe manner. in prime location beneath the building or when buried pipes. Moisture content. and other agricultural products are added. y y Slurry Processes Hazardous wastes in liquid-solid contact are treated in a closed reactor or open pit or lagoon. In 1992. If dewatering. more complete degradation. in situ treatments. and thus water treatment. After treatment solids are allowed to settle. Waste is taken in the reactor in form of slurry and mixed with micro-organisms. naphthalene. To facilitate aeration and heat control. cost will increase. Soil Slurry orbioslurry technology is used for remediating explosives-contaminated soils. and temperature are easily monitored. Consideration of disposed soil placement is affected by risk-based goals and future land use. polycyclic aromatic hydrocarbons. Disposition of the treated soil is an important part of bioslurry process costs. Windrow composting is costeffective technology with modest equipment and monitoring requirements. USAEC demonstrated windrow composting to reduce explosive concentrations over 99% and toxicity over 90-98%. y y y y y Pump and Treat type of processes Other ex-situ technologies include a wide variety of bioreactors.

control of biofouling or excessive microbial growth. The method gives ultimate on-site solution to completely remove contaminant from the concerned environment. inorganic nutrients and oxygen were added and it was allowed to reinfiltrate through contaminated soil. 2. refined petroleum. * Immiscible with water. In 'Bioscreening'. porous walls are inserted into the ground and soaked with micro-organisms which screen out any contamination. Brief about Oil Spills ± 1. effective mixing for contacting micro-organisms. nutrients. Bioremediation can destroy the VOCs in place and reduce the limitations of mass transport associated with VOC adsorption/desorption to sediments and dissolution into the groundwater that occur in pump-and-treat technologies. and adequate tools for designing. The time and cost of clean-up could be substantially reduced if bioremediation could be effectively employed alone. and oil mixed with waste. adequate nutrient delivery systems. and monitoring the performance of in situtechnologies in heterogeneous subsurface environments. including crude oil. contaminants. Biotechnology in treating petroleum sludge and oil spills A. Within 14 weeks concentration of styrene was reduced to less than 0. effectively isolating and treating the area simultaneously. Effects of oil spills are: . The main limitation of this technology when compared with baseline methods is the difficulty in designing and implementing an effective subsurface treatment system for highly heterogeneous media. Characteristics of oil spills are: * Unintentional discharge of liquid hydrocarbons into environment. * Can greatly affect the environment and biological web in location where spill occurred. Styrene spillage in sandy soil which had also reached groundwater could be cleaned by in situ treatment in which groundwater was extracted. or in conjunction with other bulk-contaminant removal technologies. The in situ treatment is carried out for fuel spills. These are.y y y is more. predicting.05 mg/kg. Several key technical issues have prevented widespread use of bioremediation for organic and inorganic subsurface contaminants. * Contain many types of oil. chlorinated solvents and pollutant mixtures. so oil can remain on surface for lengthy periods of time.

Clean-up of oil spills: a. corporations. * Oil is collected into confined area to prevent spread.* Smothering of plants and animals due to oil¶s stickiness and buoyancy. and other biological functions. reproduction. and governments. * In-situ burning involves igniting the oil while still on the ocean surface. * Toxicity due to water soluble portions in oil can lead to mutations in organisms that stunt growth. Possible solution to Oil Spills (Biotechnological way) ± . Mechanical response * Booms and other physical devices aid in clean up. metabolism. Chemical response * Oil broken down with chemicals into constituents. * Disruption of nutrient cycle due to large increase in hydrocarbons. so a better method is desired by environmentalists. physically recovered by skimming surface with boats or vacuums b. 3. Both approaches leave pollutants and are quite costly. dispersed to decrease harm to wildlife. B.

Following points to be noted in relation to oil spills: * Oil spills can be extremely harmful to the environment and need to be cleaned up in efficient. and in the process can clean up spilled oil in a natural manner. when eaten by humans. the main component of petroleum. but biodegrading bacteria are usually specific to one compound. * Certain marine microorganisms can break down complex hydrocarbons for energy. normally discharged into the sea from petroleum refineries. Oil sludge. Biotechnology. however. attacking persistent compounds that other techniques failed to break down. * Certain compounds are promptly absorbed by the soil or water around the spill. can protect the marine environment by breaking down various types of hydrocarbons. normally found in soil. C. The problems that plague oil spill bioremediation through microorganisms * Oil spills involve many different compounds with different chemical structures. present a serious health hazard. * First tested on a large scale to some success in the Exxon-Valdez spill in 1989. they are capable of inserting an oxygen atom into hydrocarbon to form a degradable alcohol or carboxylic acid. has shown that particular species of bacteria and fungi. * Biodegrading bacteria use the oil compounds as energy sources. however. contains toxic compoundhat are a major threat to the marine ecology. * Microorganisms are especially good at cleaning up last stages of an oil spill. . so if the concentration of the compounds is low then the bacteria are unable to produce enzymes that break the compounds down. It may be necessary to use some of the techniques of modern biotechnology to introduce these characteristics into the appropriate oil-eating micro-organisms. cheap ways.* Certain marine microorganisms have the unique ability to break down hydrocarbons to access energy stored in bonds. and contaminated fish. micro-organisms must be able to withstand the marine environment _ for example they need to survive in high salt concentrations and to grow at low temperatures. All forms of aquatic life are adversely affected. * A spill could contain other compounds that harm the biodegrading bacteria instead of providing energy. To be effective in cleaning up marine oil spills. diluting them to a concentration where they can no longer function as the energy sources of the biodegrading bacteria.

