Metals and Materials International

https://doi.org/10.1007/s12540-018-0038-2

Failure Analysis of Pitted Copper Pipes Used in Underground Water

and Preventive Measures
Gi‑ho Nam1 · Jong‑kwon Lee1 · Kyung‑ja Kim2

Received: 23 February 2017 / Accepted: 21 August 2017

© The Korean Institute of Metals and Materials 2018

Abstract
This study performed an experiment on the causes of pitting corrosion in a copper tubing used for a sprinkler system. Corro‑
sion products of a copper tubing that sustained pitting corrosion were collected and cultured in Culture medium [Luria–Ber‑
tani, Brain heart infusion, Tryptic soy broth (TSB), R2A]. Four types of bacteria were found through identification: Micro‑
coccus luteus sp (species)., Staphylococcus sp., Sphingomonas sp., and Bacillus sp. The copper toxicity test was performed
for each microorganism. Among the four microorganisms, Micrococcus luteus sp. showed good growth in the environment
containing copper ions. On the immersion test, changes in pH and Optical density were measured; On the inductively coupled
plasma optical emission spectrometry test, the copper concentration of each culture medium was measured. The surface of
each copper sample was observed using a scanning electron microscope. The corrosion potential of a copper sample, after
48 h exposure of the TSB medium containing Micrococcus luteus sp., was measured using a potentiodynamic polarization
experiment. The next experiment was conducted to prevent microbial corrosion by suppressing the growth of microorganisms.
Six 30 ml TSB culture media with controlled pH value of 4, 5, 6, 7, 8, and 9 through HCl and NaOH were manufactured.
Then the microorganisms were cultured in 37 °C 133 rpm, of which the growth status was checked every 24 h for 3 days. It
was found that microorganisms did not grow on culture media with the pH value of 6 and lower. The same experiment con‑
ducted on culture media controlled with acetic acid, nitric acid, and sulfuric acid, also showed no growth of microorganisms
on media with pH value of 6 and lower. Six 5 ml TSB culture media each containing 0.5, 0.25, 0.125, 0.0625, 0.0312%, and
0.0156% NaOCl and NaOBr as germicides were manufactured. 0.01 μl of microorganisms were inoculated on the media and
cultured in 37 °C for 48 h. It was found that microorganisms did not grow in media with NaOCl and NaOBr concentration of
0.0625% and higher. Therefore, it can be suggested that in environments with pH value of 6 or lower, or NaOCl and NaOBr
concentration of higher than 0.0625% suppresses microbial growth, thereby preventing microbial corrosion.

Keywords  Copper pipes · Pitting corrosion · Microbiologically influenced corrosion · Bacteria · Micrococcus luteus

1 Introduction

Copper has high thermal conductivity, and also has excel‑

lent ductility and malleability, making itself easy to work
with. Also, its excellent erosion resistance makes it possible
* Jong‑kwon Lee
jklee@sch.ac.kr for use in various types of water pipes used in daily lives,
including boiler pipes and drinking water pipes. Recently,
Gi‑ho Nam
giho1001@sch.ac.kr pitting corrosion outbreaks on copper sprinkler pipes with
stagnant tapwater or groundwater emerged as an issue. Dep‑
Kyung‑ja Kim
kyoungjakim@hotmail.com osition of oxide, and the presence of chloride ion are origi‑
nally blamed for the causes of pitting corrosion of copper
1
Department of Materials Engineering, Soonchunhyang [1]. And Lee’s recent study suggests that another cause of
University, 22 Soonchunhyang‑ro, Shinchang‑myun, pitting corrosion of copper pipes with stagnant groundwater
Asan 31538, Korea
is the residual carbon film after extrusion [2–5]. In addition,
2
Department of Life Science, Soonchunhyang University, 22 microbiologically influenced corrosion is also mentioned as
Soonchunhyang‑ro, Shinchang‑myun, Asan 31538, Korea

