CONTENT

• INTRODUCTION

• PRINCIPLE OF BIOREMEDIATION

• FECTOR OF BIOREMEDIATION

• MICROBIAL POPULATIONS FOR

BIOREMEDIATION PROCESSES
• ENVIRONMENTAL FACTOR OF
BIOREMEDIATION
• ADVANTAGES OF BIOREMEDIATION

• DISADVANTAGES OF BIOREMEDIATION

• USE OF BIOREMEDIATION

• BIOREMEDIATION OF SOIL

• BIOREMEDIATION OF OIL POLLUTANTS

• BIOREMEDIATION OF AIR POLLUTANTS

• REFERENCE
INTRODUCTION
The quality of life on Earth is linked inextricably to the overall quality of the
environment. In early times, we believed that we had an unlimited
abundance of land and resources; today, however, the resources in the world
show, in greater or lesser degree, our carelessness and negligence in using
them. The problems associated with contaminated sites now assume
increasing prominence in many countries. Contaminated lands generally
result from past industrial activities when awareness of the health and
environmental effects connected with the production, use, and disposal of
hazardous substances were less well recognized than today. The problem is
worldwide, and the estimated number of contaminated sites is significant. It
is now widely recognized that contaminated land is a potential threat to
human health, and its continual discovery over recent years has led to
international efforts to remedy many of these sites, either as a response to
the risk of adverse health or environmental effects caused by contamination
or to enable the site to be redeveloped for use.
The conventional techniques used for remediation have been to dig up
contaminated soil and remove it to a landfill, or to cap and contain the
contaminated areas of a site. The methods have some drawbacks. The first
method simply moves the contamination elsewhere and may create
significant risks in the excavation, handling, and transport of hazardous
material. Additionally, it is very difficult and increasingly expensive to find
new landfill sites for the final disposal of the material. The cap and contain
method is only an interim solution since the contamination remains on site,
requiring monitoring and maintenance of the isolation barriers long into the
future, with all the associated costs and potential liability. A better approach
than these traditional methods is to completely destroy the pollutants if
possible, or at least to transform them to innocuous substances. Some
technologies that have been used are high-temperature incineration and
various types of chemical decomposition (e.g., base-catalyzed
dechlorination, UV oxidation). They can be very effective at reducing levels
of a range of contaminants, but have several drawbacks, principally their
technological complexity, the cost for small-scale application, and the lack of
public acceptance, especially for incineration that may increase the exposure
to contaminants for both the workers at the site and nearby residents.
Bioremediation is an option that offers the possibility to destroy or render
harmless various contaminants using natural biological activity. As such, it
uses relatively low-cost, low-technology techniques, which generally have a
high public acceptance and can often be carried out on site. It will not always
be suitable, however, as the range of contaminants on which it is effective is
limited, the time scales involved are relatively long, and the residual
contaminant levels achievable may not always be appropriate. Although the
methodologies employed are not technically complex, considerable
experience and expertise may be required to design and implement a
successful bioremediation program, due to the need to thoroughly assess a
site for suitability and to optimize conditions to achieve a satisfactory result.
Because bioremediation seems to be a good alternative to conventional
clean-up technologies research in this field, especially in the United States,
rapidly increasing.
Bioremediation has been used at a number of sites worldwide, including
Europe, with varying degrees of success. Techniques are improving as
greater knowledge and experience are gained, and there is no doubt that
bioremediation has great potential for dealing with certain types of site
contamination. Unfortunately, the principles, techniques, advantages, and
disadvantages of bioremediation are not widely known or understood,
especially among those who will have to deal directly with bioremediation
proposals, such as site owners and regulators. Here, we intended to assist by
providing a straightforward, pragmatic view of the processes involved in
bioremediation, the pros and cons of the technique, and the issues to be
considered when dealing with a proposal for bioremediation.
Some tests make an exhaustive examination of the literature of
bioremediation of organic [2–4] and inorganic pollutants, and another test
takes a look at pertinent field application case histories.

