2nd International Conference on Chemical, Ecology and Environmental Sciences (ICCEES'2012) Singapore April 28-29, 2012

A Study of Pseudomonas Aeruginosa Bacteria

in Microbial Corrosion
Hamidreza Mansouri, S.A Alavi, Mehdi Yari

controversial. Pipeline corrosion was estimated in 1996 to cost

the gas industry about $840 million/year [2] and in 2001 it
was estimated that the annual cost of all forms of corrosion to
the oil and gas industries was $13.4 billion, of which
microbially influenced corrosion accounted for about $2
billion [3]
While it is well recognized that chemical and microbial
mechanisms both contribute to corrosion, it is uncertain what
the relative contribution of microbial activity to overall pipe
corrosion is. It has been estimated that 40% of all internal
pipeline corrosion in the gas industry can be attributed to
microbial corrosion [4] but data are needed to confirm or
revise this estimate. Basic research to increase our
understanding of the microbial species involved in microbial

Abstract microbially influenced corrosion has been the subject

of extensive studies for the past five decades and several models have
been proposed to explain Mechanisms governing biocorrosionone of
the most famous bacteria in this field is Pseudomonas aeruginosa, a
Gram-negative motile rod bacteria and widely distributed in nature
and can grow in distilled water, laboratory hot water baths, hot tubes,
wet IV tubing, and other water containing vessels. Some of these
organisms may be primary pathogens .These Bacteria are capable of
sensing surfaces.Contacting.the surface initiates a complex
differentiation program resulting in e.g. synthesis of alginate. Metal
surfaces are rapidly colonized by microorganisms in contact with
natural or industrial aquatic environments, giving rise to a complex
and strongly adhering microbial community, termed as biofilm
Pseudomonas genus are acknowledged to be the pioneer colonizers in
the process of biofilm formation and often found in the primary stage
of biofilm formation in aquatic environments The biofilm
accumulation not only protects microbial cells from the external
environment, but it is also detrimental to the underlying substratum
thereby causing physical degradation or biodeterioration of the metal
surface.

KeywordsPseudomonas

aeruginosa, microbial corrosion,

microbiologically influenced corrosion (MIC), biocorrosion

I. INTRODUCTION

WING TO its economic and environmental importance,

especially for the oil and gas industry, microbially
influenced corrosion has been the subject of extensive
studies for the past five decades and several models have been
proposed to explain Mechanisms governing biocorrosion [1]
Corrosion is a leading cause of pipe failure and is a main
component of the operating and maintenance costs of gas
industry pipelines. Quantifying the cost of corrosion generally,
and more specifically the cost associated with microbial
corrosion, in the gas industry is not easily done and is

Fig.1 Pseudomonas aeruginosa [19]

corrosion and their interactions with metal surfaces and with
other microorganisms will be the basis for the development of
new approaches for the detection, monitoring, and control of
microbial corrosion. A thorough knowledge of the causes of
microbially influenced corrosion and an efficient and effective
means of detecting and preventing corrosion are lacking. It is
well recognized that microorganisms are a major cause of
corrosion of metal pipes, but despite decades of study it is still
not known with certainty how many species of
microorganisms contribute to corrosion, how to reliably detect
their presence prior to corrosion events, or how to rapidly
assess the efficacy of biocides and mitigation procedures [5].
Investigations of microbial species present in gas industry
pipelines have traditionally relied upon the use of samples
obtained from pipelines to grow bacterial cultures in the
laboratory [6]. Laboratory growth media cannot accurately

Hamidreza Mansouri is with Azad University Science and Research

Branch of Tehra. Email: mansoorihamidreza2012@gmail.com, Tel:
+989171825738
S.A.Alavi Department of Chemical engineering, Azad University Science
and Research Branch of Tehran.
Mehdi Yari, Department of Metallurgy and Materials, Azad University
Science and Research Branch of Tehran.

42

2nd International Conference on Chemical, Ecology and Environmental Sciences (ICCEES'2012) Singapore April 28-29, 2012

reflect the true conditions within pipelines, and

microbiologists have recognized that the vast majority of
microbial species cannot currently be grown in the laboratory
[7] thus; culture-dependent approaches underestimate the
biocomplexity of microbial communities.
.

cells and growth of the population, while keeping cell-to-cell

contacts, result in the formation of micro colonies.
Mature biofilms often possess a hallmark architecture where
micro colonies are surrounded by a network of water channels
allowing the flow of nutrients into the interior of the biofilm.
Hydrodynamics as well as cell-to-cell signaling molecules
play a regulatory role in the development of the biofilm
architecture.
Stoodley et al. (2002) speculated that also the maintenance
of open water channels in multi-species biofilms requires
interspecies signalling to inhibit growth and EPS production
into the channels. Some cells can be released to a planktonic
lifestyle ensuring the occupation of new niches. The release of
cells occurs due to physical detachment caused by shear
forces, or due to a programmed set of events controlled by
signaling molecules and leading to a local hydrolysis of EPS
matrix, e.g. P. aeruginosa can cleave its own EPS by alginate
lays enzyme. [18].

