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FRAS Technology AS, Industrial Microbiology Reviews nr 1. 2013
Current knowledge on microbial induced problems and biofouling in lubrication
systems of ships and marine installations
Christer Moe Fjeld
FRAS Technology AS
Kongeveien 30 1430 Ås, Norway
Summary
Microbial induced problems (MIP) and biofouling in lubrication oil systems are probably rare and usually
an occult event. MIP and biofouling are in most cases assumed to be of mechanical origin and mitigation
is undertaken without knowledge of the underlying cause. Because of this, the extent of such problems
is not known, but assumed to be rare. Based on the ubiquitous nature of microorganisms and their
versatile arsenal of metabolic properties, it is not surprising that they are capable of causing detrimental
effects in a wide variety of industries. Industries using oils for lubrication purposes are no exception. If
water is present and the water activity is sufficient, the systems are prone to microbial colonization,
which may in turn result in several adverse effects. Lubricants and especially spent lubricants, are toxic
to many organisms, thus environmental challenges are indeed a legitimate concern. These days, several
industries that are using oils for lubrication purposes are shifting to a “green” line in their choice of
fluids. In industries such as forestry and other terrestrial applications, these lubricants are excellent
replacements to the petroleum based lubricants. However, care should be exerted in maritime
industries since water will usually be found in various amounts in the lubrication systems. The
Environmentally Acceptable Lubricants (EAL) are more biodegradable than the traditional oils based on
petroleum. Therefore, it is reasonable to assume that these fluids are more prone to biodeterioration in
the lubrication systems. Because of the higher biodegradability of the EAL, there is a risk that microbial
induced problems and biofouling will increase in the future. This emphasizes the importance of
increased monitoring of such problems in the marine industry. In this review, I used reports from FRAS
technology AS and the current literature with the aim of trying to explain some of the reasons for
unknown machinery breakdown. In addition, I present some cases where microorganisms are the cause
of machinery breakdown, either directly or indirectly.
Key words: Petroleum based lubricants, environmentally acceptable lubricants, additives, biofouling
Correspondence:
Christer@fras.no
Kongeveien 30, 1430 Ås, Norway
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FRAS Technology AS, Industrial Microbiology Reviews nr 1. 2013
1. Introduction
Microbial growth is not as solitary and simple as one would believe. Microbial growth and
colonization is complex, involving social interactions, synergistic cooperation, intense
competition for nutrients and a very large microbial diversity over a very small area. The
presence of active and proliferating microorganisms are ubiquitous in nature, including sodium
chlorine brine water (cryopegs) at sub zero temperatures in the Antarctic (Gilichinsky, et al.,
2005) and in the vicinity of hydrothermal vents on the oceans floor where microorganisms are
proliferating in temperatures at around 121o
C (Kashefi & Lovley, 2003). The strain identified by
Kashefi and Lovley was also able to grow at an astonishing 130o
C. Microorganisms are also
found in acidic and alkaline environments. The general requirements for microbial growth to
occur is the presence of free water (or a water activity aw > 0.85 for most microbes), a carbon
source (glucose, cellulose, hydrocarbons, lipids, proteins etc), for the heterotrophs and CO2 for
the autotrophs, and various amounts of nitrogen and phosphorous depending on microbial
species (oligotrophic organisms are able to grow in very pure water e.g. Caulobacter sp.) In
addition microorganisms require micronutrients such as sulfur, iron, manganese etc for optimal
growth. The key factor involved in biodegradation is usually the bioavailability of compounds
with a few exceptions such as polycyclic aromatic hydrocarbons (PAH) due to higher water
solubility than many long chain alkanes (Huesemann, et al., 2004). In general, hydrophilic
compounds have a higher bioavailability than hydrophobic compounds (Bosma, et al., 1996).
When microorganisms are in contact with a moist surface, they will attach. After some time
(hours to days depending on environmental factors and species present) they will form a
complex 3D structure composed of extracellular polymeric substances (EPS; carbohydrates,
lipids, proteins and DNA), cells and water (Costerton, 1995, Sutherland, 2001, Hall-Stoodley, et
al., 2004). This is referred to as a biofilm. Dental plaque is a good example of a typical biofilm.
Cells that are imbedded in the EPS matrix are provided with both shelter from predation,
antimicrobial agents, chemical stress and UV radiation (Carpentier & Cerf, 1993, Lewis, 2008,
Flemming, 2009).
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FRAS Technology AS, Industrial Microbiology Reviews nr 1. 2013
Another benefit of living in close proximity in biofilms is the facilitation of metabolic
cooperation between different species (Costerton, 1995), However, strong competition also
occur (Elias & Banin, 2012). Biodegradation of complex organic compounds such as
hydrocarbons is often more efficient in biofilm cultures compared to planktonic degradation of
hydrocarbons (Jain, et al., 2011). With this short introduction into microbial processes in nature
it is easy to understand that microorganisms are able to cause adverse effects in a wide variety
of manmade applications. When biofilms are established in undesired locations in manmade
applications it is referred to as biofouling, i.e. a biofilm in the wrong place. Biofouling has been
identified as a cause of severe adverse effects in a wide variety of industries (Table 1).
Table 1. Industries and applications known to be adversely affected by microbial induced
problems and biofouling
Industry or application Manifestation of problems References
Petroleum industry Microbial influenced
corrosion (MIC) and souring of
applications and crude oil
respectively.
(Neria-González, et al., 2006,
Gieg, et al., 2011)
Shipping industry MIC of ships hulls, Increased
drag and spread of non
indigenous species through
ballast water.
(Holm, et al., 2004, Drake, et
al., 2007)
Paper industry Product withdrawal,
increased maintenance costs
and machinery downtime.
(Klahre & Flemming, 2000)
Metal working industry Reduced properties of fluids
and, the development of
hypersensitive pneumonitis in
workers and various
dermatological symptoms
(Kreiss & Cox-Ganser, 1997,
van der Gast, et al., 2001,
Lucchesi, et al., 2012)
Hydroelectric power plants Increased friction in lower
vane bearing bushings.
(Schneider, et al., 2006)
Medical applications Biofilms on prosthetic joints
and other implants causing
infection. Biofilms in hospital
environments may cause
nosocomial infections.
(Zimmerli, et al., 2004,
Lindsay & von Holy, 2006)
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FRAS Technology AS, Industrial Microbiology Reviews nr 1. 2013
Biofouling and microbial induced problems (MIP) in lubrication systems in any industry are
directly associated with the presence of free water in the systems (aw > 0.85). This is the main
reason why emulsion type metal working fluids (mixture between oil and water) are especially
prone to severe microbial contamination and the resulting detrimental effects (Mattsby-
Baltzer, et al., 1989).
Water may enter the hydraulic/ lubrication systems via three independent routes:
1. Oil storage barrels are often stored outdoors and on the deck of ships. Water spray from
the sea will definitely contain hydrocarbon degrading organisms, independent of
geographic location. Ambient temperature fluctuations will cause pressure and vacuum
shifts of the air in the barrel’s head space. Water with hydrocarbon degrading organism
will enter the barrels through the sealing if water is present on top of the barrels.
2. Humidity in the air may precipitate in lubrication oil tank and humid air could also be
dissolved in the oil and precipitate in cooler areas of the system, such as in ships
thrusters or in stern tubes.