PRINCIPLES OF BIOREMEDIATION
Environmental biotechnology is not a new field; composting and wastewater
treatments are familiar examples of old environmental biotechnologies.
However, recent studies in molecular biology and ecology offer opportunities
for more efficient biological processes. Notable accomplishments of these
studies include the clean-up of polluted water and land areas.
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.
By definition, bioremediation is the use of living organisms, primarily
microorganisms, to degrade the environmental contaminants into less toxic
forms. It uses naturally occurring bacteria and fungi or plants to degrade or
detoxify substances hazardous to human health and/or the environment.
The microorganisms may be indigenous to a contaminated area or they may
be isolated from elsewhere and brought to the contaminated site.
Contaminant compounds are transformed by living organisms through
reactions that take place as a part of their metabolic processes.
Biodegradation of a compound is often a result of the actions of multiple
organisms. When microorganisms are imported to a contaminated site to
enhance degradation we have a process known as bioaugmentation.
For bioremediation to be effective, microorganisms must enzymatically
attack the pollutants and convert them to harmless products. As
bioremediation can be effective only where environmental conditions permit
microbial growth and activity, its application often involves the manipulation
of environmental parameters to allow microbial growth and degradation to
proceed at a faster rate. Like other technologies, bioremediation has its
limitations. Some contaminants, such as chlorinated organic or high aromatic
hydrocarbons, are resistant to microbial attack. They are degraded either
slowly or not at all , hence it is not easy to predict the rates of clean-up for a
bioremediation exercise; there are no rules to predict if a contaminant can
be degraded. Bioremediation techniques are typically more economical than
traditional methods such as incineration, and some pollutants can be treated
on site, thus reducing exposure risks for clean-up personnel, or potentially
wider exposure as a result of transportation accidents. Since bioremediation
is based on natural attenuation the public considers it more acceptable than
other technologies.
Most bioremediation systems are run under aerobic conditions, but running a
system under anaerobic conditions may permit microbial organisms to
degrade otherwise recalcitrant molecules.

FACTORS OF BIOREMEDIATION

The control and optimization of bioremediation processes is a complex

system of many factors. These factors include: the existence of a microbial
population capable of degrading the pollutants; the availability of
contaminants to the microbial population; the environment factors (type of
soil, temperature, pH, the presence of oxygen or other electron acceptors,
and nutrients).

MICROBIAL POPULATIONS FOR BIOREMEDIATION

PROCESSES

Microorganisms can be isolated from almost any environmental conditions.

Microbes will adapt and grow at subzero temperatures, as well as extreme
heat, desert conditions, in water, with an excess of oxygen, and in anaerobic
conditions, with the presence of hazardous compounds or on any waste
stream. The main requirements are an energy source and a carbon source.
Because of the adaptability of microbes and other biological systems, these
can be used to degrade or remediate environmental hazards. We can
subdivide these microorganisms into the following groups:
Aerobic. In the presence of oxygen. Examples of aerobic bacteria recognized
for their degrada-
tive abilities are Pseudomonas, Alcaligenes, Sphingomonas, Rhodococcus,
and Mycobacterium. These microbes have often been reported to degrade
pesticides and hydrocarbons, both alkanes and polyaromatic compounds.
Many of these bacteria use the contaminant as the sole source of carbon and
energy.
Anaerobic. In the absence of oxygen. Anaerobic bacteria are not as
frequently used as aerobic
bacteria. There is an increasing interest in anaerobic bacteria used for
bioremediation of polychlorinated biphenyls (PCBs) in river sediments,
dechlorination of the solvent trichloroethylene (TCE), and chloroform.
Ligninolytic fungi. Fungi such as the white rot fungus Phanaerochaete
chrysosporium have the
ability to degrade an extremely diverse range of persistent or toxic
environmental pollutants. Common substrates used include straw, saw dust,
or corn cobs.
Methylotrophs. Aerobic bacteria that grow utilizing methane for carbon and
energy. The initial
enzyme in the pathway for aerobic degradation, methane monooxygenase,
has a broad substrate range and is active against a wide range of
compounds, including the chlorinated aliphatics trichloroethylene and 1,2-
dichloroethane.
An overview of the microbiological aspects of the application of
microorganisms is given in [9].
For degradation it is necessary that bacteria and the contaminants be in
contact. This is not easily achieved, as neither the microbes nor
contaminants are uniformly spread in the soil. Some bacteria are mobile and
exhibit a chemotactic response, sensing the contaminant and moving toward
it. Other microbes such as fungi grow in a filamentous form toward the
contaminant. It is possible to enhance the mobilization of the contaminant
utilizing some surfactants such as sodium dodecyl sulphate (SDS).

