Microorganisms &

Biocide treatment
Program
By : Safaa a. Al-hulfy
Laboratory & Operation Development Lead / Basra-Iraq
 What is Bacteria !

• Bacteria: are single cell Microbes there is no nucleus or

membrane bound organelles, it can find in a very wide
range conditions.
• Bacteria: could float in the fluid known as planktonic or
settling as sessile. Bacteria could be found at (fluid
stream, filter, bottom tank and dead leg).
Microorganisms types:

Microorganisms: require water, nutrients, and electron

acceptors.
Algae : it is microorganism need for sunlight for
photosynthesis, and it contain chlorophyll
Fungi : it is micro organisms does not contain chlorophyll so
it is not able to carry out photosynthesis.
Bacteria : it has intermediate properties between Algae
and fungi.
 Bacteria classification:
Aerobic bacteria: it require Oxygen to grow.
Anaerobic bacteria: grow in absent of Oxygen.
Macrophallic: growing best in the presence of small mounts
of atmospheric oxygen.
Facultative Bacteria : grow either the presence or absent of
Oxygen.
Strict anaerobic: requires rigorous exclusion of oxygen.

The majority of bacteria are aerobic or facultative

anaerobic.
The most important groups of Bacteria :
Sulphate Reducing Bacteria (SRB)

 Thiosulphate Reducing Bacteria (TSRB)

Acid Producing Bacteria (APB)

Iron Related Bacteria (IRB)

General Aerobic also called Slime Forming Bacteria.

 SRB ‘s
• Sulphate Reducing Bacteria (SRB): is bacteria that cause
some problems in oil and gas facilities. These bacteria are
nonpathogenic with various shape (Desulfobacter,
Desulfococcus, Desulfobulbus, Desulfosarcina and
Desulfonema).

SRB is one of the microorganism can cause corrosion(MIC),

because it reduce the sulfate or sulfite ions presence in the
media to sulfide ions and resulting in H2S as by – product.
 Biological corrosion rather than a corrosion mechanism is a
deterioration of a metal by corrosion processes that occur directly
or indirectly as a result of the activity of living organisms.
 These organisms include micro forms such as bacteria – microbial
corrosion “MIC” – and macro types such as algae and barnacles.
Microscopic and macroscopic organisms have been observed to
live and reproduce in mediums with :
 pH values between 0.5 and 11.
temperatures between 0°C and 80°C.
and resisting extremely high pressures.
 Biological activity may influence corrosion in a variety of
environments including natural water and sea water as well as
natural petroleum products.
What is the MIC !

MIC: is a form of corrosion associated with the activity of

microorganisms.

Microorganisms: are able to interact with the metal surface

environment initiating, facilitating and accelerating
reactions.
MIC is believed to account for 20% of the damage caused
by corrosion.
MAIN SYMPTOMS AND DIAGNOSIS OF MIC:
1-The presence of deposits.
Sessile bacteria responsible for corrosion problems grow in bio-film.
The increasing of the bio-film creates an evident mass deposition on the
interested surface.
In general the biological produced masses present a soft, slimy and wet
aspect, unlike inorganic mineral scale that are hard and closely adhering
deposits.
Depending on the predominant bacterial group the deposited masses could
have different colors:
- Black deposits are typically due to the activity of SRB. (iron sulfides)
- Brown gelatinous, or sometimes filamentous, slimes are typical of IRB growth.
- Yellow deposits are due to the presence of elemental sulfur in the deposits
and they are caused by bacteria such sulphur oxidizing bacteria (SOB).
2- The presence of bad smell.
Bacterial deposits, water contaminated by bacteria could have
different bad odours, depending on the chemical compounds
produced by the colonies :

- SRB deposits have the typical smell of rotten eggs, due to the
presence of sulfides
- Decaying material smell, is due to the activity of various types of
bacteria.
Detection of MIC
1- Pitting corrosion rater then lateral surface corrosion.
2- Presence of biofilm-biomass on corroded surfaces.
3- Hydrogen sulfide in anaerobic environment.
4- Ferric(ous) hydroxides in aerobic environments.
5- Excessive bacterial population in the bulk water.
6- Corrosion in stagnant or “dead flow” areas.
7- Corrosion in system with non-aggressive water chemistry.