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the cause of accelerating the pitting [6, 7]. Microbiologi‑
cally influenced corrosion (MIC) refers to the phenomenon
when the corrosion accelerates due to the direct and indirect
effects of metabolism of microorganisms in the environment
[8, 9]. Microorganisms are generally tiny with several μm in
size, and are survivable in diverse temperature, pH, and oxy‑
gen levels, existing in almost every environments on earth.
Proliferating in various conditions based on their surround‑
ings, they metabolize using various resources including
simple inorganic substances to complex organic substances,
and eve metallic ions [10–12]. Through metabolism these
microorganisms create metabolic wastes including various
organic and inorganic acids, or oxidize and reduce metal or
metallic ions to directly accelerate the corrosion. They may
also create highly viscous biofilm consisting of extracellular
polymer (EPS, or exopolymer) to capture nutriments, caus‑
ing corrosion on differential aeration cells. There has been
reported cases of MIC on power plant [13], water supply and
sewerage pipes [14], maritime structures [15], and under‑
ground buried structures [16], which indicate the exposure
of metallic structures to microorganisms, and are focused
by ongoing research. A. Reyes’ study on the copper pipes
of urban and rural areas shows that pitting was discovered
in copper pipes located in rural areas where biofilm existed,
while the copper pipes located in urban and some rural areas
where biofilm did not exist did not show any signs of pit‑
ting [17]. Valcarce confirmed that Pseudomonas bacteria
increases the chance of corrosion of copper and copper
alloys, and dezincification occurs in case of brass. Also,
after observing the anodic polarization curve, it was found
that the presence of bacteria increases the pitting potential,
but at the same time increase the difference between repas‑
sivation and the pitting potential [18]. Also, according to
Vargas, the adsorbated biofilm generates minute changes to
the metal surface, and the organic substances in the biofilm
absorbs and desorbs unstable copper attached to itself on
the interface between the biofilm and the metal, in certain
moisture and chemical conditions. This absorption/desorp‑
tion, in turn, transmits reactive chemical substances, thereby
directly posing impact on the corrosion [19, 20]. Therefore,
the study aimed to confirm the presence of microbiologi‑
cally influenced corrosion pitting on copper, and if so, which
microorganisms are responsible for the acceleration of the
pitting. First, the corrosive substances were collected from
copper sprinkler pipes where the pitting occurred, and were
cultured. The colonies of cultured bacteria were categorized
depending on their size, shape, and color, and then re-culti‑
vated. The strains of bacteria were identified, and the degrees
of proliferation for each bacteria in copper ion-existing cir‑
cumstances were checked to confirm the most proliferative
bacteria on copper materials. Then, to assess the impacts of
bacteria on copper, copper specimen (10 × 10) was immersed
in the culture fluid, and the bacterium proliferating in copper
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ion-existing circumstances was inoculated, and the degree
of proliferation of the bacterium and the pH changes were
measured. After the culture was finished, the changes in the
weight of the copper specimen, and the copper concentra‑
tion of the culture fluid was measured, and the surface of the
copper specimen was observed. Also, the anodic polariza‑
tion measurement was performed using the copper speci‑
men immersed in bacterium-inoculated culture fluid, the
specimen immersed only in culture fluid, and the specimen
immersed in groundwater. The polarization potentials and
the current distributions of each specimen were measured
and compared to assess the impact of the bacterium on the
corrosion rate. Also, to prevent MIC occurrence, measures
to restrain the proliferation of microorganisms were studied.
Research on the effects of different pH levels and different
types of acids on microorganisms was conducted, and the
minimum inhibitory concentration of NaBrO and NaClO,
the components of bactericides, on the growth of micro‑
organisms was assessed. However, it should be noted that
acid significantly increases corrosivity. To inhibit the cor‑
rosion of copper, the amount of inhibitor is important in the
sense of expense and corrosivity. The smaller the required
inhibitor concentration is, the better. Even though NaBrO
and NaClO could decrease the initiation of pitting, those
chemicals can increase the corrodibility of copper. Since
those chemicals are strong oxidizer and can be applied as
disinfection and water treatment agents, high dosage is not
recommended. However, they can be dissociated naturally
without remaining any harmful effect if they added in small
amounts. In this reason, the addition of NaBrO and NaClO
as biocide are frequently employed in the water treatment
of cooling system.
2 Experiment Procedures
2.1 Cultivation and Identification of Corrosion
Products in Copper Tubing
Microorganisms were cultured and identified on corrosion
products of a copper tubing in which pitting corrosion had
occurred. Four different types of culture media were used for
the incubation, and the culture media are shown in Table 1.
100 µl of suspension of corrosion products was collected
and put into the four different culture media (2 ml each).
Then, the media were cultivated in the incubator at 36 °C,
132 rpm for 48 h. To ensure complete isolation, the suspen‑
sion was smeared on the plates coated with each of the cul‑
ture media, and these were cultivated for 48 h at 30 °C. After
48 h, colonies cultured in the media were isolated by color,
shape and size; Solgent co. was then requested to perform
identification.
Metals and Materials International