ENVIRONMENTAL FACTORS
Nutrients
Although the microorganisms are present in contaminated soil, they cannot
necessarily be there in the numbers required for bioremediation of the site.
Their growth and activity must be stimulated. Biostimulation usually involves
the addition of nutrients and oxygen to help indigenous microorganisms.
These nutrients are the basic building blocks of life and allow microbes to
create the necessary enzymes to break down the contaminants. All of them
will need nitrogen, phosphorous, and carbon. Carbon is the most basic
element of living forms and is needed in greater quantities than other
elements. In addition to hydrogen, oxygen, and nitrogen it constitutes about
95% of the weight of cells Phosphorous and sulfur contribute with 70% of the
remainders. The nutritional requirement of carbon to nitrogen ratio is 10:1,
and carbon to phosphorous is 30:1 Microbial growth and activity are readily
affected by pH, temperature, and moisture. Although microorganisms have
been also isolated in extreme conditions, most of them grow optimally over a
narrow range, so that it is important to achieve optimal conditions. If the soil
has too much acid it is possible to rinse the pH by adding lime. Temperature
affects biochemical reactions rates, and the rates of many of them double for
each 10 °C rise in temperature. Above a certain temperature, however, the
cells die. Plastic covering can be used to enhance solar warming in late
spring, summer, and autumn. Available water is essential for all the living
organisms, and irrigation is needed to achieve the optimal moisture level.
The amount of available oxygen will determine whether the system is
aerobic or anaerobic. Hydrocarbons are readily degraded under aerobic
conditions, whereas chlorurate compounds are degraded only in anaerobic
ones. To increase the oxygen amount in the soil it is possible to till or sparge
air. In some cases, hydrogen peroxide or magnesium peroxide can be
introduced in the environment. Soil structure controls the effective delivery
of air, water, and nutrients. To improve soil structure, materials such as
gypsum or organic matter can be applied. Low soil permeability can impede
movement of water, nutrients, and oxygen; hence, soils with low
permeability may not be appropriate for in situ clean-up techniques.

BIOREMEDIATION STRATEGIES

Different techniques are employed depending on the degree of saturation

and aeration of an area. In situ techniques are defined as those that are
applied to soil and groundwater at the site with minimal disturbance. Ex situ
techniques are those that are applied to soil and groundwater at the site
which has been removed from the site via excavation (soil) or pumping
(water). Bioaugmentation techniques involve the addition of microorganisms
with the ability to degrade pollutants.

In situ bioremediation

These technique are generally the most desirable options due to lower cost
and less disturbance since they provide the treatment in place avoiding
excavation and transport of contaminants. In situ treatment is limited by the
depth of the soil that can be effectively treated. In many soils effective
oxygen diffusion for desirable rates of bioremediation extend to a range of
only a few centimeters to about 30 cm into the soil, although depths of 60
cm and greater have been effectively treated in some cases.
The most important land treatments are:
Bioventing is the most common in situ treatment and involves supplying air
and nutrients through wells to contaminated soil to stimulate the indigenous
bacteria. Bioventing employs low air flow rates and provides only the amount
of oxygen necessary for the biodegradation while minimizing volatilization
and release of contaminants to the atmosphere. It works for simple
hydrocarbons and can be used where the contamination is deep under the
surface.
In situ biodegradation 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.
Biosparging- Biosparging 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. Biosparging increases the mixing in the saturated zone and there
by increases the contact between soil and groundwater. The ease and low
cost of installing small-diameter air injection points allows considerable
flexibility in the design and construction of the system.
Bioaugmentation. Bioremediation frequently involves the addition of
microorganisms indigenous or exogenous to the contaminated sites. Two
factors limit the use of added microbial cultures in a land treatment unit: 1)
nonindigenous cultures rarely compete well enough with an indigenous
population to develop and sustain useful population levels and 2) most soils
with long-term exposure to biodegradable waste have indigenous
microorganisms that are effective degrades if the land treatment unit is well
managed.

