Electrochemical corrosion behavior of carbon steel exposed to low

concentration agar with and without nutrients

Amy Spark, RMIT University & CSIRO, Melbourne, Australia; Ivan Cole, CSIRO,
Australia; Donavan Marney, CSIRO, Australia; David Law, RMIT University,
Australia; Liam Ward, RMIT University, Australia

Summary

A novel technique for the study of microbial corrosion in soil is currently being devel-
oped. This technique uses microbiologically graded agar to simulate more closely the
conditions experienced in soil than is possible with an aqueous solution. Traditionally,
most soil based corrosion testing is conducted in the laboratory with solutions made
to mimic the chemical composition of soil. This does not take into account the physi-
cal structure of soil and resultant electrochemical interactions between the soil, steel
and microbes. Agar is commonly used as a solidly setting gelling agent. However, in
lower concentrations it forms a sludge like consistency, similar to the consistency of
saturated soil.

Utilizing low concentration agar in an electrochemical cell, the corrosion behavior of

carbon steel has been investigated under varying nutrient conditions.
Potentiodynamic scans, potential hold measurements, optical microscopy and SEM
imaging were used to monitor the steel surface both during and after exposure of the
steel to the agar. These measurements will make the basis of a control for later ex-
periments when the response of Pseudomonas to nutrient changes under a range of
potentials when in an electrochemical set up will be investigated.

This work is part of a continuing investigation into the mechanisms of microbial cor-
rosion at the soil-steel interface of buried potable water pipes.