3. Water is frequently introduced to systems through heating and cooling events during
operation, pressure fluctuations as a result of propeller movement and as a result of the
vessel’s movement in the water column. I.e. stern tubes and thrusters will experience
pressure variation due to waves, resulting in a shifting water depth around the
propeller. To compensate for this pressure variation and thus to prevent severe water
ingress, pressure is applied to the systems. This overpressure leads to lubricant loss
when the surrounding pressure is below the pressure applied on the system. This occurs
for example when the propeller is surfacing during rough seas. However, no water
ingress is difficult to achieve.
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FRAS Technology AS, Industrial Microbiology Reviews nr 1. 2013
To my knowledge, scientific literature on biofouling in systems using oils for lubrication
purposes has been absent for nearly two decades. Some work, however, was undertaken in the
1970`s, 80`s and early 90`s regarding microbial growth in lubrication systems (Summers-Smith,
1982, Okpokwasili & Okorie, 1988, Stuart, 1994-1995). The extent of such problems these days
is therefore unknown. One of the reasons could be that the manifestation of biofouling in such
systems is easily confused with mechanical issues. Previously, the industries were more post
active, after problems occurred, with water drainage of systems and other maintenances
necessary to mitigate the occurring detrimental effects. Upon opening of systems they would
probably observe large quantities of slime, notice foul odor etc, which cannot be explained by a
standard mechanical approach. Such observations would lead to suspicion towards microbial
contamination. Nowadays, however, systems are under more thorough surveillance, thus the
industries are more proactive in avoiding adverse occurrences, especially with emphasis on
water contamination and increased wear (commonly measured by particle count according to
the SAE AS 4059 standard). Therefore biofouling and the following MIP in such systems are
most likely evolving more slowly, based on the small body of water compared to the bulk
amount of lubricant. I.e. the amount of water present is not sufficient to promote extensive
growth with rapid manifestation of detrimental effects. On the other hand, biofouling could
speed up the expected mechanical wear of systems, by facilitating emulsification of oils,
foaming and sludge formation. Relevant questions are, therefore: Have any shipping companies
had a higher maintenance cost than predicted in some vessels? Are there any plausible
explanations for this possible increase in maintenance cost? Does any company report
malfunctions or incidences which cannot be explained by a standard mechanical approach? If
no mechanical or technical explanation can explain the condition, microbial causes should be
considered. Fuels on the other hand are well known to contain harmful microorganisms and
their associated problems. Therefore, biofouling and MIP have for a long time been known in
the fuel industry and accepted as a cause of numerous machinery breakdowns and are thus
especially monitored in the aviation fuel industry (Gaylarde, et al., 1999, Itah, et al., 2009,
McFarlane, 2011).
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FRAS Technology AS, Industrial Microbiology Reviews nr 1. 2013
Microbial growth in jet fuel has even been identified as the primary source of a plane crash in
1958. This was caused by fuel filter clogging in a B- 52 bomber (Rauch, et al., 2006).
This review highlights the current literature on biofouling and MIP, reports from FRAS
technology AS and my own experience working with biofouling and MIP in lubrication and
hydraulic applications from the maritime and offshore industry. Also included in this review is
the new paradigm in lubricant technology, namely the EAL, also referred to as Ecolabel
lubricants. From a microbial point of view, the EAL could be more prone to biodeterioration in
systems, during operation, based on their environmentally friendly chemistries. Therefore, are
there any reasons for additional concern emphasizing systems and fluid integrity using the EAL?
2. A wide variety of organisms are capable of colonizing lubrication systems.
The marine environment is abundant in microorganisms, typically in the range of 107
- 109
organisms L-1
. Microbial numbers in the marine environment will vary greatly depending on a
variety of factors. E.g. microbial numbers will be relatively low in oligotrophic (nutrient
deprived) waters such as in the middle of the Pacific Ocean to very high in eutrophic (nutrient
rich) basins or river outlets. In this review, the focus is on organisms which are able to degrade
petroleum hydrocarbons, the EAL and additives. However, other organisms unable to degrade
the lubricants (petroleum based and environmentally acceptable chemistries) or additives could
also be able to colonize such systems. This requires that they are able to utilize metabolites
derived from lubricant and/ or additive degradation, and display tolerance against toxic
petroleum hydrocarbons and additives. Since the EAL are significantly less toxic and more
biodegradable, it is reasonable to assume the presence of a higher microbial diversity, capable
of degrading the EAL, additives and the metabolites produced.
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FRAS Technology AS, Industrial Microbiology Reviews nr 1. 2013
2.1. Chemistries and biodegradation of petroleum based lubricants.
The petroleum based lubricants are a mixture of hydrocarbons derived from the refining of
crude oil. The hydrocarbon groups found in lubricants are alkanes (paraffins), branched alkanes
(isoparaffins), cycloalkanes (Naphthenes) and aromatics. The abundance of the different groups
varies considerably, depending on the specific application where the lubricant is to be used
(Anderson, et al., 2003). The abundance of aromatics, however, is usually minimized, due to
some undesired properties of aromatics, such as poor oxidation stability and carcinogenicity.
Because of the different compounds in crude oils, base stocks used in the formulation of
lubricants are classified according to the American Petroleum Institute (API).
The criteria according to API are summarized in table 2. API group II and III are the most
oxidation resistant, with the latter being the most resistant (Sharma & Stipanovic, 2003).
Table 2. API classification of lubricant base stocks
API group Saturates (%) aromatics (%) Viscosity index sulfur (%)
I < 90 > 10 < 120 > 0.03
II > 90 < 10 80- 120 < 0.03
III > 90 < 10 > 120 < 0.03
Adapted from Sharma & Stipanovic, 2003
In general, the biodegradability of hydrocarbons follows in decreasing order; alkanes,
isoalkanes, low molecular weight aromatics, cycloalkanes and polycyclic aromatic hydrocarbons
(PAH`s) (Huesemann, 1995). Another important factor in the biodegradability of lubricants is
the viscosity. Low viscosity oils are in general more biodegradable than the high viscosity oils
(Haus, et al., 2004). Hydrocarbon degrading microorganisms are ubiquitous in nature (found in
soil, water and sediments all over the globe) and include bacteria, archaea, fungi and some
algae (Röling, et al., 2003, Antić, et al., 2006, Head, et al., 2006).
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FRAS Technology AS, Industrial Microbiology Reviews nr 1. 2013
Hydrocarbon degradation also occurs deep down in oil reservoirs if the temperature is below
90o
C (Philippi, 1977, Magot, et al., 2000). Also, evidence indicates that heavy crude oil (rich in
resins and asphaltenes) is a result of biodegradation of the lighter hydrocarbon fractions (Head,
et al., 2003). Hydrocarbon degradation also occurs at subzero temperatures (Rike, et al., 2003).
