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Stina Wallstrm


Som med tillstnd av Kungliga Tekniska Hgskolan i Stockholm framlgges till

offentlig granskning fr avlggande av teknisk doktorsexamen torsdagen den 21 april
2005, kl. 13.00 i Kollegiesalen, Valhallavgen 79, KTH, Stockholm. Avhandlingen
frsvaras p svenska.
To Anders
Silicone rubber high voltage insulators are sometimes colonised by microorganisms which
form a biofilm on the surface of the infected unit. In this work insulators exposed to the
outdoor environment in Sweden, Sri Lanka and Tanzania respectively have been studied. The
biofilms colonising the insulators were shown to be of roughly the same composition
regardless of their origin. Algae in association with bacteria dominated the biofilms and
provided nutrition to mold growth. The isolated microorganisms were further used to study
the effect of a biofilm on different silicone rubber materials. New tools for diagnosing
biological growth on polymeric materials were developed and used to analyse the silicone
rubber samples.

No evidence of biodegradation of the polydimethylsiloxane (PDMS) molecule has been found

in this work. However, this does not mean that PDMS rubbers used in high voltage insulators
can be called bioresistant. Silicone insulating materials always contain additives and these
may promote or hinder growth. For this reason, an extensive test program was developed, in
order to evaluate the effect of different additives on the degree of biological growth. The
program spanned from fast and easy methods, useful for screening large amount of samples,
to the construction of specially designed microenvironment chambers in which mixed
biofilms, similar to those formed on the surface of silicone rubber insulators in the field, were
successfully grown.

The test program showed that the flame retardant zinc borate protected the materials, whereas
alumina trihydrate (ATH) did not hinder biological growth. On the contrary, environmental
scanning microscopy (ESEM) in combination with X-ray energy dispersive spectroscopy
(EDS) showed that the surface roughening caused by the addition of ATH to the silicone
rubber matrix made the materials more difficult to clean.

Furthermore when the hydrophobic surface of a silicone rubber insulator is covered by a

hydrophilic biofilm this leads to a reduction of the surface hydrophobicity of the material.
This may alter the electrical properties of the insulator. It is therefore important to develop
methods to identify biofouled units. In this work, laser-induced fluorescence (LIF)
spectroscopy was explored as a tool for the detection of biofilms on silicone rubbers. The
experiments revealed that weak traces of algae or fungal growth, even those not visible to the
naked eye, could be detected by this technique. In addition, it was shown that photography
and subsequent digital image analysis could be utilised to estimate the area covered by
biofilm growth. The results obtained indicate that LIF spectroscopy in combination with
image analysis could be used for field diagnostics of biological growth on insulators in
This Thesis is a summary of the following papers

Paper I Wallstrm, S., Dowling, K., and Karlsson, S., Development and comparison of
test methods for evaluating formation of biofilms on silicones. Polymer
Degradation and Stability, 2002. 78(2): p. 257-262.

Paper II Wallstrm, S. and Karlsson, S., Biofilms on silicone rubber insulators; microbial
composition and diagnostics of removal by use of ESEM/EDS. Polymer
Degradation and Stability, 2004. 85(2): p. 841-846.

Paper III Wallstrm, S., Strmberg, E., and Karlsson, S., Microbiological growth testing of
polymeric materials: an evaluation of new methods. Polymer Testing, 2005:
In press

Paper IV Wallstrm, S., Dernfalk, A.D., Bengtsson, M., Krll, S., Gubanski, S.M., and
Karlsson, S., Image analysis and laser induced fluorescence combined to
determine biological growth on silicone rubber insulators. Polymer Degradation
and Stability, 2005: 88(3): p 394-400

Paper V Wallstrm, S. and Karlsson, S., Effects of microbiological growth on PDMS

materials. Manuscript, to be submitted to polymer degradation and stability.

Paper VI Bengtsson, M., Wallstrm, S., Sjholm, M., Grnlund, R., Andersson, M.,
Larsson, A., Karlsson, S., Krll, S., and Svanberg, S., Fungus covered insulator
materials studied with laser induced fluorescence and principal component
analysis. Applied Spectroscopy, 2005: Submitted.

Paper VII Bengtsson, M., Grnlund, R., Sjholm, M., Abrahamsson, C., Dernfalk, A.D.,
Wallstrm, S., Larsson, A., Weibring, P., Karlsson, S., Gubanski, S.M., Krll, S.,
and Svanberg, S., Fluorescence lidar imaging of fungal growth on high-voltage
outdoor composite insulators. Optics and Lasers in Engineering, 2005. 43(6): p.
The author was responsible for and carried out the major part of the work in paper I-V

In paper VI the author was responsible for sample preparation and assisted in the
measurement of fluorescence, the analysis of the data and the writing of the paper.

In paper VII the author was responsible for sample preparation and microscopy and assisted
in the analysis of the data.

Mathematical processing of data from fluorescence measurements and image analysis were
performed by Magnus Bengtsson and Andreas Dernfalk respectively in all studies performed.
1 PURPOSE OF THE STUDY ..........................................................................................1
2 INTRODUCTION ...........................................................................................................3
2.1 HIGH VOLTAGE INSULATORS ......................................................................................3
2.2 PROPERTIES OF SILICONE RUBBER ..............................................................................4
2.3 DEGRADATION MECHANISMS......................................................................................5
2.4 DIAGNOSTIC METHODS ...............................................................................................6
2.4.1 Accelerated ageing.............................................................................................6
2.4.2 Analytical tools ..................................................................................................6
2.4.3 On-line measurements .......................................................................................7
2.5 COMPOSITION OF BIOFILMS.........................................................................................8
2.6 DAMAGES CAUSED BY BIOFILMS .................................................................................9
2.6.1 Fouling...............................................................................................................9
2.6.2 Degradation of leaching components ................................................................9
2.6.3 Biotic degradation ...........................................................................................10
2.6.4 Hydration and penetration...............................................................................11
2.6.5 Color and odor.................................................................................................11
2.7 PREVENTION OF MICROBIOLOGICAL GROWTH ...........................................................11
2.7.1 Biocides............................................................................................................11
2.7.2 Cleaning...........................................................................................................12
2.7.3 Surface modification ........................................................................................12
3 EXPERIMENTAL.........................................................................................................13
3.1 MATERIALS ..............................................................................................................13
3.1.1 Silicones ...........................................................................................................13
3.1.2 Insulators .........................................................................................................14
3.1.3 Microorganisms ...............................................................................................14
3.2 GROWTH TESTS.........................................................................................................15
3.2.1 Powder test, mold.............................................................................................16
3.2.2 Powder test, algae............................................................................................16
3.2.3 IEC 68-2-10, mold ...........................................................................................16
3.2.4 ASTM G21-90, mold ........................................................................................16
3.2.5 ASTM D 5589-97, algae ..................................................................................16
3.2.6 Quick test, mold ...............................................................................................17
3.2.7 Microenvironment chambers ...........................................................................17
3.3 LASER-INDUCED FLUORESCENCE ..............................................................................18
3.4 IMAGE ANALYSIS ......................................................................................................19
3.5 DEGRADATION STUDIES ............................................................................................20
3.5.1 Enzymatic degradation ....................................................................................20
3.5.2 Degradation by Fenton mechanism .................................................................21
3.6 INSTRUMENTATION...................................................................................................21
3.6.1 Oxygen plasma ageing.....................................................................................21
3.6.2 Light microscopy..............................................................................................21
3.6.3 SEM..................................................................................................................21
3.6.4 ESEM/EDS.......................................................................................................21
3.6.5 IR......................................................................................................................21
3.6.6 GC/FID ............................................................................................................22
3.6.7 SEC ..................................................................................................................22
4 RESULTS AND DISCUSSION ....................................................................................23
4.1 COMPOSITION OF BIOFILMS.......................................................................................23
4.2 EFFECT OF ADDITIVES ...............................................................................................24
4.2.1 Powder tests.....................................................................................................24
4.2.2 Tests of silicone rubbers ..................................................................................25
4.3 DETERMINATION OF BIOFOULING .............................................................................28
4.3.1 Image analysis .................................................................................................29
4.3.2 Laser-induced fluorescence .............................................................................30
4.3.3 Hydrophobicity ................................................................................................33
4.3.4 ESEM/EDX ......................................................................................................34
4.4 BIODEGRADATION OF THE PDMS MOLECULE...........................................................37
5 CONCLUSIONS ............................................................................................................41
6 FUTURE WORK...........................................................................................................43
7 ACKNOWLEDGEMENTS ..........................................................................................45
8 REFERENCES ..............................................................................................................47
Purpose of the study


Silicone rubber high voltage outdoor insulators are sometimes colonised by microorganisms
which form a biofilm on the surface of the infected units. This problem was first noted in
tropical climates[1-8], but it has now also been observed in temperate environments [9-11].
Since silicone rubbers are considered to be bioresistant materials, the biofouling problem was
not expected and little reported research could be found in relevant areas [12-14]. This project
was initiated to develop knowledge of the effects of a biofilm on silicone rubbers used as high
voltage insulating materials.

Areas of interest for this study included the identification of the microorganisms causing the
observed fouling problem, the examination of their effect on the chemical and mechanical
properties of infected materials, and the evaluation of different silicone rubber formulations
with respect to their ability to hinder biological growth. In addition, potential test methods for
the identification of infected silicone rubber insulators in use were explored.

Purpose of the study


An electrical insulator in a high voltage system is designed to support a charged conductor
and electrically isolate it. Insulators made from glass or porcelain have been used for this
purpose in power transmission lines for over 120 years. Over the last 40 years new designs
have been developed and used for the construction of high voltage insulators. A modern
composite insulator, as shown in Fig 2.1, consists of a mechanically supporting structure
covered by a housing. The main functions of the housing are to protect the core from
environmental stresses and to minimise leakage currents between the energised end and
ground [14, 15]. The core often consists of a glass-fibre reinforced resin-bonded rod onto
which two metal end-fittings are attached. The housing is made from polymeric materials
such as EPDM or silicone rubber. Today it is widely agreed that regarding service
performance and ageing, silicone rubber is the best housing material available [16].

a) b)

Figure 2.1 a) Schematic drawing of a composite insulator. b) Picture of two silicone rubber



Silicone elastomers made from crosslinked polydimethylsiloxane (PDMS) (Fig. 2.2) mixed
with suitable fillers and additives are used in many applications such as medical devices,
textile coatings, foams, seals and electrical insulation. The silicone elastomers used as high
voltage insulation materials generally contain at least two kinds of fillers, reinforcing fillers to
improve the mechanical properties and a flame retardant filler to dissipate heat from energetic
events in the high voltage system. Silica is the most common reinforcing filler and the most
commonly used flame retardant is alumina trihydrate (ATH) [17]. Furthermore, additives such
as antioxidants, colorants, crosslinking agents, processing aids and stabilisers are often used.
The types and amounts of fillers are of major importance for the properties of the rubber.