II. PSEUDOMONAS AERUGINOSA BACTERIA

P. aeruginosa is a Gram-negative motile rod capable of
causing infection in almost all of the bodys tissues. P.
aeruginosa utilises a variety of virulence factors and can
produce biofilms to aid attachment and dispersal and uses its
flagella and pili to establish infections in the host. [19] See
figure 1.
P. aeruginosa infections are often associated with
blue/green pus or exudates with a characteristic musky odour
P. aeruginosa widely distributed in nature (soil, water, plants,
animals). it can grow in distilled water, laboratory hot water
baths, hot tubes, wet IV tubing, and other water containing
vessels. This explains why the organism is responsible for so
many nosocomial infections. [8]. Pseudomonas genus are
acknowledged to be the pioneer colonizers in the process of
biofilm formation and often found in the primary stage of
biofilm formation in aquatic environments [9].

IV.

THE INFLUENCE OF PSEUDOMONAS AERUGINOSA ON

ALLOYS

Bacteria are capable of sensing surfaces [13].Contacting the

surface initiates a complex differentiation program resulting in
e.g. synthesis of alginate. Genes essential for production of
this major EPS component of Pseudomonas aeruginosa were
shown to be up-regulated already 15 min after attachment.
[14]
Figure 3 show the SEM images of the Ni-Co discs surfaces
immersed in the medium without P. aeruginosa and with P.
aeruginosa[20] after incubation for 5 h. in fig. 3a the oxide
components of Ni and Co can be clearly seen.

III. BIOFILM AND BIOCORROSION

Metal surfaces are rapidly colonized by microorganisms in
contact with natural or industrial aquatic environments, giving
rise to a complex and strongly adhering microbial community,
termed as biofilm .[16] The biofilm accumulation not only
protects microbial cells from the external environment, but it
is also detrimental to the underlying substratum, thereby
causing physical degradation or biodeterioration of the metal
surface This phenomenon is widely recognized as
biocorrosion or microbiologically influenced corrosion
(MIC).[17]

Fig.3 (a) the oxide components of Ni and Co

As displayed in Fig.3b-e after 5 hours of immersion with
medium containing bacterium, it can be seen that the Ni-Co
surface partially covered with clusters of microbial cells, EPS
and metabolism products. In addition, from the Fig. 3e,
pitting corrosion and crevice corrosion were observed on the
surface of the inoculated medium rather than on the surface
exposed to only medium. The energy dispersive X-ray spectra
(EDS) analysis of the corrosion products on discs was
illustrated in Fig.3d.

Fig.2 Model of biofilm development as a part of bacterial life

cycle [13]
Model of biofilm development as a part of bacterial life
cycle is shown in figure 2.[13] Individual planktonic cells can
reach the surface by active or passive means. The initial cellto-surface contact is often reversible, but if environmental
cues and possible signaling molecules favor surface-attached
growth, then the cells attach irreversibly with the aid of
excreted extracellular polymeric substances (EPS). Division of

43

2nd International Conference on Chemical, Ecology and Environmental Sciences (ICCEES'2012) Singapore April 28-29, 2012

Fig.3 (e) pitting corrosion and crevice corrosion

Fig.3 (b) medium with P. aeruginosa

The corrosivity of Pseudomonas aeruginosa and sulfatereducing bacteria (SRB) in seawater was evaluated in aerobic
and anaerobic conditions [20]. The Pseudomonas species are
potential producers of extracellular polymers (EPS), the
production of which is intensified in the presence of oxygen.
In turn, the presence of EPS in the biofilms generates a
different aeration gradient, creating favorable conditions for
the development of the SRB. So that, although the SRB are
strict anaerobes, a higher number of this group of bacteria is
observed in the biofilms formed under aerobic conditions [21]
Two experiments were carried out (under aerobiosis and
anaerobiosis) in electrochemical cells, in duplicate, as
presented in figure 4. In these systems, aseptically, AISI 1020
carbon steel coupons were placed, of which 5 (about 1 cm
area) for polarization experiments and another 4 (about 5 cm
area) for both sessile microorganism quantification and weight
loss measurements. Subsequently, the system was filled with
sterile seawater and some nutrients in order to guarantee the
viability of the microorganisms. Each experiment lasted for 28
days and was kept under slight magnetic agitation and at room
temperature. In the experiment under anaerobiosis, nitrogen
purges were performed, before and after the inoculation, as
well as during the entire experiment in order to establish
dissolved oxygen concentration lower than 3 ppm. [20].

