BIOREMEDIATION

 Man is a rational being and is curious to

exploit/discover new things in the
environment. In order for him to understand
the life in a better aspect and try to solve
the mystery of the nature/environment.

 While exploiting the environment, human

activities resulted to pollution and
RELEASE OF certain HAZARDOUS
MATERIALS.
 Bioremediation came as an option that
offers the possibility to destroy or render
harmless various contaminants using
natural biological activity.
 Bioremediation

 the
branch of biotechnology that
uses biological process to
overcome environmental problems.
 Bioremediation can be defined
as any process that uses:
microorganisms, fungi, green
plants or their enzymes to return the
natural environment altered by
contaminants to its original condition.
 History

 Bioremediation technology using

microorganisms was reportedly
invented by George M. Robinson.
 He was the assistant county
petroleum engineer for Santa Maria,
California. During the 1960s, he spent
his spare time experimenting with dirty
jars and various mixes of microbes.
 Bioremediation may be
employed:
 to attack specific soil contaminants, such as
degradation of chlorinated hydrocarbons by
bacteria.

 An example of a more general approach is the

cleanup of oil spills by the addition of nitrate
and/or sulphate fertilisers to facilitate the
decomposition of crude oil by indigenous or
exogenous bacteria.
How might microorganisms
attack hazardous organic
wastes?
 BIODEGRADATION

 The breakdown of organic compounds by

microorganisms

 Mineralizecompound directly,
compound converted to harmless
inorganic molecules such as
carbon dioxide and salts
 Cont.
 Ofprime importance are microorganisms
capable of producing enzymes that will
degrade the hazardous chemical (target
compound) as enzymes degrade
compounds through exploitation of the
organism’s energy need.

 Converting compound to some other

compound, which may also be toxic and
recalcitrant to further degradation
REQUIREMENTS FOR BIOREMEDIATION

MICROORGANISMS

ENERGY ELECTRON
SOURCE ACCEPTOR

MOISTURE pH

NUTRIENTS TEMPERATURE

ABSENCE OF REMOVAL OF ABSENCE OF

TOXICITY METABOLITIES COMPETITIVE
ORGANISMS

BIOREMEDIATION
 THE BASIC PROBLEM:
RELEASE OF HAZARDOUS MATERIALS

 Enormous quantities of organic & inorganic

compounds are released into the environment each
year as a result of human activities.

 The release may be:

 Deliberate and well regulated (industrial
emissions)
 Accidental and largely unavoidable (chemical/oil
spills)
 US EPA estimated that in 1980 at least 57 millions
metric tons of the total waste can be categorized
into three general groups:
 Heavy metal, Pb, Hg, Cd, Ni and Be can
accumulate in various organs, interfere with
normal enzymatic reactions and cause disease
including cancer

 Chlorinated hydrocarbons, also known as

organochlorides including pesticides and other
organic compounds such as PCB (polychlorinated
biphenyls)
 Research proven a positive correlation
between cancer in lab animals and
organochlorides.

 Nuclear waste including radioactive material such

as plutonium which are dangerous for thousands
of years
 BIOREMEDIATION

 Bioremediation is the application of biological process

principles to the treatment of groundwater, soil and sludges
contaminated with hazardous chemicals.

 It requires the control and manipulation of microbial

processes in surface reactors or in the subsurface.

 The contaminants can be biodegraded in situ or removed

and placed in bioreactor (at or off the contamination sites).

 Idea:
 To isolate microbes that can degrade or eat a particular
contaminant
 To provide the conditions whereby it can do this most
effectively, thereby eliminating the contaminant
CONCEPTS:

 Heterotrophic microorganisms are the principal user

of organic matter in the biosphere and are key in
cycling carbon from the organic to the inorganic
state. Provided that sufficient inorganic nutrients as
an energy source and a terminal electron acceptor
for metabolism are present, all naturally occurring
organic material can be biodegraded eventually.
CONCEPTS:

 Simple organic compounds such as acetate may

persist under condition that do not favor microbial
activity. These conditions include extremes in
temperature or pH, the presence of toxicants or
antimicrobial agents, the inhibition or exclusion of
microbial enzymes, and the lack of water and an
electron acceptor.
Microbial Divisions
 Two kinds of cells are recognized, the procaryotic and
eucaryotic.