Development of root zone treatment Technology for sewage Reeds. nearly 2. The process in a root zone system to treat the sewage is very simple to explain yet complex in nature. which harbor around roots get oxygen form the weak membranes of the roots and aerobically oxidize the organic matter of the effluent.500 types of bacteria and 10. * A bacterium like Alcanivorax borkumensis could provide the base for such an organism.000 types of Fungi. making it hard for one microorganism to clean up a mix of oil.* These bacteria are structure specific. The characteristics of plants . * A genetically modified organism that can break down multiple compounds could be used to clean spills very efficiently. * This GMO could be created by inserting the gene that codes for different hydrocarbon breakdown intoAlcanivorax through a plasmid and then tested for efficiency. The effluent percolates through the bed that has all the roots of the wetland plants spread very thickly . Raw effluent (after removing grit or floating material is passed horizontally or vertically through a bed of soil having impervious bottom. Phragmites australis is considered to be the best plant because of its roots form horizontal rhizome that guarantee a perfect root zone filter bed.

the sewage percolates through bed and that has all roots of the wetland plants spread very thickly nearly with 2500 types of bacteria and 10.TOC and Total coliforms . It is though an effluent treatment plant. It is recommended to combine vertical flow and then horizontal flow of sewage with a soil having impervious bottom. It achieves the standard for tertiary level treatment standard with no operating cost. installed with a drain pipe in a bed of pieces of lime stones and filled up with pebbles. From the reed bed the treated water is collected and used for irrigation by gradient flow. It has no sludge handling problem such as scraping of slurry from the sludge drying beds and its disposal twice in a week. it does not have mosquitoes problem. usually containing common reeds. anaerobic digestion also takes place. The root zone system has low maintenance cost since it involves no machinery and its associated maintenance. Hence the system becomes maintenance free and can run upto 50 to 60 years without any loss of efficiency as has been described.000 types of fungi and aerobically oxidized organic matter of the effluent. iron filings and graded sand. it does not have odour problem and though It is a green zone. the action with fungi etc.of absorbing oxygen through their leaves and passing it down to roots through their stems which are hollow. Away from the roots. Generally reed bed is made in shallow pits. compared to the conventional treatment processes such as activated sludge. y y y y y y y y It enhances the landscape and gives the site a green appeal. aerated lagoons. In the horizontal flow system. The sludge gets mineralized in the vertical zone of the reed bed . waste stabilization pond etc the performance of the root zone treatment system is good with regard to the removal or reduction of BOD. Reed bed is considered as an effective and reliable secondary and tertiary treatment method where land area is not a major constraint.The sludge mineralizing beds needs to be disposed once in 10 or 15 years. There is no chemical used for pH adjustment or for flocculation. The reeds are not grazed by ruminants.COD. industrial and agricultural wastes. reed plants (with hollow root which bring oxygen into the filter bed) are planted It is advantageous to treat the sewage by root zone system. The filtering action of the soil bed. Salinity may not be a problem for a survival or operations of reed beds. In this sandy body. y y However. is utilized as a bio-pump. Reed bed is one of the natural and attractive methods of treating domestic. Root zone system gives a very good performance of removing 90% BOD and 63% Nitrogen. Phragmites australis has been found more efficient in nitrogen removal compared to Typha latifolia. It provides natural habitat for birds and after a few years gives an appearance of a Bird´s sanctuary. Low electricity is consumed for pumping treated water from the collection tank to the reed bed. It requires negligible attendance for operation and monitoring. A reed bed is an engineered method of purifying polluted water as it passes through artificially constructed wetland area. The system of plants regenerates itself as the old plants die and form useful humus. and chemical action with certain existing or added inorganic chemicals help in finally obtaining a very clear and clean water.