1 Introduction
Localized corrosion initiated on the external surface of pipelines has been identified
as the main cause of failure for metal pipelines [1]. With Australia’s dependence on
an extensive pipeline network for the provision of drinking water to the bulk of its
population, seventy percent of which is ferrous based and thus susceptible to corro-
sion, this is of great concern. Bacteria and other microbes form biofilms in most natu-
ral environments, incorporating extracellular polymeric substances and water, to in-
crease the microbes ability to survive under the given conditions [2]. On metal sur-
faces, biofilms are known to contribute to corrosion in a process known as Microbially
Influenced Corrosion (MIC). Biofilms are also known to increase the in-homogeneity
of soil along pipelines. MIC due to biofilms leading to increased localized corrosion/
rate of localized corrosion along potable water pipelines is an area of interest that
requires further exploration as limited work in soils has been undertaken to date.
Soil is a complex environment containing solids (soil particles), liquids (soil water)
and gases [3]. The physical structure of soil is very important as it can restrict the
flow of liquids, which restricts the replenishment of oxygen to different processes and
causes in-homogeneities along a metal structure buried in the soil, such as a pipe-
line. This impacts on both the corrosion of metal structures and the formation of bio-
films. Soil is a typical liquid/gas/solid multiphase corrosion system as the solid soil
particles form pores within which water is trapped resulting in liquid dispersion across
a metal surface [4]. Jiang et al have modelled the importance of the geometry of tri-
ple phase boundary (TPB) zones in multiphase systems, such as soil, on corrosion
rates [5]. As the length of total TPB zones on a metal surface increases, the liquid
dispersion across the surface increases and the corrosion current density increases
in a linear fashion. The geometry of TPB zones is affected by soil type, soil compres-
sion and water content. Similarly, the physical structure of soil affects the ability of
microbes to move through soil and form biofilms. Bacteria can only move through soil
in water, so are restricted to water filled pores as planktonic bacteria or form biofilms
along the surface of soil particles where they can remain encased in a hydrated gel
[3]. Studies were conducted in the late 1950s which showed increased levels of mi-
crobial activity in disturbed soil and along pipelines where water was free flowing
compared with undisturbed soil [6, 7]. This was believed to be due to higher levels of
moisture in these areas. Where soil is less compacted, such as along pipelines, wa-
ter is able to penetrate and flow more rapidly leading to increased aeration [8]. This
has the potential to increase both microbial activity and corrosion rates.
To date, studies of MIC in soil have been commonly undertaken in solutions or soil
samples. Nutrients are added to solutions to mirror the mineral content of soils [9-11]
but this does not take into account the physical structure of soils and the resultant
electrochemical interactions that are dependent on soil geometry. The alternative is
taking soil samples from a site of interest or using artificial soils such as sand [12,
13]. This approach also has limitations as with the removal of soils from their natural
environment the geometry of the soil is altered which can impact both corrosion rates
and the microbe communities. If microbes are to be introduced to the soil, the sample
must be first sterilized which also impacts on the native soil composition. An added
disadvantage of soil samples is that it is difficult to ensure reproducible conditions
across experiments.
Nutrients are added to solutions to mimic the mineral content of soil and to encour-
age the growth of bacteria. The effect of these nutrients on electrochemical perfor-
mance is not well understood. Two studies which used liquid media specially devel-
oped for SRBs took into consideration the effects of nutrients on electrochemical
properties [14, 15] One of the studies found different effects between liquid media
with and without yeast extract present [15]. Initially yeast extract was included in the
media, but in later tests it was removed. It was found that while the rate of growth of
bacteria was reduced, the interference noted in previous electrochemical measure-
ments was eliminated when yeast extract was removed indicating the yeast extract
was the cause of the interference. The second study took a sulfate based medium
and built on this to produce a sulfate/ lactate medium designed to be highly support-
ive of the growth of Desulfovibrio desulfuricans [14]. As distinct from the first study,
both media were analyzed with and without bacteria. They were able to obtain repro-
ducible results from potentiodynamic scans and indicated no problems with the yeast
extract in their medium.
Nutrients other than yeast extract have also been shown to impact on corrosion. It is
well documented that chloride ions are very important for localized corrosion and that
other anions can inhibit corrosion by competing with chloride ions at the metal inter-
face [16]. In MIC control of this balance is made difficult by bacteria consuming and
fixing various nutrients such as sulfate, so what was initially an inhibiting solution can
become a corrosive solution as anions other than chloride are removed from the so-
lution. Webster & Newman included sodium chloride to ensure initiation of corrosion
in stainless steel and removed ferrous ions and reducing agents as there were indi-
cations that both these groups interfered with electrochemical measurements [15].
This suggests that more information is required about the electrochemical properties
of common bacterial growth media without the presence of bacteria to more readily
identify the effect of bacteria.
A novel system utilizing agar as a gelling agent for the study of MIC in soil is being
developed which will take into account the physical composition of soil as well as the
chemical composition and be reproducible and controllable under laboratory condi-
tions. Silica and polyacrymalide gels have been used previously to replicate sludge
for the investigation of corrosion in wastewater sludge [17, 18]. It was determined
that steel exposed to the gels had a more negative corrosion potential than when
exposed to equivalent solutions, but that the gels lowered the corrosion current. The
corrosion behavior of carbon steel exposed to solid agar under a range of oxygen
conditions has been investigated. From this it has been determined that the oxygen
level of the agar impacts on the ability to obtain replicable results with solid agar and
the mode of corrosion [19]. A significant level of crevice corrosion was observed, the
severity of the crevicing being dependent on the oxygen level [20]. This made it very
difficult to compare results. In later work this has been remedied with the aid of a dif-
ferent seal on the sample area. It has also been determined that solid agar (15g/L) is
not an ideal medium for the study of MIC in soil.
This work investigates the corrosion properties of carbon steel exposed to semi solid
agar (4g/L) with a range of nutrient conditions. Future work will incorporate the addi-
tion of Pseudomonas to the system under investigation. Pseudomonas is a gram
negative facultative anaerobic genus of bacteria commonly found in soil that has
been implicated in contributing to corrosion along buried steel pipelines. Before the
effect of bacteria can be analyzed, a complete understanding of the performance of
carbon steel exposed to semi solid agar is required. The effect of nutrients other than
sodium chloride on this system must also be understood.

2 Methodology
Three sets of agar with different nutrient conditions, one with minimal nutrients, one
with 12.5g/L of basic nutrients added and one with 25g/L of basic nutrients added,
were compared.

2.1 Sample preparation

Hot rolled carbon steel sheet 1mm thick was cut into 25mm 2 coupons. Based on the
manufacturer’s specifications the steel was composed of 0.04-0.07% carbon, 0.03%
silicon, 0.5% manganese, 0.03% phosphorus, 0.03% sulphur, 0.1% aluminium,
0.04% titanium with iron composing the remainder (99.3%). Each sample was pol-
ished using a combination of SiC paper and diamond paste to a final finish with 1µm
diamond paste. For the final stages analytical grade ethanol lubricant was used. Fol-
lowing polishing the samples were rinsed in ethanol and heat dried.