Table 1  Compositions of culture media

Culture medium Composition

Luria–Bertani (LB) Tryptone 10.0 g/l, yeast extract 5.0 g/l, NaCl 10.0 g/l

Brain heart infusion (BHI)
Tryptic soy broth (TSB)
R2A
brain infusion solids 12.5 g/l, beef heart infusion solids 5.0 g/l, proteose peptone 10.0 g/l, sodium
chloride 5.0 g/l, glucose 2.0 g/l, disodium phosphate 2.5 g/l
tryptone 17 g/l, soytone 3 g/l, dextrose 2.5 g/l, sodium chloride 5 g/l, dipotassium phosphate 2.5 g/l
yeast extract 0.5 g/l, proteose peptone 0.5 g/l, casein acid hydrolysate 0.5 g/l, glucose 0.5 g/l, starch
0.5 g/l, di-potassium phosphate 0.3 g/l, magnesium sulphate 0.024 g/l, sodium pyruvate 0.3 g/l,
agar 15.0 g/l

2.2 Copper Toxicity Test every 24 h to check the proliferation of microorganisms. To

To examine the growth potential of microorganisms—con‑
firmed by the identification—on the copper surface, an Agar
diffusion test (paper disc method) was performed. First of
all, the plates were kept for 2–3 h in the incubator at 37 °C
to determine whether the plates had been contaminated for
other microorganisms to grow. The culture media were put
onto the pure plates on which microorganisms did not exist,
and then hardened. 200 μl of microorganisms was smeared
on the plates coated with the culture media, and a paper disc
(Ф8 mm) was put in the middle of the plates. To test the
toxicity of copper, 1, 10, and 100 mM of C ­ uCl2 and ­CuSO4
were injected into the discs, respectively, and incubated for
48 h until the clear zone appeared in the incubator at 37 °C.
2.3 Cultivation of Microorganisms in Various pH
Solution
100 µl of each microorganism—which grew very well in the
presence of copper ion—was inoculated into 25 ml of each
sterilized TSB (Tryptic soy broth) medium in pH 4.0, 5.0,
6.0, 7.0, 8.0 and 9.0. These were cultivated in the incubator
at 37 °C and 133 rpm for 3 days. Optical density (OD) and
changes in pH were measured every 24 h for the period.
The OD values were measured in the wavelength of 600 nm,
which minimizes the absorbance on components of the cul‑
ture media; an UV–Vis spectrophotometer by Shimadzu was
used. The pH level of TSB medium was 7.2 and was adjusted
with 2 N HCl and 1 N NaOH.
examine the effects of microorganisms, 30 ml of distilled
water with a copper sample and 30 ml of TSB medium were
sterilized in the same manner without microorganisms inoc‑
ulated, and then were cultured in the incubator for 6 days.
A copper sample after the process of aging was immersed
with microorganisms, taken out, and placed into a flat speci‑
men holder. The test solution was ground water. The satu‑
rated calomel was used as the reference electrode, and the
high-density graphite stick as the relative electrode. The test
equipment was DC105B by Gamry set to range between
– 300 and 400 mV from the corrosion potential and scan at
0.167 mV/s. OD and pH values measured for the period were
compared; the weight loss of each copper sample as well as
the concentration of copper in each solution were measured.
The concentration was measured by ICP-OES optic7000DV.
3.1 Inhibition the Growth of Microorganisms
by Acid Type
A copper sample was cut out to a size of
12 mm × 12 mm × 0.3t. The pH levels of the sample were
adjusted using nitric acid, hydrochloric acid, sulfuric acid
and acetic acid to pH 4, 5 and 6. Samples were put into 30 ml
of TSB medium and sterilized in an autoclave at 1 atm and
121 °C for an hour. 100 µl of microorganisms was inoculated
into the sterilized TSB medium (30 ml), and cultured in
the incubator at 37 °C and 133 rpm for 7 days. Changes in
OD and pH values were measured every 24 h for the period
to check the growth of microorganisms. After 7 days, the
weight reduction of the sample was calculated.