Procaryotic cell Eucaryotic cell

Bacteria Plants
Blue-green bacteria or Animals
cyanobacteria Rotifers
Protozoa
Fungi
Most algae
 The most important groups to bioremediation are
bacteria and fungi.
 Microorganisms destroy organic
contaminants in the course of using
the chemicals for their own growth
and reproduction.
 Organic chemicals provide:

carbon, source of cell building

material, electrons, source of energy
 Metabolism is defined by the nature of the redox
reaction
 Metabolism modes are divided into two; aerobic
and anaerobic
 Cells catalyze oxidation of organic chemicals
(electron donors), causing transfer of electrons
from organic chemicals to some electron
acceptor

 Electron acceptors:
In aerobic oxidation, acceptor is oxygen
In anaerobic, acceptor is:
-nitrate
-manganese
-iron
-sulfate
TYPES OF BIOREMEDIATION

 The two main types of bioremediation

are in situ bioremediation and ex situ
bioremediation. In addition, another
offshoot of bioremediation is
phytoremediation.
 In Situ Bioremediation
 In situ bioremediation is when the
contaminated site is cleaned up exactly
where it occurred. It is the most commonly
used type of bioremediation because it is
the cheapest and most efficient, so it’s
generally better to use. There are two main
types of in situ bioremediation: intrinsic
bioremediation and accelerated
bioremediation.
 Intrinsic Bioremediation
 Intrinsic bioremediation uses
microorganisms already present in the
environment to biodegrade harmful
contaminant. There is no human
intervention involved in this type of
bioremediation, and since it is the cheapest
means of bioremediation available, it is the
most commonly used. When intrinsic
bioremediation isn’t feasible, scientists turn
next to accelerated bioremediation.
Accelerated Bioremediation

 In accelerated bioremediation, either

substrate or nutrients are added to the
environment to help break down the
toxic spill by making the microorganisms
grow more rapidly. Usually the
microorganisms are indigenous, but
occasionally microorganisms that are
very efficient at degrading a certain
contaminant are additionally added.
 Main advantage is that site disturbance is
minimized, which is particularly important
when the contaminated plume has moved
under permanent structures.

 Biggest limitation of in situ treatment has

been the inability to deal effectively with metal
contaminants mixed with organic compounds.

 The goal of in situ treatment is to manage and

manipulate the subsurface environment to
optimize microbial degradation.
 In Situ Bioremediation
 Land treatments:

Bioventing is the most common in situ

treatment and involves supplying air
and nutrients through wells to
contaminated soil to stimulate the
indigenous bacteria.
In situ biodegradation involves
supplying oxygen and nutrients by
circulating aqueous solutions through
contaminated soils to stimulate
naturally occurring bacteria to
degrade organic contaminants.

Bioaugmentation Bioremediation
frequently involves the addition of
microorganisms indigenous or
exogenous to the contaminated sites.
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
thereby increases the contact
between soil and groundwater.
 Ex Situ Bioremediation
 Another type of bioremediation is ex situ
bioremediation, which is when
contaminated land are taken out of the
area to be cleaned up by the organisms.
This type of bioremediation is generally
used only when the site is threatened for
some reason, usually by the spill that
needs to be cleaned up. Ex situ
bioremediation is only used when
necessary because it’s expensive and
damaging to the area, since the
contaminated land is physically removed.
 Ex Situ Bioremediation
Landfarming is a simple technique in which
contaminated soil is excavated and spread
over a prepared bed and periodically tilled
until pollutants are degraded.

