ABSTRACT
INTRODUCTION
METHODOLOGY
BIOREMEDIATION OF OIL SPILLS
CASE STUDY
CONCLUSION
Subtopics
Bio remediation in hot and cold environments
Use of Nitrogen fixing Bacteria
Bio remediation using fungi from soil samples
Bio remediation using bacteria and case studies
4. INTRODUCTION
An oil spill is the release of a liquid petroleum hydrocarbon
into the environment, especially the marine ecosystem, due
to human activity.
Bioremediation is emerging technology for the
treatment of soil and ground water oil spills.
Different microorganisms are used in bioremediation
such as bacteria, fungi and algae etc. It is a cost-effective
and environmentally sound remediation technology
5. Oil spills occur frequently
throughout the world. The
production and
transportation of crude oil
causes the incidents of oil
spills.
Bioremediation is the most
important technology used
in restoration of oil-
polluted environments by
the use of natural microbial
biodegradation activities..
6. Bioremediations of many
hydrocarbon-contaminated
environments are dependent on
various factors such as temperature,
pH, salt concentrations or pressure.
Microorganisms, adapted to grow and
thrive in these environments, play an
important role in the bioremediation
of polluted extreme habitats.
7. BIOREMEDIATION IN LOW AND
HIGH TEMPERATURE
COLD ADAPTED
MICROORGANISMS
– Psychrophiles have an
optimum growth temperature
of ≤15°C and do not grow
above 20°C
– psychrotrophs (cold-tolerant)
have optimum and maximum
growth temperatures above
15°C and 20°C.
PSYCHROPHILES
PSYCHOTROPHS
8. BIOREMEDIATION IN LOW AND
HIGH TEMPERATURE
THERMOPHILIC
MICROORGANISMS
– Thermophiles are microorganisms
that can grow optimally above
40°C. Most thermophiles show an
upper temperature border of
growth between 50 and 70°C.
Optimal growth of extreme
thermophiles and
hyperthermophiles occurs at 70–
80°C and above 80°C.
THERMOPHILES
HYPERTHERMOPHILES
9. BIOREMEDIATION OF SOILS
BY USING
BIOAUGMENTATION,
BIOSTIMULATION AND
FREE-LIVING NITROGEN
FIXING BACTERIA
Presentd by
10. INTRODUCTION
The production and
transportation of
crude oil can cause
incidents of oil spills.
Aging phenomenon
is one of the well-
known phenomenon
in oil contamination
Due to aging
phenomenon,
biodegradation
efficiency is reduced.
11. Continue…
Bio augmentation is
most efficient method
for biodegradation
and if we use NFB
for degradation of
hydrocarbons, we
can increase the
efficiency of
biodegradation.
12. Bio-Stimulation
In this process, surviving bacteria that has
ability to degrade hydrocarbon are stimulated.
Stimulation can be done by adding some
nutrients e.g; oxygen, nitrogen etc.
Biostimulative agents are oleophilic, it means
they bond completely to hydrocarbons.
13. BIOAUGMENTATION:
Bio Augmentation is the
practice of adding cultured
microorganisms into the
subsurface for the
purpose of biodegrading
specific soil and
groundwater
contaminants.
14. Bioaugmentation vs. biostimulation
If we compare those processes
then bioaugmentation is best one
for bioremediation, because
nutrients are loaded in
biostimulation but
bioaugmentation involves natural
microbes which make
bioremediation more effieicient.
15. NITROGEN FIXING BACTERIA
Its growth rate is faster in hydrocarbons than
other bacteria. This makes NFB more efficient
and effective than other bacteria.
For example; After oil spill, Harmful effects of
oil spill lower the organic material and
nitrogen. After its treatment with NFB, its
texture becomes less oily and less sticky. NFB
count increased from 13 × 104 CFU to 2 × 109
CFU after its minimum time of treatment (16
months).
16. SELECTION OF
CONSORTIUM MEMBERS
In this method, many
species of bacteria were
present in the
contaminated soil, but
few of them are chosen
out for degradation.
Two characteristics are
analyzed, first is the
growth rate of bacteria in
hydrocarbons and second
is the production of
biosurfactants.
18. WHY USE FUNGI FOR BIOREMEDIATION OF
OIL?
The fungi are
unique among
microorganisms
because they
secrete a variety
of extracellular
enzymes
19. STEPS OF USING FUNGI FOR
BIOREMEDIATION OF OIL
STEP 1
The
isolation of
fungi
STEP 2
The
identification of
fungi from
contaminated
oil samples
STEP 3
Appliance of an
effectual
method as
biodegradation
of oil spill
STEP 4
Testing the
capacity of
isolated fungi
and showing
their ability in
bioremediation
20. BIOREMEDIATION OF OIL SPILL IN KINGDOM OF SAUDI
ARABIA
BY USING FUNGI ISOLATED FROM POLLUTED SOILS
BY A.M. MUSTAFA(SAUDI ARABIA)
Soil samples were collected from Shay bah
field and Dammam ports of Saudi Arabia.