Ex situ bioremediation

These techniques involve the excavation or removal of contaminated soil

from ground.
Landfarming is a simple technique in which contaminated soil is excavated
and spread over a prepared bed and periodically tilled until pollutants are
degraded. The goal is to stimulate indigenous biodegradative
microorganisms and facilitate their aerobic degradation of contaminants. In
general, the practice is limited to the treatment of superficial 10–35 cm of
soil. Since landfarming has the potential to reduce monitoring and
maintenance costs, as well as clean-up liabilities, it has received much
attention as a disposal alternative.
Composting 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 elevated temperature characteristic of composting.
Biopiles are a hybrid of landfarming and composting. Essentially, engineered
cells are con-
structed 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 aerobic and anaerobic microorganisms.
Bioreactors. Slurry reactors or aqueous reactors are used for ex situ
treatment of contaminated soil and water pumped up from a contaminated
plume. 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. In general, the rate and extent of biodegradation are greater
in a bioreactor system than in situ or in solid-phase systems because the
contained environment is more manageable and hence more controllable
and predictable. Despite the advantages of reactor systems, there are some
disadvantages. The contaminated soil requires pre treatment (e.g.,
excavation) or alternatively the contaminant can be stripped from the soil
via soil washing or physical extraction (e.g., vacuum extraction) before being
placed in a bioreactor

Advantages of bioremediation

•Bioremediation is a natural process and is therefore perceived by the public

as an acceptable waste treatment process for contaminated material such as
soil. Microbes able to degrade the contaminant increase in numbers when
the contaminant is present; when the contaminant is degraded, the
biodegradative population declines. The residues for the treatment are
usually harmless products and include carbon dioxide, water, and cell
biomass.
•Theoretically, bioremediation is useful for the complete destruction of a
wide variety of contaminants. Many compounds that are legally considered
to be hazardous can be transformed to harmless products. This eliminates
the chance of future liability associated with treatment and disposal of
contaminated material.
•Instead of transferring contaminants from one environmental medium to
another, for example, from land to water or air, the complete destruction of
target pollutants is possible.
•Bioremediation can often be carried out on site, often without causing a
major disruption of normal activities. This also eliminates the need to
transport quantities of waste off site and the potential threats to human
health and the environment that can arise during transportation.
•Bioremediation can prove less expensive than other technologies that are
used for clean-up of hazardous waste.

Disadvantages of bioremediation

•Bioremediation is limited to those compounds that are biodegradable. Not

all compounds are susceptible to rapid and complete degradation.
•There are some concerns that the products of biodegradation may be more
persistent or toxic than the parent compound.
•Biological processes are often highly specific. Important site factors
required for success include the presence of metabolically capable microbial
populations, suitable environmental growth conditions, and appropriate
levels of nutrients and contaminants.
•It is difficult to extrapolate from bench and pilot-scale studies to full-scale
field operations.
•Research is needed to develop and engineer bioremediation technologies
that are appropriate for sites with complex mixtures of contaminants that are
not evenly dispersed in the environment.
Contaminants may be present as solids, liquids, and gases.

Bioremediation often takes longer than other treatment options, such as

excavation and removal of soil or incineration.
•Regulatory uncertainty remains regarding acceptable performance criteria
for bioremediation. There is no accepted definition of “clean”, evaluating
performance of bioremediation is difficult, and there are no acceptable
endpoints for bioremediation treatments.

PHYTOREMEDIATION
Although the application of microbe biotechnology has been successful with
petroleum-based constituents, microbial digestion has met limited success
for widespread residual organic and metals pollutants. Vegetation- based
remediation shows potential for accumulating, immobilizing, and
transforming a low level of persistent contaminants. In natural ecosystems,
plants act as filters and metabolize substances generated by nature.
Phytoremediation is an emerging technology that uses plants to remove
contaminants from soil and water [14–16]. The term “phytoremediation” is
relatively new, coined in 1991. Its potential for encouraging the
biodegradation of organic contaminants requires further research, although
it may be a promising area for the future.
We can find five types of phytoremediation techniques, classified based on
the contaminant fate:
phytoextraction, phytotransformation, phytostabilization, phytodegradation,
rhizofiltration, even if a combination of these can be found in nature.
Phytoextraction or phytoaccumulation is the process used by the plants to
accumulate contami-
nants into the roots and aboveground shoots or leaves. This technique saves
tremendous remediation cost by accumulating low levels of contaminants
from a widespread area. Unlike the degradation mechanisms, this process
produces a mass of plants and contaminants (usually metals) that can be
transported for disposal or recycling.
Phytotransformation or phytodegradation refers to the uptake of organic
contaminants from soil, sediments, or water and, subsequently, their
transformation to more stable, less toxic, or less mobile form. Metal
chromium can be reduced from hexavalent to trivalent chromium, which is a
less mobile and noncarcinogenic form.
Phytostabilization is a technique in which plants reduce the mobility and
migration of contami-
nated soil. Leachable constituents are adsorbed and bound into the plant
structure so that they form a stable mass of plant from which the
contaminants will not reenter the environment.
Phytodegradation or rhizodegradation is the breakdown of contaminants
through the activity existing in the rhizosphere. This activity is due to the
presence of proteins and enzymes produced by the plants or by soil
organisms such as bacteria, yeast, and fungi. Rhizodegradation is a
symbiotic relationship that has evolved between plants and microbes. Plants
provide nutrients necessary for the microbes to thrive, while microbes
provide a healthier soil environment.

