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BIOREMEDIATION

Introductory concept
CONVENTIONAL METHOD: Concept of dig n dump Insufficient method The toxic materials from these dig n dump sites have begun to leak into water sources and into areas that sustain human life. 1891 the first biological sewage treatment plant was created in Sussex, UK. The word bioremediation did not appear in peer-reviewed scientific literature until 1987. CONCEPT: Bioremediation is the transformation or degradation of contaminants into nonhazardous or less hazardous chemicals. Bacteria are generally used for bioremediation, but fungi, algae and plants have also been used. Bioremediation is not a new technology.

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Increases in environmental contamination lead to a progressive deterioration of environmental quality. The increasing size of the human population requires the need for a biological approach to improve environmental conditions. Exposure to pest control and environmental chemical remediation is increasing because environmental release of biotechnology agents is becoming moreof biotechnology agents becoming accepted. Therefore, it is important to have reliable data on potential health effects of these agents.

H y m a a u e m y in st p e s a p ts

WHAT IS CONTAMINATION ?
Any substance causing harm directly or indirectly to any living form ,process is called contamination and substance causing contamination are called contaminants. Human population releases toxicants such as detergents, fertilizers, propellants, pesticides, refrigerants, and many other chemicals. Feces and urine contain biological nitrogen waste and are produced by animals. Radioactive compounds yield other radioactive metabolites, which can be as hazardous or more so than the parent

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Xeno occurrin compou biotic the s environm that are ch present em unnatur ica concent lly sy nth esi ze d c metals o m p o u n d s t h a t h a v e

never occurred in nature. Examples:

pesticides herbicides plastics

What is Bioremediation?
By definition, bioremediation is the use of living
organisms, primarily microorganisms, to degrade the environmental
contaminants into less toxic forms. The use of bacteria and

fungi and plants to break down or degrade toxic chemical

compo nds that ha e acc m lated in the en ironment.


Bioremediation is the productive utilization of living systems to degrade, detoxify, transform, immobilize or stabilize toxic environmental contaminants.

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certain harmful chemicals, such as those found in gasoline and oil spills. When microbes completely digest these chemicals, they change them into water and harmless gases such as carbon dioxide.

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General equation of bioremediation


Microbes + contaminants + electron acceptor = CO + H 0 + biomass
2 2

Thus, bioremediation is defined as the process


whereby organic wastes are biologically degraded under controlled conditions to an innocuous state, or to levels below concentration limits

established by regulatory authorities.

Principles of bioremediation
In year 2000, Tate summarized the principles of bioremediation: 1 The relationship ofof comparative biochemistry applies equally to cultures of microbes and those present in the soil. 2. The microbial growth requirements are the same whether in the laboratory culture or in the field. 3. The ecological factors need to accommodate the presence of other microbes and adapt to the physical and chemical properties of the soil.
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Aerobic and Anaerobic bioremediation

Various factors controls bioremediation


1. Availability of microorganisms such as bacteria, fungi etc which are capable of bioremediation 2. Availability of contaminants 3. Environmental factors such as:
i. Type of soil ii. Temperature iii. pH iv. Oxygen v. Electron acceptors vi. Nutrients

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Factor Microbial population Oxygen

Desired Conditions Suitable kinds of organisms that can biodegrade all of the contaminants Enough to support aerobic biodegradation (about 2% oxygen in the gas phase or 0.4 mg/liter in the soil water) Soil moisture should be from 5070% of the water holding capacity of the soil Ni h h lf ad h n d i

Water

Nutrients

to support good microbial growth Temperature Appropriate temperatures for microbial growth (0 40C) pH Best range is from 6.5 to 7.5

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Important organisms involved in bioremediation


Metabolizing Microbes Indigenous microbes those found naturally at a polluted site Bacteria Pseudomonas E.coli Algae and fungi Fusarium oxysporum Mortierella hyaline

Bioremediation Technology
Technologies can be generally classified as in situ or ex situ.
IN SITU: It involves treating the contaminated material at

the site. It is an application of biological treatment to the


cleanup of hazardous chemicals in the soil and surface or

subsurface waters.
EX SITU: It involves the removal of the contaminated material to be treated elsewhere. In this process

generally the 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.