Fig.3 (c) medium with P. aeruginosa

Fig.3 (d) The EDS analysis of the corrosion products on discs

Fig.4 Test cell

44

Fig.5 Variation
of the corrosion potential
throughout the time

2nd International Conference on Chemical, Ecology and Environmental Sciences (ICCEES'2012) Singapore April 28-29, 2012

The weight loss was proportional to the exposure time

presenting practically linear behavior in the aerated system see
Figure 5.In this Figure variation of the corrosion potential
throughout the time is shown. [20].
Another study on Copper and galvanized steel has
implemented these materials are frequently used in the
construction of cooling towers because of their well-known
antifouling property. Biofilm in cooling water systems is a
fairly common problem. When the biofilm layer could not be
cleaned, pathogen bacteria living there can lead to fatal
diseases. Copper and galvanized steel coupons (50 x 25 x 1
mm) were prepared according to guidelines in ASTM G1-72
(ASTM 1975).Copper coupons: Surface of copper coupons
(99.9% purity) were sanded. The total surface area of each
coupon was determined. Galvanized steel coupons: The
thickness of the zinc coating covering the stainless steel was
5m. The cut areas of all the coupons were coated with epoxy
zinc phosphate primer (Moravia Turkey) and then covered
with epoxy finish coating (Moravia, Turkey) to avoid the
initiation of corrosion at these disturbed areas. The total
surface area of each coupon was determined.[21] Three
coupons of each material were removed monthly from the
cooling water system during ten months. Biofilms on their
surfaces were scraped by sterile swab, suspended in sterile tap
water and vortexes for 60 s [22]. Biofilm was confirmed by
SEM on copper and galvanized coupons in experiments in
Figure 7, SEM micrograph of the biofilm formed on copper
(A,B) and galvanized steel (C,D) surfaces, 1th month (A,C),
10th month (B,D) are shown.

All isolates were Gram negative and non-spore forming

bacteria. [23]
TABLE I :PREDOMINANT VIABLE MICROORGANISMS IN THE
BIOFILM ON COPPER AND GALVANIZED STEEL SURFACES

Fig.8 TAMHB in the biofilm on copper and galvanized steel surfaces.

Error bars represent the standard deviation. CFU: Colony forming
unit

V. HEALTH RISKS FROM MICROBIAL GROWTH AND BIOFILMS

2B

Microbes can enter distribution systems through a wide

range of avenues, including treatment processes or through
deficiencies of the distribution system infrastructure.
Microbial presence in the distribution system can result in
colonization of the distribution system infrastructure. Once
biofilm development begins, subsequent material, organisms
and contamination introduced to the distribution system can
become entrained in the biofilm. The biofilm can protect
microbes from disinfection and allow microbes injured by
environmental stress and disinfectants to recover and grow. In
addition, biofilms may increase pipe corrosion, adversely
affect pipe hydraulics and reduce the utility of total coli forms
as indicator organisms. Microbial growth in biofilms may
result in deterioration of water quality, generation of bad tastes
and odors, and proliferation of macroinvertebrates.
Contamination and material in the biofilm may subsequently
be released into the flowing water under various
circumstances. As a result, biofilms can act as a slow-release

Fig.7 SEM micrograph of the biofilm formed on copper (A, B)

and galvanized steel (C, D) surfaces, 1th month (A, C), 10th
month (B, D)
During ten months the result of total aerobic mesophilic
heterotrophic bacteria (TAMHB) on copper and galvanized
steel surfaces are shown in Figure 8. Statistical analysis
demonstrated that TAMHB counts were significantly (P<0.01)
higher on the surface of galvanized steel coupons than on the
surface
of
copper
coupons.
Predominant
viable
microorganisms isolated on copper and galvanized steel
biofilms were shown in Table I. [23]
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2nd International Conference on Chemical, Ecology and Environmental Sciences (ICCEES'2012) Singapore April 28-29, 2012

[9] Toder, DS. 1998. Pseudomonas aeruginosa: ubiquitous pathogen. pp.

mechanism for persistent contamination of the water. The

organisms and their products may decrease disinfectant levels
(by increasing disinfectant demand), pose a direct public
health risk, or create taste and odor problems. Biofilms likely
exist in all distribution systems, and are recognized as a
normal part of the distribution system.[12]
P. aeruginosa is widespread in environmental waters,
especially in those waters associated with human activity. The
organism is often found in finished waters and in pipe
biofilms. Although P.aeruginosa has not been conclusively
implicated in a reported waterborne disease outbreak, it has a
significant role in nosocomial illness, including outbreaks. It is
a pathogen of concern for people with severe burns and
wounds, diabetes, and is the primary cause of injury and death
in people with cystic fibrosis.[10] Some strains cause
pneumonia in general intensive care units (ICUs) and pediatric
ICUs. Clinically significant strains have been found in the
hospital plumbing system, suggesting that drinking water may
contribute to nosocomial infections. A list of nosocomial
outbreaks associated with P. aeruginosa in contaminated
drinking water appears in Highsmith et al. (1986). However,
the linking between the water distribution system (as opposed
to the hospital plumbing system) and the presence of clinically
important strains of P. aeruginosa in the nosocomial setting is
still open to question [11]. Numerous changes in gene
regulation cause the biofilm cells to become phenotypically
and metabolically different from their planktonic counterparts
[15]

[10]
[11]

[12]
[13]
[14]
[15]
[16]
[17]
[18]
[19]
[20]

[21]

ACKNOWLEDGMENT

[22]

The authors thank to Mr.Reza Javaherdashti for his valuable

comments in this research

[23]

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