Various hydrocarbons enter the environment naturally through reservoir seepage, incomplete
combustion of wood and plant secretions of waxes. These natural sources have been
introducing hydrocarbons into the biosphere over millions of years, and microorganisms have
adapted to metabolize these energy rich substances. Microbial hydrocarbon degradation is
initiated via four principle routes. The most efficient way of degrading hydrocarbons is to
enhance its surface and water solubility. This is achieved through the production of
biosurfactants, resulting in emulsification of hydrocarbons, thus increasing its bioavailability
(Bouchez Naïtali, et al., 1999). This way of initiating biodegradation of petroleum hydrocarbons
in addition to the sorption capacity and high cell density is probably one of the reasons for the
increased extent of biodegradation seen in biofilm cultures (Singh, et al., 2006, Jain, et al.,
2011). Organisms in the biofilm matrix may generate higher amounts of biosurfactants
compared to the planktonic population, thus some biosurfactants are important components of
the EPS in the biofilm matrix (Flemming & Wingender, 2010). Oil emulsions are consequently
one of the leading causes of MIP. Emulsions formed will easily clog filters nozzles and valves.
Also, if such emulsions are transported between load carrying surfaces it could lead to loss of
lubrication film resulting in excessive friction. Other strategies to access hydrocarbons are;
micrometer sized droplets dispersed in the water column, direct attachment to oil slick/
droplets and degradation of the water soluble hydrocarbon fraction. The latter yields low
amounts of biomass as the water solubility of hydrocarbons is very low. As an example, the
solubility of hexadecane in water is 0,029mg L-1
(Bai, et al., 1997). The water solubility of
saturated hydrocarbons decreases with increasing molecular weight. Reasons for the well
recognized problems with biofouling and MIP in the fuel industry is that diesel, for example,
consists of a lighter fraction of hydrocarbons and is thus more biodegradable than lubricants
which consists of a heavier petroleum fraction.
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FRAS Technology AS, Industrial Microbiology Reviews nr 1. 2013
On the other hand, the lightest petroleum fraction used in fuels e.g. petrol, is not as
biodegradable due to membrane toxicity of these low molecular weight hydrocarbons
(Sikkema, et al., 1995). Also, gasoline separates quickly from water, whereas diesel fuels tend to
retain and disperse more water than gasoline, thus there is a higher surface/ volume ratio in
which microorganisms can initiate their attack (unpublished observation). Historically, the
fraction that has been reported to be the most vulnerable for microbial attack is the fraction
with boiling range between approximately 150- 400o
C (middle distillate) (McFarlane, 2011).
2.2. Chemistries and biodegradation of additives.
To compensate for weaknesses or enhance other properties of the fluids, a wide variety of
different chemicals are added to the lubricating fluids. The additives of such fluids include pour
point depressant, antioxidants, anti wear agents, viscosity index improvers, rust inhibitors,
defoamants, detergents and dispersants. The additives are typically organic and metallo-
organic compounds. This class of compounds in lubricants has received very little attention
regarding microbial degradation. In fuels, however, additives have been shown to be vulnerable
to microbial attack (Gaylarde, et al., 1999). Many of these compounds contain nitrogen,
phosphorous and sulfur, which are important elements for growth. As an example, anti wear
agents are important additives in lubricants whose role is to form a protecting film on
components of the system. These anti wear agents are usually based on organophosphates,
such as triphenyl phosphorothionate (Mangolini, et al., 2011). This group of compounds is
readily degraded by a wide variety bacteria and fungi. In general, organophosphates (e.g. tri- n-
butyl phosphate) are toxic to algae but not bacteria (Michel, et al., 2004). Antioxidants are
another important additive, which typically contains amines and phenols. Emphasizing the
important role of additives and their vulnerability to microbial attack during usage, the
biodegradation of these compounds should be studied in more detail. The additives of the EAL
must also have a higher biodegradability than the additives used in petroleum based fluids. It
raises additional concerns using even more degradable additives.
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FRAS Technology AS, Industrial Microbiology Reviews nr 1. 2013
This is especially true when the role of the additives is to compensate for weaknesses in the
base fluid. For example, the degradation of viscosity index improvers would certainly reduce
the properties of the fluid during either cold or hot operational temperatures. Deterioration of
additives could therefore lead to severe reduction of important properties of the fluids
(lubricity, hydrolytic and oxidative stability etc). Also, based on their chemistry the additives
might serve as a source of phosphorus, sulfur and nitrogen, which could speed up the process
of hydrocarbon degradation or the deterioration of EAL.
2.3. Biodegradation of EAL
The whole idea of the EAL is their low- toxicity and their high biodegradability compared to the
petroleum based lubricants. For the lubricant to be considered as environmentally acceptable it
has to be more than 60% biodegradable in 28 days at 25o
C. The biodegradability is determined
using the OECD 301B and 301D, CEC -L-33-A-94, EPA 560/6-82-003 and ASTM D-5864 test
protocols. The biodegradability is measured as a mean of ultimate (substances is converted
CO2, water and biomass) or primary degradation (modification of physical chemical properties
of the substance). Because of the lower toxicity and higher biodegradability compared to
petroleum based fluids, it is likely that a larger microbial diversity is able to degrade these
fluids. Also, microbial degradation of the EAL will most likely occur at a higher rate and extent,
thus causing problems earlier than microbial degradation of petroleum based lubricants. That
is, if and only if, water is present in sufficient quantities. In addition, the new criteria for the
next generation EAL are that it must contain a minimum of 50% renewable materials, according
to the EPA 800-R-11-002 standard. In this review, however, I will discuss the base stocks for EAL
approved at present time. It is also worth mentioning that fully formulated EAL could be a mix
of different base stocks. A common blend of base stocks is for example polyalphaolefins mixed
with vegetable oil.
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FRAS Technology AS, Industrial Microbiology Reviews nr 1. 2013
2.3.1. Polyalphaolefins (PAO`s)
This class of lubricant is synthesized from olefins (alkenes) found in petroleum and usually with
ethylene to form saturated branched hydrocarbons i.e. isoalkanes. The PAO`s are therefore not
biobased and It has the lowest bioavailability of the EAL based lubricants. The PAO`s are
equivalent and even have some superior properties over the petroleum based fluids, such as
excellent oxidation and hydrolytic stability. Compared to the petroleum based lubricants, the
PAO`s are significantly more biodegradable (at least the lower viscosity PAO`s) even though
alkanes are in general more biodegradable than branched alkanes. Based on the chemistry of
the PAO, the initial biodegradation process involves enzymes related to hydrocarbon
degradation such as monooxygenases and dioxygenases (van Beilen & Funhoff, 2005, van
Beilen, et al., 2006). The microbial diversity capable of degrading the PAO`s may therefore be
somewhat similar to the diversity able to degrade petroleum hydrocarbons, especially those
organisms that are able to degrade alkanes, isoalkanes and cycloalkanes.
2.3.2. Triglycerides (plant oils)
These EAL are based on plant oils, such as rapeseed or castor oils. The triglyceride based
lubricants are the most biodegradable base stock. This is primarily due to a higher
bioavailability of such oils, and a larger diversity of microorganisms able to degrade triglycerides
compared to petroleum hydrocarbons. After all, fatty acids are found in every cell in the
biosphere, and most species possess pathways for fatty acid degradation e.g. β- oxidation.
Another challenge regarding vegetable oils is their low oxidation stability and low hydrolytic
stability. Therefore, vegetable based lubricants is suitable only in low temperature applications
and in open machinery such as chainsaws (Nagendramma & Kaul, 2012). Interestingly, from a
totally environmental point of view, from crop production, using fertilizers, harvesters etc to
use in machinery, with higher turnover of lubricants and spare parts, vegetable oils could have
a higher environmental impact than petroleum based lubricants, except for CO2 emissions
(McManus, et al., 2003).
