Petroleum Microbiology Lecture 5 Oil Spill

MCB 410
PETROLEUM MICROBIOLOGY
LECTURE 5
BY:
DR. M.A. ADEFISOYE
EFFECTS OF OIL SPILLS ON
MICROBIAL ECOLOGY OF SEA AND
SOILS
EFFECTS OF OIL SPILLS ON MICROBIAL ECOLOGY
OF SEA AND SOILS
Oil Spillage
• When crude oil or refined oil is spilt into any
environment deliberately or accidentally most
especially if it is on a large quantity, it is
referred to as Oil Spillage.
Sources of Oil Spill
 Leakages from tankers- probably due to corrosion, some spot on the
tankers are leaking.
 Leakages from oil-rig platforms: may happen during offshore oil
drilling.
.
 Accidental spills e.g. collision of oil tankers on the

sea or by road tankers on land.
Sources of Oil Spill
 Pipeline breaks: through corrosion or damage by
bunkering.
 Used motors and industrial oils: in garages and
mechanic shops we find a lot of grease.
 Normal tanker operations: Oil has to be loaded and
during this we find oil being spilt.
 Refinery waste: Heavy distillate found at the bottom
of refinery machine which are dumped off.
Sources of Oil Spill (contd.)
• Normal off-shore production.
• Ballast water:
 When a tanker is loaded with crude or refined
petroleum a certain weight must be achieved to
make the tanker buoyant on the sea.
 When you add the weight of the tanker (w1) and the
weight of the ship (w2), w1 + w2 must make ship
buoyant.
 When oil has been off-loaded from the tanker, to
keep w1 so that ship will be buoyant, the tanker is
loaded with water from the sea, that is the water
required to keep the ballast floating.
 This is emptied into the sea when tanker is to be
reloaded, thus constitute a source of spill
environment.
 It has been observed that roughly between 2.1 - 4.9

metric tons pollute the sea water annually.
What is Oil Pollution?
When oil is spilt into an environment and it
brings about an undesirable effect it is referred
to as Oil Pollution.
Effect of Pollution
• Oil spill can affect the environment both as a
result of physical smothering and toxic effects.
• An effect can be either direct damage to a

resource or damage to the ability of an
environment to support a resource.
Effect of Pollution
Oil can
• kill marine organisms,
• reduce their fitness through sublethal effects, and
• disrupt the structure and function of marine
communities and ecosystems.
Effect of Pollution
• sublethal long-term effects on populations,

communities and ecosystems at low doses and
in the presence of other contaminants poses
significant scientific challenges.
Ecological Effects:
• Ecological effects of polluting oil depend largely on the
response of the microorganism in that ecosystem.
• There are evidences to show that hydrocarbons are

deleterious to some microorganisms while some of
them are capable to use hydrocarbon as a source of
energy.
• therefore, when oil is spilt into an environment

• the microorganism that can utilize the hydrocarbon
will survive, multiply and increase their population e.g.
 the fungus Fucus have some tolerance to crude oil.
Ecological Effects:
 Fucus vesiculosus is resistant to light and heavy

exposure to fuel oil in laboratory experiments. This
may have been due to the mucilage layer found in
Fucus.
 Cross et al. (1987) found that biomass of two of

three algal species tested at Baffin Island, Canada
were not affected by an experimental subsurface
release of oil.
 The other species, Dictyosiphon foeniculaceus,

showed an increase in growth in the year following
the release of oil.
Ecological Effects:
 On the other hand, those microorganisms that would

not utilize the hydrocarbons will not survive and
probably go into extinct.
 Oil pollution may therefore result in what is called

Shifts in Ecological Balance.
Ecological Effects (contd.)
• The degradation of spilt petroleum hydrocarbons can
be mediated
 by specific enzyme system.
Other mechanisms involved are;

 attachment of microbial cells to the substrates and
 production of biosurfactants.
Ecological Effects (contd.)
• The uptake mechanism linked to the attachment of

cell to oil droplet is still unknown but production of
biosurfactants has been well studied.
• Surfactants enhance solubilization and removal of

contaminants.
• Biodegradation is also enhanced by surfactants due

to increased bioavailability of pollutants.
• biosurfactants also help in bioremediation of oil

sludge using
Biosurfactants
Biosurfactants
• (BS) are heterogeneous group of surface active
chemical compounds produced by a wide variety of
microorganisms.
• Amphiphilic (Hydrophobic and hydrophilic) compounds

and
• Main structural types are lipopeptides and glycolipids,

being the fatty acid molecule, the hydrophobic part.
•BS are compounds produced on living surfaces,
mostly microbial cell surfaces, or excreted
extracellularly and
•contain hydrophobic and hydrophilic moieties

that reduce surface tension (and interfacial
tensions between individual molecules at the
surface and interface respectively.
Biosurfactants
•BS structure may be that of Glycolipids,