Si O
Fig 2.2 The repeating unit of PDMS

PDMS molecules display low intermolecular forces, high intramolecular siloxane bond
energy, high backbone flexibility, and a partially polar backbone. These characteristics give
PDMS-rubber a number of properties that make the material suitable for the production of
high-voltage insulators [18]:

Good dielectric properties

High surface mobility
Excellent resistance to weathering
Low surface free energy
Hydrophobic surface properties
Non-adhering surface
Insolubility in water
Low glass transition temperature
High thermal and oxidative stability
Low reactivity, toxicity and combustibility

The low surface free energy of a silicone rubber means that the material has excellent surface
hydrophobicity. In addition, silicone rubber has the potential to recover this feature after
exposure to ageing factors in the environment. Several plausible mechanisms for the
hydrophobic recovery are suggested in the literature However, most researchers believe that
the migration of low molar mass PDMS is the dominant mechanism for the hydrophobic
recovery [19].


Other advantages of silicone rubber insulators over conventional porcelain insulators are that
they are less attractive targets to vandals, and their lightweight and minor vulnerability make
them easy to store, transport and install [20]. The possibility of producing insulators with a
smaller surface area and a long leakage path is also advantageous, and this improves the
function of the insulator under wet and contaminated conditions [15, 21]


Even though silicone rubbers (SIR) have a number of characteristics that make the materials
suitable for high voltage applications, rubbers are more sensitive than inorganic materials to
degradation under exposure to discharges and arcing [22, 23]. Surface-eroding climatic
factors such as wind, rain, temperature, and UV radiation, also contribute to the degradation
of the rubber, despite the fact that silicones are known to exhibit excellent resistant to
weathering. [19, 22, 24, 25].

Electric fields at the edges of high voltage insulators or around water droplets on a
hydrophobic silicone rubber surface can give rise to corona discharges [26]. These discharges
produce a temporary loss of hydrophobicity [27, 28]. When hydrophobicity is lost, dry-band
arcing capable of decomposing the polymeric material may occur [29]. Dry- band arcing
causes tracking and erosion and the heat generated can seriously damage the insulating
material [30, 31]. Low molecular weight (LMW) silicones, which contribute to the
hydrophobic recovery, have relatively low boiling temperatures (<400C), which means that
LMW silicones evaporate from the rubber at the elevated temperatures reached [32].
However, the generated heat also form new LMW-chains by inducing chain scission reactions
in the material [25, 33].

In addition to electrical ageing factors, silicone rubbers used in high voltage insulators are
exposed to severe environmental stresses such as UV-radiation. Even though PDMS-rubber is
known to be very stable towards UV-radiation from sunlight, prolonged exposure can lead to
crosslinking reactions in the surface of the material [34, 35]. It is also known that exposure of
fouled SIR surfaces to UV increases the rate of hydrophobic recovery; indicating that chain
scission reactions producing LMW-silicones are induced [36].

All PDMS-materials are highly insoluble in water, but this does not mean that organosilicones
do not interact with humid surroundings. For example, water on the surface of silicone rubber
gives a temporary loss of hydrophobicity by reorientation of methyl groups and by
solubilisation of LMW chains [37]. Acid rain may also affect silicone rubbers. Artificial acid
rain with pH 1.5-2.5 has been shown to cause chemical changes in the rubber matrix [38].
Even though natural rainfall never reaches such a low pH, acid rain may still harm an
insulator. Fillers such as ATH are more sensitive to acidic pH and can be eroded by acid rain
with a pH of about 5 [39]. However, diffusion of LMW-silicones helps silicone rubber
materials to recover their original surface properties even after severe exposure to acid rain


Degradation of PDMS through clay-catalysed hydrolysis is a well studied phenomenon [41,

42]. Clay minerals in soil are able to catalyse the hydrolysis of PDMS to dimethylsilanediol, a
substance that is accessible to microorganisms and can be biodegraded to CO2, SiO2 and H2O
[43-45]. However, organosilicones of higher molecular weight do not provide an easily
accessible carbon source for microorganisms. Silicones are artificial materials, and no
biological process is known that produces or degrades the covalent bonds of a PDMS-
molecule [46]. Therefore, PDMS-materials always respond as bioresistant in biodegradability
tests [42]. Nevertheless microorganisms can affect silicone rubber by several different
mechanisms, as will be discussed further.


2.4.1 Accelerated ageing

The diverse interactions of a silicon rubber insulator with its surrounding environment mean
that it is complicated to create relevant procedures to simulate the ageing of insulating
materials. Even the most ambitious test programs often fail to represent true service
conditions [10, 47]. In spite of this, several methods have been designed to test the
performance of an insulator or a silicone rubber under the influence of accelerated ageing.

Insulators or test rods can be placed in a fog chamber, were they are energized and exposed to
artificial fog, often contaminated with salts or other eroding substances [48, 49]. Tracking
wheel tests where samples are dipped in a water-bath and subsequently exposed to a high
voltage are also common. [50]. Weatherometer ageing is another method often used to
simulate the combined effects of several climatic ageing factors such as UV-radiation,
elevated temperature, temperature cycling, thermal shock, high humidity and gaseous
contamination. The electrical performance of the insulators or materials is recorded during the
tests in order to determine the resistance of the samples to the applied stresses [51].

To reduce the risk of artefacts, insulators can be aged under field conditions. This is time-
consuming, but the results obtained are adequate and useful [52, 53]. An unforeseen ageing
factor discovered during field ageing was the biofouling of silicone rubber. Insulators aged
outdoors in a tropical climate were found to be covered with a greenish-black film of
biological origin [2]. None of the techniques for accelerated ageing address the problem of
biological growth on the surface of an insulating material.

2.4.2 Analytical tools

Numerous analytical techniques are suitable for studying the changes in the chemical and
morphological characteristics of silicone rubber. One often used tool is scanning electron
microscopy (SEM), with which it is possible to monitor the surface microstructure of a rubber
and thus study morphological changes. After corona exposure, small cracks can be observed
at the sample surface. Thermal stress also causes cracks, and UV-radiation makes ATH filler
particles appear on the surface of the material [25, 54]. If chemical changes in the silicone


material are to be observed, techniques such as Fourier transform infrared spectroscopy

(FTIR) or Electron spectroscopy for chemical analysis (ESCA) are needed. FTIR is
commonly used to verify scission or recombination of chemical bonds in the surface layer of
silicone rubber, such as the silanol groups formed by oxidation reactions induced by UV-
exposure, corona or dry-band arcing [25]. ESCA can be used to study the proportions of
different elements at the surface of a silicone rubber insulator. The of C/Si and Si/Al ratios
indicate the magnitude of ageing in PDMS and the amount of ATH filler respectively [25,

Another feature that is often affected by ageing is the surface hydrophobicity of the silicone
rubber [25, 54]. The most common method used to study the loss and recovery of
hydrophobicity is water contact angle measurement. By measuring the angle between the
silicone rubber surface and a drop of water, the hydrophobicity of the material can be assessed
[22]. Low molecular weight silicones, important for the hydrophobic recovery of a silicone
rubber, can be detected by gas chromatography (GC) [56, 57]. If PDMS-oils of higher
molecular weights are to be studied, size exclusion chromatography (SEC) is a reliable and
often used method [33, 56, 58].

2.4.3 On-line measurements

Today there is no standardised or widely used technique for the inspection of composite
insulators in service [21]. Nevertheless a majority of utilities examine the condition of their
insulators on the line [59]. The most common inspection technique used is visual
examination. An experienced inspector can detect surface damage as well as evidence of
internal faults just by looking at an insulator. Night vision equipment also enables discharge
activity on the surface of the insulator to be distinguished [21, 59, 60].

Manual inspection is however operator-dependent and more objective techniques would be

preferred. Infra-red thermography exploits the fact that the degradation caused by electrical
events in the high voltage system is associated with heat. The infra-red radiation produced can
be used to locate defects in composite insulators from the ground. Electrical field
measurement is another promising method for the detection of faulty insulators. At the
location of a defect, the electrical field is changed and this change can be recorded. However
it is necessary to climb the tower to perform the measurement [59-61].

One disadvantage with the described techniques is that they only detect large defects like
puncture and cracks, not fouling or other changes in surface properties of the insulator. New
diagnostic methods are needed to detect smaller defects. Digital image analysis is one
potentially useful technique. Information extracted from digital images of aged insulators
enables features such as loss of hydrophobicity or extent of biofouling to be assessed. [62-65].
Laser-induced fluorescence offers another solution to the problem. By measuring the
spontaneous emission of radiation by which electrons in atoms or molecules relaxes from an
upper energy level to ground state level, changes in material properties of an insulator can be
observed and biological growth detected [66-69].



Microorganisms colonising a bioresistant substrate tend to form a film on the surface of the
material. Such a biofilm consists of microorganisms embedded in a highly hydrated matrix of
extracellular polymeric substances (EPS), mainly polysaccharides and proteins. Mixed
populations of bacteria, fungi, protozoa and algae often coexist in the film [70]. However, in a
mature biofilm, the fraction of living cells is often small. Water is the most abundant element
in most biofilms and the organic portion of the film is often dominated by EPS. In addition,
particulate matter such as clay, humic substances, corrosion products etc. can be included in
the mature biofilm. When the cause of a biofouling problem is being sought, it is easy to
overlook the small content of microorganisms in the biofilm; thus neglecting the cause of the
fouling [71].

Biofilm formation is an interfacial process, and surface features of the microorganisms and
the substrate material are important for biofilm adhesion and growth. Interaction forces
between adhering cells and substrate, such as van der Waals forces, electrostatic forces and
acid base interactions, are of great importance for the initial microbial adhesion [73]. These
forces, together with several other factors, some of them summarised in Tab. 2.1, also
influence the growth rate and strength of adhesion of a biofilm [72].

Table 2.1
Factors affecting the adhesion and growth of a biofilm
Genotypic factors Expression of genes encoding surface properties,
formation of EPS, growth rate etc.
Physio-chemical factors Substrate composition and roughness, temperature, pH,
water potential, oxygen supply, etc.
Stochastic processes Initial colonisation and random changes in biotic and
abiotic factors.
Deterministic phenomena Specific interactions between organisms, competition,
co-operation, predation, etc.
Mechanical Processes Shear from water or airflow over the substrate

The structure of the biofilm formed varies, depending on the interactions of the
microorganisms with the surrounding environment. However, most biofilms have a sponge-
like structure containing a system of pores and channels in which a liquid phase is free to
move and transport oxygen and nutrients between different parts of the biofilm [72, 74]

From an ecological point of view, life in a biofilm offers many advantages to a cell; such as
protection against desiccation, accumulation of nutrients from the bulk water phase,
protection against toxic substances, facilitated exchange of genetic material and opportunities
for co-metabolism [70, 72] The incitement for biofilm formation is therefore strong and
biofilms are abundant in nature.



