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ASSIGNMENT EX-SITU BIOREMEDIATION

SUBMITTED TO: SUBMITTED BY: ROLL NO :

SIR SEGHUM HAFIZ UBAID ULLAH 021-BH-2007

TABLE OF CONTENTS INTRODUCTION CONTAMINANTS PHASES OF EX-SITU BIOREMEDIATION

SOLID PHASE
LAND FARMING COMPOSTING BIOPILES

SLURRY PHASE
AERATED LAGOONS AIR-LIFTED BIOREACTORS FLUDIZED-BED-BIOREACTORS

ADVANTAGES OF EX-SITU BIOREMEDIATION DISADVANTAGES OF EX-SITU BIOREMEDIATION

INTRODUCTION DEFINITION Ex-situ bioremediation is a biological process in which excavated soil is placed in a lined above-ground treatment area and aerated following processing to enhance the degradation of organic contaminants by the indigenous microbial population. Under aerobic conditions, specific micro-organisms can utilize organic contaminants such as petroleum hydrocarbon mixtures, polycyclic aromatic hydrocarbons (PAH), phenols, cresols and some pesticides as a source of carbon and energy and degrade them ultimately to carbon dioxide and water. it is unusual to require the addition of microbial populations but usual to assess the nutrient requirement and amend the basic nutrients and organic substrate of the soil if any of these elements are deficient or absent. Oxygen (via the introduction of air) is essential to allow the microbial population to develop cultures capable of sustaining degradation. CONTAMINANTS Ex-situ bioremediation can remediate a wide range of hydrocarbon contaminants including but not limited to: General hydrocarbons Kerosene Phenols Cresols Polycyclic aromatic hydrocarbons Semi-volatile organic compounds Diesel range hydrocarbons Lubricating oils

Straight chain aliphatic Non-chlorinated hydrocarbons within the carbon chain lengths C6 to C14 are readily treatable, non-chlorinated hydrocarbons with carbon chain lengths C15-C32 are treatable but require longer time periods to degrade. Chlorinated hydrocarbons and other more complex chains can be degraded but require detailed assessment and analysis to determine suitability. The steps involved include excavation, screening and fractionation, mixing, homogenizing and final disposal, all of which are very costly. On the basis of phases of contaminated materials, ex-situ bioremediation is of two types. Solid phase treatment The system includes organic wastes (like leaves, animal manures and agricultural wastes) and problematic wastes (domestic and industrial wastes, sewage sludge, municipal solid wastes, etc). The traditional clean-up practice involves the informal processing of the organic materials and production of composts which may be used as soil amendment. The solid phase treatment can be done through three methods which are land forming, soil biopile and composting. Land Farming The simplest form of bioremediation, contaminated soil is excavated and spread out in layers approximately 0.3m in thickness on a lined treatment area. Bioremediation can be enhanced by periodic turning of the bed and addition of nutrients. Due to the limiting thickness of soil layers (0.3m) land farming techniques require large areas and are not generally suitable for small sites, but can be the cheapest and most basic form of bioremediation. Indusial soil contains a huge amount of phenols

and polycyclic aromatic hydrocarbons. These compounds were treated through land farming and concentration of these contaminants decrease 40mg/kg and 290mg/kg to 2mg/kg and less than 200mg/kg respectively. The high molecular weights PAH were degraded slowly. The strategy applied was biostimulation (Guerin, 1998). Another application of land farming is the remediation of the hydrocarboncontaminated polar soils. The ex-situ is the better approach for this work because this can be done by controlling low and fluctuating soil temperature, low level of nutrients, alkalinity and low moisture content otherwise the excess level can inhibit the hydrocarbon degradation decreasing soil water moisture. So, the land farming may be the better approach for bioremediation of the polar soil (Aislabie j., Saul DJ., et all.2006). Land farming also used for the degradation of the polynuclear aromatic hydrocarbons PAH -contaminated soil. The experiment was performed in a 100m2 field area and treated with chemical, microbiological and ecotoxicological level strategies. The concentration of PAH was 1140mg/kg and decreased up to 63% in three month time period (Picado A., Baetta-Hall L., et all 2001).