3 Experiment on Copper Corrosion 3.2 Minimum Inhibitory Concentration of NaBrO

in the Culture Medium and NaClO Against Microorganisms

A copper sample was cut out to a size of
12 mm × 12 mm × 0.3t. This was put into 30 ml of TSB
(Tryptic soy broth) medium, and sterilized in an autoclave
at 1 atm and 121 °C for an hour. 100 µl of microorgan‑
isms was inoculated into the sterilized TSB medium with
the copper sample, and cultivated in the incubator at 37 °C
and 133 rpm for 6 days. OD and pH values were measured
Acid increases the corrosivity of copper. NaBrO and NaClO
were used to reduce the risk of corrosivity of acids and
at the same time suppress the growth of microorganisms
which may induce microbial corrosion. However, it should
be noted that appropriate usage of NaBrO and NaClO are
needed as the substances may also increase corrosivity.
Therefore, Minimum Inhibitory Concentration (MIC) test

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was conducted to identify the MIC of NaBrO and NaClO
on microorganisms. Using the sterilized TSB medium (auto‑
clave, 1 atm, 121 °C, 1 h), 5 ml of each TSB medium with
the concentrations of 0.5, 0.25, 0.125, 0.0625, 0.0312 and
0.0156% of NaBrO and NaClO were made. 0.01 µl of micro‑
organisms was inoculated and then the growth of microor‑
ganisms was confirmed after 48 h of incubation at 37 °C. A
copper sample (12 mm × 12 mm × 0.3t) was added to 30 ml
of TSB medium with the minimum inhibitory concentration
of NaBrO and NaClO, and incubated at 37 °C and 133 rpm
for 48 h. After the incubation, it was confirmed that micro‑
organisms had grown.
4.2 Copper Toxicity
Each of the four types of microorganisms—Bacillus sp.
Staphylococcus sp. Sphingomonas sp. and Micrococcus
sp.—was tested for copper toxicity by applying 1, 10, and
100 mM of ­CuCl2 and C ­ uSO4. The results are presented
in Figs. 1, 2. No clear zone was found around all of the
four types of microorganisms after the application of 1 and
10 mM of ­CuCl2 and ­CuSO4. However, the application of
100 mM of C ­ uCl2 and ­CuSO4 showed a clear zone around
Bacillus sp. Staphylococcus sp. and Sphingomonas sp., and
did not show any around Micrococcus sp. This confirms that
Micrococcus sp. grows well even in an environment contain‑
ing copper ions.

4 Results 4.3 Cultivation of Microorganisms in Various pH

4.1 Identification of Corrosion Products in Copper
Tubing
Corrosion products that had formed around pits of copper
tubing were cultivated using four types of culture media. In
the LB medium, three types of microorganisms which had
different sizes, shapes, and colors were found. In the TSB
medium, one type of microorganism was found. The identifi‑
cation of these four types of microorganisms was performed
using SolGent, and the results are presented in Table 2. The
three types of microorganisms from the LB medium were
identified as Bacillus sp., Staphylococcus sp. and Sphingo‑
monas sp., while the one type of microorganism from the
TSB medium was identified as Micrococcus sp.
Solution
While general bacteria proliferate in conditions of pH value
ranging from 5.0 to 8.0, the ideal pH value range of patho‑
genic bacteria is 7.2–7.6. However, the preferred acidity
may vary among bacteria, as lactobacillus prefers acidic
conditions, while Vibrio cholera prefers alkaline condi‑
tions. Inoculation of Escherichia coli on liquid media sig‑
nificantly changes the acidity of the media, with the initial
measurement of pH 7.3 acidulated to pH 5.1 in 5 h, and
slightly neutralized to pH 5.3 in 19  h. Various types of
yeast or fungus proliferate on subacid culture media of pH
5–6. Although bacteria usually proliferate in neutral pH or
alkalescent environments, there exist bacteria which prefer
acidic conditions, including Thiobacillus thiooxdance, an

