SPE 157388

New Test for Material Resistance against Microbiologically Influenced

Corrosion
Jeppe Jensen, SPE, Ketil Bernt Sorensen, Susanne Juhler, Thomas Lundgaard, SPE, DTI Oil & Gas, Danish
Technological Institute, Kongsvang all 29, DK-8000 Aarhus C, Denmark.

Copyright 2012, Society of Petroleum Engineers

This paper was prepared for presentation at the SPE International Production and Operations Conference and Exhibition held in Doha Qatar, 1416 May 2012.

This paper was selected for presentation by an SPE program committee following review of information contained in an abstract submitted by the author(s). Contents of the paper have not been
reviewed by the Society of Petroleum Engineers and are subject to correction by the author(s). The material does not necessarily reflect any position of the Society of Petroleum Engineers, its
officers, or members. Electronic reproduction, distribution, or storage of any part of this paper without the written consent of the Society of Petroleum Engineers is prohibited. Permission to
reproduce in print is restricted to an abstract of not more than 300 words; illustrations may not be copied. The abstract must contain conspicuous acknowledgment of SPE copyright.

Abstract

A material test for microbiologically influenced corrosion (MIC) is presented. This laboratory based test evaluates the material
resistance to MIC caused by corrosive, methane-producing microorganisms. Material testing and corrosion modeling is a
natural and important step to ensure material integrity during oil production. Currently, good models for physical and chemical
parameters exist, but MIC remains very hard to predict in spite of many years of research, and failures continue to occur
prematurely.

MIC can to some degree be mitigated by biocide treatment. However, the effect can be limited because responsible
microorganisms are part of a protective biofilm of the metal surface. Parts of the biofilms often survive even the best biocide
program, and the surviving microorganisms will quickly re-establish their activity in the system. Metal alloys react differently
to these biofilms, and the choice of material is a crucial parameter in minimizing MIC.

Biofilms formed by methane-producing microorganisms are known to cause MIC. These microorganisms are not detected in
standard monitoring tests, such as bottle incubations, widely used by the industry. However, several recent studies employing
Molecular Microbiology Methods (MMM) have shown that these microorganisms are numerous, widespread, and highly
corrosive in off- as well as on-shore oil fields. The new material test described in this paper uses methane-producing
microorganisms to test the MIC resistance of metal alloys. This enables petroleum companies to choose a suitable material for
an environment with high MIC potential and to predict the corrosion rate at close to in situ conditions.

Introduction

Here we present a new method for testing materials against Microbiologically Influenced Corrosion (MIC) caused by
methane-producing biofilms.

Alloys and metallic coatings are usually tested in standardized set-ups to evaluate the resistance to different types of chemical
and physical corrosion before installation. For example, there are international standards for the testing of pipeline and
pressure vessel material towards hydrogen induced cracking (NACE TM0284) and sulfide stress corrosion cracking (NACE
TM0177). It is recognized that corrosion resistance of alloys and coatings varies in flowing seawater and many have dual
surface behavior and can become active after a passive behavior, particularly in low-velocity or poorly aerated water and at
shielded areas (Winston, 2011). In addition to the physical and chemical corrosion processes, the phenomenon of
Microbiologically Influenced Corrosion (MIC) is gaining increasing recognition as a widespread problem in the oil, gas and
fuel sector (NACE 31205, 2006), and is thought to be the main cause of many of the failures and unpredictable corrosion
incidents that continue to haunt the industry. Thus, it has been estimated that 10% to 30% of all serious corrosion cases have
been attributed to MIC, and 75% to 90% of all internal pipeline leaks are reportedly due to localized corrosion of which MIC
is a major cause (Jenneman, 2012). Since no standards exist for testing the resistance of materials towards MIC, and because
the factors that trigger the process are poorly understood, MIC remains a serious unaccounted for risk factor in material
durability predictions.
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MIC is caused by the growth of corrosive biofilms on the surface of vulnerable materials such as metal alloys. Different types
of microorganisms can contribute to MIC, but in particular methane-producing and sulfate-reducing bugs have been shown to
cause corrosion problems (Boopathy and Daniels, 1991, Daniels et al. 1987, Sorensen et al 2012). The growth of these
troublesome microorganisms in oil production systems is usually reduced to an acceptable level by a combination of
mechanical and chemical cleaning and extensive biociding programs. These treatments are tedious and costly, and there is a
large potential for reducing operational costs by selecting materials that are less vulnerable to MIC under the given
environmental conditions in the field.