Phytoremediation is well suited for use at very large field sites where other
methods of remediation are not cost effective or practicable; at sites with a
low concentration of contaminants where only polish treatment is required
over long periods of time; and in conjunction with other technologies where
vegetation is used as a final cap and closure of the site. There are some
limitations to the technology that it is necessary to consider carefully before
it is selected for site remediation: long duration of time for remediation,
potential contamination of the vegetation and food chain, and difficulty
establishing and maintaining vegetation at some sites with high toxic levels.

Methanotrophic Bacteria: Use in Bioremediation

Microbial communities are highly diverse and capable of conducting an extensive range of
metabolic activities. Irrespective of depth or geological formation, subsurface microorganisms
carry out all the major nutrient cycling, i.e., carbon, sulfur, nitrogen, manganese, iron and
phosphorus. Although each geological formation appears to have its own microbial structure,
sandy formations that are highly permeable to air or water flow have higher microbial activity.
Considering a generally large subsurface microbiota there is considerable interest for the
prospect of degrading hazardous contaminants in situ by stimulating selective bacterial
populations (biostimulation) or by the addition of organisms to contaminated sites
(bioaugmentation). Stimulation of an indigenous population of methanotrophs by methane is
likely to enrich for species that are well adapted to their environment, whereas the deliberate
addition of more microorganisms into such an environment may be compromised since the
introduced organisms are not as likely to be able to compete.

Defining of temporal and spatial relationships and population dynamics/interactions of selected

microorganisms such as methanotrophs in the natural setting is important for the evaluation of
bioremediation potential and its effectiveness. It has become increasingly evident that indigenous
microbial systems are able to facilitate the degradation and mineralization of a wealth of
compounds that twenty years ago were thought to be biologically recalcitrant. This realization
has necessitated technologies whereby defined microbial types can be followed in situ in real
time by techniques that are designed to be selective, sensitive and easily applicable to soils,
sediments and groundwater.
Methanotrophs are physiologically versatile in their ability to exist in a variety of habitats and
live in hostile environments having a wide range of pH, temperature, heavy metal concentrations,
oxygen concentrations, barometric pressures, salinity and radiation. Under these diverse
conditions a number of methanotrophs have been isolated that facilitate the degradation of TCE
and its daughter products.

Evaluation, characterization, and utilization of microbial communities associated with in situ

bioremediation of subsurface and groundwater contamination is a technological necessity for
environmental restoration and assessment. In the past sixty years both industrial and government
nuclear production and waste management facilities have generated a significant quantity of
organic wastes. These wastes have found their way into the vadose zones and groundwater
resulting in unacceptable environmental impacts. The adaptability and manageability of
indigenous microorganisms such as methanotrophs make them ideal for the remediation of
hazardous environmental wastes under a diverse range of habitats.
 Figure 1. Oxidation of methane by the methane monooxygenase enzyme
and associated cometabolism of trichloroethylene.

