Lecture 31: Biofouling of Titanium Biofilm Studies

NPTEL Web Course

Lecture 31
Biofouling of Titanium Biofilm Studies
Keywords: Biofouling, Sea Water, Titanium.

Titanium has many attractive properties such as light weight, good mechanical
strength and above all excellent corrosion resistance even in sea water. It will then
be very useful to assess biofouling tendency of titanium in sea water environments in
order to understand its utility in several nuclear power generation plants.

Biofouling refers to undesirable accumulation of a biofilm deposit on an engineered

surface. An organic film composed of microorganisms embedded in a polymeric
matrix forms the basis for the attachment of various macro-organisms, inorganic and
corrosion products. Such formation of biofilms on material surfaces poses a threat to
the successful utilization of materials in industrial environments.
Energy losses reduced heat transfer efficiency resulting from insulating
biofilm
Increased capital costs.
Enhanced maintenance costs and replacements.
Shutdowns .
Stages in marine biofouling:
a) Successive coverage of a surface by growth of primary colonizing bacteria
and other organisms. Aerobic followed by anaerobic organisms colonise in
succession.
b) A transient stage where multilayers of cells become embedded in their own
polymer matrix.
c) The final stage of development of a mature biofilm with high density of
macrobial population (diatoms, protozoa etc).

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Course Title: Advances in Corrosion Engineering
Course Co-ordinator: Prof. K. A. Natarajan, IISc Bangalore

Lecture 31: Biofouling of Titanium Biofilm Studies

NPTEL Web Course

Biofilms initially formed can be easily removed by physical or chemical means.

However, those formed for extensive periods of time are difficult to be removed due
to incorporation of inorganic and organic products formed by the metabolic activities
of attached microorganisms. Biomineralization such as oxidized manganic and iron
oxides occur within biofilms.
Two fundamentally different forms of biomineralization are recognized. Biologically
induced mineralization in which organisms generate conditions suitable for
precipitation of extracellular mineral phases. The second one called boundary
organized biomineralization involves growth of inorganic particles within organic
biomatrix.
Titanium is used as the tube material for seawater-cooled condensers in nuclear
reactors and also, in plate-type heat exchangers of the auxiliary cooling water
system. Titanium is known to be prone to microbial attachment and biofouling.
Biofouling of heat exchanger surfaces may result in the deposition of biogenic MnO2
on the surfaces severely compromising efficiency.

Investigations were carried out to study mineralization of manganese on titanium

surfaces immersed in seawater due to the activities of marine bacteria. Microbial
fouling and formation of inorganic deposits on titanium grade-2, commonly used for
condenser tubes was established.

Bacterial isolation, culturing, growth and

attachment on titanium surfaces were studied. Total viable counts of culturable

marine bacteria on titanium surface were monitored as shown in Fig. 31.1.

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Course Title: Advances in Corrosion Engineering
Course Co-ordinator: Prof. K. A. Natarajan, IISc Bangalore

Lecture 31: Biofouling of Titanium Biofilm Studies

NPTEL Web Course

Viable counts / cm

Marine organisms
Manganese oxidizers

10

10

10

10

10

12

14

16

18

20

Exposure (Months)
Fig. 31.1 Viable counts of marine bacteria and manganese oxidizers on titanium exposed to sea water.

The percentage of Manganese oxidizing bacteria (MOB) in the total bacterial counts
showed an increase from an initial value of about 3040% to almost 100% at the end
of about 20 months.
Epifluorescence studies showed increased biofilm formation on titanium surface
exposed to seawater with exposure time. Throughout the study the biofilm was
always patchy and non-uniform. Epifluorescence and confocal images indicated
highly irregular biofilms (Fig. 31.2).

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Course Title: Advances in Corrosion Engineering
Course Co-ordinator: Prof. K. A. Natarajan, IISc Bangalore

Lecture 31: Biofouling of Titanium Biofilm Studies

NPTEL Web Course

Fig. 31.2 Epifluorescence micrographs of biofilm development on titanium.

Surface coverage of the biofilm considerably increased with time.

The EPS content, which is the other major component of the biofilm besides the
microbial constituents, was determined as a function of exposure time. The Protein
and carbohydrate constituents of the EPS were analyzed. Contribution of
carbohydrate was more than that of protein in the biofilm.
Confocal scanning laser microscope image of an year old biofilm showing mature
biofilm is illustrated in Fig. 31.3.

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Course Title: Advances in Corrosion Engineering
Course Co-ordinator: Prof. K. A. Natarajan, IISc Bangalore

Lecture 31: Biofouling of Titanium Biofilm Studies

NPTEL Web Course

Fig. 31.3 confocal image of an year old biofilm on titanium.

Protein levels in the biofilm however increased rapidly after a year of exposure and
almost equaled the carbohydrate levels.
The ability of the isolated bacteria to oxidize manganese was determined both in
liquid as well as solid media.
The formation of visible brown colonies on a solid agar plate containing 50 ppm of
Mn (II) was confirmation of bacterial manganese oxidation. Solid agar plates
inoculated with MOB and manganous ions are shown in Fig. 31.4. Bacterial
manganese oxidation is evident as growth of coloured colonies. Presence of
brownish colonies on filter membranes is a confirmatory test for the manganese
oxidation ability of marine bacteria isolated from biofilms formed on titanium.
Biomineralization of manganese by the isolated bacteria was further confirmed by
studying manganese oxidation in liquid media. Various manganese oxidizing
bacteria were categorized with respect to their ability to oxidize manganous ions at
different concentration levels. The isolates exhibiting highest manganese oxidation
rates were shortlisted for further detailed investigations.

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Course Title: Advances in Corrosion Engineering
Course Co-ordinator: Prof. K. A. Natarajan, IISc Bangalore

Lecture 31: Biofouling of Titanium Biofilm Studies

NPTEL Web Course

Fig. 31.4 Agar plates containing manganous ions and manganese oxidizing bacteria (MOB) showing growth of
coloured colonies.

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Course Title: Advances in Corrosion Engineering
Course Co-ordinator: Prof. K. A. Natarajan, IISc Bangalore

Lecture 31: Biofouling of Titanium Biofilm Studies

NPTEL Web Course

Optical micrographs of some manganese (II) oxidizing bacteria isolated from the
biofilms grown on titanium surfaces are illustrated in Fig. 31.5.

Pseudomonas

Vibrio

Bacillus

Micrococcus

Bacillus

Fig. 31.5 Predominant manganese oxidizing bacteria isolated from marine biofilms on titanium.

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Course Title: Advances in Corrosion Engineering
Course Co-ordinator: Prof. K. A. Natarajan, IISc Bangalore

Lecture 31: Biofouling of Titanium Biofilm Studies

NPTEL Web Course

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Course Title: Advances in Corrosion Engineering
Course Co-ordinator: Prof. K. A. Natarajan, IISc Bangalore