A biofilm can affect the properties of a polymeric material by several different mechanisms.
Undesired effects range from discoloration to complete degradation of the polymeric material.
Some of the major damaging mechanisms are summarised in figure 2.3 [70].

Proces Fouling Degradation of Biotic Hydration Colour

leaching degradation Penetration


Effect Change in Loss of Loss of Conductivity Change in

surface stability stability Swelling appearance
Figure 2.3 Undesired effects caused by a biofilm

2.6.1 Fouling
Many polymeric materials are considered to be inert to microbial attack. However, these
materials may still function as a support for a biofilm. The unwanted deposition of biological
matter is referred to as fouling [75]. If the colonised material resists degradation,
microorganisms can use pollutants from the surroundings to gain nutrition [76-78]. This was
the case at a Florida-based transmission station where silicone bushings were trapping
airborne pine pollen that served as a feedstock for a severe mould growth [8]. This ability of a
biofilm to use an external carbon source makes laboratory testing difficult. Materials that
seem to be inert in degradation studies may nevertheless encourage growth when exposed to
outdoor conditions [79].

A fouling problem does not change the bulk properties of the infected material; it can
however disturb the surface properties of the colonised object. When a silicone rubber
insulator is covered by a hydrophilic biofilm, the hydrophobic surface properties are reduced.
This has been shown to cause an increase in leakage current under wet conditions, compared
to clean insulators [8].

2.6.2 Degradation of leaching components

Plastics and rubbers often contain additives and impurities that may leak from the matrix to
the surroundings and interact with a biofilm. Impurities such as unreacted low molecular
weight material and residual processing aids can provide nutrients for an attached biofilm
[77]. Fillers such as adiapates, epoxidised fatty acids, oleates, stearates and polyesters are also
known to serve as nutrients for biological growth [80]. The same is true of carbon-based
plasticizers and many other additives [70, 77, 80]. There are indications that some silicone


rubber materials used in high voltage insulators contain additives susceptible to

microbiological attack [7, 10].

In contrast, accelerators such as dithiocarbamates and sulphur-based components may inhibit

growth, as may residual heavy metal catalysts and many other additives [77, 80]. Field studies
have shown that not all insulators are affected by biological growth. This may be due to the
protective effect of additives used in the production of the silicone rubber.

2.6.3 Biotic degradation

If a polymer is to be used as a source of nutrition for biological growth, it has to be degraded
to segments small enough to be transported across the cell membrane [81]. This degradation
can be mediated by degrading factors in the surrounding environment, such as wind, rain,
oxygen, UV-radiation etc [82]. However, degradation can also be directly initiated by the
living cell through the production of degrading substances such as enzymes, radicals or
organic acids [82-86].

Surface degradation is an interfacial process depending on parameters such as pH, redox

potential and the concentration of oxygen and salts. Biofilms can strongly influence these
parameters [70]. Microorganisms sometimes excrete organic acids as by-products of
metabolism. The excretion of organic compounds is usually the result of an unbalanced
growth, a surplus or a limited supply of some compound essential for the metabolism of the
microorganism. Anions, that are final products of microbial metabolism, react with cationic
components and form salts. An increase in the salt concentration or a drop in the pH may
facilitate the breakdown of the polymer [85].

Microorganisms can also incite the breakdown of an insoluble polymeric material by the
production of exoenzymes, capable of mediating degradation outside the cell. It has been
suggested that siloxane oils degrade under the influence of enzymes produced by bacteria
[87], and there are indications that silicone rubber voice prostheses degrade under the
influence of a mixed biofilm [88, 89]. The three most probable processes by which enzymes
influence polymer degradation are [84]:

1. Hydrolysis by lysosomal enzymes of susceptible polymers

2. The oxidation of polymers by oxidase enzymes
3. The breakdown of natural biopolymer structures by enzymes such as collagenase

Many molds develop powerful enzyme systems with the ability to degrade highly stable
polymeric structures. Among these, the white rot fungi hold a unique position. This class of
molds has developed non-specific mechanisms for degrading the complex polymeric structure
of lignin. The enzyme system responsible for the degradation includes peroxidases that
promote the reduction of peroxides to free radicals [90-93]. It has been suggested that the
white rot fungus Phanerochaete chrysosporium is able to degrade PDMS as well as lignin


2.6.4 Hydration and penetration

Fungal hyphae can penetrate their support and thereby cause cracks and pores in the material.
The mechanism of the penetration is unclear, but it is known that hyphae are able to create
great turgor pressures that force the cells through the material. This leads to a decrease in
mechanical stability and offers a way for water to enter the polymeric material [70, 95]. The
latter may increase the electrical conductivity of an insulating material.

2.6.5 Color and odor

Many microorganisms produce pigments. These pigments are often lipophilic and therefore
tend to diffuse into the matrix of a polymeric material. The stains produced are impossible to
remove by simple cleaning [70, 78]. In addition, metabolites from biodegradation are
sometimes sources of odour. A small concentration of a substance can be enough to cause
problems [80]. However, this problem is of an aesthetic nature and does not affect the
insulating properties of the silicone rubber.


2.7.1 Biocides
Active substances that eliminate microorganisms or inhibit their reproduction are denoted
biocides. Numerous additives of this kind exist on the market, and it is a demanding task to
choose a biocide able to protect a high voltage insulator from biological growth. It is
important that the substance is compatible with the rubber and does not affect its properties. It
is also necessary that the biocide effectively kills unwanted microorganisms, while leaving
people and the environment unharmed. In addition, to be effective in an outdoor environment,
inhabited by an enormous variety of microorganisms, a biocide has to have a broad effectivity
spectrum and offer small probability for microorganisms to develop resistance to the active
substance [78, 80].

High voltage insulators are meant to function in an outdoor environment for many years with
no need for service or replacement. This put demands on biocides that often have to be
present on the surface of the silicon rubber in a certain minimum concentration to be
effective. The active substance consequently has to be effective in low concentrations and
have a low washout if it is required to be active over a long period of time [78]. It is important
to take the washout effect into account when testing the effectiveness of a biocide. It has been
shown that a biocide with a high initial activity can become inactivated after a period of
outdoor storage. This may be due, for example, to washout or inadequate light stability [80].

It is quite common to use a biocide not as an additive to prevent growth but as a surface
treatment to kill a mature biofilm. This is not to be recommended since the biocide kills the
microorganisms, but it does not remove the dead biomass from the system. The accumulation
of biodegradable organic material often give rise to a rapid microbial regrowth [71, 96].


2.7.2 Cleaning
Cleaning an infected material removes dead biomass from the system and thus makes the
surface less attractive for microorganisms. A rational cleaning often includes two steps, one
where the physical stability of the biofilm is weakened and a second in which the biofilm is
removed from the system. Oxidisers or detergents are useful for the first step, while flushing,
brushing or similar physical techniques are useful for the second step. [71]. Tests performed
on silicon rubber insulators shows that water repellent properties and other properties
disturbed by the biofilm were partially restored after cleaning with alcohol and high pressure
water [8].

2.7.3 Surface modification

The hydrophobic nature of a silicone rubber contributes to its resistance to utilisation by
microorganisms. Water and nutrients are essential for all kinds of biological growth and both
are limited on a hydrophobic surface where water forms droplets that run off the material
washing nutrients away [77]. In addition, the hydrophobicity substantially decreases the rate
of hydrolysis and microbial attack of the polymer [82].

The low surface free energy of silicone rubber makes it useful in the production of non-stick
coatings [97]. Silicone rubber is used in applications were biofouling cannot be avoided, such
as in the coating of fishing nets. The weak adhesion between the adhering biofilm and the
silicone-coated net facilitates cleaning [98]. Changing the microstructure of the rubber surface
can further strengthen the non-stick characteristics of the silicone rubber. Micro-scale riblets
or pyramids printed on to the surface of a silicone rubber sample may protect the material
from biofouling [99, 100].



3.1.1 Silicones
PDMS rubber base with 20% SiO2 filler and 0,52 % di(4-methyl benzoyl)peroxide added as
crosslinker was mixed with Martinal OL-104 S vinylsilane surface treated aluminium
trihydrate (ATH) and Firebrake 290 zinc borate delivered by Wacker Silicone as shown in
table 3.1.

Table 3.1
Type and amount of flame retardant added to the silicone rubber materials
No: Material Flame retardant (pph*) Flame retardant %
1 Base No flame retardant added No flame retardant added
2 Base + ATH 100 pph ATH 50% ATH
3 Base + ATH + zinc borate 90 pph ATH + 10 pph zinc 45% ATH + 5% zinc
borate borate
4 Base + zinc borate 10 pph zinc borate 9% zinc borate
5 Commercial mix Unknown Unknown
Parts per hundred resin

Materials were kneaded at 12 rpm in a Brabender internal mixer for 15 min prior to curing.
The curing was performed in a Schwabentan Polystat press for 10 min at a temperature of
180C and a pressure of 10 MPa. The resulting discs had a diameter of 200 mm and a
thickness of 2 mm. Samples were post-cured for 4 h in a hot air oven at 200C before use. The
commercial material was pressed and cured in accordance with instructions from the supplier.

To study the degradation of PDMS mediated by enzymes or radicals more defined materials
were needed. Silicone oil with a viscosity of 100 cSt and decamethyltetrasiloxane from Sigma
Aldrich were used together with a platina catalysed PDMS rubber. The rubber was prepared
by mixing vinyl terminated PDMS oil with crosslinker (6 % of methylhydro-
dimethylsiloxane) and catalyst (35 ppm of a platinum divinyltetramethyldisiloxane complex).


The curing was performed in a Pasadena Hydraulics Inc. 0230 press for 15 min at 135C
under a pressure of 2 MPa [101].

3.1.2 Insulators
Insulators infected by biological growth were collected from the Bagamoyo test station,
Tanzania (tropical climate), the University of Peradeiya , Sri Lanka (tropical climate) and the
Anneberg test station, Sweden (temperate climate). The insulators had been in service for
several years and no information about the composition of the materials used for production
of the housings was available.

The worst infected parts of the insulators were analysed with ESEM/EDX before and after
gentle cleaning with a soft cloth dampened with distilled water. Small pieces of rubber cut
from the centre of the insulators were used as reference.

3.1.3 Microorganisms
Microorganisms were isolated from the studied insulators by inoculating different growth
media with finely grained biofilm suspended in water. Algae were grown on BG 11, a
medium designed for the isolation of green and blue-green algae, and Jaworskis medium
with added NaSiO3 for the isolation of green algae and diatoms [102]. Fungal growth was
observed on potato dextrose agar and malt extract agar; i.e. two commonly used undefined
media designed for the growth and isolation of fungi [103]. Isolation was than made from
malt extract agar diluted to one tenth of the recommended concentration. This medium is poor
in nutrients and thus resembles the natural habitat of the isolated fungi better than the nutrient-
rich medium used for observation.