BIOREMEDIATION OF CONTAMINATED SOIL

Many substances known to have toxic properties have been introduced into
the environment through human activity. These substances range in degree
of toxicity and danger to human health. Many of these substances either
immediately or ultimately come in contact with and are sequestered by soil.
Conventional methods to remove, reduce, or mitigate toxic substances
introduced into soil or ground water via anthropogenic activities and
processes include pump and treat systems, soil vapor extraction,
incineration, and containment. Utility of each of these conventional methods
of treatment of contaminated soil and/or water suffers from recognizable
drawbacks and may involve some level of risk.
The emerging science and technology of bioremediation offers an alternative
method to detoxify contaminants. Bioremediation has been demonstrated
and is being used as an effective means of mitigating:
• hydrocarbons
• halogenated organic solvents
• halogenated organic compounds
• non-chlorinated pesticides and herbicides
• nitrogen compounds
• metals (lead, mercury, chromium)
• radionuclides
Bioremediation technology exploits various naturally occurring mitigation
processes: natural attenuation, biostimulation, and bioaugmentation.
Bioremediation which occurs without human intervention other than
monitoring is often called natural attenuation. This natural attenuation
relies on natural conditions and behavior of soil microorganisms that are
indigenous to soil. Biostimulation also utilizes indigenous microbial
populations to remediate contaminated soils. Biostimulation consists of
adding nutrients and other substances to soil to catalyze natural attenuation
processes. Bioaugmentation involves introduction of exogenic
microorganisms (sourced from outside the soil environment) capable of
detoxifying a particular contaminant, sometimes employing genetically
altered microorganisms (Biobasics, 2006).
During bioremediation, microbes utilize chemical contaminants in the soil as
an energy source and, through oxidation-reduction reactions, metabolize the
target contaminant into useable energy for microbes. By-products
(metabolites) released back into the environment are typically in a less toxic
form than the parent contaminants. For example, petroleum hydrocarbons
can be degraded by microorganisms in the presence of oxygen through
aerobic respiration. The hydrocarbon loses electrons and is oxidized while
oxygen gains electrons and is reduced. The result is formation of carbon
dioxide and water (Nester et al., 2001). When oxygen is limited in supply or
absent, as in saturated or anaerobic soils or lake sediment, anaerobic
(without oxygen) respiration prevails. Generally, inorganic compounds such
as nitrate, sulfate, ferric iron, manganese, or carbon dioxide serve as
terminal electron acceptors to facilitate biodegradation (State of Mississippi,
Department of Environmental Quality, 1998).
Three primary ingredients for bioremediation are: 1) presence of a
contaminant, 2) an electron acceptor, and 3) presence of microorganisms
that are capable of degrading the specific contaminant. Generally, a
contaminant is more easily and quickly degraded if it is a naturally occurring
compound in the environment, or chemically similar to a naturally occurring
compound, because microorganisms capable of its biodegradation are more
likely to have evolved (State of Mississippi, Department of Environmental
Quality, 1998). Petroleum hydrocarbons are naturally occurring chemicals;
therefore, microorganisms which are capable of attenuating or degrading
hydrocarbons exist in the environment. Development of biodegradation
technologies of synthetic chemicals such DDT is dependent on outcomes of
research that searches for natural or genetically improved strains of
microorganisms to degrade such contaminants into less toxic forms.
Microorganisms have limits of tolerance for particular environmental
conditions, as well as optimal conditions for pinnacle performance. Factors
that affect success and rate of microbial biodegradation are nutrient
availability, moisture content, pH, and temperature of the soil matrix.
Inorganic nutrients including, but not limited to, nitrogen, and phosphorus
are necessary for microbial activity and cell growth. It has been shown that
“treating petroleum-contaminated soil with nitrogen can increase cell growth
rate, decrease the microbial lag phase, help to maintain microbial
populations at high activity levels, and increase the rate of hydrocarbon
degradation” (Walworth et al., 2005). However, it has also been shown that
excessive amounts of nitrogen in soil cause microbial inhibition. Walworth et
al. (2005) suggest maintaining nitrogen levels below 1800 mg nitrogen/kg
H2O for optimal biodegradation of petroleum hydrocarbons. Addition of
phosphorus has benefits similar to that of nitrogen, but also results in similar
limitations when applied in excess (State of Mississippi, Department of
Environmental Quality, 1998).
All soil microorganisms require moisture for cell growth and function.
Availability of water affects diffusion of water and soluble nutrients into and
out of microorganism cells. However, excess moisture, such as in saturated
soil, is undesirable because it reduces the amount of available oxygen for
aerobic respiration. Anaerobic respiration, which produces less energy for
microorganisms (than aerobic respiration) and slows the rate of
biodegradation, becomes the predominant process. Soil moisture content
“between 45 and 85 percent of the water-holding capacity (field capacity) of
the soil or about 12 percent to 30 percent by weight” is optimal for
petroleum hydrocarbon degradation (US EPA, 2006, “Landfarming”).
Soil pH is important because most microbial species can survive only within a
certain pH range. Furthermore, soil pH can affect availability of nutrients.
Biodegradation of petroleum hydrocarbons is optimal at a pH 7 (neutral); the
acceptable range is pH 6 – 8 (US EPA, 2006, “Landfarming”; State of
Mississippi, Department of Environmental Quality, 1998).
Temperature influences rate of biodegradation by controlling rate of
enzymatic reactions within microorganisms. Generally, “speed of enzymatic
reactions in the cell approximately doubles for each 10 oC rise in
temperature” (Nester et al., 2001). There is an upper limit to the
temperature that microorganisms can withstand. Most bacteria found in soil,
including many bacteria that degrade petroleum hydrocarbons, are
mesophiles which have an optimum temperature ranging from 25 degree C
to 45 degree C (Nester et al., 2001). Thermophilic bacteria (those which
survive and thrive at relatively high temperatures) which are normally found
in hot springs and compost heaps exist indigenously in cool soil
environments and can be activated to degrade hydrocarbons with an
increase in temperature to 60 degree C. This finding “suggested an intrinsic
potential for natural attenuation in cool soils through thermally enhanced
bioremediation techniques” (Perfumo et al., 2007).
Contaminants can adsorb to soil particles, rendering some contaminants
unavailable to microorganisms for biodegradation. Thus, in some
circumstances, bioavailability of contaminants depends not only on the
nature of the contaminant but also on soil type. Hydrophobic contaminants,
like petroleum hydrocarbons, have low solubility in water and tend to adsorb
strongly in soil with high organic matter content. In such cases, surfactants
are utilized as part of the bioremediation process to increase solubility and
mobility of these contaminants (State of Mississippi, Department of
Environmental Quality, 1998). Additional research findings of the existence
of thermophilic bacteria in cool soil also suggest that high temperatures
enhance the rate of biodegradation by increasing the bioavailability of
contaminants. It is suggested that contaminants adsorbed to soil particles
are mobilized and their solubility increased by high temperatures (Perfumo
et al., 2007).
Soil type is an important consideration when determining the best suited
bioremediation approach to a particular situation.
In situ bioremediation refers to treatment of soil in place. In situ
biostimulation treatments usually involve bioventing, in which oxygen
and/or nutrients are pumped through injection wells into the soil. It is
imperative that oxygen and nutrients are distributed evenly throughout the
contaminated soil. Soil texture directly affects the utility of bioventing, in as
much as permeability of soil to air and water is a function of soil texture.
Fine-textured soils like clays have low permeability, which prevents
biovented oxygen and nutrients from dispersing throughout the soil. It is also
difficult to control moisture content in fine textured soils because their
smaller pores and high surface area allow it to retain water. Fine textured
soils are slow to drain from water-saturated soil conditions, thus preventing
oxygen from reaching soil microbes throughout the contaminated area (US
EPA, 2006, “Bioventing”). Bioventing is well-suited for well-drained, medium,
and coarse-textured soils.
In situ bioremediation causes minimal disturbance to the environment at
the contamination site. In addition, it incurs less cost than conventional soil
remediation or removal and replacement treatments because there is no
transport of contaminated materials for off-site treatment. However, in situ
bioremediation has some limitations: 1) it is not suitable for all soils, 2)
complete degradation is difficult to achieve, and 3) natural conditions (i.e.
temperature) are hard to control for optimal biodegradation.