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FRAS Technology AS, Industrial Microbiology Reviews nr 1. 2013
The latter citation is nearly 10 years old, thus new additives could have been developed which
could reduce some of the environmental impacts described in their work.
2.3.3. Synthetic esters
Synthetic esters (SE) have been used as lubricants for more than fifty years and have many
desirable properties, including good low temperature characteristics, high viscosity index, high
thermo- oxidative stability, good hydrolytic stability and good anti-wear properties. As base
stock for lubricants, two types of SE are available. These are the diesters and polyol esters and
are composed of dicarboxylic acids and fatty acids attached to an alcohol, respectively
(Nagendramma & Kaul, 2012). SE are readily degraded by microorganisms due to the
ubiquitous nature of esters in the biosphere. The biodegradation involve esterase enzymes
which splits the esters into an acid and an alcohol through hydrolysis (Kawai, 2012).
2.3.4. Poly glycols
Poly glycols or polyalkylene glycols (PG), could be either water soluble or water insoluble based
on molecular weight of the polymer i.e. what precursor it is derived from (EPA 800- R- 11- 002).
The water soluble glycols are the most biodegradable. On the other hand, it is also the most
toxic of the PG. If the recipient is small and lubricant loss is large, the concentration of glycols in
water could be very high and toxic to organisms in its proximity. There is a wide variety of
organisms able to degrade poly glycols, with the low molecular weight glycols being the most
biodegradable (Kawai, 2012). PG is being successfully used in vessels using stern tubes for
propulsion. This type of EAL can also tolerate significant higher percentage water
contamination because it can be water soluble (Sada et al., 2008). However, for thruster
application this type of EAL does not provide sufficient lubricity (Sada et al., 2009).
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FRAS Technology AS, Industrial Microbiology Reviews nr 1. 2013
3. Cases where microorganisms are a probable cause of machinery malfunctioning in
lubrication and hydraulic systems using mineral based lubricants.
Case 1.
Hydraulic control valves from an oil drilling platform (fig 1) were sent to FRAS technology AS for
failure investigation. The unit was expected to have a duration period of approximately 10
years. After just 9 months the crane which controlled the drilling cone started to respond in an
unpredictable manner.
Upon arrival the unit was opened and the unit interior was found to be completely destroyed
with the presence of elemental sulfur (S0
) and aluminum oxide. Obviously, strong galvanic
corrosion had taken place between the steel and aluminum parts of the valve, indicating the
presence of a liquid with good electric conductivity in the valve (in lieu of the hydraulic oil),
such as seawater. The presence of S0
must be considered as a hallmark of an underlying
microbial process in such systems (Tang, et al., 2009). Most likely the sulfur originated from
sulfate via sulfide, which is present in high amounts in seawater.
Figure 1. A. Hydraulic control unit from an oil drilling platform, where microbial processes has
deteriorated its interior. The unit was supposed to endure ten years of operation but this
photograph was taken only after nine months of operation (photo taken by FRAS tech AS). B.
Scanning electron micrograph, showing both coccoid and bacilli shaped microorganisms (with
permission from Elin Ørmen at the Norwegian University of life sciences).
A B
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FRAS Technology AS, Industrial Microbiology Reviews nr 1. 2013
The mechanism behind the detrimental effects seen in this control unit is beyond the scope of
this review, but it involves very complex chemical reactions initiated by bacteria. However, it is
tempting to propose a short explanation, sulfate reducing prokaryotes (SRP) are strictly
anaerobic organisms, and do not grow in the presence of oxygen. Interestingly, the microbial
conversion of sulfide to S0
, requires oxygen. One of the probable scenarios therefore follows;
SRP in anaerobic conditions causes the interior to deteriorate, sealing material is compromised
and the organisms in the unit are exposed to oxygen due to air ingress. This leads to a shift in
the microbial community where sulfide is oxidized to S0
by colorless sulfur bacteria, such as
Thiobacillus sp (Tang et al., 2009). Also possible is the presence of a biofilm which contain both
anaerobic and aerobic zones. This would sustain the simultaneous growth of both the
anaerobic SRP and the aerobic colorless sulfur bacteria.
Case 2.
A ship’s thruster had a breakdown after a long period of downtime. According to the failure
report (pers com CEO Sølve Fjerdingstad at FRAS technology AS and Dr. John Olav Nøkleby at
DNV GL), the malfunctioning was caused by the lack of lubrication, resulting in massive stress
on the mechanical parts. The damaged parts were sent in for mechanical inspection and a slimy
substance was literally pouring out upon opening (fig 2). This slimy material is most likely not of
mechanical origin, but most likely a result of microbial growth. Mechanically produced sludge is
often highly viscous or semi solid and does not behave like a slimy substance, due to the
presence of wear debris, oxidation products and sometimes water. A microbial survey was not
initiated due to late recognition that this could be a microbial problem rather than a technical
issue.
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FRAS Technology AS, Industrial Microbiology Reviews nr 1. 2013
Figure 2. Slime of microbial origin and emulsified oil pouring out from the bearing part of a
thruster (photo taken by FRAS tech AS)
Case 3.
A supply vessel had been in dry dock for maintenance due to unknown machinery breakdown.
During this maintenance the lubrication oil was most likely not changed, nor was the system
checked for the presence of any free water. Shortly after the vessel was put into operation after
the maintenance procedure, the ship’s lubrication system stopped and the reason was found to
be filter clogging. Since the vessel had been in dry dock for maintenance and no reasons for the
previous breakdown were found, the lubrication oil was drained and the oil sump was
thoroughly cleaned using a steamer. During steaming, flakes of paint loosened from the oil
sump surface (fig 3).
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FRAS Technology AS, Industrial Microbiology Reviews nr 1. 2013
Surface of the paint flake Yeasts Fungal spores Fungal hyphae
EPS
Bacteria
Figure 3. Scanning electron micrograph showing various microorganisms and a thin biofilm, covering the
surface of a paint flake sample (scale bar indicate 2µm).
The engineers responsible for cleaning and maintenance observed material on the surface
which they did not believe had any mechanical or chemical origin. The flakes of paint were sent
to The Norwegian University of life sciences via Fras technology AS for microbial analysis. After
cultivation, nine different bacterial genera were identified in addition to three fungal genera
(Table 3). I will here give a short argumentation for the resulting filter clogging. During dry dock,
the water in the lubrication system was stagnant. This leads to the formation of lose and patchy
biofilms covered with oil emulsions, on surfaces and in the oil/ water interface (Fjeld, 2012
unpublished results). A possible reason for this unexpected occurrence of filter clogging is that
these biofilms with low mechanical stability and bound emulsions will be disrupted when the oil
again is in circulation. Consequently biofilm material and emulsified oil will follow the liquid
flow, until it reached the filters, leading to filter clogging.
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FRAS Technology AS, Industrial Microbiology Reviews nr 1. 2013
Table 3. Organisms identified from the cultivated oil phase, water phase and biofilm, size of
query sequence, similarity with the closest match in the NCBI BLAST database and the proposed
taxonomic affiliation of the organisms.