The test presented here employs a methane-producing test organism incubated in the laboratory under simulated field
conditions. The role of this type of microorganism in MIC is well documented, and three examples are briefly described in the
following:
1) Caliper investigations of a producing well in the North Sea revealed severe corrosion and several holes in the tubing
from 10,000-13,000 ft. (3,048-3,962 m) MD and from 15,500-16,000 ft. (4,724-4,877 m) MD. CO2 (sweet corrosion)
could not account for the observed corrosion. An investigation of solid downhole material concluded that the
corrosion issues were due to MIC and that methanogens were the most abundant microorganism present (Larsen et al.
2011).

2) Topside production facilities are often subject to MIC and high numbers of methanogens are usually associated with
high corrosion rates. A water leg from a separator tank was inspected by ultrasonic measurements from 2002 to 2008.
High corrosion rates were found throughout the period; the water leg was replaced and a MIC investigation
commenced. It was concluded through a combination of Molecular Microbiology Methods (MMM) and chemical
tests that the observed corrosion was promoted by microbial activity and that methanogens were present in highest
numbers (Skovhus et al. 2010).

3) Pipelines are notoriously susceptible to corrosion and some pipelines have been shown to contain methanogens in
very high numbers. The Rolf A to Gorm E pipeline in the North Sea was classified as a high MIC risk pipeline due to
high methanogen numbers. Intelligent pigging of the pipeline conducted in 2009 showed significant corrosion events
which corresponded to a corrosion rate of 0.7 mm/year. A total of 900 features were found and the deepest pit was
19% of the nominal wall thickness (Sorensen et al, 2012).

The process by which the methanogens oxidize iron and produce CH4 has been known for a long time (Daniels et al. 1987,
Dinh et al. 2004). In methanogenic MIC, CO2 is reduced to CH4 under the oxidation of metals. Through this corrosion process,
the methanogens gain energy for their cell metabolism and growth. The redox reaction is exemplified here by the oxidation of
iron (Fe0) to ferrous iron (Fe2+):

4Fe0 + 8H+ + CO2 4Fe2+ + 2H2O + CH4 (equation 1), iron oxidation by reduction of CO2 to methane by methanogens.

The reaction can also be indicated as two coupled half-reactions:

Fe0 4Fe2+ + 8e- (anode) (equation 2)

CO2 + 8e- + H+ CH4 (cathode) (equation 3)

The exact means of electron transfer between the two half-reactions is not yet fully understood. However, the reduction of CO2
to CH4 is the crucial step in methanogenic MIC, and this process does not occur without the activity of methanogens.
Figure 1 below shows the oxidation of metal by methanogens and how it may cause pit formation.

Figure 1. Pit generation by methanogenic MIC. Redrawn from Daniels et al. 1987. The cathodic reaction is accelerated by methanogenic
activity and elemental iron is oxidized to ferrous iron and dissolved at the anodic reaction centre.

As methane can only be produced from microbial activity, the concentration of methane in a closed system is a robust measure
of the microbial activity.
SPE 157388 3

Methodology experimental set-up

The experiment presented here describes the generic experimental setup for metal material MIC testing. The presentation is
exemplified by carbon steel, which is commonly used in the oil industry for many purposes. The test is, however, directly
transferrable to any other metal alloys and coatings applied in water or multiphase conducting parts of an oil production
system. The test conditions presented are designed to simulate the marine environment present in an injection water system,
but can be modified as well. The only energy source added for microbial growth is the test material.