Ex situ bioremediation, in which contaminated soil is excavated and treated

elsewhere, is an alternative.
Ex situ bioremediation approaches include use of bioreactors, landfarming,
and biopiles. In the use of a bioreactor, contaminated soil is mixed with
water and nutrients and the mixture is agitated by a mechanical bioreactor
to stimulate action of microorganisms. This method is better-suited to clay
soils than other methods and is generally a quick process (US EPA, 2006,
“Guide”).
Landfarming involves spreading contaminated soil over a collection system
and stimulating microbial activity by allowing good aeration and by
monitoring nutrient availability (US EPA, 2006, “Landfarming”).
Biopiles are mounds of contaminated soils that are kept aerated by pumping
air into piles of soil through an injection system (US EPA, 2006, “Biopiles”).
In each of these methods, conditions need to be monitored and adjusted
regularly for optimal biodegradation. Use of landfarming and biopiles also
present the issue of monitoring and containing volatilization of contaminants.
Like in situ methods, ex situ bioremediation techniques generally cost less
than conventional techniques and apply natural methods. However, they can
require a large amount of land and, similar to in situ bioremediation,
complete degradation is difficult to achieve, and evaporation of volatile
components is a concern (US EPA, 2006, “Landfarming”; US EPA, 2006,
“Biopiles”).
If the challenges of bioremediation, particularly of in situ techniques, can be
overcome, bioremediation has potential to provide a low cost, non-intrusive,
natural method to render toxic substances in soil less harmful or harmless
over time. Currently, research is being conducted to improve and overcome
limitations that hinder bioremediation of petroleum hydrocarbons. On a
broader scope, much research has been and continues to be developed
enhance understanding of the essence of microbial behavior as microbes
interact with various toxic contaminants. Additional research continues to
evaluate conditions for successful introduction of exogenic and genetically
engineered microbes into a contaminated environment, and how to translate
success in the laboratory to success in the field (US DOE, 2006).