Composting is a technique that involves

combining contaminated soil with non-
hazardous organic compounds such as
agricultural wastes. The presence of these
organic materials supports the development
of a rich microbial population and elevated
temperature characteristic of composting.
 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.
 Phytoremediation

 Phytoremediation is the use of plants

to clean up potentially damaging
spills. The plants work with soil
organisms to transform contaminants,
such as heavy metals and toxic
organic compounds, into harmless or
valuable forms.
 Biodegradation
• Biodegradation – microbial catalyzed
reduction in complexity of chemicals

• Involves the breakdown of organic

compounds either through biotransformation
into less complex metabolites or through
mineralization into inorganic minerals, H2O,
CO2 or CH4.

• Mineralization - conversion of an organic

substrate to inorganic end products
• Growth-linked metabolism biodegradation
provides carbon and energy to support growth.

• Maintenance metabolism biodegradation not

linked to multiplication, but to obtaining carbon for
respiration to maintain cell viability; take place only
when organic carbon concentrations very low.

• The extent and rate of biodegradation depend on

many factors including pH, temperature, oxygen,
microbial population, degree of acclimation,
accessibility of nutrients, chemical structure of the
compound, cellular transport properties and
chemical partitioning in growth medium.
 BIODEGRADATION SYSTEM IN BIOREMEDIATION

MICROORGANISMS
•Growth
•Physiology
•Genetic competence
•Metabolic diversity
•Enzymology
•metabolites

CONTAMINANTS ENVIRONMENTAL
•Mass transfer FACTORS
•Bioavailability •pH
•Hydrophobicity •Temperature
•Recalcitrance •Moisture
•Structure •Oxygen
•Toxicity •Nutrients
•Soil type
Microorganisms
 Microorganisms can be isolated from almost
any environmental conditions.
 Microbes will adapt and grow at different
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.
 Because of the adaptability of microbes and
other biological systems, these can be used
to degrade or remediate environmental
hazards.
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.
 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.
 The nutritional requirement of carbon to
nitrogen ratio is 10:1, and carbon to
phosphorous is 30:1.
Environmental requirements
Limitations to biodegradation
 Adequate bacterial concentrations (although
populations generally increase if there is
food present)
 Electron acceptors
 Nutrients (e.g., nitrogen and phosphorus)
 Non-toxic conditions
 Minimum carbon source
Relative biodegradability
Advantages of bioremediation
 Bioremediation is a natural process and is
therefore perceived by the public as an
acceptable waste treatment process
 Many compounds that are legally
considered to be hazardous can be
transformed to harmless products.
 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.
Advantages of bioremediation

 Bioremediation can often be carried

out on site, often without causing a
major disruption of normal activities.
 Bioremediation is less expensive
Disadvantages of bioremediation

 Bioremediation is limited to those

compounds that are biodegradable.
 There are some concerns that the
products of biodegradation may be
more persistent or toxic than the
parent compound.
 It is difficult to extrapolate from bench
and pilot-scale studies to full-scale
field operations.
Disadvantages of bioremediation
 Bioremediation often takes longer
than other treatment options
 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.
Case Study: Oil spill Bioremediation

 As a result of the petroleum industry millions

of tons of these compounds enter the oceans
every year. Many hydrocarbons dissolve
slowly in water. Others such as the aromatic
compounds like benzene are more soluble,
and these are toxic to living cells.

 While accidental releases may contribute to

only a small percentage of the oil released
into the marine environment large accidental
oil spills receive much attention and evoke
considerable public concern because they
can result in contamination of ocean and
shoreline environments.
 Oil spill!!

The biggest spill ever occurred during the

1991 Persian Gulf war when about 240
million gallons spilled from oil terminals and
tankers off the coast of Prince William
Sound, Alaska. The Exxon Valdez accident
at Bligh Reef in 1989 discharged 40 million
litres….
Bioremediation to the rescue?
 Initial studies showed that the number of oil
degrading microorganisms on oiled beaches in
comparison with untreated controls increased by
as much as 10,000 times.

 Oleophilic fertilizer enhanced biodegradation of

oil.

 Bioremediation was a useful cleanup alternative

that was used by Exxon on large scale.
 Oleophilic fertilizer proven to be an
effective nutrient source for oil degrading
microbial communities.

 The beaches are more compatible with

local wildlife (less tendency for fur and
feathers to become oiled).