BIOREMEDIATION

OF OIL SPILLS
 INTRODUCTION
An oil spill is the release of a liquid petroleum hydrocarbon
into the environment (water or soil habitats), 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
 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
 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.
 BIOREMEDIATION IN LOW
AND HIGH TEMPERATURE
COLD ADAPTED – Psychrophiles have an
MICROORGANISMS optimum growth
temperature of ≤15°C and do
not grow above 20°C
PSYCHROPHILE
PSYCHOTROPH – psychrotrophs
S
S (cold-tolerant) have
optimum and maximum
growth temperatures above
 BIOREMEDIATION IN LOW
AND HIGH TEMPERATURE
THERMOPHILIC – Thermophiles are
MICROORGANISMS microorganisms that can grow
optimally above 40°C. Most
thermophiles show an upper
THERMOPHILES
temperature border of growth
HYPERTHERMOPHILES between 50 and 70°C. Optimal
growth of extreme thermophiles
and hyperthermophiles occurs at
70– 80°C and above 80°C.
 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.
BIOAUGMENTATION
:
Bio Augmentation is the
practice of adding cultured
microorganisms into the
subsurface for the purpose
of biodegrading specific
soil and groundwater
contaminants.
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.
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).
 SELECTION OF
CONSORTIUM
MEMBERS
In this method, many Two characteristics are
species of bacteria were analyzed, first is the
present in the growth rate of bacteria in
contaminated soil, but hydrocarbons and second
few of them are chosen is the production of
out for degradation. biosurfactants.
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.
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.
conti.
.
Borkumensis creates enzymes AlkB1
and AlkB2.
AlkB1 degrade alkanes with 5 to
12 carbons
and AlkB2 degrade alkanes with 8 to
16 carbons
BIOREMEDIATION OF OILL SPILLS
USING FUNGI
WHY USE FUNGI FOR BIOREMEDIATION
OF OIL?

The fungi are

unique among
microorganisms
because they
secrete a variety
of extracellular
enzymes
 STEPS OF USING FUNGI FOR
BIOREMEDIATION OF OIL

STEP 3 STEP 4
STEP 1 STEP 2
Appliance of an Testing the
The
capacity of
The identification of effectual
isolated fungi
fungi from method as
isolation of contaminated biodegradation
and showing
their ability in
fungi oil samples of oil spill bioremediation
 BIODEGRADABILITY TEST – DCPIP
INDICATOR

Hydrocarbon contaminated Soils were to evaluate

fungal ability for oil biodegradation
An electron acceptor such as DCPIP was added
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.
 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%

 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
 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

increased the efficiency of the fungi to reduce contaminants
BIODEGRADATION OF PETROLEUM
HYDRO COMPOUNDS

n-alkanes of Longer chain alkanes Highly condensed

intermediate known as waxes (C20 aromatic
Branched chain
length (C10 –C20) structures have
–C40) are alkanes are also
are the preferred the highest
hydrophobic and, degraded more
substrates and boiling points and
slowly.
most readily are difficult to exhibit greatest
degradable. degrade resistance
due their poor
water solubility
CHALLENGES OF BIOREMEDIATION IN
OIL CONTAMINATED
ENVIRONMENT
Heterogeneity
of the
contaminants

Variable Extreme
environmental concentrations
Conditions of hydrocarbons
 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)
 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%).
 FEASIBLE AND
COST
EFFECTIVE

SIGNIFICANCE OF
EXTREMOPHILIC
NATURALL MICROORGANISMS.
Y NITROGEN FIXING
CONDITIONS
OCCURING

PROMISING
CANDIDATES TO
LESS TOXIC REMOVE
PETROLEUM
THAN HYDROCARBON
DISPERSANTS FROM
CONTAMINATED
ENVIRONMENTS.