The test setup consists of three different bottle incubations (each performed in triplicates) with sterile anoxic artificial seawater
and nitrogen and carbon dioxide filled headspace, as illustrated in figure 2 below. The carbon dioxide is added as an electron
acceptor to feed the methanogenic test organism. No organic growth substrates are added to the medium and the only energy
source (electron donor) for the methanogenic microorganisms is the test metal.

The MIC-causing test organism applied is a methanogen species, which is commonly found in oil production facilities, and
which has been shown to cause MIC in laboratory setups. The metal to be tested is incubated with and without the
methanogenic organisms, as illustrated in table 1 and figure 2 below (incubation A and B). This is to separate the corrosion
rate directly related to MIC from any chemical corrosion occurring in the incubations. The third incubation contains only the
methanogens and no metal (incubation C). This is to verify that the methanogens will only grow when the metal is added as an
energy source, i.e. that any microbial growth in the incubations is related to MIC.

Table 1. The experimental setup. Each of the three different incubations is performed in triplicates.
Presence of Presence of
Test incubation
methanogen test Material
A
B
C

Figure 2. Diagram of the experimental setup. Three incubations with artificial seawater and nitrogen/carbon dioxide headspace are incubated
with: A) Test material and methanogen, B) Test material only, and C) methanogen. The methanogen uses electrons and CO2 to produce
methane and energy for growth.

The experiment runs for four weeks, after which the rate of MIC is evaluated. The test parametres includes an array of direct
and indirect measurements of MIC (table 2), which are described in more detail below.

Table 2. Test parametres for MIC analysis.

Parameters Week 1 Week 2 Week 3 Week 4
2+
Dissolved ions (e.g. Fe )
Total weight loss of the material
Microbial activity (methane production)
Planktonic population of methanogens
Sessile population of methanogens
pH
Evaluation (SEM) of material surface

Corrosion of test material

The corrosion of the test material is determined by analysis for dissolved metal ions, weight loss of the test material, and
Scanning Electron Microscopy (SEM). The dissolution of ions from the test material is calculated from the concentration of
metal ions in the solution, which is analyzed weekly throughout the test period. The total weight loss of the test material is
4 SPE 157388

measured by weighing the material before and after the incubation period, after removal of corrosion products. At the end of
experiment, the surface of the test material is furthermore investigated for pit formation by SEM.

Microbial activity and growth

As the methanogenic microorganisms produce methane gas as a result of their energy metabolism, their activity can be
determined directly as the methane production rate in the incubations. Weekly measurements of the methane concentration in
the headspace and liquid phase of the incubation give a measure of the integrated production of methane within the test period.
The growth of the methanogenic microorganisms is determined by enumeration of cell numbers by quantitative polymerase
chain reaction (qPCR) and fluorescent microscopy (DAPI), as described elsewhere (Skovhus et al. 2007, Larsen et al. 2010).
Both planktonic cell numbers (suspended in the liquid phase) and sessile cell numbers (growing on the surface of the test
material) is determined.

Interpretation of the results

The developed setup for testing the materials resistance to MIC has been experimentally verified in the laboratory. In the
following section, the possible outcomes of the material test and interpretation of the test results is discussed. For this purpose,
results are exemplified as simplified graphical drawings, describing the two scenarios where a test material is either resistant or
susceptible to methanogenic MIC.

Microbial cell numbers

The number of sessile and planktonic cells in each of the incubations is determined at the end of the incubation period. Cell
numbers in incubations lacking either metal or methanogens are expected to be much lower than in incubations where metal is
present.