2.2 Agar preparation

Three different nutrient conditions were used to continue the investigation into the
electrochemical properties of steel exposed to agar. The first contained 4g/L of agar
and 5g/L of sodium chloride which are fully dissolved when mechanically mixed with
distilled water on high heat until boiling. The second nutrient set contained 25g/L of
Oxoid Nutrient Broth No. 2 (10.0g/L Lab Lemco powder, 10.0g/L peptone and 5g/L
sodium chloride) with 4g/L agar which was prepared in the same manner. The third
was the same as the second but with 12.5g/L of Oxoid Nutrient Broth No 2 instead of
25g/L. Once the components were dissolved, the solutions were then adjusted with
0.1M sodium hydroxide and hydrogen chloride to the desired pH (7.0±0.1). Nitrogen
gas is then bubbled through the agar for 30 minutes to de-aerate it. The agar is then
cooled to below 60°C with nitrogen gas and the dissolved oxygen (DO) levels meas-
ured with a Mettler Toledo portable meter and probe. The DO was brought to below
1.0mg/L before air is bubbled through the solution to bring the DO to maximum (ap-
proximately 5.0mg/L). All samples are set in the presence of bubbled air to ensure
the maximum oxygen levels are maintained.

2.3 Electrochemical characterisation

A standard three electrode cell was constructed with a 5cm 2 platinised titanium mesh
counter electrode, standard calomel reference electrode and a polished carbon steel
sample as the working electrode. A circular area with a 1cm diameter was exposed
of the working electrode. To limit crevice corrosion a viton fluoroelastomer o-ring seal
was used instead of the standard Teflon ring at the working electrode and the edges
of the seal coated with Vaseline. Agar was poured into the completed cell as a solu-
tion and then allowed to set for 2.5 hours before measuring the open circuit potential
(OCP) for three hours. Following OCP measurement, a cyclic polarisation scan was
conducted from -1.0V vs SCE relative to the Eoc to 1.4V vs SCE relative to the refer-
ence electrode. A scan rate of 0.167mV/s was used. For each set of nutrient condi-
tions another experiment was conducted. Potential hold measurements at -600mV vs
SCE were held for 48 hours following a 2.5 hour setting period. Triplicate scans were
conducted under all conditions.

2.4 Electrochemical measurements

Surface analysis of the steel samples after scanning was conducted with optical mi-
croscopy and scanning electrical microscopy (SEM). A FEI Quanta 200 Environmen-
tal SEM was used at 15.0kV or 30.0kV and 5.0 spot size. This enabled the surface to
be examined without a conductive coating being applied.

3 Results
The open circuit potential (OCP) under each of the nutrient conditions was measured
for three hours. With nutrients the OCP of -0.745±0.009mV vs SCE was approxi-
mately 50mV vs SCE more negative than the samples without nutrients (-0.696
±0.001mV vs SCE and -0.694±0.002mV vs SCE) and the more nutrients present, the
higher the OCP.

Table 1: Open circuit potential values for three nutrient conditions

Average OCP (V vs SCE)
No nutrients -0.745 ±0.009
12.5 g/L nutrients -0.696 ±0.001
25g/L nutrients -0.694 ±0.002
3.1 Potentiodynamic scans
As nutrient levels increase, there is a clear trend in the anodic regions of the
potentiodynamic scans, as seen in Figure 1. The transpassive regions are occurring
at lower potentials and as the corrosion potential is not changing significantly across
the three nutrient conditions, the anodic region is becoming more compact with in-
creasing nutrient concentration. The sample exposed to agar with no nutrients (red)
has two active regions and a relatively stable passive region from approximately -
600mV to -300mV. It can be noticed that within this region, a slight protrusion is evi-
dent at around -400mV, characterized by a slight increase and subsequent decrease
in current. This may be indicative of a breakdown and subsequent re-formation of the
passive layer. Further, as the concentration of nutrients is increased, this breakdown
and subsequent reformation of the passive layer becomes more noticeable and se-
vere, as evidenced by the protrusions occurring at -520 and -720 mV in the green
(12.5g/L) and blue (25g/L) curves respectively. There is also a shift to the right and
lowering of the passive regions as nutrient concentration increases. This suggests
that within the regions of passivity, the system is thermodynamically more active (in-
creased negative potential range) and exhibits higher corrosion rates.
0
-0.2
-0.4
-0.6
E (V vs SCE)

-0.8
-1
-1.2
No nutrients
-1.4 12.5 g/L nutrients
-1.6 25 g/L nutrients
-1.8
1E-07 1E-06 1E-05 1E-04 1E-03 1E-02 1E-01 1E+00 1E+01
|i| (mA/cm2)

Figure 1: Potentiodynamic scans for three different nutrient conditions

3.2 Potential hold measurements

Potential hold measurements were taken at -600mV for all three nutrient conditions
and can be seen in Figure 2. There was a considerable drop in current with the intro-
duction of nutrients, the measured current initially one third of that without nutrients
and over time this decreased to a fifth. The sample with 25g/L of nutrients added has
a much less undulating curve than that with no nutrients as well as the curve with
12.5g/L of nutrients. The spacing of the undulations in the 12.5g/L nutrient sample is
closer than that without nutrients.
 Figure 2: Current over time for three different nutrient conditions at -600mV vs SCE

4 Discussion
Nutrients have been shown in the past to interfere with electrochemical measure-
ments [15], though this has been mostly attributed to yeast extract. Due to this a nu-
trient broth without yeast extract was selected to investigate the electrochemical per-
formance of semi-solid agar with nutrients when compared to that without nutrients.
Consistent results are important under any given set of parameters to enable trends
to be determined. With the nutrient combination used here in different concentrations,
reproducible results were obtained.