Table 2  Result of Bacteria identification

Culture medium Gene

1. Luria–Bertani (LB) Bacillus subtilis strain J-18 16S ribosomal RNA gene, partial sequence
Bacillus subtilis strain ATCC 19217, complete genome
Bacillus amyloliquefaciens gene for 16S ribosomal RNA, partial sequence, strain: RNB-113
Bacillus amyloliquefaciens SQR9, complete genome
Bacillus sp. BAB-3426 16S ribosomal RNA gene, partial sequence
2. Luria–Bertani (LB) Staphylococcus hominis partial 16S rRNA gene, isolate B6
Uncultured bacterium partial 16S rRNA gene, clone MD01A10
Uncultured bacterium clone PE08 16S ribosomal RNA gene, partial sequence
Select seq emb|AM084016.1| Staphylococcus sp. R-25050 16S rRNA gene, strain R-25050
Staphylococcus sp. H292 gene for 16S ribosomal RNA, isolate: H292
3. Luria–Bertani (LB) Sphingomonasyabuuchiae strain R2A 25-5 16S ribosomal RNA gene, partial sequence
Sphingomonaspseudosanguinis strain J1-q 16S ribosomal RNA gene, partial sequence
Uncultured bacterium clone B11-159 16S ribosomal RNA gene, partial sequence
Sphingomonas sp. Iso-85 16S ribosomal RNA gene, partial sequence
Bacterium CAGY7 strain CAGY7 16S ribosomal RNA gene, partial sequence
4. Tryptic soy broth (TSB) Micrococcus sp. 3409 16S ribosomal RNA gene, partial sequence
Micrococcus sp. 3723 16S ribosomal RNA gene, partial sequence
Micrococcus sp. 3498 16S ribosomal RNA gene, partial sequence
Micrococcus luteus strain OH4847 16S ribosomal RNA gene, partial sequence
Micrococcus luteus strain INBI-1 16S ribosomal RNA gene, partial sequence

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Fig. 1  a Bacillus sp. b Staphylococcus sp. c Sphingomonas sp. d Micrococcus sp. Toxicity Test in C
zone: about a 15 mm, b 7 mm, c 10 mm, d None
extreme acidophile which proliferates around pH 0 condi‑
tions. Majority of acidophiles grow in neutral conditions,
but extreme acidophiles cannot. In this way, the ideal range
of pH value for growth differ in various microorganisms.
Also, as they each produce different types of metabolites,
the existence of these microorganisms may change the pH
value of the surroundings, thereby impacting its own prolif‑
eration. Therefore, the ideal pH value for microbial growth
was measured in order to control the pH value, to suppress
microbial growth and corrosion. 100 µl of Micrococcus sp.
­ uCl2 (100  mM). The radius of the clear
was inoculated into each sterilized TSB medium of pH 4, 5,
6, 7, 8, and 9. It was then cultivated in the incubator at 37 °C
and 133 rpm for 3 days, during which OD at different pH
levels and changes in pH levels were measured. The meas‑
urements of OD are presented in Table 3. Originally, OD at
each pH level was 0.104. After 24 h, it showed no change
at pH 4, 5, and 6 but increased to 2.249 at pH 7 and to over
2.4 at pH 8 and 9. After 48 h, it showed no change at pH 4,
5, and 6 but increased to over 2.7 at pH 7, 8, and 9. After
72 h, it showed no change at pH 4, 5, and 6 but increased
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Fig. 2  a Bacillus sp. b Staphylococcus sp. c Sphingomonas sp. d Micrococcus sp. Toxicity Test in ­CuSO4 (100 mM). The radius of the clear
zone: about a 16 mm, b 8 mm, c 10 mm, d None
to 2.839 at pH 7, which was the highest value. It increased
to 2.781 at pH 8, which was similar to the previous value,
but decreased to 2.384 at pH 9. Changes in pH levels are
presented in Table 4. The TSB media at pH 4, 5, and 6 did
not show any change in pH. After 72 h, the TSB medium
at pH 7 reached pH 8.442, and the TSB medium at pH 8
reached pH 8.859. The TSB medium at pH 9 decreased to
pH 8.046 after 24 h and increased to pH 8.342 after 72 h.
This indicates that Micrococcus sp. does not proliferate at
pH 4, 5, and 6, yet proliferates at pH 7, 8, and 9. The reason
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for the decrease from pH 9 to pH 8.064 after 24 h was the
rapid proliferation of Micrococcus sp. in the early stage. The
increase to pH 8.342 after 72 h was due to the depletion of
nutrients in the TSB medium, which inhibited the prolifera‑
tion of the microorganism.
4.4 Copper Corrosion in Culture Medium
Micrococcus sp. was cultured for 6 days in a TSB medium
to which a copper sample had been added. OD and changes
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Table 3  Opticla density of Micrococcus sp. in TSB medium for Micrococcus luteus. In comparison, it increased by 60% to
3 days 863.64 μg/g in the TSB medium with Micrococcus luteus.
OD (600 nm) This indicates that Micrococcus sp. accelerates the dissolu‑
tion of copper in the medium. Figure 3 shows a photo of the
Initial 1 day 2 days 3 days
copper sample that underwent aging for 6 days in the TSB
pH4 0.103 0.112 0.099 0.104 medium with Micrococcus sp. After being cultured in the
pH5 0.099 0.104 0.094 0.099 medium with the microorganism, the surface of the cop‑
pH6 0.102 0.112 0.103 0.1 per sample was examined. The pits presented in Fig. 3 was
pH7 0.104 2.249 2.777 2.839 confirmed. There were crazes over the entire surface, and
pH8 0.101 2.479 2.726 2.781 thin layers peeled off easily. Figure 4 shows the result of the
pH9 0.104 2.449 2.726 2.383 anodic polarization test, which used 1 l of ground water as
the test solution, for each copper sample that was put into the
TSB medium containing Micrococcus sp., the TSB medium
Table 4  pH of Micrococcus sp. in TSB medium for 3 days without Micrococcus sp., and ground water for 48 h. The
pH
corrosion potential of Micrococcus sp. has lowest value.
When converted in mmpy using tafel extrapolation method,
Initial 1 day 2 days 3 days
the corrosion rate was 0.1093 for the TSB medium contain‑
pH4 4.014 4.025 4.019 4.032 ing Micrococcus sp., 0.0386 for the TSB medium without
pH5 5.032 5.028 5.034 5.047 Micrococcus sp., and 0.0257 for ground water. Micrococcus
pH6 6.011 6.024 6.018 6.013 sp. increased the corrosion rate in the test solution by 4.2
pH7 7.045 7.248 7.915 8.442 times. This confirms that that Micrococcus sp. accelerates
pH8 8.031 7.948 8.383 8.859 corrosion. It was inevitable that Micrococcus luteus caused
pH9 9.014 8.046 8.342 8.818 serious damage to the copper surface.