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Examples of bioremediation technologies are 1. Phytoremediation 2. Bioventing 3. Bioleaching 4. Landfarming 5. Bioreactors 6. Composting 7. Bioaugmentation 8. Rhizofiltration 9. Biostimulation.

Types of in situ bioremediation


1. 2. 3. 4. 5. 6. BIOVENTING IN SITU BIODEGRADATION BIOSPARGING BIOLEACHING BIOAUGMENTATION PHYTOREMEDIATION

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1. BIOVENTING:

Bioventing is an In-Situ remediation technology Uses indigenous microorganisms to biodegrade organic constituents adsorbed to soils in the unsaturated zone. In Bioventing, the activity of the indigenous bacteria is enhanced by inducing air (or oxygen) flow into the unsaturated zone (using extraction or injection wells) and, if necessary, by adding nutrients.

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2. IN SITU BIODEGRADATION/ BIOSTIMULATION: It involves supplying oxygen and nutrients by circulating aqueous solutions through contaminated soils to stimulate
naturally occurring bacteria to degrade organic contaminants. It can be used for soil and groundwater Generally this

technique includes conditions such as the infiltration of water containing nutrients and oxygen or other electron acceptors for groundwater treatment. Biostimulation is the addition of nutrients, oxygen or other electron donors and acceptors to the coordinated site in order to increase the population or activity of naturally occurring microorganisms available for bioremediation.

3. BIOSPARGING: It involves the injection of air under pressure below


the water table to increase groundwater oxygen concentrations

and enhance the rate of biological degradation of contaminants


by naturally occurring bacteria. It can be used to reduce concentrations of petroleum constituents that are dissolved in

groundwater.

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4. BIOLEACHING: It is the extraction of specific metals from their ores through the use of living organisms.
Benefits of bioleaching Simple and inexpensive process. Ecofriendly process No need for high pressure or temperature Leaching residues less active than in physico-chemical processes Pretreatment of refractory gold in ores where high arsenic content concentrares. Operating the bioleaching plant is very simple, as bacteria do most of the work Relatively low cost

Thiobacillus ferrooxidans, Leptospirillum ferrooxidans and Thermophilic species of Sulfobacillus, Acidianus and Sulfolobus These bacteria tolerate acids and metabolize sulphur. The particular bacteria use a chemical reaction known as Oxidation reaction to convert metal sulphide crystals into sulphates and sheer metals. 1 Th b t i idi f i (F 2+) d l h (S)

to produce ferric iron (Fe3+) and sulphate (SO42-) 2. The Fe3+ in turn reacts with the sulphide minerals to produce Fe2+ and S

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5 . BIOAUGMENTATION:
It frequently involves the addition of microorganisms indigenous or
exogenous to the contaminated sites. Microorganisms can biotransform or biodegrade contaminants. The microorganisms added can be a completely new species or more members of a

species that already exists at the site. For this approach to be successful in the field, the seed microorganisms b bl d d l

maintain genetic stability and viability during storage, survive in foreign and hostile environments, effectively compete with indigenous microorganisms, and move through the pores of the sediment to the contaminants

6. PHYTOREMEDIATION
Phytoremediation is the use of living green plants for reduction and/or removal of contaminants from contaminated soil, water, sediments, and air. Advantages: Low cost The possibility of phytomining Eco friendly Limitations: Phytoremediation is limited to the surface area and depth occupied by the roots. Time consuming The survival of the plants is affected by the toxicity of the contaminated land and the general condition of the soil. Bio-accumulation of contaminants, especially metals, into the plants which then pass into the food chain, from primary level consumers upwards or requires the safe disposal of the affected plant material.

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How does it work:

Modes of phytoremediation:
PHYTODEGRADATION: The process PHYTOEXTRACTION: The use of plants where plants are able to metabolically to extract contaminants from the degrade organic pollutants environment

of soil microbial activity for the degradation of contaminants, typically by organisms that associate with roots.

PHYTOSTIMULATION: Enhancement PHYTOSTABILISATION: Use of plants to

reduce bioavailability and migration of contaminants

PHYOTVOLATILISATION:

The use of plants to volatilise pollutants from polluted soils and water

The uptake of contaminants by the roots of plants which are immersed in water

RHIZOFILTRATION:

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In situ bioremediation:
Advantages: 1. 2. 3. 4. It does not requires removal of the contaminants from the contaminated site. Less time consuming. Less costly. No potential danger involved.