With no nutrients present in the semi-solid agar, the OCP was determined to be
50mV vs SCE more negative than those measured with nutrients present. The differ-
ence of 2mV vs SCE between the two conditions with nutrients suggests that above
a certain concentration that has not yet been determined, the concentration of nutri-
ents has less of an effect than the presence of nutrients at all on the OCP. The posi-
tive increase in potential measured indicates that the nutrients are reducing the drive
for corrosion to occur.

This is supported by the trends seen when a steady potential of -600mV was applied
and the current changes with time monitored as seen in Figure 2. With nutrients pre-
sent the current measured was five times less than that without nutrients. There was
also very little difference between the 12.5g/L nutrient samples and the 25g/L nutrient
samples, though the 12.5g/L samples has slightly higher initial peaks in current. This
significant decrease in current could be due to a number of reasons. With nutrients
the conductivity of the agar could be reduced or the nutrients could be interfering with
the activation characteristics of the metal surface. Either of these explanations would
lead to a reduction in corrosion rates as seen here. The nutrients could be altering
the activation characteristics at the metal surface by acting as a retarding catalyst by
interfering with the interaction of chloride ions with the surface. They could also be
binding the chloride ions before they can interact with the surface.

A second effect was seen with the potential hold measurements. With no nutrients
present, the current undulates with time with an amplitude of approximately 0.05mA.
This occurs across the 48 hours. With nutrients present there is some modulation
initially, though after 15 hours both samples are very stable and the amplitude is
never greater than 0.02mA. The modulation without nutrients has been associated
with an unstable passive film forming, then breaking down and then reforming. With
nutrients corrosion is believed to be retarded overall with less likelihood of a passive
layer forming.

The increased instability of passive layer formation with nutrients present is support-
ed by the potentiodynamic scans seen in Figure 1. As nutrient concentration increas-
es, the transpassive regions shift to more negative potentials and the anodic regions
become more compressed. The passive regions are also shifting to higher current
densities and more negative potentials while becoming shorter and more unstable.
Of the three conditions, this data suggests that the sample is most inert with no nutri-
ents present and that corrosion is thermodynamically more favorable across the po-
tential range tested the more nutrients are present. Nutrients appear to be interfering
with the ability for oxide layers to form. It is suggested that with less stable oxide
films, when they form the current density is not decreased to the same extent as with
more stable oxide films and less potential needs to be applied for chloride ions to
penetrate the surface and cause pitting. In essence, the critical pitting potential is
reduced with increased nutrient concentration.

Nutrients competing with chloride ions at the metal surface have been previously
shown to decrease corrosion rates [16]. Initially, corrosion rates with nutrients were
seen to be lower than without nutrients, but with a lower tendency for corrosion to
occur, passivation is thermodynamically less favorable [21]. This means that as the
potential becomes more positive, corrosion rates are increased with nutrients as they
are less protected. Without an external potential applied or when a low potential be-
low the passivating potential of the samples without nutrients is applied, as was the
case with the potential holds shown here, the corrosion rate of the samples with nu-
trients is lower than those without. Above the passivating potential it is hypothesized
based on the potentiodynamic scans seen here that this would be reversed.

5 Conclusions
The electrochemical effect of nutrients added to semi solid agar has begun to be ex-
plored. Initial observations suggest that the nutrients have a retarding effect on the
ability of chlorides to attack the steel surface and that corrosion rates are reduced.
However, this also decreases the ability of the steel to form a protective oxide layer
and thus passivation when nutrients are present is much less stable and at these
potentials a lower current density is seen for samples exposed to agar without nutri-
ents than those exposed to agar with nutrients. Using this nutrient and agar concen-
tration combination results were consistent and replicable. This work is aiming to en-
able a better understanding from MIC tests to be gathered as a more complete un-
derstanding of the electrochemical effects without bacteria will increase the ability for
the precise effect of bacteria to be seen. Future work will further test the system be-
ing developed with and without bacteria.

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