Lipoproteins/lipopeptides, Lipopolysaccharides, mycolic
acid, Phospholipids or the microbial cell surface.
•They vary in chemical properties and molecular size.
•While the low molecular weight BS are often glycolipids,
•the high mol. wt ones are generally either polyanionic

heteropolysacharides or complexes with both
polysaccharides and proteins.
able 1: Biosurfactants produced by microorganisms
Biosurfactants Microorganisms
Corynomycolates and Mycolates Corynobacterium sp. Mycobacterium
sp., Nocardia sp. and Rhodococcus sp.
Glycolipid Aeromonas sp., Bacillus sp.

Lipomannan Candida tropicalis
Lipoproteins/lipopeptides e.g Bacillus subtilis
surfactin, subtilisin
Lipopolysaccharides e.g. Acinetobacter sp.
emulsan
Phospholipids Candida sp., Micrococcus sp.,
Thiobacillus sp.
Rhamnolipids Pseudomonas aeruginosa, P.
fluorescens
Sophorolipids Candida bombicola, Torulopsis sp.
Surfactin Bacillus subtilis
Toxic Effects of Petroleum Hydrocarbons
The responses of organisms to petroleum hydrocarbons
can be manifested at four levels of biological
organization;
(1.) Biochemical and cellular:
 losses in membrane integrity resulting from the
partitioning of lipophilic compounds.
 For example, membrane fluidity can be decreased
through increased membrane ordering.
(2.) Organismal:
including the integration of physiological, biochemical
and behavioral responses:
In terms of general stress responses, bacteria may
form biofilms, alter their cell surface hydrophobicity to
regulate their partitioning with respect to hydrocarbon-
water interfaces.
Toxic Effects of Petroleum Hydrocarbons
The responses of organisms to petroleum
hydrocarbons can be manifested at four levels
of biological organization;
(3.) Population:
including alterations in population dynamics.
(4.) Community:
resulting in alterations in community structure
and dynamics.
Toxic Effects of Petroleum Hydrocarbons (contd.)
• Impairment of behavioral, developmental, and
physiological processes may occur at concentrations
significantly lower than acutely toxic levels;
• such responses may alter the long-term survival of

affected populations.
• Thus, the integration of physiological and behavioral

disturbances may result in alterations at the
population and community levels.
•The effects of petroleum hydrocarbons in the
marine environment can be either acute or
chronic.
•Acute toxicity is defined as the immediate

short-term effect of a single exposure to a
toxicant.
•Chronic toxicity is defined as either the effects

of long-term and continuous exposure to a
toxicant or the long-term sublethal effects of
acute exposure.
• Acute and chronic toxicity of petroleum hydrocarbons to marine
organisms is dependent upon:
 concentration of petroleum hydrocarbons and length of exposure,
 persistence and bioavailability of specific hydrocarbons,
 the ability of organisms to accumulate and metabolize various

hydrocarbons,
 the fate of metabolized products,
 the interference of specific hydrocarbons (or metabolites) with

normal metabolic processes that may alter an organism’s
chances for survival and reproduction in the environment, and
 the specific narcotic effects of hydrocarbons on nerve

transmission.
EFFECTS OF OIL SPILLS ON MICROBIAL ECOLOGY OF SEA AND
SOILS (Contd.)
• The acute toxicity of individual hydrocarbons is largely related to their
water solubility.
• The acute toxicity of a specific oil type is the result of the additive toxicity
of individual compounds, especially aromatic compounds.
• Narcotic effects of individual petroleum compounds are an important

component of acute toxicity and are most closely related to low molecular
weight volatile compounds.
• Sublethal effects following acute or chronic exposure to petroleum

hydrocarbons include;
 disruption in energetic processes,
 interference with biosynthetic processes and structural development,
and
 direct toxic effects on developmental and reproductive stages.
Mans Effort to Control Oil Pollution
• The most satisfactory way of control is to prevent
occurrence.
• Prevention – this is to prevent willful discharge of oil,

as in ballast water and oil industry.
• This requires self discipline or legal restriction.
• These methods operate in the US.
• The international convention to prevent oil pollution

of 1954 zoned coastal waters.
Some of the decisions were;
 Zones where oil is discharged must be regulated.
 All records of oil losses must be well kept i.e.