Hyphal tips, colonies or spores were selected for isolation. 50 fungal cultures and 20 algae
cultures from each biofilm were isolated. Each fungal culture was subcultured three times on
malt extract agar and algae were subcultured three times on BG 11 (as agar and in liquid
dilution series). All isolates were visually examined and studied under a light microscope to
roughly determine their class. The three most common fungal species isolated from each
biofilm were sent to Centraalbureau voor Schimmelcultures, the Netherlands, for
identification, while algae cultures were submitted to the Department of Agricultural
Microbiology, Agricultural University of Wroclaw, Poland.

Standard test organisms

The molds prescribed in standard tests and the molds used in the degradation studies
performed are listed in Tab. 3.2. All microorganisms were grown and kept in accordance with
instructions given by the supplier.


Table 3.2
Fungal strains used in standard tests and degradation studies
Fungal specie Supplier
Aspergillus niger DSMZ Strain DSM 1957
Aspergillus terreus DSMZ Strain DSM 1958
Aureobasidium pullulans DSMZ Strain DSM 2404
Chaetomium globosum DSMZ Strain DSM 1962
Paecilomyces variotii DSMZ Strain DSM 1961
Penicillium funiculosum DSMZ Strain DSM 1944
Scopulariopsis brevicaulis DSMZ Strain DSM 9122
Trichoderma virens DSMZ Strain DSM 1963
Phanerochaete chrysosporium DSMZ Strain DSM 1556
Phanerochaete chrysosporium CBS Strain CBS 481.73
Phanerochaete chrysosporium ATCC Strain 34541


The resistance of silicone rubber to biological growth was tested using standard test
procedures in combination with new methods developed for this study. All the samples used
were washed with 70% ethanol and dried in a sterile air-flow overnight before inoculation
with microorganisms. Inoculation was mediated by spraying the surface of the samples with
suspensions of microorganisms, at the same time controlles were inoculated with sterile
medium. Unless otherwise stated, tests were performed in triplicate on circular samples with a
diameter of 30 mm and a thickness of 2 mm. In most cases, the extent of biological growth
was judged according to the scale in Tab. 3.3.

Table 3.3
Scale for determining the extent of growth on the specimens.
0 No growth apparent under a nominal magnification of 60
Growth not, or hardly visible to the naked eye, but clearly visible under the
Growth plainly visible to the naked eye, but covering less than 25% of the test
Growth plainly visible to the naked eye and covering more than 25% of the test
Growth plainly visible to the naked eye and covering more than 50% of the test

A stereomicroscope with 20x ocular and 3x objective was used to evaluate the samples. In
cases where the contrast between biological growth and sample surface was low, SEM was
used for the evaluation.


3.2.1 Powder test, mold

Zinc borate and ATH were mixed into malt extract agar in concentrations from 20 pph to 100
pph. The resulting agar was sprayed with suspensions of fungal spores in distilled water.
Three different suspensions were prepared, one for each country studied, by pouring 15 ml of
distilled water onto the surface of the fungal isolates. The wetted surface of each culture was
scraped with a sterile platinum wire and the suspended spores were filtered through a thin
layer of glass wool to an Erlenmeyer flask containing glass beads. Suspensions were shaken
to break clusters of spores and mix the suspension. The resulting liquid was used for
inoculation of samples. Samples were incubated in room temperature for 28 days prior to

3.2.2 Powder test, algae

Zinc borate and ATH were mixed into BG11 agar [102] in concentrations from 20 pph to 100
pph. The resulting agar was sprayed with suspensions of algae cells in BG11 liquid growth
medium. Suspensions were prepared by pouring 15 ml of BG 11 onto the surface of the algae
isolates. The wetted surfaces were scraped with a sterile platinum wire and the resulting liquid
was used for inoculation of the samples. Samples were incubated at room temperature in a
window for 28 days prior to evaluation.

3.2.3 IEC 68-2-10, mold

The silicone rubber materials described in Tab. 3.1 were tested in accordance with the basic
environmental testing procedure IEC 68-2-10, part 2, test J, mold growth. However, in some
of the experiments, the microorganisms stipulated in the standard test were replaced by the
fungal isolates. Two variations of the test were carried out; the first one specified inoculation
of the specimen with a suspension of mold spores in distilled water, while the second
variation specified inoculation with mold spores suspended in a nutrient solution. Samples
were incubated for 84-days in 98% humidity at 28C prior to evaluation.

3.2.4 ASTM G21-90, mold

The silicone rubber materials described in Tab. 3.1 were tested in accordance with the ASTM
G21-90 standard practice for determining the resistance of synthetic polymeric materials to
fungi. However, in some of the experiments, the microorganisms stipulated in the standard
test were replaced by the fungal isolates. Samples of virgin materials and materials aged with
oxygen plasma were incubated for 84-days in 98% humidity at 28C prior to evaluation.

3.2.5 ASTM D 5589-97, algae

The silicone rubber materials described in Tab. 3.1 were tested in accordance with the ASTM
D 5589-97 standard test method for determining the resistance of paint film and related
coatings to algal defacement. The algal isolates were used as test organisms. Samples were
incubated in room temperature under a fluorescent lamp with a cycle of 14 h light and 10 h
darkness for 21-days at a relative humidity of 85% prior to evaluation.


3.2.6 Quick test, mold

10 ml of water was added to each of the following mold cultures: Aspergillus niger,
Trichoderma virens, Chaetomium globosum, Penicillium funiculosum and Aureobasidium
pullulans. The surfaces of the wetted cultures were scraped gently with a flame-sterilised
platinum wire. The suspended spores were filtered through a thin layer of glass wool to an
Erlenmeyer flask prepared with 20 solid glass beads. The mixture was then diluted with
distilled water to a volume of 100 ml and the flask was shaken to mix the spores. Samples of
the silicone rubber materials described in Tab. 3.1 were put on malt extract agar. Virgin
materials as well as materials aged with oxygen plasma were tested. The mounted samples
were sprayed with the mixed spore suspension and the containers were sealed to maintain a
moist atmosphere. The sealed Petri discs were stored in room temperature for 28 days prior to

3.2.7 Microenvironment chambers

Discs of the materials described in Tab. 3.1 were cut into plates shaped as a quarter of a circle
with a radius of 10 cm. Three plates of each material were hung in inclined positions using
nylon thread. The lowest plate of each material was divided into two sections. One was
mechanically aged with sandpaper; the other was aged with oxygen plasma just before
inoculation with microorganisms.

The assemblies of the five studied materials were placed in four closed microenvironment
chambers as shown in Fig 3.1. Three chambers represented the studied countries respectively.
The fourth was used as a reference chamber with negative control specimens. All chambers
were maintained at a temperature of approximately 25C and a relative humidity of 85%. A
daylight lamp with a cycle of 12 h light and 12 h darkness was used to provide the UV
radiation necessary for algae growth.

1. Base + zinc borate
2. Base + ATH + zinc borate
3. Base
2 4 7
4. Commercial mix
5. Base + ATH
1 6. Water, about 5 cm deep
3 5
7. Thermometer/Hygrometer
8. Tyburski, Euro-Light,
LT15W/010 daylight lamp

Figure 3.1 Schematic drawing of a microenvironment chamber.

30 day old mixed cultures, containing algae bacteria and fungi from the different biofilms,
were harvested from BG11 agar by pouring 10 ml of BG11 liquid broth onto the agar surface
and gently scraping the microorganisms with a sterile platinum wire. Plates were inoculated


with a solution of microorganisms from the specific country by spraying. All specimens were
reinoculated with microorganisms suspended in BG 11 once a week during the first five
months of the experiment, to create a self-sustaining biofilm; thereafter samples were
reinoculated once every month to support the growing biofilm. Reference samples were
sprayed with sterile BG11 liquid broth.

Samples were photographed and evaluated by image analysis and laser-induced fluorescence
after 6 months of incubation. After one year, the effects of the biofilms on the materials tested
were studied using ESEM/EDS and IR. At the same time, the hydrophobicity of the second
sample in each assembly of materials were evaluated in accordance with instructions given in
STRI Guide 8, were the drop pattern formed after wetting of a surface is studied in order to
assess surface hydrophobicity [104, 105]. Hydrophobicity was recorded before and after
gentle cleaning of the tested surface with a cloth damped with distilled water.


In order to study the effect of mold growth on the laser-induced fluorescence of different
silicone rubber materials a series of samples, listed in Tab. 3.4, was prepared by spraying the
materials described in Tab. 3.1 with suspensions of mold spores. Fungal spore suspensions
were prepared by pouring 20 ml of IEC 68-2-10 fungal nutrient medium on the agar surface
of each of the fungal isolates. The surfaces of the wetted cultures were scraped gently with a
flame-sterilised platinum wire. The resulting suspensions were filtered through thin layers of
glass wool to Erlenmeyer flasks prepared with 30 ml of IEC 68-2-10 nutrient medium and 10
solid glass beads. The flasks were shaken and the suspensions were sprayed onto the silicone
rubber samples. All samples were incubated for 45 days at 27C and 95% relative humidity
before measurements were conducted.

Table 3.4
Samples grown for laser-induced fluorescence analysis
Materials tested
Fungal strain used 1 2 3 4
Epicoccum nigrum x
Microsphaeriopsis x
Cladosporium cladosporioides x x x x
Fusarium semitectum x
Polyscytalum fecundissimum x
Stagonospora x
Curvularia lunata x
Cladosporium tenuissimum x
Clean materials x x x x

A laser generating radiation in pulses (3 ns, ~15 Hz, ~1 J) at a wavelength of 337 nm was
used to study samples from the microenvironment chamber as well as the samples listed in


Tab. 3.4. Each spectrum was accumulated from 100 laser pulses and 20 spectra were collected
from each sample. The laser light was guided via a telescope through a fused silica optical
fibre, 600 m in diameter, to the sample under investigation. The tip of the fibre was placed
about 1 mm above the sample. The laser-induced fluorescence was spatially emitted in all
directions and a fraction of it was collected by the same fibre and guided back to an optical
multi-channel analyser system. The resolution of the system, set by the 100 m slit width, was
2.2 nm, and the spectrum could be recorded up to 805 nm. A detailed description of the
system can be found in Ref. [106].

Data obtained from the analysis were pre-processed by mean normalisation and mean centring
in order to produce a fluorescence spectrum representing the sample. In some cases, this
spectrum was simplified using principal component analysis (PCA) that projects the large
data set of the spectra analysed into a smaller set of data that is easier to overview. The first
principal component (PC1) is the combination of variables that explains the greatest amount
of variation between spectra. The second PC (PC2) describes the second greatest amount of
variation orthogonal to the first PC and so on. Higher order PCs correspond to small variances
and can be seen as noise [107].