Proposed taxonomic
affiliation
Query
sequence
(bases)
Closest match in the BLAST database Max
identity
(%)
Taxonomic rank
Bacteria
Acinetobacter 1500 Acinetobacter sp GXA5 (AY902243.1) 98% Gamma
proteobacteria,
pseudomonadales
Shewanella ** 1493 Shewanella sp. W3-18-1 (CP000503.1) 99% Gamma
proteobacteria,
alteromonadales
Delftia 1495 Delftia sp. LFJ11-1 (DQ140182.1) 99% Beta proteobacteria,
burkholderiales
Comamonadaceae
Achromobacter ** 1492 Achromobacter sp. MT-E3 (EU727196.1) 99% Beta proteobacteria,
burkholderiales
Pseudomonas 1497 Pseudomonas sp. VKM B-2265 (DQ264636.1) 94% Gamma
proteobacteria,
pseudomonadales
Variovorax ** 1488 Variovorax sp.RKS7-5 16S (EU934231.1) 96% Beta proteobacteria,
burkholderiales
Comamonadaceae
Chryseobacterium * 1384 Chryseobacterium sp. B2 (EU109732.1) 96% Flavobacteria,
Flavobacteriales,
Flavobacteriaceae
Serratia * 1492 Serratia sp. GU124497.1 95% Gamma
proteobacteria,
Enterobacteriales
Bradyrhizobium * 1427 Uncultured Bradyrhizobium sp. clone PSC8 99% Alpha
proteobacteria,
Rhizobiales
Fungi
Phialophora ** 625 Phialophora sp. WRCF- AB8 (AY618680.1) 100% Ascomycota,
Pezizomycotina,
Sordariomycetes
Fusarium ** 973 Fusarium solani strain ATCC 56480
FJ345352.1
98% Ascomycota,
Pezizomycotina,
Sordariomycetes
Phoma ** 1024 Phoma sp. GS9N1a (AY465466.1) 100%
Ascomycota
Dothideomycetes
Pleosporomycetidae
* Organisms only found in the biofilm samples
** Organisms only found in the liquid (oil and water) samples.
18
FRAS Technology AS, Industrial Microbiology Reviews nr 1. 2013
Case 4.
This case is not from the marine or offshore industry but still, it is important in the
documentation of microorganisms that cause problems in lubrication oil systems. A project was
initiated in 1996, called the baktiol (Bakterier I olje, or Bactiol; Bacteria in oil) project, ISBN 82-
576-9804-0. Senior researcher Jon Fredrik Hanssen and Dr. Aaslaug Lode at the Agricultural
University of Norway (NLH), (now the Norwegian University of Life Sciences (NMBU)) initiated a
thorough microbial investigation (Financed by the Norwegian research council and Statkraft
engineering) based on traditional microbial techniques. During that time, new ash less
lubricant additives (additives not containing metals) came into the market and industries were
encouraged to use the more environmentally friendly lubricants. The hydroelectric industry in
Norway noted a sudden increase in maintenance cost due to more frequent oil replacements
and maintenance of machinery in some installations. The hydroelectric industry rarely replaces
their turbine lubricants (pers com Dr. John Olav Nøkleby at DNV). Samples with high particle-
and water content were received from Det Norske Veritas (DNV). The samples were heavily
contaminated (opaque appearance, a lot of wear debris, high water content etc) oil samples
from many different hydroelectric power plants in Norway. Of 67 oil samples, 24 contained
microorganisms based on microscopic examination and culturing. The oil samples that
contained microorganisms also contained free water. If polymerase chain reaction (PCR) based
techniques had been used, the number of samples testing positive for microorganisms would
probably be higher. The organisms isolated included hyphael fungi, yeasts and bacteria. Most of
the isolates were able to grow between 20 and 40o
C and some isolates grew well at 4o
C. This
study provided recommendations concerning reduction of especially water content but also the
input of air in open oil systems. One drawback of their study was the lack of establishing the
threshold of interference in such machinery. This threshold is very important if correct
conclusions are to be drawn, thus microorganisms does not cause problems until they exceed a
certain number of cells. Microorganisms will for sure be present in water that enters the
systems and hydrocarbon degrading organisms are ubiquitous in nature. Therefore, the finding
of microorganisms in water phase or in oil does not necessarily imply biofouling and MIP.
19
FRAS Technology AS, Industrial Microbiology Reviews nr 1. 2013
Additional factors that should be addressed if MIP is to be elucidated are the amount of water
in systems, cellular numbers and the presence of events that cannot be explained by
mechanical causes. Nevertheless, biocide treatment and short term follow up resulted in
remission and maintenance cost was reduced, indicating that microbial growth was the cause
of the problems experienced by the industry.
4. Future challenges with emphasis on environmentally acceptable lubricants
EAL are now considered to be the new paradigm in lubrication technology. A large amount of
lubricants are lost to the environment annually, which may have serious ecotoxicological effects
(Michel, et al., 2004). The change into using such EAL has been pivotal for industries such as
forestry and other terrestrial industries where lubricant loss is extensive. Because of
environmental regulations, industries are encouraged into adapting EAL (forced in US waters).
From a microbial and ecological point of view, they are, as already discussed, more
biodegradable and could have less toxicological impact on their surroundings. This fact raises
the question of whether these lubricants will deteriorate at a higher rate in systems during
operation, if the systems have been contaminated with water. It is feasible to assume so, with
reduced operational time and increased maintenance costs (downtime, replacement of spare
parts, cleaning and disinfection) as a result. Biofouling and MIP in the marine industry are easily
confused with mechanical causes and the incidence of MIP is therefore unknown, even with the
use of petroleum based lubricants. Case nr 2 and 3 are good candidates for such confusion as
they were initially reported as machinery malfunctioning of unknown causes. Therefore, there
are reasons to believe that such problems could be underestimated even with the use of
petroleum based lubricants. Bacterial infection of systems could coincide with other
operational and mechanical occurrences. This could lead to a more rapid reduction in
operational time of the systems and compromising of the integrity of the lubricating fluids, than
if mechanical causes were the sole cause for detrimental effects.
20
FRAS Technology AS, Industrial Microbiology Reviews nr 1. 2013
If the EAL are going to successfully replace the petroleum based lubricants, the industries
should implement a microbial surveillance protocol to monitor and detect any possible
detrimental effects caused by microorganisms. If microbial problems are increasing
dramatically in the future and future environmental regulations are not making an exception
for the maritime industry, new systems design and construction materials should be
considered. I would also recommend a higher degree of cross disciplinary cooperation where
microbiology should be considered if reasons for machinery malfunctioning or other adverse
events cannot be explained by a traditional approach. Indeed, the EAL have replaced petroleum
based lubricants with great success in industries such as forestry with emphasis on direct
environmental impact. Also, synthetic esters have been used extensively as jet engine
lubricants with great success for several decades, due to many superior properties over
petroleum based lubricant. In Forestry a rapid and complete degradation are desirable in
combination with some superior properties of the EAL. It should be noted that care should be
exerted regarding systems in which water is a common contaminant.