The incubations are supplied with CO2 in elevated levels and the test material will consequently be subject to chemical
(abiotic) oxidation:

H+ + HCO3- + Fe0 FeCO3 + H2 (equation 4)

As seen in the above equation, the oxidation of iron produces hydrogen and iron carbonates. The iron carbonates serves a
protective layer against further corrosion, but are easily removed by mechanical stress (water flow etc.) (Winston, 2011). This
is interesting, as many types of methanogens are capable of consuming the abiotic formed hydrogen and thereby sustain
growth (Schwartz and Friedrich, 2006). The theoretical methane production would in that case be in concordance with the
hydrogen production in sterile conditions (i.e. formed by abiotic corrosion) and the corrosion rates would consequently not be
higher.

At the end of the experiment, the number of cells present will either correspond to the original inoculum (i.e. the start-up cells
added at the beginning of the experiment), have increased slightly due to microbial growth from abiotic produced hydrogen
(equation 4), or have increased due to growth from biocorrosion. The three scenarios are depicted in the figure 3 below.

Figure 3. Cell numbers present in the incubations at the end of the test period, in case of 1) MIC-susceptible test material, and 2) MIC-resistant
test material, respectively. Results are depicted for incubation A: Test material + methanogens, incubation B: Test material, and incubation C:
Methanogens.

Consequently, if the test material is resistant to MIC, but susceptible to abiotic corrosion, the cell numbers in incubations
including metal will increase only slightly more than in incubations without the metal. If the test material is susceptible to
MIC, the cell numbers will increase more than is made possible by the abiotic hydrogen formation. Furthermore, the cells will
mainly be associated with the metal surface and the planktonic cells will mostly originate from the slough-off of sessile cells.
SPE 157388 5

Since methanogens are capable of growth supported by abiotic produced hydrogen alone, more test parameters are necessary
to evaluate the material resistance against methanogenic MIC.

Material weight loss

The material weight loss during incubation of MIC-resistant and MIC-susceptible material is depicted in figure 4 below. If the
material is MIC-resistant, the material weight loss will not depend on the presence of methanogens (i.e. the weight loss
occurring in incubation A and B will be equal). The opposite applies if the material is susceptible to methanogenic MIC, and
the weight loss will consequently increase in the presence of methanogens. I.e. the weight loss occurring in incubation A will
be higher than in incubation B. From the weight loss differences between the two incubations, the microbiologically
influenced corrosion rate can be estimated. Hence, different test materials can be ranked based on their respective MIC rates.

Figure 4. Test material weight loss during the incubation period, in case of 1) MIC-susceptible test material and 2) MIC-resistant test material.
Results are depicted for incubation A: Test material + methanogens, and incubation B: Test material.

Methane production rates

If the test material is susceptible to methanogenic MIC, methane will be generated throughout the experimental period in
incubations containing both methanogens and metal, but not in incubations lacking either methanogens or iron. The rate will
reflect the increasing cell numbers in the incubation and will consequently increase along with cell numbers. However, if the
material is susceptible to abiotic corrosion only, and hydrogen is formed, the methanogens will produce methane from
hydrogen, but the production rate will be stable and in accordance with the abiotic corrosion. The two scenarios are depicted in
figure 5 below.

Figure 5. Methane production rates during incubation of 1) MIC-susceptible and 2) MIC-resistant test material, in the presence of methanogens.

Dissolved iron in solution

The concentration of dissolved metal ions in the solution will increase during the incubation period as a result of the material
corrosion process. The susceptibility of the test material to MIC will be reflected in the speed with which the ion concentration
increases in the presence of methanogens (incubation A) relative to in the abscense of methanogens (incubation B). As
illustrated in figure 6, the dissolution of metal ions caused by abiotic corrosion processes can be determined from incubation
B. The relative susceptibility of the test material to MIC can be determined from the additional increase in dissolved ions
occurring in incubation A relative to incubation B.
6 SPE 157388

2+
Figure 6. Graph showing the dissolution of Fe during incubation of 1) MIC-susceptible and 2) MIC-resistant test material, for incubation A:
Test material + methanogens, and incubation B: Test material only.