BIODEGRADABILITY TEST – DCPIP
INDICATOR
To evaluate fungal ability for oil
biodegradation
An electron acceptor such as DCPIP was
21. Aspergillus Niger and Fusarium
oxysporum showed the highest
frequency represented by (9.8
%)
The results showed that these
two were the most efficient
fungi in reducing of oil spill
from the environment.
22. BIOREMEDIATION OF CRUDE OIL BY WHITE
ROT FUNGI POLYPORUS SP. S133
KRISTANTI(JAPAN)
The bioremediation potential of crude oil by polyporus sp.
S133 was investigated in two separate experiment trials;
liquid medium and soil.
The effect of nutrients, oxygen flow, and some absorbents
on efficiency of bioremediation was also evaluated.
The highest degradation rate of crude oil was 93%
23. ABILITY OF SOME SOIL FUNGI IN
BIODEGRADATION OF PETROLEUM
HYDROCARBON
JAWAHRI (IRAQ)
• This study investigated the abilities of fungi isolated
from polluted soil to utilize petroleum hydrocarbon
• The highest percentage loss of petroleum hydrocarbon
concentration by the cultures of fungi was 95% with a.
Niger after 28 days of treatment
24. MYCOREMEDIATION (BIOREMEDIATION WITH
FUNGI)
BY CHRISTOPHER J. RHODES
For the use of white-rot fungi successfully for bioremediation
• Knowledge must be taken from different fields of fungal physiology
,biochemistry, enzymology, ecology and other similar subjects.
• It is critical to attain the correct nitrogen/carbon ratio in the
substrates used to avoid any impeding effect on the efficiency of the
fungi
• Fungal inoculum coated with alginate, gelatin, agarose etc ,may
26. BIODEGRADATION OF PETROLEUM HYDRO
COMPOUNDS
n-alkanes of
intermediate
length (C10 –C20)
are the preferred
substrates and
most readily
degradable.
Longer chain alkanes
known as waxes (C20
–C40) are
hydrophobic and, are
difficult to degrade
due their poor water
solubility
Branched chain
alkanes are also
degraded more
slowly.
Highly condensed
aromatic
structures have
the highest
boiling points and
exhibit greatest
resistance
27. CHALLENGES OF BIOREMEDIATION IN OIL
CONTAMINATED ENVIRONMENT
Heterogeneity of
the
contaminants
Extreme
concentrations
of hydrocarbons
Variable
environmental
Conditions
28. Sand samples contaminated with oil spill
were collected from Pensacola beach (Gulf of
Mexico) and tested to isolates fungal diversity
associated with beach sands.
Species were identified by using DNA
sequence method.
The biodegradability of isolated fungi was
verified using the modified technique based
on the redox indicator 2, 6-dichlorophenol
indophenol (DCPIP)
29. Cont …..
Fungi use crude oil as a substrate for
their survival growth using extra cellular
enzymes to break down the recalcitrant
hydrocarbon molecules by converting
petroleum into simpler forms or
products that can be absorbed for the
growth and nutrition of the fungi
From sixteen fungal strains, four strains
were confirmed for biodegradation ability
of crude oil, the isolated fungi belongs to
Aspergillus niger with higher activity
weightloss of 8.6% followed by Penicillium
documbens (7.9%), Cochliobolus
lutanus(4.7%) and Fusarium solani (1.9%).
31. GULF OIL SPILL
• On April 20, 2010, the Deepwater Horizon mobile drilling
unit exploded, caught fire, and eventually sank, resulting in
a massive release of oil and other substances.
• For 87 days after the explosion, the well blasted oil and
natural gas continuously into the northern Gulf of Mexico.
• Approximately 3.19 million barrels of oil was released into
the ocean, by far the largest offshore marine oil spill in U.S.
history .
32. Cont….
Oil under pressure gushed into the deep ocean from BP’s
Macondo well.
Oil moved with deep-sea currents, creating a plume of oil
within the deep sea.
More buoyant oil traveled up through about a mile (1.6
kilometers) of water column and formed large surface slicks.
33. RESPONSE ACTIONS:
CHEMICAL ACTIONS
• Chemical dispersants were used
such as corexit.
• Purpose was to break the oil
into smaller droplets.
• Approximately 1.84 million
gallons of chemical dispersant
were used
PHYSICAL ACTIONS
• Deploying of containment boom.
• Burning of Floating oil
• Physical removal
34.
35. BIOREMEDIATION
• The microbe used for cleaning oil was Alcanivorax borkumensis
bacteria.
• Alcanivorax borkumensis is a marine bacteria.
• Can absorb and digest linear and branched alkanes.
• It is a gram-negative bacteria, has an outer membrane of
lipopolysaccharides.
• It occurs naturally in unpolluted waters all over the world.
36. Alcanivorax borkumensis
It naturally flourishes after an oil spill because
there is a more abundant source of energy that
can sustain a larger population.
It apart the bonds in hydrocarbons in oil that have
been exposed to the sea, using enzymes and
oxygen found in the seawater.
37. conti..
Borkumensis creates enzymes AlkB1 and
AlkB2.
AlkB1 degrade alkanes with 5 to 12
carbons
and AlkB2 degrade alkanes with 8 to 16
carbons