logging.
 Adequate measure for apprehending and

prosecuting violators must be provided.
 Obliged signatory governments to install in their

ports oil receiving facilities.
 The convention defined oil discharges as those

containing more than 100 parts per million of oil.
Mans Effort to Control Oil Pollution (contd.)
Some effective measures given as advice were;

 The “load on top” technique.
 Oily water should be pumped into what is known

as ‘Slop tank’ to recover part of the oil so that oily
top can be returned to port and fresh oil can be
pumped into tankers.
 Care and safety measures to be installed when

pumping or loading.
 Stations connecting oil pipes to be constantly

checked.
Mans Effort to Control Oil Pollution (contd.)
Some effective measures given as advice were;

 Electronic navigating equipments are to be
preferred.
 Strict observance of shipping lanes can minimize

collision on the seas.
 New ship designs were recommended so that the

holding capacities of the tanker are increased
without affecting the structural safety of the tanker.
 The design is in such a way that even when there

is an accident or oil spill, the spill is minimized.
 This is known as ‘Double Containment Design”.

Physical and Chemical Removal of Oil
• When oil is spilt far from the land (offshore), the
probability of the shore land being polluted is low,
hence,
• Oil slicks may remain untreated, thus allowing for

natural degradation by the autochtonous microflora.
• If the oil threatens the shore line, the choice of

cleaning will depend on the following;
 The quantity of oil spilt.
 The nature of the oil.
 The location of the spill (distance).
 The prevailing weather condition.
Methods available for Physical Removal of Oil
• Containment and Collection:
• From the environmental point of view, the best way to deal with
an oil spillage is by collection.
• To facilitate collection, the oil has to be contained and in

practice, this is achieved by;
 using booms (a temporary floating barrier use to contain an
oil spill). Various types of booms are available depending on
the choice of the buyers.
 Oil can also be contained by using chemicals
such as oleic acids which spread rapidly on water
and pushes back the oil.
 However, use of such chemicals are hazardous

with respect to effectiveness under rough sea
conditions or their toxicity to marine life is not
sufficiently understood.
 It has also been reported that some commercial oil

harders stimulates biodegradation of the oil.

 Another method of containment is by spraying
the oil with Ferro fluid and
 the oil slicks are magnetized together (the ferro –

responsible for the magnetism).
 The Ferro fluid is oleophilic and confers magnetic

properties on the oil.
(contd.)
After containment, there are various ways of removing the oil. Some
of the removal methods include;
•Suction;
i.Scheming into storage tank or oil barges.
ii.Oil can be absorbed on materials such as straw, rice husks or
36
plastic foams.
i. The absorbed oil can then be recovered physically with
a conveyor system (pressing out).
(contd.)
iii. Sinking: the oil can be sunk into the bottom of the sea.
 This removes oil from the surface but not from the aquatic
system.
 Sinking agents are used, this include carbonized sand,
gypsum, stearate coated chalk can also be used as sinking agent.
 Once the oil is sunk, the oil ideally remains there.
 However, physical sinking of oil has a lot of disadvantages;

 The oil can resurface again on the beach.
 It may also cover benthic organisms and the sessile ones can be
exterminated because they cannot escape.
 This will retard the natural degradation of the oil limiting the
surface area and the availability of O 2 required for
biodegradation.
 The degradation of oil at the benthos is slow, it is therefore not
advisable to sink oil in areas with frequent oil pollution because of
the danger of building up thick layers of oil.
 Sinking of oil may jeopardize fishing.
(contd.)
v. Dispersion:
v. It involves breaking oil slicks without removing or degrading the
oil with dispersing agent.
vi.Many commercial dispersant are available in the market.
vii. Some of the disadvantages of this observed is toxicity to marine

life itself or it potentiates the toxicity of the oil.
viii.It has been concluded that all dispersing agents are toxic to
marine life to a high or low degree.
ix.Dispersing agents are cosmetic aids and should not be used