Disadvantages: 1. As the site is not contained,therefore it is harder to control conditions and monitor the progress.

Types of Ex situ bioremediation


These techniques generally involve the excavation or removal of contaminated soil/water from the site.

1. 2. 3. 4.

LANDFARMING/ BIOFARMING COMPOSTING BIOPILES BIOREACTORS

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3. LAND/ BIO FARMING: It is a simple technique in which


contaminated soil is excavated and spread over a prepared

bed and periodically tilled until pollutants are degraded.

Contaminated soils are mixed with soil amendments such as soil bulking agents and nutrients, and then they are tilled into the earth. The material is periodically tilled for aeration. LIMITATIONS: Large space requirements Time consuming Leaching problem

2. COMPOSTING: It is a technique that involves combining

contaminated soil with nonhazardous organic amendants


such as manure or agricultural wastes. The presence of these

organic materials supports the development of a rich microbial population and characteristic of composting.
3. BIOPILES are a hybrid of landfarming and composting. Essentially, engineered cells are constructed as aerated composted piles. Typically used for treatment of surface contamination with petroleum hydrocarbons they are a refined version of landfarming that tend to control physical losses of the contaminants by leaching and volatilization. Biopiles provide a favorable environment for indigenous

elevated temperature

aerobic and anaerobic microorganisms.

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4. Bioreactors.
A bioreactor may refer to any manufactured or engineered device or system that supports a biologically active environment. It is a vessel in which a chemical process is carried out. Bioreactors are also designed to treat sewage and wastewater. Bioremediation in reactors involves the processing of contaminated solid material (soil, sediment, sludge) or water through an engineered containment system. A slurry bioreactor may be defined as a containment vessel and apparatus used to create a three-phase (solid, liquid, and gas) mixing condition to increase the bioremediation rate of soil bound and water-soluble pollutants as a water slurry of the contaminated soil and biomass (usually indigenous microorganisms) capable of degrading target contaminants.

Ex situ bioremediation:
Advantages: 1. Removes the contaminants and place them in a contained environment. 2. This contained environment allows easy monitoring and maintenance and thus makes whole process faster. Disadvantages: 1. Removal of contaminants from the contaminated sites is time consuming,costly and potentially dangerous. 2. Increased exposure to the toxic material.

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Advantages of Bioremediation: 1) Bioremediation is a natural process. 2) It is cost effective. 3) T iToxic h from environment and not just merely separated. 4) Low capital expenditure. 5) Less energy is required as compared to other technologies 6) Less manual supervision. il are d t d or removedd

Disadvantages 1) The process of bioremediation is slow. Time required is in day to months. 2) Heavy metals are not removed. 3) For in situ bioremediation site must have soil with high permeability. 4) It does not remove all quantities of contaminants.

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Recombinant DNA technology and bioremediation


Genetically engineered microorganisms (GEMs) have shown potential for the bioremediation of soil and l d treatment
Example: 1. Enterobacteragglomerans DK3 containing plasmid RP4 Tn 4371 , encodes for Biophenyl degradation. This donor strain when added to the soil disappears quickly, but the plasmid is transferred to the other microbes present, which carry out the biophenyl degradation. 2. The bacterium Deinococcus radiodurans (the most radioresistant organism known) has been modified to consume and digest toluene and ionic mercury from highly radioactive nuclear waste

Conclusion
Bioremediation is costeffective and is a beneficial addition to chemical and physical methods of managing wastes and environmental pollutants. Biodegradative treatment offers a savings of 60 to 90% over landfill disposal costs. New tools and techniques for use of bioremediation in situ in biofilters, and in bioreactors are contributing to the rapid growth of this field.
It is clear that new bioremediation technologies that can better monitor

and control many types of societal wastes are emerging. Genetically engineered microbes will require further study to clarify issues of safety and containment. Perhaps the larger problem facing policy makers in the future is how to decide where available bioremedial dollars will benefit human and environmental health the most. Target cleanup goals have been judged to be highly unrealistic in some cases. At most of these sites remedial efforts are as yet incomplete, and some efforts have had little effect. Bioremediation will play an increasingly important role as a result of new and emerging techniques and processes.

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