The theoretical ratio between the amount of iron dissolved from the metal surfaces and the amount of methane produced by the
methanogens is given by the stoichiometry of equation 1 described above. Thus, 4 molecules of Fe should be removed from
the metal for every molecule of methane produced. The Fe2+: CH4 ratio is consequently a way to determine the electron source
for the methane production, as seen in figure 7. If the ratio is lower than 4, methane is produced partly from a different source
than the corrosion of iron. If the ratio is higher than 4, more iron is oxidized than can be explained by biocorrosion alone.

2+ :
Figure 7. Diagram showing three different Fe CH4 ratios.

In laboratory experiments, this relationship may be slightly higher. The reason for this is probably that additional electron
equivalents are used by the autotrophic methanogens to reduce CO2 to organic material during cell growth. However, in a real-
life scenario the methanogens have access to organic material (VFAs etc.) from the environment, and they may need fewer
electrons for fixation of CO2. Therefore, the Fe2+: CH4 ratio would most likely be closer to 4 than measured in the
experimental setup described here when iron is the sole source of electrons.

pH
The consumption of protons and hence the increase in pH per molecule Fe2+ dissolved due to methanogenic corrosion is equal
to the proton consumption in abiotic CO2 corrosion (equation 1 and 4). However, as seen in Figure 8, the chemical
consumption of protons can be considered negligible compared to the consumption of protons, if the test material is
susceptible to methanogenic MIC. According to the stoichiometry (equation 1), 8 protons are consumed per methane molecule
produced, and the pH level in the closed system is therefore expected to increase concurrently with methanogenic activity.
Figure 8 below shows the expected pH development during incubations with MIC susceptible and MIC resistant test material.
Thus, the pH is an indication of MIC activity in this test setup.
SPE 157388 7

Figure 8. Increase in pH during the test period in case of MIC susceptible material, for incubation A: Test material + methanogens, and
incubation B: Test material.

Pit formation
Corrosion may occur as a uniform general corrosion or a localized pitting corrosion. As the formation of pits will lead to a
more severe type of MIC with higher risk of system failures, the susceptibility of the test material to pit formation is an
important factor in evaluating the material resistance to MIC. The potential pit generation will be revealed and quantified by
SEM analysis of the test material surface. The MIC-resistance of different test materials can thus be ranked based on their
respective pit density. In case no pitting occurs, the corrosion rate obtained from the material weight loss (described above)
can be used as an estimate of the average yearly decrease in metal wall thickness in a system coated with the tested metal.

Compiled data evaluation

As described in detail above, the material test consists of a range of measurements that in combination evaluates the material
resistance to methanogenic MIC. The parameters are summarized in table 3 below.

Table 3. Summarized test parameters. Incubation A: Test material + methanogens, incubation B: Test material, and incubation C:
Methanogens.
Test parameter MIC susceptible test material MIC resistant test material
Markedly higher in incubation A compared to Equal or only slightly higher in incubation A compared to
Cell numbers
incubation C incubation C
Material weight loss Higher in incubation A than B Equal for incubation A and B
Methane production rate High in incubation A None or very low in all incubations
Concentration of dissolved
High in incubation A compared to incubation B Equal for incubation A and B
metal ions
pH increase High in incubation A compared to incubation B Equal for incubation A and B
8 SPE 157388

Conclusions and Perspectives

The new test for material resistance against microbiologically influenced corrosion estimates the MIC rates of the test material
by direct and indirect measurements under conditions very close to full-scale water-containing systems in oil production. The
test organism is a commonly occurring MIC-causing methanogen that is usually overlooked by normal bacterial monitoring
programs, but nevertheless have been shown to be the main cause of corrosion-related failures in pipelines, downhole tubings,
and topside facilities.