4.5 Inhibition of the Growth of Microorganisms

in pH were measured every 24 h. For comparison, distilled
water containing a copper sample and a TSB medium con‑
taining a copper sample without Micrococcus sp. were left
for 6 days. OD and changes in pH observed in the three are
presented in Table 5. For the TSB medium with Micrococ‑
cus sp., the OD rapidly increased to 2.638 and continued
the upward trend, reaching 3.072 after 6 days. The pH level
increased from 7.229 to 9.371. Micrococcus sp. prolifer‑
ated well in the environment containing a copper sample.
For the TSB medium without Micrococcus sp., the OD
increased from 0.139 to 0.591 yet the pH level remained
nearly the same. Changes in OD were due to C­ l− in the TSB
medium that was increased by the dissolution of the cop‑
per sample. In distilled water, the OD and pH showed no
change. The weight of each copper sample was measured.
The copper sample in distilled water decreased from 0.615
to 0.614 g, which barely showed any change in weight. In
the TSB medium without Micrococcus sp., it decreased
from 0.6154 to 0.6096 g. The copper sample in the TSB
medium with Micrococcus sp. showed the biggest change
in weight, decreasing from 0.6097 to 0.5984 g. The copper
concentration was 523.81 μg/g in the TSB medium without
by Acid Type
100 µl of Micrococcus luteus was inoculated into TSB media
whose pH levels were adjusted using nitric acid, sulfuric
acid, hydrochloric acid, and acetic acid to pH 4, 5 and 6.
Changes in OD and pH values were measured over 7 days.
As time passed, OD values in Micrococcus luteus increased
on all four acids with the highest OD value at pH 4 after
7 days. Figure 5a shows a OD value increased in pH 4 media
controlled with sulfuric acid by 0.75. Figure 5b reflects the
observation of OD value of TSB culture media, after con‑
trolling the pH value to 4, 5, and 6 using sulfuric acid and
immersing copper specimen, while not inoculating Micro‑
coccus sp. The highest OD value was retrieved in media
with pH value of 4, which is identical to the experiment
with Micrococcus sp. inoculation. The OD value of pH4
media controlled with sulfuric acid increased by 0.73. As
can be seen from the two figures, the changing trends in OD
values were found to be similar between the media inocu‑
lated with and without Micrococcus sp. Therefore, it can
be identified that the increase in OD value resulted from
the dissolution of copper on acid, rather than from bacterial

Table 5  OD and pH (24 h) Initial 1 day 2 days 3 days 4 days 5 days 6 days
change of TSB solution
containing Micrococcus sp Optical dansity 0.139 2.638 2.823 2.976 2.88 2.833 3.072
pH 7.229 7.576 8.231 8.647 9.156 9.283 9.371