Composting
Composting is a self-heating, substrate-dense, managed microbial system, and one solid-phase biological treatment technology which is suitable to the treatment of large amount of contaminated solid materials. However, many hazardous compounds are resistant to microbial degradation due to complex chemical structure, toxicity and compound concentration that hardly support growth. Microbial growth is also affected by moisture, pH, inorganic nutrients and particle size. Be cause composting of hazardous wastes typically involves the bioremediation of contaminated substrate-sparse soils, support of microbial self-heating needs incorporation of proper amount of supplements. The hazardous compounds reported to disappear through composting includes aliphatic and aromatic hydrocarbons and certain halogenated compounds. The possible routes lead

ing to disappearance of hazardous compounds include volatilization, assimilation, adsorption, polymerization and leaching (Hogan, 1998). Composting can be done in open system i.e. land treatment, and in closed system. The open land system can be inexpensive treatment method, but the temperature fluctuates from summer to winter. Therefore, rate of biodegradation of waste materials declines. Secondly, land treatment system may become oxygen limited, depending on amount of substrate, depth of waste, application, etc. However, efficiency of open treatment system can be increased by passing air. This approach is referred to as engineered soil piles and forced aeration treatment. The closed treatment system is preferred over the open land treatment system because controlled air is supplied to maintain the microbial activity. As a result of microbial growth and volatilization of hazardous compounds, internal temperature gradually rises. Therefore, use of blowers for air circulation and exhaust for removal of toxic volatiles are set up in closed treatment system. Ventilators supply oxygen and remove heat through evaporation of water.

Soil biopiles
Soil biopiles, also known as biocells, is a biodegradation technique used for the remediation of excavated soil contaminated with petroleum products. This technology involves the accumulation of contaminated soil into piles and the stimulation of microbial activity either aerobically or through the addition of nutrients, minerals or moisture. The biopiles are typically from three to ten feet high. This technique is similar to land farm method due to the fact that it also uses oxygen as a way to stimulate bacteria growth. However, while tilling or plowing is used to aerate land farms, biopiles are aerated by injecting the air through perforated piping placed throughout the pile. The oil is usually mixed with a bulking agent (straw) to improve aeration and thus enhance the growth of microbial population. Since air is also injected into the soil there is also the possibility of the evaporation or volatilization of contaminants. To encounter this problem, the system also incorporates the monitoring and containment of soil vapors.

The bioremediation of chlorobenzene through biocells was carried out in which the level of this industrial contaminant was reduced from 6mg/kg to 0.2mg/kg in soil. The target was obtained in 2-3 weeks through stimulating the natural microfolra by providing aeration and nutrients (Guerin, 2007). Another experiment was carried out in Taiwan in which the diesel oil and fuel oil were degraded up to 70% and 63% respectively by using biopile. This success was achieved in 28 days through the combine process of biostimulation and gioaugmentation (lin, Pan et all. 2009). The biopiles also had been used for degradation of total petroleum hydrocarbons in present in drilling mud-polluted site. The biopiles was full off one ton soil in each and nutrients and bulking agent were added to carry the treatment. Moisture content was containing around 30% to 35%. Result showed that after 180 days total TPH concentration decreased from 23000mg/kg to 700mg/kg (rojas-averlizapa., roldan-carillo et all. 2007).

Slurry-phaseTreatment
The contaminated solid materials (soil, degraded sediments, etc), microorganisms and water formulated into slurry are brought within a bioreactor i.e. fermenter. Thus slurry-phase treatment is a triphasic system involving three major components: water, suspended particulate matter and air. Here water serves as suspending medium where nutrients, trace elements, pH adjustment chemicals and desorbed contaminants are dissolved. Suspended particulate matter includes a biologically inert substratum consisting of contaminants (soil particles) and biomass attached to soil matrix or free in suspending medium. Air provides oxygen for bacterial growth. Slurry-phase reactors are new design in bioremediation. The objectives of bioreactor designing are to (i) alleviate microbial growth limiting factors in soil environment such as substrate, nutrients and oxygen availability, (ii) promote suitable environmental conditions for bacterial growth such as moisture, pH, temperature,

and (iii) minimize mass transfer limitations and facilitate desorption of organic material from the soil matrix (Christodoultos and Kontsospyros, 1998).