Samples were laid flat on a blue background and photographed using a standard Canon IXUS
V3 digital colour camera. The image analysis was than performed in six steps using Matlab.

1. In the first step, the background was separated from the sample. This was done by
determining differences in colour between sample and background. First, each image
was represented as a set of three matrices, containing intensity values of the red,
green and blue components, respectively. The blue component was then divided by
the sum of the three colour components. The quotient of this operation differs
between sample and background; hence the sample can be separated from the
background through intensity thresholding [108]. The result of this operation is a
black and white image of the sample alone.
2. In the second step, the possible influence of shading at the sample edge was removed
by erosion of the image by a circular element, removing object pixels at its
3. The third step produced a colour image of the sample from the black and white
image obtained in the two first steps by multiplication of this image by the original
colour image.
4. The fourth step compensated for illumination variations by dividing the sample into a
set of sub-images in which the maximum values are identified and used to calibrate
the remaining operations.
5. The fifth step separated biological growth from silicone rubber. Again, the blue
component of the image was used for segmentation of the sample into covered and
non-covered areas by means of intensity thresholding [109]. Blue was selected since


the contrast between the rubber and the biofilm was highest in this component. The
resulting black and white image separates regions of growth from clean regions
6. Finally, the covered area was calculated by counting the numbers of pixels classified
as growth. To avoid the need for calibration, the covered area was always related to
the area classified as sample surface.


In order to decide whether the chemical bonds of the PDMS molecule can be affected by
biologically mediated processes, an unfilled PDMS rubber, PDMS oil and a DMS oligomer
were treated with lignin-degrading enzymes or radicals produced through the Fenton
mechanism. In each case 0,020 g of the sample was weighed into a 100 ml Erlenmeyer flask
where the reaction was performed. Samples and reference samples were set in triplicates.

After treatment, PDMS rubber was removed from the reaction vessel and studied with SEM
and IR. Oils and oligomers were extracted to chloroform according to the following

1. The content of the Erlenmeyer flask was filtered through a finely meshed strainer
into a separation funnel, and solid matter caught by the mesh was returned to the
Erlenmeyer flask.
2. 10 ml of chloroform was then added to the separation funnel and the funnel was
manually agitated for 5 min.
3. After 5 min equilibration the chloroform phase was emptied into the Erlenmeyer
4. Steps two and three were then repeated with fresh chloroform.
5. Finally, the Erlenmeyer flask was treated in an ultrasonic bath for 10 min.

After extraction PDMS oil was analysed using SEC and GC/FID, while PDMS oligomers
were analysed with GC/FID only.

3.5.1 Enzymatic degradation

According to United States Patent No 6,020,148, the lignin-degrading fungus Phanerochaete
chrysosporium is capable of degrading the siloxane bond of PDMS [94]. The growth medium
prescribed in the patent is known to promote the formation of peroxidases [110]. The test
described was repeated twice, using fungal strains from DSMZ and CBS respectively, in order
to study the degradation process. In addition, the trial was repeated a third time using an
optimised growth medium and an ATCC strain of Phanerochaete chrysosporium [111, 112].

Optimised growth medium was prepared by dissolving 0.1 g (NH4)2SO4, 20 g glucose, 1,0 g
KH2PO4, 0,2 g NaHPO4, 0,5 g MgSO4*7H2O, 0,017 g MnSO4*H2O, 10 g ZnSO4*7H2O, 20
g CuSO4*5H2O, 100g CaCl2, 100 g FeSO4*7H2O and 5 g thiamine in 50 mM Malonate


buffer spiked with 0,1% Tween 80. The pH was adjusted to 4,5 before the addition of fungal

Samples were incubated as stationary cultures in 20 ml liquid growth medium at 30C for a
period of 60 days before extraction and analysis.

3.5.2 Degradation by Fenton mechanism

When hydrogen peroxide and Fe2+ react, OH radicals are formed through Fenton mechanism.
These radicals may be able to degrade the chemical bonds of PDMS [113]. A simple Fenton
reaction was performed as follows: 10 ml of a 10 mmol/L solution of FeSO4*7H20 was added
to each sample. Reaction was then initiated by drop wise addition of 10 ml of a 1 mol/L
solution of H2O2. The reaction was completed at room temperature overnight before
extraction and analysis.


3.6.1 Oxygen plasma ageing

Oxygen plasma aging was performed in a V15-G microwave frequency reactor from Plasma-
Finish GmbH, operated at 2.45 GHz and 100 W for 180 s at an oxygen pressure of 28 Pa and
a gas flow of 50 ml/min.

3.6.2 Light microscopy

A Leitz Ortholux II POL-BK optical microscope was used to study the biofilms and the
isolated microorganisms.

3.6.3 SEM
A Jeol JSM-5400 Scanning Electron Microscope was used at an acceleration-voltage of 10
kV. Samples were dried in vacuum at room temperature overnight and sputtered with
palladium/gold before analysis.

3.6.4 ESEM/EDS
An XL TMP (W) Environmental Scanning Electron Microscope from FEI/Philips was used in
the low vacuum mode at an acceleration voltage of 10 kV. Microanalysis for determination of
atomic composition in the surface layer was carried out with an EDS-system from EDAX.

3.6.5 IR
Infrared spectra were recorded on a Perkin Elmer Spectrum 2000 FTIR equipped with a
golden gate normal single reflection ATR accessory. Infrared absorption was recorded from
600 to 4000 cm-1.


3.6.6 GC/FID
A Varian 3400 Gas Chromatograph equipped with a flame ionisation detector and a CP8752
column, 30 m x 0,32 mm, film thickness 0,25 m, from Varian was used. The column
temperature was raised from 40C to 250C at a rate of 10C/min. The injector was used in
split mode at 250C, while the detector was operated at 275C.

3.6.7 SEC
A 510 Size Exclusion Chromatograph from Waters equipped with three PLgel 10m mixed-B
columns, 300x7,5 mm, from Polymer Labs was used at room temperature and a flow rate of 1
ml/min. Spectra were recorded with a PL-ELS 1000 evaporative light scattering detector.

Results and discussion


Biofilms are complex structures in which a multitude of microorganisms from different
biological classes may coexist, compete or co-operate. The composition of the biofilm affects
its interaction with the support material [70]. Therefore it is important to know the
composition of the biofilm colonising the surface of interest, in this case silicone rubber

In this work, three different biofilms, collected from silicone rubber insulators exposed to the
outdoor environment in Sweden, Sri Lanka and Tanzania, have been analysed. Filamentous
fungi and micro algae in association with bacteria were isolated from the biofilms under
study. Even though the insulators were collected from three different continents, the
compositions of the biofilms studied were remarkably similar. Small unicellular green algae
of the Chlorella family, living in symbiosis with bacteria, dominated the biofilms and traces
of filamentous fungi were spread through the algae matrix. The similarity between the
different biofilms indicates that the mechanism of biofouling of silicone rubber insulators is
the same all over the world. The identified species, listed in Tab. 4.1, were therefore used to
test the resistance of several silicone rubber formulations to biological growth.

Table 4.1
Microorganisms used for inoculation of silicone rubber samples.
Country Fungi Algae
Epicoccum nigrum Chlorella saccharophila
Sweden Microsphaeriopsis1 +6 bacterial strains
Cladosporium cladosporioides
Fusarium semitectum Chlorella vulgaris var. Autotrophica
Sri Lanka Polyscytalum fecundissimum
Stagonospora2 +5 bacterial strains
Curvularia lunata Chlorella vulgaris var. Autotrophica
Cladosporium tenuissimum +2 bacterial strains
Not possible to identify on species level. 2 This anamorph has not been given a formal name.

Results and discussion


Not all insulators in the field are affected by biological growth, indicating that some
formulations are protected. In an attempt to identify the source of the observed differences in
sensitivity, it was decided to analyse some of the materials used. The investigation was soon
focused on the effect of the flame-retardants. A series of tests, designed to determine the
sensitivity of a polymeric material and its additives to microbiological growth, was developed
and used to evaluate the effect of zinc borate and alumina trihydrate (ATH) on biological
growth. Results of the tests were evaluated according to the scale given in Tab. 3.3, in which
0 denotes a clean surface and 4 a surface covered to more than 50 % by biological growth.

4.2.1 Powder tests

Tests with flame-retardants, zinc borate and ATH, as powder mixed in growth medium were
performed in order to study the immediate influence of these chemicals on microbiological
growth. As can be seen in Tab. 4.2, mold growth was not hindered on any of the samples
containing ATH. Zinc borate on the other hand seems to have a preventive effect on the
growth of mold, as can be seen in Tab. 4.3. The extent of growth on samples with added zinc
borate is dependent on the concentration of additive as well as on the types of microorganisms
tested. It seems that while molds from Tanzania is very sensitive to the toxic effects of zinc
borate, molds from Sri Lanka and Sweden are able to tolerate higher concentrations of the

Table 4.2
Extent of mold growth on 3 samples for each concentration of ATH.
Concentration Sweden Sri Lanka Tanzania
of ATH (pph) 1 2 3 1 2 3 1 2 3
20 4 4 4 4 4 4 4 4 4
40 4 4 4 4 4 4 4 4 4
60 4 4 4 4 4 4 4 4 4
80 4 4 4 4 4 4 4 4 4
100 4 4 4 4 4 4 4 4 4

Table 4.3
Extent of mold growth on 3 samples for each concentration of zinc borate.
Concentration of Sweden Sri Lanka Tanzania
zinc borate (pph) 1 2 3 1 2 3 1 2 3
20 3 3 3 3 3 3 0 0 0
40 3 3 3 2 2 2 0 0 0
60 3 3 3 1 1 1 0 0 0
80 2 2 2 0 0 0 0 0 0
100 2 2 2 0 0 0 0 0 0

Results and discussion

Further testing showed that zinc borate powder could prevent algal growth as well as mold
growth. As can be seen in Tab. 4.4, the addition of ATH to the algae culture medium did not
affect the growth of algae. On the other hand, addition of zinc borate inhibited algal growth as
shown in Tab. 4.5. The living algae cells sprayed on the surface of the agar could not
reproduce when zinc borate was added to the medium. When the concentration of zinc borate
was high, all the algae cells died and lost their green colour.

Table 4.4
Extent of algae growth on 3 samples for each concentration of ATH.
Concentration Sweden Sri Lanka Tanzania
of ATH (pph) 1 2 3 1 2 3 1 2 3
20 4 4 4 4 4 4 4 4 4
60 4 4 4 4 4 4 4 4 4
100 4 4 4 4 4 4 4 4 4

Table 4.5
Extent of algae growth on 3 samples for each concentration of zinc borate.
Concentration of Sweden Sri Lanka Tanzania
zinc borate (pph) 1 2 3 1 2 3 1 2 3
20 2 2 2 0 0 0 2 2 2
60 0 0 0 0 0 0 0 0 0
100 0 0 0 0 0 0 0 0 0

The powder tests developed for this study are fast and easy to use. A clear trend was observed
already one week after incubation, and after 28 days it could definitely be concluded that zinc
borate is toxic to the microorganisms tested, while ATH is not. However, for some of the
species, a very high concentration of zinc borate was needed to effectively hinder growth.
This should not be interpreted as meaning that zinc borate is not effective as a growth
suppressor in silicone rubber materials. Growth conditions are very poor on the surface of a
silicone rubber material compared to those in the agar used in the powder tests, where
nutrients are abundant.