The main outcomes of the MIC material test are:

- Ranking of different metal materials and/or alloys based on their relative resistance to MIC.
- An estimated MIC rate for each test material at close to in situ conditions. This includes an estimate of the average
yearly wall-thickness reduction of the metal.
- Surface scanning of the test material and evaluation of its pitting tendency when subjected to MIC

These outcomes will enable petroleum companies to rank materials in a best-in-class test, and select the most MIC-resistent
materials for on- and offshore constructions. This will be of special relevance with respect to system integrity maintenance at
critical or difficult reachable locations, where mechanical MIC mitigation is not an easy option. Material-specific MIC rates
can furthermore be included in existing corrosion prediction models for the improvement of corrosion risk assessments and
system integrity lifetime calculations.

References

E. Schwartz and B. Friedrich, The H2-Metabolizing Prokaryotes. The Prokaryotes vol 2, 2006, p. 496 564,. Publisher: Springer, 2006-07-
13. ISBN-13: 9780387254920

G. Jenneman, MIC: Managing Risk and Uncertainties. Conference paper (114-MIC-17), 14th MiddleEast Conference & Exhibition,
February 12-15, 2012, Manama, Kingdom of Bahrain.

H.T Dinh, J. Kuever, M. Mussmann, A.W. Hassel, M. Stratmann, F. Widdel,Iron corrosion by novel anaerobic microorganisms, Nature,
427 (2004): p. 829-832.

J. Larsen, K. Rasmussen, H. Pedersen, K. Srensen, T. Lundgaard, T.L. Skovhus, Consortia of MIC Bacteria and Archaea causing Pitting
Corrosion in Top Side Oil Production Facilities, Corrosion 2010, paper 10252, (Houston, TX: NACE International, 2010).

J. Larsen, K. Srensen, L. Holmkvist, T.L. Skovhus, Identification and Quantification of Microorganisms involved in Downhole MIC in a
Dan Oil Producing Well, Corrosion 2011, paper ID 17792, (Houston, TX: NACE International 2011).

K.B. Srensen, U.S. Thomsen, S. Juhler, J. Larsen, Cost Efficient MIC Management System based on Molecular Microbiological
Methods, NACE International, 2012, Conference paper ID: C2012-0001111

L. Daniels, N. Belay, D. S. Rajagopal, P. J. Weimer, , Bacterial Methanogenesis and Growth from CO2 with Elemental Iron as the Sole
Source of Electrons. Science, 31 (1987): p. 509-511
NACE 31205 (2006), Selection, Application, and Evaluation of Biocides in the Oil and Gas Industry (Houston, TX: NACE)

NACE Standard TM0284 - 2011, Standard Test Method - Evaluation of Pipeline and Pressure Vessel Steels for Resistance to Hydrogen-
Induced Cracking, National Association of Corrosion Engineers, 28-Oct-2011, 24 pages, ISBN: 1575901633

NACE Standard TM0177 2005, Laboratory Testing of Metals for Resistance to Sulfide Stress Cracking and Stress Corrosion Cracking in
H2S Environments, National Association of Corrosion Engineers, 03-Dec-2005, 43 pages, ISBN: 157590036X

R. Boopathy and L. Daniels, Effect of pH on Anaerobic Mild Steel Corrosion by Methanogenic Bacteria, Applied and Environmental
Microbiology, 1991, p. 2104-2108

R. R. Winston, Uhlig's Corrosion Handbook, Third Edition, Copyright 2011 John Wiley & Sons, Inc., Published Online: 19 APR 2011,
Print ISBN: 9780470080320, Online ISBN: 9780470872864, DOI: 10.1002/9780470872864

T. L. Skovhus, B. Hjris, A.M. Saunders, T.R. Thomsen, M. Agerbk, J. Larsen, Practical Use of New Microbiology Tools in Oil
Production, SPE 109104, SPE Offshore Europe Conference 2007, Aberdeen, U.K., 4-7 September 2007.

T.L. Skovhus, K.B. Srensen, J. Larsen, K. Rasmussen and M. Jensen, Rapid Determination of MIC in Oil Production Facilities with a
DNA-based Diagnostic Kit, SPE 130744, SPE International Conference on Oilfield Corrosion held in Aberdeen, United Kingdom, 2425
May 2010.