5. Discussion and Conclusions
To understand MIP and biofouling in lubrication oil systems we need understanding of the
underlying processes and to acknowledge that microorganisms, regardless of their small size,
are able to cause heavy machinery to break down. The big unknowns in this respect are the
threshold of interference and the extent of such problems in the maritime industry. The
amount of free water is the driving force in exceeding the limit needed for interference.
Microbial growth in systems must be avoided. The best practice to minimize biofouling and MIP
is to have appropriate procedures for oil storage and minimize the amount of free water in
systems. The four cases described were initially reported as unknown cause of machinery
breakdown. This report illustrates that there is a pertinent need for cross disciplinary
cooperation if correct mitigation strategies are to be adapted.
21
FRAS Technology AS, Industrial Microbiology Reviews nr 1. 2013
To my knowledge, microorganisms are not generally considered as a possible cause of adverse
events in lubrication systems. However, Cases 3 and 4 are the few exceptions where microbial
identification techniques were employed, after other causes had been ruled out. Some
hallmarks of microbial activity in industrial applications include; the presence of slime, the odor
of sulfide (H2S) produced by sulfate reducing prokaryotes (SRP), or other foul odor and
precipitation of sulfur aided by for example Thiobacillus sp (Tang, et al., 2009). From the
experiences with MIP and biofouling in systems using petroleum based lubricants, the problems
usually occur after maintenance or other reasons for machinery downtime. I.e. biofilms with
low mechanic stability is formed during low shear forces, and are easily disrupted when the
system are put into operation with increased shear forces (Stoodley, et al., 2002). Biofilm
material and emulsified oil will follow liquid flow and could lead to clogging of nozzles, valves
and filters and friction increase when trapped between load carrying surfaces.
Also, before adapting the EAL, I would recommend to determine the threshold of interference
for any given system, i.e. how many organisms are required for problems to occur and how
much free water does this require.
Establishing early warning systems is also important to avoid MIP due to biofouling, especially
in systems using EAL. Water control and water prevention actions will therefore be even more
important with the use of the eco label lubricants.
22
FRAS Technology AS, Industrial Microbiology Reviews nr 1. 2013
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Current knowledge on microbial induced problems and biofouling in lubrication systems of ships and marine installations (2)

  • 1. 1 FRAS Technology AS, Industrial Microbiology Reviews nr 1. 2013 Current knowledge on microbial induced problems and biofouling in lubrication systems of ships and marine installations Christer Moe Fjeld FRAS Technology AS Kongeveien 30 1430 Ås, Norway Summary Microbial induced problems (MIP) and biofouling in lubrication oil systems are probably rare and usually an occult event. MIP and biofouling are in most cases assumed to be of mechanical origin and mitigation is undertaken without knowledge of the underlying cause. Because of this, the extent of such problems is not known, but assumed to be rare. Based on the ubiquitous nature of microorganisms and their versatile arsenal of metabolic properties, it is not surprising that they are capable of causing detrimental effects in a wide variety of industries. Industries using oils for lubrication purposes are no exception. If water is present and the water activity is sufficient, the systems are prone to microbial colonization, which may in turn result in several adverse effects. Lubricants and especially spent lubricants, are toxic to many organisms, thus environmental challenges are indeed a legitimate concern. These days, several industries that are using oils for lubrication purposes are shifting to a “green” line in their choice of fluids. In industries such as forestry and other terrestrial applications, these lubricants are excellent replacements to the petroleum based lubricants. However, care should be exerted in maritime industries since water will usually be found in various amounts in the lubrication systems. The Environmentally Acceptable Lubricants (EAL) are more biodegradable than the traditional oils based on petroleum. Therefore, it is reasonable to assume that these fluids are more prone to biodeterioration in the lubrication systems. Because of the higher biodegradability of the EAL, there is a risk that microbial induced problems and biofouling will increase in the future. This emphasizes the importance of increased monitoring of such problems in the marine industry. In this review, I used reports from FRAS technology AS and the current literature with the aim of trying to explain some of the reasons for unknown machinery breakdown. In addition, I present some cases where microorganisms are the cause of machinery breakdown, either directly or indirectly. Key words: Petroleum based lubricants, environmentally acceptable lubricants, additives, biofouling Correspondence: Christer@fras.no Kongeveien 30, 1430 Ås, Norway
  • 2. 2 FRAS Technology AS, Industrial Microbiology Reviews nr 1. 2013 1. Introduction Microbial growth is not as solitary and simple as one would believe. Microbial growth and colonization is complex, involving social interactions, synergistic cooperation, intense competition for nutrients and a very large microbial diversity over a very small area. The presence of active and proliferating microorganisms are ubiquitous in nature, including sodium chlorine brine water (cryopegs) at sub zero temperatures in the Antarctic (Gilichinsky, et al., 2005) and in the vicinity of hydrothermal vents on the oceans floor where microorganisms are proliferating in temperatures at around 121o C (Kashefi & Lovley, 2003). The strain identified by Kashefi and Lovley was also able to grow at an astonishing 130o C. Microorganisms are also found in acidic and alkaline environments. The general requirements for microbial growth to occur is the presence of free water (or a water activity aw > 0.85 for most microbes), a carbon source (glucose, cellulose, hydrocarbons, lipids, proteins etc), for the heterotrophs and CO2 for the autotrophs, and various amounts of nitrogen and phosphorous depending on microbial species (oligotrophic organisms are able to grow in very pure water e.g. Caulobacter sp.) In addition microorganisms require micronutrients such as sulfur, iron, manganese etc for optimal growth. The key factor involved in biodegradation is usually the bioavailability of compounds with a few exceptions such as polycyclic aromatic hydrocarbons (PAH) due to higher water solubility than many long chain alkanes (Huesemann, et al., 2004). In general, hydrophilic compounds have a higher bioavailability than hydrophobic compounds (Bosma, et al., 1996). When microorganisms are in contact with a moist surface, they will attach. After some time (hours to days depending on environmental factors and species present) they will form a complex 3D structure composed of extracellular polymeric substances (EPS; carbohydrates, lipids, proteins and DNA), cells and water (Costerton, 1995, Sutherland, 2001, Hall-Stoodley, et al., 2004). This is referred to as a biofilm. Dental plaque is a good example of a typical biofilm. Cells that are imbedded in the EPS matrix are provided with both shelter from predation, antimicrobial agents, chemical stress and UV radiation (Carpentier & Cerf, 1993, Lewis, 2008, Flemming, 2009).