When several of the mentioned criteria are observed it

may indicate that the corrosion is related to uncontrolled
growth of microorganisms.
In order to confirm or exclude MIC it is essential to obtain data
regarding the presence of microorganisms on corrosion site.
At this purpose liquid and solid samples must be collected and
analyzed.
Analysis of water samples collected from the plant and
preferentially from the failure nearest sampling point gives
indication about the actual contamination of the plant by
planktonic bacteria, Although this is an important information,
unfortunately it is not essential in the evaluation process, for the
following reasons:
− it does not demonstrate the microbiological nature of the
corrosive event. − it is not representative of the entity of surface
contamination as planktonic population is usually less concentrated
than sessile.
 Finally MIC is defined as :

The degradation of material under the influence of

environmental factors complicated by the
metabolic activities of micro-organisms.
SRB activities consequences :
 Slime in the inner surface.
 Rotten egg odor.
 Black water.
 Deposit at filters and increase delta pressure.
 Increase iron sulfide in lines or equipment.
 Souring by increase H2S levels in sweet system.
 Increase emulsion stabilization.
 Microbially Induced Corrosion (MIC) in equipment.
Forms of SRB
There are two forms of SRB

Planktonic SRB:
Usually found free in the water and not attached to any surrounding
surface.

Sessile SRB:
Found attached to the surrounding surfaces and may be found under
deposits, they are directly responsible for pitting attack on metal
surface.
The Requirements for SRB Growth:

1- Nutrients (mainly sulfate ions, SO4).

2- Moderate temperature(5-50˚C) & most common strains of
SRB grow best at temperatures from 25° to 35°C, some are
thermophilic grow up to 80˚C.
3- Moderate pH(5-10).
4- Pressure less than 500 bar.
5- Absence of poisoning species such as oxygen or biocides.
6- Suitable flow rate to establish colonies on pipe wall.
Biofilm

Biofilm: is a colony of bacteria residing in a porous organic

material, form at low velocity (<3 ft/s) then start produce sticky
material EPS (Extracellular Polymeric Substances) Sessile
bacteria start to grow and starting a colony, the colony may
be a mixed colony made up of two or more types, In the
process generate (H2S, acid, slime and other materials). Pitting
will typically be occurring beneath these colonies.
Biofilm Formation
Biofilm formation consists of a sequence of steps and
begins with:

1- Adsorption of macromolecules (proteins,

polysaccharides, and acids) and smaller molecules
(fatty acids and lipids) at surfaces.
2- Adsorbed molecules form conditioning films that alter
physiochemical characteristics of the interface,
including surface hydrophobicity and electrical charge.
3- The amount of adsorbed organic material is a
function of ionic strength and can be enhanced on
metal surfaces by polarization.
Surface conditioning: as a metal surface comes in contact with water, it will be
rapidly covered by an organic layer, due to the presence of organic molecules in the water
source;

Attachment:
some planktonic cells adsorb to the surface, thus starting to grow as sessile bacteria; − some
of the adsorbed cells, because of different physical forces acting on the surface, can desorb
so returning to the bulk liquid phase;

Colonization:
the cells adsorbed on the surface start to grow, doubling their number in short time. In this
phase bio-film increases both in width and thickness (multi-layers structure);
sessile bacteria will produce large amount of EPS molecules, used to strongly attach at the
surface.
At the same time, EPS act as a protective shield, covering the growing bacterial colonies;
EPS have a sticky nature. For this reason, particulate matter and living organisms present in
the water phase could be trapped in EPS contributing to net bio-film mass increase.

Detachment:
as bio-film grows, the action of different physical forces on the outer layers leads to the
detachment of material from the bio-film. The detached material returns to the water phase
and it is able to colonize other surface areas.
Growing as sessile organisms, bacteria produce extracellular
polymeric substances (EPS). By means of EPS bacteria are able to
tightly stick to the metal surface, thus leading to the formation of
a structure named bio-film.
(1) Micro-organisms adhere to the surfaces of solids
(2) Micro-organisms produce sticky matter around their
bodies
(3) Inorganic suspended solids adhere due to the binding
action of the sticky matter of micro-organism
(4) Slime adhesion proceeds Biofilm
SRB Corrosion Mechanism
Electrochemical Reactions
Step one:

Fe → Fe++ + 2e- (Anodic Reaction).

4H2O ↔ 2H+ + 2OH- (Water Dissociation).
2H+ + 2e- → H2 (Cathodic Reaction).
SRB Corrosion Mechanism

Step two:

SO4-- + 8H ↔ S2- + 4H2O (Bacterial Consumption).