4.2.2 Tests of silicone rubbers

The difference in growth conditions between powder tests and tests performed on polymeric
materials was clearly shown when silicone rubber materials containing ATH or zinc borate
were evaluated. The silicone rubber formulations described in Tab. 3.1 were subjected to a
series of tests to determine their resistance to biological growth.

Two variations of the standard procedure IEC 68-2-10, designed for the determination of the
resistance of a material to mold growth, were used in the present work; one in which nutrients
that support mold growth were added to the silicone rubbers together with the spores and one
in which the specimens were inoculated with nothing but mold spores and water. In the latter
case, the fungi must use the supporting rubber as the single source of nutrient for growth.

Results and discussion

Tests were carried out with standard test organisms and with organisms isolated from the
insulators studied in this work. Results of the tests are shown in Tab. 4.6 and Tab. 4.7.

Table 4.6
IEC 68-2-10 Extent of mold growth on silicone rubber treated with spores and nutrients.
Sweden Sri Lanka Tanzania Standard
Material 1 2 1 2 3 3 1 2 3 1 2 3
Base 3 3 3 1 1 1 2 2 2 3 3 3
Base + ATH 3 3 3 1 1 1 3 3 3 3 3 3
Base + ATH + ZnBO3 3 3 3 0 0 0 2 2 2 3 3 3
Base + ZnBO3 2 2 2 0 0 0 2 2 2 3 3 3
Commercial mix 3 3 3 2 2 2 3 3 3 3 3 3

Table 4.7
IEC 68-2-10 Extent of mold growth on silicone rubber treated with spores and water.
Sweden Sri Lanka Tanzania Standard
Material 1 2 1 2 3 3 1 2 3 1 2 3
Base 1 0 0 0 0 0 0 0 0 0 0 0
Base + ATH 0 0 0 0 0 0 1 1 1 0 1 0
Base + ATH + ZnBO3 0 0 0 0 0 0 0 0 0 0 0 0
Base + ZnBO3 0 0 0 0 0 0 0 0 0 0 0 0
Commercial mix 2 1 1 1 1 1 2 2 2 2 2 2

The results of these standard tests show clearly that, if no nutrients are added to the samples,
none of the silicone rubber materials mixed in the laboratory support growth. However, the
commercial silicone rubber mix supports mold growth. These results strongly suggest that
some component or combination of components in the commercial mix promotes the ability
of the material to support mold growth.

When nutrients were added to the silicone rubbers, most of the samples supported mold
growth. In an out-door environment, nutrients from the surroundings may adhere to the
surface of a silicone rubber insulator and provide nutrients for growth of a biofilm. However,
growth is less severe on samples with zinc borate added. The low concentration of flame
retardant used in the materials is sufficient to partially protect the silicone rubber from mold
growth. This could not be predicted from the powder tests.

The sensitivity of the fungal species to the addition of zinc borate also differs between the
standard tests and the powder tests. The fungal cultures from Tanzania seemed very sensitive
when powders were tested, but standard tests show that, when these molds are grown on
silicone rubber materials, they can tolerate higher concentrations of zinc borate. Instead,
fungal cultures from Sri Lanka, that were easy to grow on the agar used in the powder test,
were very hard to grow on silicone rubber and impossible to grow when zinc borate was
added to the rubber. Nevertheless, both types of tests show clearly that the sensitivity to zinc
borate differs between different fungal species. This emphasises that care is needed in the

Results and discussion

selection of test organisms. If possible, species likely to infect the material under normal
service conditions should be chosen.

On the other hand, valuable information can also be gained from standard tests performed as
recommended. Microorganisms recommended in standard test procedures are chosen for their
ability to negatively affect the tested materials, and they can therefore be said to represent a
worst-case scenario. In this work, the advantages of using standard test organisms was shown
when samples were subjected to the ASTM G21-90 standard for determining the resistance of
synthetic polymeric materials to fungi. Samples were treated with mold spores and nutrient
salts, but no source of carbon was added to the solution. If the spores were to germinate, they
would thus have to use the test material as the source of carbon. The results of the test, when
performed with organisms isolated from the insulators studied, are shown in Tab. 4.8 and they
resemble the results of the IEC 68-2-10 standard tests.

Table 4.8
ASTM G 21-90 Extent of mold growth on silicone rubber treated with spores and salt.
Sweden Sri Lanka Tanzania
Material 1 2 3 1 2 3 1 2 3
Base 0 0 0 0 0 0 0 0 0
Base + ATH 0 0 0 0 0 0 0 0 0
Base + ATH + zinc borate 0 0 0 0 0 0 0 0 0
Base + zinc borate 0 0 0 0 0 0 0 0 0
Commercial mix 2 2 2 1 0 0 3 3 3

However, when the test was carried out with the organisms stipulated in the standard, more
information was gained. In Fig. 4.1, results from the unmodified ASTM standard test are
shown together with results from a new test method called the Quick Test. In this method,
growth was initiated and well established on highly nutritive agar before it started to colonise
the sample. The fact that a material is easily colonised may be important, not for the initiation
but for the advance of biological growth. Both tests were performed on virgin material as well
as on materials aged with oxygen plasma prior to inoculation with microorganisms.

2,5 Figure 4.1

Results from the Quick Test and
2 the ASTM G21-90 standard test
when performed using the
microorganisms stipulated in the
Extent of growth

ASTM unaged
ASTM aged standard.
Quick unaged
1 Quick aged Tests were performed in
triplicates and results were
evaluated according to the scale
described in Tab 3.3. The average
of this evaluation is plotted.
Base Base+ATH Base+ATH+Zn Base+Zn Commercial

Results and discussion

The results obtained again indicate that the commercial silicone rubber is the most sensitive
material, while the silicone rubber formulations with added zinc borate seem to be more
protected. However, when the very potent fungal species stipulated in the standard test were
used, the extent of growth varied among the materials tested. Apart from the fact that zinc
borate protects the samples; the results indicate that ATH promotes growth to some extent.
This could bee due to the surface treatment of the ATH particles, the vinylsilanes used contain
carbon that may provide a source of nutrients for microorganisms. However, for the degree of
colonisation measured in the Quick Test, the surface roughening caused by the addition of
ATH-filler is probably more significant. A rough surface offers better support for an
advancing growth than a smooth material. Results from the Quick test also indicate a
difference between virgin materials and materials aged with oxygen plasma. Growth
advanced faster and further on the aged samples when compared to unaged rubber of the same
type. This is probably due to the partially hydrophilic nature of the plasma treated rubbers.
The wetted surface formed by the treatment provided water and support to the advancing

The sensitivity to algae growth of the silicone materials under investigation was tested
according to the procedure described in the ASTM D 5589-97 standard practice. Algae use
CO2 as source of carbon through photosynthesis, therefore no source of carbon was added to
the growth medium used in these experiments. The results shown in Tab. 4.9 indicate that
zinc borate is toxic to the studied algae. Growth was not affected on pure silicone, the mix
with ATH or the commercial mix, but algal growth was hindered or inhibited on materials
with added zinc borate. This can be very important for the performance of the materials when
they are used outdoors. Algae growth is not expected to degrade its support material directly;
but in an outdoor environment mixed biofilms are formed. Algae growth may attract other
kinds of microorganisms that can use the colonised material as a carbon source.

Table 4.9
ASTM D 5589-97 Extent of algal growth on silicone rubber treated with algal cells.
Sweden Sri Lanka Tanzania
Material 1 2 3 1 2 3 1 2 3
Base 2 2 2 2 2 2 2 2 2
Base + ATH 2 2 2 2 2 2 2 2 2
Base + ATH + zinc borate 1 1 1 2 2 2 1 1 1
Base + zinc borate 0 0 0 0 0 0 0 0 0
Commercial mix 2 2 2 2 2 2 2 2 2


In order to simulate biofilm growth on high voltage insulators in an outdoor environment,
microenvironment chambers were prepared. In these chambers, mixed biofilms resembling
those formed on insulators in field use were grown on the silicone rubber materials. Algae
cells immediately started to grow on materials not containing zinc borate. After several

Results and discussion

weeks, fungal growth started to develop on the materials infected by algae growth, and
biofilms similar to those formed on insulators in the field were formed. The zinc borate-filled
materials where algae refused to grow seemed protected from fungal growth as well.
Microorganisms in a biofilm tend to live in symbiosis. It is known that the algae used in this
study often grow in association with bacteria, and it seems probable that the fungal species
isolated from the studied biofilms need algae as source of nutrients for growth [114-116]. This
means that the protective effect of zinc borate against algal growth also strengthens the
protection against mold growth.

Materials incubated in the microenvironment chambers were not used only to study the
resistance of the silicone rubbers to the formation of a biofilm. ESEM/EDX and IR analysis
were performed on the materials in order to detect changes in chemical composition of the
sample surfaces, while image analysis and laser-induced fluorescence spectroscopy were used
to study the development of the biological growth.

4.3.1 Image analysis

Manual inspection is the most commonly used way to estimate the area covered by biological
growth. However, the method is operator-dependent and therefore suffers from a lack of
precision. Samples from the microenvironment chambers were therefore subjected to image
analysis in an attempt to determine the covered area more precisely. Six months after
inoculation, samples were photographed and the resulting digital images were subjected to
image analysis in order to determine the extent of growth on the different materials. This was
done by segmentation of the photographs into regions of similar chromatic appearance. A
typical result obtained through the application of this segmentation technique is shown in Fig
4.2, where regions classified as growth have been marked in red. As can be seen, the
highlighted regions correspond well to the areas covered by growth.

a) b)

Figure 4.2 A typical result obtained through image analysis. a) Photograph of an

ATH-filled silicone rubber sample infected by a biofilm from Tanzania. b) Regions
identified as growth by using image analysis are marked in red.