  • 3. 3 FRAS Technology AS, Industrial Microbiology Reviews nr 1. 2013 Another benefit of living in close proximity in biofilms is the facilitation of metabolic cooperation between different species (Costerton, 1995), However, strong competition also occur (Elias & Banin, 2012). Biodegradation of complex organic compounds such as hydrocarbons is often more efficient in biofilm cultures compared to planktonic degradation of hydrocarbons (Jain, et al., 2011). With this short introduction into microbial processes in nature it is easy to understand that microorganisms are able to cause adverse effects in a wide variety of manmade applications. When biofilms are established in undesired locations in manmade applications it is referred to as biofouling, i.e. a biofilm in the wrong place. Biofouling has been identified as a cause of severe adverse effects in a wide variety of industries (Table 1). Table 1. Industries and applications known to be adversely affected by microbial induced problems and biofouling Industry or application Manifestation of problems References Petroleum industry Microbial influenced corrosion (MIC) and souring of applications and crude oil respectively. (Neria-González, et al., 2006, Gieg, et al., 2011) Shipping industry MIC of ships hulls, Increased drag and spread of non indigenous species through ballast water. (Holm, et al., 2004, Drake, et al., 2007) Paper industry Product withdrawal, increased maintenance costs and machinery downtime. (Klahre & Flemming, 2000) Metal working industry Reduced properties of fluids and, the development of hypersensitive pneumonitis in workers and various dermatological symptoms (Kreiss & Cox-Ganser, 1997, van der Gast, et al., 2001, Lucchesi, et al., 2012) Hydroelectric power plants Increased friction in lower vane bearing bushings. (Schneider, et al., 2006) Medical applications Biofilms on prosthetic joints and other implants causing infection. Biofilms in hospital environments may cause nosocomial infections. (Zimmerli, et al., 2004, Lindsay & von Holy, 2006)
  • 4. 4 FRAS Technology AS, Industrial Microbiology Reviews nr 1. 2013 Biofouling and microbial induced problems (MIP) in lubrication systems in any industry are directly associated with the presence of free water in the systems (aw > 0.85). This is the main reason why emulsion type metal working fluids (mixture between oil and water) are especially prone to severe microbial contamination and the resulting detrimental effects (Mattsby- Baltzer, et al., 1989). Water may enter the hydraulic/ lubrication systems via three independent routes: 1. Oil storage barrels are often stored outdoors and on the deck of ships. Water spray from the sea will definitely contain hydrocarbon degrading organisms, independent of geographic location. Ambient temperature fluctuations will cause pressure and vacuum shifts of the air in the barrel’s head space. Water with hydrocarbon degrading organism will enter the barrels through the sealing if water is present on top of the barrels. 2. Humidity in the air may precipitate in lubrication oil tank and humid air could also be dissolved in the oil and precipitate in cooler areas of the system, such as in ships thrusters or in stern tubes. 3. Water is frequently introduced to systems through heating and cooling events during operation, pressure fluctuations as a result of propeller movement and as a result of the vessel’s movement in the water column. I.e. stern tubes and thrusters will experience pressure variation due to waves, resulting in a shifting water depth around the propeller. To compensate for this pressure variation and thus to prevent severe water ingress, pressure is applied to the systems. This overpressure leads to lubricant loss when the surrounding pressure is below the pressure applied on the system. This occurs for example when the propeller is surfacing during rough seas. However, no water ingress is difficult to achieve.
  • 5. 5 FRAS Technology AS, Industrial Microbiology Reviews nr 1. 2013 To my knowledge, scientific literature on biofouling in systems using oils for lubrication purposes has been absent for nearly two decades. Some work, however, was undertaken in the 1970`s, 80`s and early 90`s regarding microbial growth in lubrication systems (Summers-Smith, 1982, Okpokwasili & Okorie, 1988, Stuart, 1994-1995). The extent of such problems these days is therefore unknown. One of the reasons could be that the manifestation of biofouling in such systems is easily confused with mechanical issues. Previously, the industries were more post active, after problems occurred, with water drainage of systems and other maintenances necessary to mitigate the occurring detrimental effects. Upon opening of systems they would probably observe large quantities of slime, notice foul odor etc, which cannot be explained by a standard mechanical approach. Such observations would lead to suspicion towards microbial contamination. Nowadays, however, systems are under more thorough surveillance, thus the industries are more proactive in avoiding adverse occurrences, especially with emphasis on water contamination and increased wear (commonly measured by particle count according to the SAE AS 4059 standard). Therefore biofouling and the following MIP in such systems are most likely evolving more slowly, based on the small body of water compared to the bulk amount of lubricant. I.e. the amount of water present is not sufficient to promote extensive growth with rapid manifestation of detrimental effects. On the other hand, biofouling could speed up the expected mechanical wear of systems, by facilitating emulsification of oils, foaming and sludge formation. Relevant questions are, therefore: Have any shipping companies had a higher maintenance cost than predicted in some vessels? Are there any plausible explanations for this possible increase in maintenance cost? Does any company report malfunctions or incidences which cannot be explained by a standard mechanical approach? If no mechanical or technical explanation can explain the condition, microbial causes should be considered. Fuels on the other hand are well known to contain harmful microorganisms and their associated problems. Therefore, biofouling and MIP have for a long time been known in the fuel industry and accepted as a cause of numerous machinery breakdowns and are thus especially monitored in the aviation fuel industry (Gaylarde, et al., 1999, Itah, et al., 2009, McFarlane, 2011).
  • 6. 6 FRAS Technology AS, Industrial Microbiology Reviews nr 1. 2013 Microbial growth in jet fuel has even been identified as the primary source of a plane crash in 1958. This was caused by fuel filter clogging in a B- 52 bomber (Rauch, et al., 2006). This review highlights the current literature on biofouling and MIP, reports from FRAS technology AS and my own experience working with biofouling and MIP in lubrication and hydraulic applications from the maritime and offshore industry. Also included in this review is the new paradigm in lubricant technology, namely the EAL, also referred to as Ecolabel lubricants. From a microbial point of view, the EAL could be more prone to biodeterioration in systems, during operation, based on their environmentally friendly chemistries. Therefore, are there any reasons for additional concern emphasizing systems and fluid integrity using the EAL? 2. A wide variety of organisms are capable of colonizing lubrication systems. The marine environment is abundant in microorganisms, typically in the range of 107 - 109 organisms L-1 . Microbial numbers in the marine environment will vary greatly depending on a variety of factors. E.g. microbial numbers will be relatively low in oligotrophic (nutrient deprived) waters such as in the middle of the Pacific Ocean to very high in eutrophic (nutrient rich) basins or river outlets. In this review, the focus is on organisms which are able to degrade petroleum hydrocarbons, the EAL and additives. However, other organisms unable to degrade the lubricants (petroleum based and environmentally acceptable chemistries) or additives could also be able to colonize such systems. This requires that they are able to utilize metabolites derived from lubricant and/ or additive degradation, and display tolerance against toxic petroleum hydrocarbons and additives. Since the EAL are significantly less toxic and more biodegradable, it is reasonable to assume the presence of a higher microbial diversity, capable of degrading the EAL, additives and the metabolites produced.
  • 7. 7 FRAS Technology AS, Industrial Microbiology Reviews nr 1. 2013 2.1. Chemistries and biodegradation of petroleum based lubricants. The petroleum based lubricants are a mixture of hydrocarbons derived from the refining of crude oil. The hydrocarbon groups found in lubricants are alkanes (paraffins), branched alkanes (isoparaffins), cycloalkanes (Naphthenes) and aromatics. The abundance of the different groups varies considerably, depending on the specific application where the lubricant is to be used (Anderson, et al., 2003). The abundance of aromatics, however, is usually minimized, due to some undesired properties of aromatics, such as poor oxidation stability and carcinogenicity. Because of the different compounds in crude oils, base stocks used in the formulation of lubricants are classified according to the American Petroleum Institute (API). The criteria according to API are summarized in table 2. API group II and III are the most oxidation resistant, with the latter being the most resistant (Sharma & Stipanovic, 2003). Table 2. API classification of lubricant base stocks API group Saturates (%) aromatics (%) Viscosity index sulfur (%) I < 90 > 10 < 120 > 0.03 II > 90 < 10 80- 120 < 0.03 III > 90 < 10 > 120 < 0.03 Adapted from Sharma & Stipanovic, 2003 In general, the biodegradability of hydrocarbons follows in decreasing order; alkanes, isoalkanes, low molecular weight aromatics, cycloalkanes and polycyclic aromatic hydrocarbons (PAH`s) (Huesemann, 1995). Another important factor in the biodegradability of lubricants is the viscosity. Low viscosity oils are in general more biodegradable than the high viscosity oils (Haus, et al., 2004). Hydrocarbon degrading microorganisms are ubiquitous in nature (found in soil, water and sediments all over the globe) and include bacteria, archaea, fungi and some algae (Röling, et al., 2003, Antić, et al., 2006, Head, et al., 2006).