because of their toxic effect on marine life.
x. Generally, it has been suggested that it may be difficult to

differentiate the toxicity of dispersants from that of the oil itself.
(contd.)
vi. Combustion: It is an attractive method that has been

reviewed by several workers. The 1st problem is;
 If the oil is very close to the shore line and facilities, the fire
may become unabated thus resulting to a fire disaster.
 Freshly spilt oil is very hazardous to ignite, adequate

protection is very necessary.
(contd.)
• Some of the advantages of combustion are;
 Low cost, rapid removal of oil is assured.
 Minimal ecological damage.
• N.B:
 The greatest limitation this method has is most oils are
difficult to ignite and to keep burning unless ignited
immediately after the spill.
 Volatile fractions evaporate leaving a residue that has a
relatively high flash point.
 Similarly, emulsifying of the oil will hamper burning.
 However, to aid or facilitate burning, commercial aiders of
burning are available.
 Some of these contain hydroigniters such as metallic sodium
(Na). Straw can be used as a wicking agent.
 Finally, the residues still have to be collected.
(contd.)
vii.Stimulated biodegradation:
• It is a new technique and its purpose to make use of the
biodegradable components of oil.
• Several species of bacteria, fungi can biodegrade petroleum.
• The ability for any microorganism to degrade petroleum depends on a

number of factors. These factors are:
 The biodegradability of the petroleum itself, putting into
consideration its specific gravity, its viscosity and also of the
components of the oil.
 Contact between the petroleum and the microorganisms.
 The environment whether it is soil or water, etc.
 The presence of the necessary enzymes and plasmids.
•It has been observed that among the aliphatic components of oil, the
n-alkanes are preferentially degraded by microorganisms.
SOILS (Contd.)
Methods available for Physical Removal of Oil (contd.)
vii. Stimulated biodegradation (contd.)
• The intermediary metabolites of alkanes include alcohol and fatty acids.
The main pathway exhibited by microorganism is the attack from the
terminal end via β-oxidation. There could be attack from both ends via Ω-
oxidation.
• Di-carboxylic acids are the intermediate products of the attack e.g.
CH3-(CH2)n-CH3
• The double arrows are to show that there are four steps before you get to
the next product.
• The biodegradation of n-alkane of which molecular mass of n = 44 has

been studied.
• The degradation proceeds by a mono-terminal attack usually a primary

alkanol is formed followed by an alkanal and a monocarboxylic acid.
EFFECTS OF OIL SPILLS ON MICROBIAL ECOLOGY OF SEA
AND SOILS (Contd.)
vii. Stimulated biodegradation (contd.)
• Further degradation of the COOH proceeds by ß-oxidation with
the subsequent formation of CO2.
• Some fatty acids which are toxic have been found to accumulate
during hydrocarbon biooxidation.
• The omega oxidation has also been reported in some

microorganisms.
• Sub-terminal oxidation may also occur with the formation of a

secondary alkanol and a subsequent ketone.
• However, this does not appear to be a primary metabolic pathway

utilized by most n-alkane degrading microorganisms.
EFFECTS OF OIL SPILLS ON MICROBIAL
ECOLOGY OF SEA AND SOILS (Contd.)
Advantage of stimulated biodegradation.
 It is not hampered by winds, waves or currents in the ocean
or seas.
 The process is self perpetuating until the biodegradable

portion of the petroleum is eliminated.
 It is a natural process therefore the possibility of unexpected

ecological side effect is low e.g. ecological shifts.
 The process would channel nutrients into the marine food

chain and may increase fish and shellfish production.
 Its cost is expected to be reasonable.

Disadvantage of stimulated biodegradation

 Biodegradation of petroleum is relatively slow even under
favourable environmental conditions.
 It may be inhibited by sub-optimal temperature or by temporary

exhaustion of dissolved O2.
 Some petroleum fractions may be passed along a food chain

without being degraded and thus may taint fish or shellfish
 Nutrient limitation may occur e.g. P and N may be limiting in

marine system.
 However all these theoretical limitations await research to

establish the validity and extent of their practical implications.
SOILS (Contd.)
Biological Cleaners or Microbial Prep in Market
How are microbes

used?
To break down oil

into a harmless
substance
 Some of the products used as biological cleaners include;