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Fig. 3  SEM microphotograph of pits (aging in the solution containing
Micrococcus sp. after 6 days at a 200×, and b 2000×
growth. An increase in OD values seemed to occur as the
result of the dissolution of copper on acids, not by bacterial
growth (Fig. 6). A slight increase in pH values on all four
acids (acetic acid, sulfuric acid, nitric acid, and hydrochloric
acid) was confirmed, which seemed to occur as a result of
the dissolution of copper. Also, the concentration of cop‑
per on each solution at pH 4 was presented in Fig. 7. Any
electrochemical reaction can be divided into oxidation and
reduction reactions. This means that corrosion process could
consist of oxidation and reduction process. Thus, in the cop‑
per corrosion process, cathodic reactions such as oxygen
reduction or hydrogen evolution reaction are subsequential.
The possible reactions in the neutral environment are as
follows.
2H2 O + 2e − H2 + 2OH−
O2 + H2 O + 4e − 4OH−
According to the above cathodic reactions, the pH of
­ H− ions. The above
solution are increased by producing O
indicates that Micrococcus luteus cannot grow at low pH
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levels, regardless of the acid type. However, the dissolution
of copper at pH 4, 5 and 6 accelerates corrosion. Therefore,
using acids is not an effective method to prevent corrosion.
5 Minimum Inhibitory Concentration
of NaBrO and NaClO Against
Microorganisms
As indicated in the above experiment results, copper was
dissolved into the medium solution. Since copper tubing
is damaged by a low pH level, an inhibitory method that
minimizes the influence of acidity and inhibits the growth of
microorganisms, which cause corrosion, was studied using
NaBrO and NaClO. After 48 h of culturing, both NaBrO
and NaClO turned white at the concentration of less than
0.0312% due to the proliferation of Micrococcus luteus.
Therefore, the minimum inhibitory concentration of NaBrO
and NaClO was 0.0625%. Micrococcus luteus was not able
to grow when the concentration of NaBrO and NaClO was
higher than 0.0625%. Thus, the growth of microorganism
could be inhibited by adding of low concentration of bio‑
cides, such as NaBrO and NaClO. However, the fact that
those biocides inhibited the growth of microorganism, does
not lead the direct measure against pitting, because those
chemicals contain pit-enhancing halides ions.
6 Discussion
Microbial corrosion is closely related to the activity of
microorganisms existing in surrounding environment.
Microbial corrosion induced by Sulfate Reducing Bacteria
(SRB) inhabiting in low-porosity and highly moist anaero‑
bic conditions, including clay, is well known [17]. Also, it
was found that MIC can occur in oxygen-abundant aerobic
conditions. In aerotropic conditions, microbial corrosion
is induced by highly corrosive metabolites (for instance,
generation of acids) generated from microbial activities,
by iron oxidizing bacteria’s effects on iron to create iron
oxide or iron hydroxide, or by generation of differential aera‑
tion cell caused by sludge or biofilm. Micrococcus sp. is a
well-known aerobic bacteria, and does not expedite corro‑
sion like anaerobic SRB. Also, it does not generate organic
acid and influence corrosion, as the experiment revealed
that the surrounding pH value was not increased. Another
major role of microorganisms on corrosion is adhering to
the metal surface to change the characteristics of metal/
electrolyte interface through metabolism, and ultimately
alter the electrochemical process. Such phenomena may
occur by microorganisms simply adsorbing onto the metal
surface [18]. Generally, microorganisms in nature exist in
sessile state (that is, adsorbed onto the surface), rather than
Metals and Materials International

Fig. 4  Polarization curves
(scan rate of 0.167 mV/s.)
ML: copper immersed in TSB
medium with Micrococcus sp.,
TSB: copper immersed in TSB
medium, UW: copper immersed
in underground water for 7 days




Optical Density


 
S+

 S+

pH

S+
  S+
         S+
Day
 S+
(a)

        
Day
 (a)
Optical Density




S+

 S+

S+
pH

  S+
         S+
Day
 S+
(b)

       
Fig. 5  Change of optical density with sulfuric acid a containing Mic‑ Day
rococcus sp., and b not containing Micrococcus sp
(b)

Fig. 6  Change of pH with sulfuric acid a containing Micrococcus sp.,

and b not containing Micrococcus sp

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 Metals and Materials International

 the componential analysis of biofilm reveals that the actual

1250
1160
microorganisms only account for 10% of the biofilm, while

exopolymer accounts for the rest [21]. Microorganisms
 protected by exopolymer is in a very safe state. Once bio‑
concentration (บ/g)