Results and discussion

As can be seen in Fig. 4.3, the image analysis results clearly show that the flame-retardant
zinc borate hinders the development of the biofilm, and that ATH has no hindering effect on
the growth. In fact, results again indicate that ATH might even support the biofilm. Another
effect observed was that the biofilm originating from Sri Lanka developed faster on the
commercial material tested, than on the other two biofilms under study. This might be due to
the composition of the biofilm. Microscopic studies showed that the biofilm from Sri Lanka
contained a larger proportion of fungi than the biofilms isolated from either Tanzania or
Sweden. The commercial rubber was, as the previous discussion shows, sensitive to fungal

Effect of filler Effect of UV-light

14,0 14,0

12,0 12,0

10,0 10,0

Covered area (%)

Covered area (%)

8,0 Sri 8,0 low

Tz medium
6,0 Swe 6,0 high

4,0 4,0

2,0 2,0

0,0 0,0
Base ATH ATH+Zn Zn Com Base ATH ATH+Zn Zn Com

Figure 4.3 The effect of material composition Figure 4.4 The effect of UV-light exposure on
on the extent of biological growth. the extent of biological growth.

Another observation was that direct UV exposure from a daylight lamp effectively hindered
the growth of the biofilm. Fig. 4.4 shows that the samples that were nearest to the daylight
lamp in the microenvironment chambers were almost completely free from growth. On the
middle plates that were partly shaded growth developed much better. This effect can also be
seen at high voltage insulators in service, where growth develop quicker on the shaded parts
of the insulators [1, 5, 13].

The plates that were close to the bottom of the environmental chambers should, according to
theory, be the worst infected samples, since they were partly shaded and hydrophilic [72].
However, this was not supported by results. This discrepancy is probably due to the method of
inoculation. In an outdoor environment, microorganisms are deposited on the silicone rubber
by wind and rain, whereas in the microenvironment chambers, microorganisms are sprayed
onto the silicone rubber samples, and droplets form on the surface of the material. On the
hydrophilic samples, these droplets pool together, form wetted traces, and run off the plate.

4.3.2 Laser-induced fluorescence

Recent studies indicate that image analysis can be used to estimate the area and distribution of
growth on both material samples and complete insulators [65]. Nevertheless, the technique

Results and discussion

has its drawbacks. For instance, the method does not discriminate between biological growth
and other coloured pollutants on the insulator surface. To overcome this problem, and
possibly increase the sensitivity compared to photography, laser-induced fluorescence (LIF)
spectroscopy was explored as a tool for the detection of growth.

In LIF surface spectroscopy, the material under study is irradiated with monochromatic light
from a laser source. Molecules in the sample surface absorb photons and electrons become
excited. Fluorescent light is emitted and detected when the electrons spontaneously return to
their ground state. The intensity of the fluorescent light is often substantially higher from a
clean surfaces than from a surface covered by biological growth [66]. However, the absolute
intensity is dependent on several factors, such as laser pulse energy, area of excited region,
and the angle between the surface under study and the laser beam. Thus, the absolute intensity
does not provide unambiguous information about the samples. A more promising approach is
to use the spectral shape of the normalised fluorescence spectrum. The fluorescence spectrum
from a fungi-covered silicone rubber is often wider than the spectrum from a clean sample
and, if algae are present on the surface, an additional peak at about 685 nm indicates
chlorophyll fluorescence [69]. These effects were clearly seen when samples from the
microenvironment chambers were studied. The broadening effect and the algae peak were
more pronounced on silicone rubbers with a severe biofilm growth than on cleaner samples.
The algae peak disappeared completely on samples with zinc borate added, as can be seen in
Fig. 4.5.
-3 -3
x 10 x 10
a) 5
b) 5
Intensity, [arb. units]

Intensity, [arb. units]


1 1

0.5 0.5

0 0
450 500 550 600 650 700 750 800 450 500 550 600 650 700 750 800
Wavelength [nm] Wavelength [nm]

Figure 4.5 Results from LIF-surface spectroscopy. a) Spectra collected from the ATH-filled
silicone rubber. The red spectrum obtained from a sample infected by a dense biofilm is wider
than the corresponding blue reference spectrum and the algae peak at 685 nm is clearly visible.
b) Spectra from the zinc borate-filled rubber. The sample looked clean to the naked eye, since no
mature biofilm had formed. The red spectrum obtained from the sample infected with
microorganisms is only slightly broader than the blue reference spectra and there is no peak
corresponding to chlorophyll fluorescence.

In addition, LIF surface spectroscopy was found to be a very sensitive method for the
detection of microorganisms; areas that looked clean to the naked eye sometimes gave a
response as if they were contaminated. When these areas were studied under the microscope it

Results and discussion

was found that traces of biological growth were in fact present. This indicates that LIF surface
spectroscopy may be used to detect microbiological attack on silicone rubber insulators before
a mature biofilm visible to the naked eye has formed.

However, the method has its drawbacks. While it is easy to detect algae defacement of a
silicone rubber surface, detection of fungal growth is more problematic. The observed
broadening of the LIF-spectra is not always easy to detect from the full spectrum obtained. A
simplified score plot based on the variations between spectra is easier to handle and it can, if
there are significant differences between samples, separate clean samples and samples
infected by fungal growth into distinctly different classes.

Fig 4.6 a) shows typical fluorescence spectra of eight different fungal cultures grown on
silicone rubber filled with ATH. Spectra obtained from the fungus-covered samples are wider
than the spectrum of the clean ATH-filled silicone rubber material used as reference.
However, for some samples these effects were small and difficult to detect in the spectra
obtained. Results are easier to interpret from a simplified score plot of the sample set. In Fig.
4.6 b) spectra obtained from the infected materials are showed and compared to spectra from
clean material in a simplified score plot. Each spectrum is reduced to a single dot in the score
plot. Dots originating from spectra of samples infected by fungal growth are spread out to the
left in the score plot, when compared with the encircled dots from the clean material. The
spread is due to the combination of fluorescence from the fungal growth and the material. A
heavily contaminated sample ends up further to the left in the score plot compared to a cleaner
sample. When results are displayed as a simplified score plot it is easy to differentiate
between a samples infected by fungal growth and a clean material. The different fungal
cultures studied all followed the same trend in the simplified score plot. In addition, further
studies showed that the method gave the same general response when other silicone rubber
materials were tested. This is desirable for the development of a general inspection technique
for composite insulators.
a) 4
b) 5
Intensity (arb. units)

PC 2

2 0

0 -5
400 500 600 700 -10 -5 0 5
Wavelength (nm) PC 1

Fig. 4.6. Fluorescence spectra of eight fungal cultures grown on silicone rubber with
added ATH a) Full fluorescence spectra of the sample set. b) Simplified score plot of
the studied samples;. results from the clean ATH-filled silicone rubber material are
located within the circle, while spectra from materials infected with fungal growth
are spread out to the left in the plot.

Results and discussion

When a silicone rubber sample infected by biological growth is subjected to a combination of

LIF surface spectroscopy and image analysis, this gives a more complete picture of the
situation than either method can give on its own. While LIF surface spectroscopy can be used
to decide the type of contaminant and reveal information of contaminants invisible to the
naked eye, image analysis can be used to estimate the severity of the growth. Both methods
have a potential for use as in-service diagnostic techniques. However, by utilizing LIF for
creating images depicting certain spectral properties, so-called imaging measurements, a
combination of LIF and digital image analysis can be obtained, which is probably even more
interesting for future applications. A mobile and self-contained light detection and ranging
systems that could be used for field measurements of this kind already exists [117].

4.3.3 Hydrophobicity
One of the reasons why silicone rubber materials are preferred to ceramics in high voltage
insulators is the hydrophobicity of the silicone rubber surface, but when a biofilm covers the
insulator surface, the hydrophobicity of the rubber is disturbed [3, 6, 7] The most common
way to measure surface hydrophobicity is by water contact angle measurements. However,
this is not practical when the effect of a biofilm on surface hydrophobicity is to be recorded.
In most cases, the biofilm is unevenly distributed over the sample surface, making it difficult
to choose representative spots for the measurement. A biofilm may also function as a sponge
withdrawing the water drop used for the contact angle measurement.

To avoid these problems, the hydrophobicity of the silicone materials incubated in the
microenvironment chambers were tested in accordance with the STRI hydrophobicity
classification guide [104]. In this method the pattern formed by the drops when a large surface
area is sprayed with water is studied and compared to a series of seven reference images with
different hydrophobicity classes (HC). HC 1 denotes a very hydrophobic sample on which
only discrete droplets are formed, while HC 7 describes a hydrophilic sample on which a
continuous water film is formed over the whole tested area. The method has a potential for
use on insulators in the field; and the results can be evaluated either by visual inspection or by
use of image analysis [62].

The worst infected samples from the microenvironment chambers were classified according to
the STRI-hydrophobicity classification guide. The results, displayed in Tab. 4.10, clearly
show that samples covered with microorganisms were more hydrophilic than virgin materials
and reference materials treated with nutrient salt solution. However, after cleaning the surface
of the infected materials with a soft cloth damped with distilled water, the hydrophobicity of
the materials were somewhat restored. However, materials containing large amounts of filler
particles were not as easy to restore as were less filled materials. Filler particles cause surface
roughening and it seemed that the gentle cleaning procedure applied was not effective enough
to completely remove the biofilm from the rough surface of the filled materials.

Results and discussion

Table 4.10
HC of silicone rubber infected by biofilms and reference materials
Samples from microenvironment chambers
Contaminated Cleaned Virgin
Material Sri Tz Sw Ref Sri Tz Sw Ref materials
Base 3 3 3 2 1 1 1 1 1
Base + ATH 4 4 4 2 2 2 2 1 1
Base + ATH + zinc borate 3 3 3 2 2 2 2 1 1
Base + zinc borate 2 2 2 2 1 1 1 1 1
Commercial mix 3 3 3 2 2 2 2 1 1

4.3.4 ESEM/EDX
If a severe growth has developed on an active silicone rubber insulator, it may be necessary to
remove the biofilm. Spraying with a biocide is not a good alternative; this may kill the
microorganisms but it does not remove them from the insulators. Dead biomass is left in the
system and this opens the way for a rapid microbiological re-growth [71]. If insulators are to
be treated in some way, cleaning is a better alternative. However, cleaning is only effective if
the adhesion between the biofilm and the silicone rubber is not too strong.

To examine the possibility of cleaning an infected silicone rubber insulator, samples from real
insulators as well as samples from microenvironment chambers were analysed by using an
environmental scanning electron microscope (ESEM) with X-ray energy dispersive
spectroscopy (EDS). ESEM/EDS not only give a microscopic picture of the infected surface
but also combine this information with an overview of the elemental composition of the
surface of the tested material. Since the molecules in the biofilm differ in chemical
composition from those of the supporting silicone rubber, the atomic composition of the
surface layer can be used to monitor the amount of contaminants before and after a washing
procedure. If traces of the biofilm remain after cleaning the surface, it can be assumed that the
adhesion between the biofilm and its support is too strong to allow the applied washing
procedure to be effective.