 “Petrodeg”,
 “Petrobac” or “Ecolo-Gest”,
 D.B.C Bacteria.
 The promotional literature describes the products as mixed hydrocardon

degrading bacteria with inoculum nutrient formulation suitable to
biodegrade oil in sea water, sewage, and sludge tanks and so on.
 For any of us to use such preparations, promotional literatures should be
subjected to experimental evaluation.
Biological Cleaners or Microbial Prep in Market
• Chemists, biologists, physicists’ altogether have formulated some
preparations to be used in case of oil spillage.
• These proponents assume that oils slick should be seeded with

microbial preparation to biodegrade them.
• Two assumptions have been made by them;

 Conditions of the oil biodegradation in sea water for e.g. are
favourable and therefore the process may occur as soon as the
right microorganisms are introduced.
 The types of the microorganisms used in the formulation are either
absent or rare in the marine system.
• Both assumptions need to be qualified.

• The properties of the seawater must also be considered.
• The number of bacteria in the formulation has to be evaluated.
• In practice, it has been reported that oil degradation is

enhanced
in the presence of N and P.
• If these two elements are added to the marine system because

of
biodegradation, it will result in eutrophication.
n conclusion,
 stimulated biodegradation cannot replace containment and
physical removal of the polluting oil especially when a large
quantity of oil has been spilt.
 stimulated biodegradation should be complementary.
 stimulated biodegradation is aimed at cleaning up of oil

slicks that has escaped primary cleaning or effluent that
has low oil content.
 Such purification is not feasible or not economically

advisable.

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MCB 410

 Accidental spills e.g. collision of oil tankers on the

 It has been observed that roughly between 2.1 - 4.9

• An effect can be either direct damage to a

• sublethal long-term effects on populations,

• There are evidences to show that hydrocarbons are

• therefore, when oil is spilt into an environment

 Fucus vesiculosus is resistant to light and heavy

 Cross et al. (1987) found that biomass of two of

 The other species, Dictyosiphon foeniculaceus,

 On the other hand, those microorganisms that would

 Oil pollution may therefore result in what is called

Other mechanisms involved are;

• The uptake mechanism linked to the attachment of

• Surfactants enhance solubilization and removal of

• Biodegradation is also enhanced by surfactants due

• biosurfactants also help in bioremediation of oil

• Amphiphilic (Hydrophobic and hydrophilic) compounds

• Main structural types are lipopeptides and glycolipids,

•contain hydrophobic and hydrophilic moieties

•BS structure may be that of Glycolipids,

•They vary in chemical properties and molecular size.

•While the low molecular weight BS are often glycolipids,

•the high mol. wt ones are generally either polyanionic

Glycolipid Aeromonas sp., Bacillus sp.

• such responses may alter the long-term survival of

• Thus, the integration of physiological and behavioral

•Acute toxicity is defined as the immediate

•Chronic toxicity is defined as either the effects

 persistence and bioavailability of specific hydrocarbons,

 the ability of organisms to accumulate and metabolize various

 the fate of metabolized products,

 the interference of specific hydrocarbons (or metabolites) with

 the specific narcotic effects of hydrocarbons on nerve

• Narcotic effects of individual petroleum compounds are an important

• Sublethal effects following acute or chronic exposure to petroleum

• Prevention – this is to prevent willful discharge of oil,

• This requires self discipline or legal restriction.

• These methods operate in the US.

• The international convention to prevent oil pollution

 All records of oil losses must be well kept i.e.

 Adequate measure for apprehending and

 Obliged signatory governments to install in their

 The convention defined oil discharges as those

Some effective measures given as advice were;

 Oily water should be pumped into what is known

 Care and safety measures to be installed when

 Stations connecting oil pipes to be constantly

Some effective measures given as advice were;

 Strict observance of shipping lanes can minimize

 New ship designs were recommended so that the

 The design is in such a way that even when there

 This is known as ‘Double Containment Design”.

• Oil slicks may remain untreated, thus allowing for

• If the oil threatens the shore line, the choice of

• To facilitate collection, the oil has to be contained and in

 However, use of such chemicals are hazardous

 It has also been reported that some commercial oil

• Containment and Collection:

 the oil slicks are magnetized together (the ferro –

 The Ferro fluid is oleophilic and confers magnetic

ii.Oil can be absorbed on materials such as straw, rice husks or

 However, physical sinking of oil has a lot of disadvantages;