Biologically there are three types of slurry-phase bioreactors : aerated lagoons, low-shear airlift reactor, and fluidized-bed soil reactor. The first two types are in use of full scale bioremediation, while the third one is in developmental stage. AERATED LAGOONS Aerated La goons system is very similar to aerated lagoon used for treatment of small common municipal waste water. Nutrients and aeration are supplied to the reactor. Mixers are fitted to mix different components and form slurry, whereas surface aerators provide air required for microbial growth. The process may be used as singlestage or multistage operation. If the waste contains volatiles, this reactor is not appropriate. Aerated lagoons are tend to have problems with high effluent ammonia concentrations during winter and early spring seasons. The factors found to affect performance were temperature and detention time (Van Dyke., Jones S., et all. 2003). Low-shear Airlift Reactors (LSARs). The LSARs are useful when waste contains volatile components; tight process control and increased efficiency of bioreactors are required. LSARs are cylindrical tanks which are made up of stainless steel. In this bioreactor pH, temperature, nutrient addition, mixing and oxygen can be controlled as desired. Shaft is equipped with impellers. It is driven by motor set up at the top. The rake arms are connected with blades which is used for resuspension of coarse materials that tend to settle on the bottom of the bioreactor. Air diffusers are placed radially along the rake arm. Airlift provides to bottom circulation of contents in reactor. Baffles make the

hydrodynamic behavior of slurry-phase bioreactors. Pre-treatment process includes size fractionation of solids, soil washing, milling to reduce particle size and slurry preparation. Certain surfactants such as anthracene, pyrene, perylene, etc. are added to enhance the rate of biodegradation. These act as co-substrate and utilize as carbon source. Co-substrates also induce the production of beneficial enzymes (Christodoultos and Kontsospyros, 1998). Another experiment was biodegradation of naphthalene present in soil. The degradation was carried out by using bacterial strain (pseudomonas putida m8) into an oxygenated bioreactor. The hydrocarbon was disappeared and the soil was remedified (collina, pitea et all., 2005). The bioreactors can be used in various ways for instance in an experiment the polymer beads were used to remedify the soil from phenols. The initial concentration of phenol was 23g/kg and was decrease to 100mg/kg absorbed by polymer beads. After that beads were placed into bioreactors inoculated with phenol degrading microbial consortium. The phenol was degraded and beads were available to use again without no loss in performance (Prpich GP., Adams RL.,et all 2006) The third category of reactors fludized-bed-reactors(FBR) also have broad applications in remedification. It has been studied in USA that FBR are the best tools for the removal of perchlorate anion present in drinking water(Sutton pm., 2006)

Advantages:
* * * * n Proven track record on UK contaminated sites Suitable for a wide range of contaminants Suitability relatively simple to assess from site investigation data Flexible with respect to volumes (Vertase FLI has treated in excess of 250,000 e s in one season on some sites) to

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Limitations:
* Not applicable to heavy metal contamination or chlorinated hydrocarbons such as trichloroethylene (TCE) * Non-permeable soils require additional processing (clays and silts etc) * Can require large areas for treatment beds and thus site size is an important consideration * Contaminants must be aerobically biodegradable, * Temperature, weather and material dependent

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References
Ta-Chen Lin., Po-Tsen Pan., 2009., Ex-situ bioremediation of oil-contaminated soil., department of biological science and technology., (pingtung country., Taiwan.) Turlough F.Gurein., 2007., Ex-situ bioremediation of chlorobenzenes in soil., (Telstra corporation limited, Melbourne, Australia.) Collina E., Bessteti G., Lasagni M., Pitea D., Naphthalene biodegradation kinetics in an aerobic slurry-phase bio reactor., 2005., (University of Milano, Italy) Rojas-Avelizapa., Roldan-Carrilo., Fernandez-lenares lc. 2007., (1nstitute of Mexico.) Aislabie j., Saul DJ., Foght JM., 2006., Bioremediation of hydrocarboncontaminated polar soils., (land care research institute,Hamilton,New Zealand.) Picado a., Nogueira A., de Fatima Rodrigues m., 2001., (National institute of engineering and technology, Lisboa, Portugal.) Van Dyke., Jones S., Ong SK., 2003., Cold weather nitrogen removal deficiencies of aerated lagoons., (Fox engineering institute, Ames, USA). Sutton PM., 2006., Bioreactor configurations for ex-situ remediation of perchlorate., (PM Sutton & Associates, inc., New Hampshir,USA). Prpich., Adams RL., Daugullis AJ., 2006., Ex-situ bioremediation of phenol contaminated soil using polymer beads., (Department of chemical engineering, Queen;s university, Kingston, Canada). Terlough F.Guerin., 1998., Bioremediation of phenols and polycyclic aromatic hydrocarbons in creosote contaminated soil using ex-situ landtreatment.,(Suite 2506, 6503 N Military Trail, Boca Raton, FL 33496, USA.)

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