840 863.64 863.64 film is created, other bacteria may adsorb onto the biofilm,
 depending on the surrounding environmental conditions,
thereby formic a microbial community (or, microbial con‑
 523.81
sortium). The impacts of biofilm on metal corrosion can be
 summarized as in Table 6. Amongst these impacts, one of
the most important factor is the inducement of local corro‑
 sion through generation of the differential aeration cell. The
existence of biofilm on metal surface leads us to believe

acetic sulfuric nitric hydrochloric TSB TSB+ML that biofilm may provide different conditions in comparison
with general electrolytes (that is, soil) including electrolyte
Fig. 7  Copper concentration of each solution in TSB medium of pH4 composition and oxygen concentration inside the biofilm.
(containing Micrococcus sp.) after 6 days Cathodic reactant should exist in order for corrosion to
take place inside the biofilm. In general cases of neutral or
Table 6  The effects of biofilm on the corrosion of metals alkalescent conditions of the soil, dissolved oxygen acts as
Physical effect Chemical effect
the cathodic reactant. The reduction of these dissolved oxy‑
gen is determined by the rate of diffusion of the oxygen.
a. diffusion barrier a. anodic reaction The oxygen inside the biofilm is to be depleted if the oxygen
b. warm currents of surface, flow b. cathodic reaction
consumption rate of aerobic bacteria exceeds the speed of
decrease c. eruption of protective film
c. heat transfer efficient decrease d. selective ion transmission diffusion onto the interface of metal and biofilm. This forms
e. pH change the differential aeration cell with the outside of the biofilm,
thereby causing local corrosion focused on the inside of the
biofilm. The possible reactions under the biofilm are as fol‑
in planktonic state. This is common in environments lack‑ lows (Fig. 8).
ing nutrition, as organic matter which microorganisms feed
on, alsotend to adsorb onto the surfacedue to surface charge 2Cu → 2Cu2+ + 4e−.
[19]. Also, it has been found that corrosion products of metal 2H2 O + O2 + 4e− → 4OH−
tend to adsorb organic matter in a stronger manner, than
that of metal itself. These adsorbing microorganisms are 2Cu2+ + 4OH− → 2Cu(OH)2
mainly composed of polysaccharide, and forms the biofilm Such phenomenon is similar to the corrosion induced by
through creasing highly viscous extracellular expolysaccha‑ the formation of differential aeration cell, found in water
ride (EPS, or exopolymer). Exopolymers not only combine table and inlet of buried pipelines. The existence of bio‑
the cells, but also protect cells from external stimulations. film is the biggest factor in explaining the locality of MIC
The biofilm sustains internal moisture, and forms polymer [18]. Micrococcus sp. was also found to expedite corrosion
matrix by accumulating nutrition. This is the reason why

Fig. 8  Oxygen concentration
cell under tubercle

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Metals and Materials International
through biofilm’s generation of differential aeration cell.
Microorganisms’ metabolic behavior vary in accordance
with the rapidly changing surrounding conditions, and is
also diverse in terms of interaction with another microorgan‑
ism. This leads us to believe that there are various factors
which expedite corrosion, and so a precise research on their
mechanism is of need.
7 Conclusion
1. Corrosion products in a copper tubing in which pitting
corrosion had occurred were collected to perform iden‑
tification. The results of the identification showed four
different types of microorganisms—Bacillus sp., Staphy‑
lococcus sp., Sphingomonas sp., and Micrococcus sp.
2. On the copper toxicity test, the clear zone appeared with
an injection of 100 mM of ­CuCl2 and ­CuSO4 for all
types except Micrococcus luteus sp. It was confirmed
that only Micrococcus luteus sp. among the found
microorganism grows well in the environment contain‑
ing copper ions.
3. On the immersion test, the presence of Micrococcus
luteus sp. helped the dissolution of copper, remaining
the pitting on surface.
4. On the minimum inhibitory concentration (MIC) test
using NaOCl and NaOBr, the growth of Micrococcus
luteus sp. was inhibited at the concentration of and
above 0.0625%. which means the MIC of NaOCI and
NaOBr were 0.0625%.
5. When pH was adjusted using acids, Micrococcus luteus
sp. could not grow at and below pH6. On the ICP test,
however, higher copper concentration was found in low
pH than by Micrococcus luteus sp., which was inad‑
equate to prevent pitting corrosion.
6. The pitting failure of copper could be ascribed to the
presence of microorganism such as Micrococcus luteus
sp., that grew in the environment containing copper ions
which could be inhibited by the addition of biocides.
The simple change of pH of solution could not affect the
growth environment of microorganism.
Acknowledgement  This research was supported by the Soonchunhy‑
ang University Research Fund.
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