Silicone rubber samples cut from insulators situated in Tanzania, Sri Lanka and Sweden were
analysed with ESEM/EDS. The results shown in Fig 4.7 and Fig 4.8 indicate that it was
possible to clean the African insulator efficiently by gently wiping the surface with a soft clot
wetted with distilled water, but that the Asian rubber was not that easy to clean from
biofouling. Traces of Mg, Na and Fe originating from the biofilm were detected after the
cleaning. ESEM pictures supported the results of the elemental analysis; the silicone rubber
from Sri Lanka showed a rough surface after cleaning, while the rubber surface from
Tanzania looked clean and smooth. The results of ESEM/EDS analysis of the Swedish
insulator resembled those of the insulator from Sri Lanka.

Results and discussion

Fig 4.7
ESEM/EDS of insulator from Tanzania

It was possible to restore the surface of the

silicone rubber by the gentle cleaning
O procedure applied. After cleaning, ESEM
C images show a smooth surface and EDX does
not show traces of molecules originating
from the biofilm.

a) EDS of virgin SIR



Na Mg S

b) EDS of SIR covered by growth c) ESEM of SIR covered by growth



d) EDS of SIR after cleaning e) ESEM of SIR after cleaning

Results and discussion

O Fig 4.8
ESEM/EDS of insulator from Sri Lanka

It was not possible to restore the surface of

Si the silicone rubber by the gentle cleaning
C procedure applied. After cleaning, traces of
Mg Na and Fe originating from the biofilm
were detected by EDS and ESEM showed a
rough surface.

a) EDS of virgin SIR


C Si

Fe Na Mg

b) EDS of SIR covered by growth c) ESEM of SIR covered by growth



Fe Na Mg

d) EDS of SIR after cleaning e) ESEM of SIR after cleaning

Results and discussion

Unlike the Tanzanian insulator, EDS analysis showed that the insulators from Sri Lanka and
Sweden contained large amounts of the flame retardant ATH. It seems that an ATH-filled
silicone rubber is more difficult to clean, than a rubber containing less filler. In order to verify
this hypothesis, samples from the microenvironment chambers were analysed with
ESEM/EDS. After one year of incubation in the microenvironment chamber representing Sri
Lanka, small samples covered with a dense biofilm were cut from ATH-filled silicone rubber
and Base rubber. These samples were analysed before and after cleaning. Results verified that
it was easy to remove the biofilm from the Base rubber, but the ATH-filled material was more
difficult to clean. This is probably due to the fact that the ATH-filled rubber had a rougher
surface structure than the Base material. A rough surface is more accessible for
microbiological growth [72].


Despite the fact that an ATH-filled silicone rubber is difficult to clean from biological growth,
no evidence was found that the biofilms examined in this study are able to degrade the
silicone rubber matrix. When samples from the microenvironment chambers were cleaned
from biological growth and analysed with infrared (IR) spectroscopy, no difference in spectra
could be observed between samples treated with microorganisms and virgin materials. This
indicates that no chemical bonds had been disrupted by the influence of the biofilm.

The PDMS molecule is known to be very stable and no biological process is known that
produces or degrades covalent bonds in organosilicones [46]. Nevertheless, silicone rubber
could still be degraded by biological processes through secondary mechanisms. Many
microorganisms secrete powerful enzymes with the ability to degrade very stable polymeric
structures. Among these, the white rot fungi hold a unique position. This class of molds has
evolved non-specific mechanisms for degrading the complex polymeric structure of lignin.
The enzyme system responsible for the degradation includes peroxidases that promote the
reduction of peroxides to free radicals [90, 93]. These radicals have been shown to attack
stable polymeric materials such as polyvinyl chloride, nylon and polyethylene [118-120].
Claims have been made that these molds could also degrade PDMS oils [94].

According to United States patent No: 6,020,184, silicone oils of high molecular weight could
be degraded by the lignin-degrading fungus Phanerochaete chrysosporium. The fungus is
grown for 60 days in a liquid medium spiked with a small amount of silicone oil. According
to the results reported this procedure degrades 17-50 % of the added silicone oil. However, we
have repeated the experiment, as described in the patent, and after modifications
recommended by experts in the field of lignin degradation. Still, no reliable proof of PDMS
degradation has been observed in any of the tests performed.

In order to study the biologically mediated degradation of the silicone molecule in detail, tests
were performed on a PDMS rubber, on a PDMS oil of high molecular weight, and on a DMS
oligomer. After treatment with the lignin-degrading fungus Phanerochaete chrysosporium,

Results and discussion

oils were analysed by SEC in order to detect main-chain scissoring reactions, oils and
oligomers were analysed with GC/FID in order to observe degradation mediated by chain end
or side chain mechanisms, and rubbers were analysed for chemical changes in the surface
layer by use of IR.

No significant difference in surface composition of the rubber or the molecular weight of the
oil was observed and no low molecular weight degradation products were found in the GC
analysis performed after treatment of the siloxane materials with the lignin-degrading fungus.
It was therefore decided that experiments with radicals generated by chemical mechanisms
should be performed as a compliment to the biodegradation study. The materials mentioned
above were exposed to hydroxyl radicals generated by the Fenton mechanism. However,
subsequent analysis showed no significant influence of the hydroxyl radicals on the silicone
materials analysed. Some results from the performed tests can be seen in fig. 4.9-4.11

Figure 4.9
No degradation of the
silicone rubber could be
Enzymes observed through IR-
spectroscopy. No
covalent bonds in the
sample surfaces had been
Radicals disrupted by the
treatment with enzymes
or radicals


Enzymes Figure 4.10 The results

from SEC analysis
showed no degradation
of the PDMS oil. The
molecular weight of the
oil was the same after
exposure to enzymes or


Results and discussion






Figure 4.11 The results from the GC analysis showed no degradation of the DMS oligomer
No new peaks denoting degradation products could be observed in the chromatograms

The results obtained from the degradation study indicate that the PDMS molecule is very
stable against biodegradation. However, it cannot be concluded, based on these results only,
that PDMS is a biologically inert material. It is possible that degradation could occur if a more
effective experimental design were developed. If, for example, the silicone oil could be
dispersed or dissolved in the reaction medium, the PDMS molecules would be more
accessible to degrading enzymes and radicals. Even so, the probability that the silicone rubber
matrix of a high voltage insulator could be degraded by the direct influence of
microorganisms is very small.

Results and discussion


Subsequent effects ranging from fouling to degradation of the polymer matrix occurs when a
polymeric material is colonised by a biofilm. For silicone rubbers in high voltage insulators
this study showed that fouling is the most severe problem. The polydimethylsiloxane (PDMS)
molecule is highly stable and no evidence of biodegradation has been found. Neither
microorganisms isolated from insulators in service nor aggressive ligninolytic fungal species
seemed to be able to break the chemical bonds of PDMS.

Biofouling can, however, seriously influence the function of an insulator. This study showed
that the hydrophobicity of a silicone rubber is reduced when the material is colonised by a
biofilm. This can alter the electrical properties of a fouled insulator. It is therefore important
to develop methods for the identification of infected units. In this work, laser-induced
fluorescence (LIF) spectroscopy was successfully used to identify biological contamination
on silicone rubber surfaces. Photography followed by image analysis was then used to
estimate the extent of growth.

The surface hydrophobicity of the silicone rubber can be restored by removing the attached
biofilm. The effectiveness of a cleaning procedure depends, however, on the composition of
the rubber. A silicone rubber containing large amounts of alumina trihydrate (ATH) has a
rougher surface structure, compared to an unmodified silicone. This makes it difficult to
remove an attached biofilm in an effective way.

In addition, surface treated ATH makes a material more accessible to microorganisms. This
was shown when different silicone rubber formulations were subjected to a series of
microbiological growth tests developed for this study. Methods useful for fast evaluation of
samples were used together with standard test methods and microenvironment chambers
constructed for long-term studies in a simulated outdoor environment. These tests showed that
while the commercial silicone rubber tested supported growth, addition of the flame-retardant
zinc borate to a material made it able to suppress the growth of mold, algae and mixed


Future work

It has been shown in a series of growth tests that addition of zinc borate to silicone rubber
protects the material from severe biofouling. However it would be advantageous if materials
could be tested outdoors under true service conditions.

The methods developed to study the biodegradation of PDMS should be perfected in order to
definitely confirm or exclude the degradation of PDMS under the influence of enzymes or

In this work laser-induced fluorescence (LIF) spectroscopy has been successfully used to
identify silicone rubbers infected by biological growth. However the causes of the observed
differences between clean and fouled materials are not yet fully understood. It would be
beneficial for the future development of the technique if the biological structures responsible
for the observed differences in fluorescence could be identified.

Finally, methods combining LIF-spectroscopy and image analysis should be developed

further and used to identify contaminated insulators in the field. One promising attempt in this
direction is the mobile light detection and ranging (LIDAR) system used by Dernfalk et al for
detection of biological growth on silicone rubber insulators. This method has the potential to
be used to diagnose energised insulators from a safe distance [66].

Future work


My heartfelt thanks to Sigbritt Karlsson for accepting me as her PhD-student, thank you for
supporting me throughout my studies. Further I would like to thank the members of the
reference group; Stanislaw Gubanski, Brbel Hahn-Hgerdal, Ulf Gedde, Hkan Wiek,
Fredrik Sahln and Sven Jansson for their interest in my research. My sincere gratitude also
goes to the leaders and administrators of the department of fibre and polymer technology.

ABB Corporate Research, Energimyndigheten and Elforsk are acknowledged for financial
support through the Elektra programme.

My fellow PhD-students Andreas Dernfalk at CTH and Magnus Begtsson at LTH are
acknowledged for interesting and fruitful collaboration. Warm thanks also to Palle Ander and
Maria del Pilar Castillo at SLU for invaluable advice in the field of lignin degradation.

To my colleagues, you were always the best part of the job. Thanks to all of you, especially
former and present members of the SK-research group, Nadja, Walker, Guillaume, Ana, Fran,
Jonas, Emma, sa and Sara

Some of you deserve special thanks for being true life savers; Jovan (San Diego and MSN),
Calle (comics and philosophy), Mattias (champagne and psychology), Tahani (my first
student), Karin (extractions and gossip), Johan (cars and sick humour), Emma (Vsters and
hot discussions) and Peter (fun food delivery). Thanks also to the coffee-room people, Clara,
Annika, Jonas, Jonas, David, Anders, Bruska, Bengt, Mari, Daniel, Alessandro, the Spanish
league and those of you I have forgotten to mention. I will miss you all.

Love to all my friends not kept in institution, especially to Jocke (breakfast soon?), Anna (the
wedding planner) and Jenz, Tobias and Lina, (cats!), Maja, Fredrik and Maria (Stayokay)
Micke (Byrdirektr Toll). Love also to Josefin (with family!), Elin and Josefin, your
friendship means a lot to me, as does kvinnokampssemestrarna.

Still, my warmest gratitude goes to my family. You have never pushed me, always helped me
and never stopped loving me. I feel safe knowing that you will always be there for me.

To Anders, for the rest of our days.



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