S2-

C COPYRIGHT PETROLITE CORP 1993

 SRB Corrosion Mechanism
Step three:
FeS (Corrosion Product).
Thus the theoretical amount of ferrous sulfide that may be formed from a given amount of sulfide
is:
ppm SO4-- / 96 (MW SO4-- ) X 88 (MW FeS) = ppm FeS

S2- + Fe++ → FeS

The Treatment & Control Program
 Biocides

A biocide is a chemical substance capable of killing

living organisms in addition to bacteria.
 Prevent fouling in water and oil pipelines.
Biocides are divided in a group of oxidizing and in a
group of non oxidizing ones.
• used to reduce SRB production and Microbiologically
induced corrosion MIC, by interfering with their activity.
Biocides Types:
Oxidizing:
❖Chlorine or hypochlorite
❖Quaternary ammonium chloride

❖Copper ions
Non Oxidizing:
❖Aldehyde (formaldehyde, glutaraldehyde)
❖Sulfur compounds (Isothiazolone, carbonates, metronidazole)
❖Quaternary phosphonium salts
❖Amines and diamines
❖Quaternary amine compounds
Classifications

Inorganic
chlorine
Organic
1- Chlorinated phenols
2- Quaternary ammonium compounds are common
examples of organic biocides.
most market available biocides are organic.
Non-Oxidizing (Organic) is commonly used in oilfields. The
non-oxidizing biocides function primarily by altering the
permeability of the cell walls of the microorganisms and
interfering with their biological processes. More compatible with
others organic chemicals, effective for planktonic and sessile
bacteria. Some manufacturer mixes more than two types of
biocide to extend the spectrum.
Factors determining the biocide selection :

1- The type of the microorganisms involved.

2- The prior operating history of the system.
3- The type of the process.
4- The chemicals being used in the system for scale and corrosion
control.
5- Chemical and physical characteristics of the water in the system.
6- Cost of the treatment.
Biocide selection and evaluation
Characteristic of effective and efficient biocide:
1. Resistant strains
2. Compatibility with water and other chemicals.
3. The bacteriostatic test.
4. Time-Kill Test.
5. Treatment Method
6. Field evaluation
7. Work at various PH.
8. Low decomposition rete.
Biocide selection and evaluation

Aspects shall be considered:

1- Resistant strains (immunity) :
Bacteria have a capability of developing strains which
build up a resistance to a particular chemical over a period
of time. Alternate use of two (or more) different chemicals
is often required to overcome this problem.
3- Bacteriostatic test .
The bacteriostatic test is a laboratory test determines the minimum concentration
of a chemical to inhibit the growth of microbial activity.) It is used for comparing
large numbers of chemicals under standard defined conditions.

4-Time-Kill Test
The time kill-test which determines the time for a chemical to kill the bacteria, is
used for the final selection of the best candidates from the bacteriostatic test.

The time-kill test defines the treatment conditions :

- Dose (Biocide concentration to be injected).
- Contact time (the suitable length of time of the treatment).
- Frequency.
 Treatment Method
Continuous injection:
At start-up : a higher concentration of biocide is usually
required for the initial control of bacterial population (total
kill).
the biocide concentration can be reduced
a smaller dosage being sufficient to keep controlled bacteria
activity (maintenance).
The optimized concentrations required for these two phases
are estimated by laboratory tests
Batch treatment.

A slug of biocide highly concentrated is periodically

disposed into the system for a total bacteria kill.
1- Slug volume
2- Concentration
contact time can be initially based on time-kill tests, or
Trials con and then adjusted by operational feedbacks.
 field evaluation
Is the selected products are working effectively ??

1- Making sure the biocide is properly injected (i.e.