BIOREMEDIATION OF OIL POLLUTANTS

An oil spill is a release of a liquid petroleum hydrocarbon into the
environment due to human activity, and is a form of pollution. The term
often refers to marine oil spills, where oil is released into the ocean or
coastal waters. Oil spills include releases of crude oil from tankers, offshore
platforms, drilling rigs and wells, as well as spills of refined petroleum
products (such as gasoline, diesel) and their by-products, and heavier fuels
used by large ships such as bunker fuel, or the spill of any oily refuse or
waste oil. Spills may take months or even years to clean up.
Oil also enters the marine environment from natural oil seeps. Public
attention and regulation has tended to focus most sharply on seagoing oil
tankers.
The oil penetrates into the structure of the plumage of birds, reducing its
insulating ability, thus making the birds more vulnerable to temperature
fluctuations and much less buoyant in the water. It also impairs birds' flight
abilities to forage and escape from predators. As they attempt to preen,
birds typically ingest oil that covers their feathers, causing kidney damage,
altered liver function, and digestive tract irritation. This and the limited
foraging ability quickly causes dehydration and metabolic imbalances.
Hormonal balance alteration including changes in luteinizing protein can also
result in some birds exposed to petroleum.
Most birds affected by an oil spill die unless there is human intervention.
Marine mammals exposed to oil spills are affected in similar ways as
seabirds. Oil coats the fur of Sea otters and seals, reducing its insulation
abilities and leading to body temperature fluctuations and hypothermia.
Ingestion of the oil causes dehydration and impaired digestions. Because oil
floats on top of water, less sunlight penetrates into the water, limiting the
photosynthesis of marine plants and phytoplankton. This, as well as
decreasing the fauna populations, affects the food chain in the
ecosystem.There are three kinds of oil-consuming bacteria. Sulfate-reducing
bacteria (SRB) and acid-producing bacteria are anaerobic, while general
aerobic bacteria (GAB) are aerobic. These bacteria occur naturally and will
act to remove oil from an ecosystem, and their biomass will tend to replace
other populations in the food chain.

REFERENCE
• Biobasics: The Science and the Issues. 9 Feb 2006. 24 Nov 2006
http://www.biobasics.gc.ca/english/View.asp?x=741
• Nester, Eugene W., Denise G. Anderson, C. Evans Roberts Jr., Nancy N.
Pearsall, and Martha T. Nester. 2001. Microbiology: A Human
Perspective. 3 rd ed. New York: McGraw-Hill.
• Perfumo, Amedea, Ibrahim M. Banat, Roger Marchant, and Luigi
Vezzulli. 2007. “Thermally Enhanced Approaches for Bioremediation of
Hydrocarbon-Contaminated Soils.” Chemosphere 66: 179-184.
• State of Mississippi. Department of Environmental Quality. 1998.
Fundamental Principles of Bioremediation. April 1998. 27 Nov 2006
<http://www.deq.state.ms.us/MDEQ.nsf/pdf/ GARD_Bioremediation/
$File/Bioremediation.pdf? Verified 12/15/2006.
• Walworth, James, Andrew Pond, Ian Snape, John Rayner, Susan
Ferguson, and Paul Harvey. 2005. “Fine Tuning Soil Nitrogen to
Maximize Petroleum Bioremediation.” ARCSACC (2005): 251-257.
• ^ "Hindsight and Foresight, 20 Years After the Exxon Valdez Spill". NOAA.
2010-03-16. Retrieved 2010-04-30.
• Dunnet, G., Crisp, D., Conan, G., Bourne, W. (1982) "Oil Pollution and
Seabird Populations [and Discussion]" Philosophical Transactions of the
Royal Society of London. B 297(1087): 413–427
• Pezeshki, S. R., Hester, M. W., Lin, Q. & Nyman, J. A. (2000). The effects
of oil spill clean-up on dominant US Gulf coast marsh macrophytes: a
review. Environmental Pollution. 108: 129-139