continuously, or with the design frequency for batch
treatment).
2- The chemical concentration (active matter).
3- At an appropriate dosage.
4- Monitoring the bacteria present with time (bacteria
count).
 N.B (if applicable .. if applicable)
 A preliminary cleaning of the pipeline is the key for biocide treatment
effectiveness.
 Solvents, acid and pigs are used for this purpose.
 For the biocide selection in seawater injection systems it should be paid
attention to suspended solids/scale which may form deposits.
 Under-deposit corrosion can occur, and in these locations corrosion inhibitors
and biocides is not effective being shielded by the deposit scale.
 Pigging operation capable to remove the complex biofilms in which sessile
bacteria grow and rendering the applied biocides ineffective is very
important.
 In order to achieve maximum effectiveness, the sodium hypochlorite is usually
followed by hydrochloric acid.
 Backflow all injection wells, If the wellbores are plugged with bacteria deposits
or slimes, strong oxidizing agent such as sodium hypochlorite is normally used
to attack them.
Note: No chemical can kill bacteria if it cannot contact them.
 Current Biocide Types:
 Glutaraldehyde Biocide: is the most commonly used oilfield organic biocide,
the kill mechanism is by cross-linking outer proteins of cells and preventing cell
permeability. Glutaraldehyde is ph. sensitive it works well in neutral to alkaline
water, it may be effective for injection waters but less effective for produced water
treatment. React with ABS (Amine Bio- Sulfate) / amines and its demand Amines in
the system. It’s in low cost.
- non-oxidizing chemical fast acting biocide
- effective to treat sulphate reducing bacteria and biofilms.
- It has a minimal effect on the environment.
- widely used in a blend with other biocides and surfactants to increase its
effectiveness. But the available type is net.
- The advantages are broad-spectrum activity, relatively insensitive to sulfide, it is
compatible with many chemicals and it tolerates salt and hardness.
Quaternary Amine biocides: are very surface active, and
because of this property, they are sometimes used in blend with
other biocides to improve the performance. The most common
quats in this class are long-chain n-alkyldimethylbenzylammonium
chlorides most effective in alkaline ph ranges, but lose their activity in
system fouled with dirt and other debris, low concentration (<250
ppm). React with polymers and demand high TDS system. Its cost is
medium.

- Quaternary Amine compounds show a broad-spectrum activity.

- A good surfactancy and persistency and low reactivity with other
chemicals.
- The disadvantages are inactivated in high brines, they are foaming
and they are slow acting.
Tetrakis Hydroxymethyl Phosphonium Sulfate:

THPS biocide: is the excellent nonoxidizing organic biocide.

 Nonfoaming biocide
 Kill bacteria mainly by cross-linking of proteins, which leads to collapse
of cells membranes (cell lysis).
 A concentration of 50-100 ppm is normally needed for planktonic
treatment, but much more for establishing biofilms.
 THPS degradation is faster under alkaline conditions.

Note: High cost biocide.

 Biocide treatment program :

 Will be conducted initially based on the samples analysis results and the
bacterial count will be obtained from the samples collected and incubated at
the lab, you can go through the corrective or preventive action plane.
 The treatment program will be depend on batch treatment to avoid the
chemical compatibility.
 We will consider the weekly bases till obtaining the results and time kill test.
 The time - kill test will be performed in lab in parallel to the system treatment.
 The dosage will depend on the source , type and severity of infection.
 The dosage will depend on the active matter concentration in the chemical
 the batch duration (contact time) will depend on the chemical type and
system pH and will be confirmed by the time-kill test.
 The frequency of batching will be based on the analysis and monitoring.
Biocide treatment injection:

1- Clean system (Bacteria population less than 10² coll/cc)

required Continuous treatment 5-25 ppm.
2- Moderate to dirty system (Bacteria population 10² - 10³
coll/cc) required Batch treatment 200-1500ppm, 1-6 hours per
treatment, twice per month.
3- Dirty system (Bacteria population > 10³ coll/cc) required
Batch treatment 300-600 ppm, 3-4 hours per treatment, once a
week and after batching do continuous injection 5-25 ppm.
Biocide Injection start up:

 The program will be conducted according to two phases :

(A) Sterilization Batch :
to clean the system and the primary receiving vessels and tanks
reference to the source of infections
 Review the chemical skids injection rates.
 Assigning the type based on the system pH, mainly start with
THPS based type It is very efficient, especially with difficult and
persistent organisms due to its good penetrating ability in a
dosage of 1200 PPM as sterilization slug during 6 Hours
(considering the active matter concentration).
 Samples will be collected after 6 hours from Batch injection (if
available) or in the next day or after treatment (NK).
 Another sample will be collected in the last day before the next
weekly batch to check number of cells growing between
treatments (Ng).
 Start the sterilization batch using the other biocide type (Ex. Quat)
type and this Batch will depend on the new results.
 Samples will be collected after 6 hours from Batch injection (if
available) or in the next day or after treatment.
 Samples will be collected in The last day before the next weekly
batch to check number of cells growing between treatments.
 We can evaluate the sterilization program and move to the
normal treatment program.
Efficiency of the batch by Monitoring:

The effectiveness of batch biocide treatment regimes

can be described by:

% efficacy = (Ng – Nk) / Ng x 100.

Where:
Ng = Number of cells growing between treatments.
Nk = Number of cells killed during treatment.
Normal treatment program:

-Will depend on the results from sterilization batch

The optimization will include :

- Dosage : The plan is to (Decrease) gradually, initially to 500
then 300 ppm based on the samples results.
- Treatment Frequency (Batches): (decrease (or) increase (or)
maintain