  • 8. 8 FRAS Technology AS, Industrial Microbiology Reviews nr 1. 2013 Hydrocarbon degradation also occurs deep down in oil reservoirs if the temperature is below 90o C (Philippi, 1977, Magot, et al., 2000). Also, evidence indicates that heavy crude oil (rich in resins and asphaltenes) is a result of biodegradation of the lighter hydrocarbon fractions (Head, et al., 2003). Hydrocarbon degradation also occurs at subzero temperatures (Rike, et al., 2003). Various hydrocarbons enter the environment naturally through reservoir seepage, incomplete combustion of wood and plant secretions of waxes. These natural sources have been introducing hydrocarbons into the biosphere over millions of years, and microorganisms have adapted to metabolize these energy rich substances. Microbial hydrocarbon degradation is initiated via four principle routes. The most efficient way of degrading hydrocarbons is to enhance its surface and water solubility. This is achieved through the production of biosurfactants, resulting in emulsification of hydrocarbons, thus increasing its bioavailability (Bouchez Naïtali, et al., 1999). This way of initiating biodegradation of petroleum hydrocarbons in addition to the sorption capacity and high cell density is probably one of the reasons for the increased extent of biodegradation seen in biofilm cultures (Singh, et al., 2006, Jain, et al., 2011). Organisms in the biofilm matrix may generate higher amounts of biosurfactants compared to the planktonic population, thus some biosurfactants are important components of the EPS in the biofilm matrix (Flemming & Wingender, 2010). Oil emulsions are consequently one of the leading causes of MIP. Emulsions formed will easily clog filters nozzles and valves. Also, if such emulsions are transported between load carrying surfaces it could lead to loss of lubrication film resulting in excessive friction. Other strategies to access hydrocarbons are; micrometer sized droplets dispersed in the water column, direct attachment to oil slick/ droplets and degradation of the water soluble hydrocarbon fraction. The latter yields low amounts of biomass as the water solubility of hydrocarbons is very low. As an example, the solubility of hexadecane in water is 0,029mg L-1 (Bai, et al., 1997). The water solubility of saturated hydrocarbons decreases with increasing molecular weight. Reasons for the well recognized problems with biofouling and MIP in the fuel industry is that diesel, for example, consists of a lighter fraction of hydrocarbons and is thus more biodegradable than lubricants which consists of a heavier petroleum fraction.
  • 9. 9 FRAS Technology AS, Industrial Microbiology Reviews nr 1. 2013 On the other hand, the lightest petroleum fraction used in fuels e.g. petrol, is not as biodegradable due to membrane toxicity of these low molecular weight hydrocarbons (Sikkema, et al., 1995). Also, gasoline separates quickly from water, whereas diesel fuels tend to retain and disperse more water than gasoline, thus there is a higher surface/ volume ratio in which microorganisms can initiate their attack (unpublished observation). Historically, the fraction that has been reported to be the most vulnerable for microbial attack is the fraction with boiling range between approximately 150- 400o C (middle distillate) (McFarlane, 2011). 2.2. Chemistries and biodegradation of additives. To compensate for weaknesses or enhance other properties of the fluids, a wide variety of different chemicals are added to the lubricating fluids. The additives of such fluids include pour point depressant, antioxidants, anti wear agents, viscosity index improvers, rust inhibitors, defoamants, detergents and dispersants. The additives are typically organic and metallo- organic compounds. This class of compounds in lubricants has received very little attention regarding microbial degradation. In fuels, however, additives have been shown to be vulnerable to microbial attack (Gaylarde, et al., 1999). Many of these compounds contain nitrogen, phosphorous and sulfur, which are important elements for growth. As an example, anti wear agents are important additives in lubricants whose role is to form a protecting film on components of the system. These anti wear agents are usually based on organophosphates, such as triphenyl phosphorothionate (Mangolini, et al., 2011). This group of compounds is readily degraded by a wide variety bacteria and fungi. In general, organophosphates (e.g. tri- n- butyl phosphate) are toxic to algae but not bacteria (Michel, et al., 2004). Antioxidants are another important additive, which typically contains amines and phenols. Emphasizing the important role of additives and their vulnerability to microbial attack during usage, the biodegradation of these compounds should be studied in more detail. The additives of the EAL must also have a higher biodegradability than the additives used in petroleum based fluids. It raises additional concerns using even more degradable additives.
  • 10. 10 FRAS Technology AS, Industrial Microbiology Reviews nr 1. 2013 This is especially true when the role of the additives is to compensate for weaknesses in the base fluid. For example, the degradation of viscosity index improvers would certainly reduce the properties of the fluid during either cold or hot operational temperatures. Deterioration of additives could therefore lead to severe reduction of important properties of the fluids (lubricity, hydrolytic and oxidative stability etc). Also, based on their chemistry the additives might serve as a source of phosphorus, sulfur and nitrogen, which could speed up the process of hydrocarbon degradation or the deterioration of EAL. 2.3. Biodegradation of EAL The whole idea of the EAL is their low- toxicity and their high biodegradability compared to the petroleum based lubricants. For the lubricant to be considered as environmentally acceptable it has to be more than 60% biodegradable in 28 days at 25o C. The biodegradability is determined using the OECD 301B and 301D, CEC -L-33-A-94, EPA 560/6-82-003 and ASTM D-5864 test protocols. The biodegradability is measured as a mean of ultimate (substances is converted CO2, water and biomass) or primary degradation (modification of physical chemical properties of the substance). Because of the lower toxicity and higher biodegradability compared to petroleum based fluids, it is likely that a larger microbial diversity is able to degrade these fluids. Also, microbial degradation of the EAL will most likely occur at a higher rate and extent, thus causing problems earlier than microbial degradation of petroleum based lubricants. That is, if and only if, water is present in sufficient quantities. In addition, the new criteria for the next generation EAL are that it must contain a minimum of 50% renewable materials, according to the EPA 800-R-11-002 standard. In this review, however, I will discuss the base stocks for EAL approved at present time. It is also worth mentioning that fully formulated EAL could be a mix of different base stocks. A common blend of base stocks is for example polyalphaolefins mixed with vegetable oil.
  • 11. 11 FRAS Technology AS, Industrial Microbiology Reviews nr 1. 2013 2.3.1. Polyalphaolefins (PAO`s) This class of lubricant is synthesized from olefins (alkenes) found in petroleum and usually with ethylene to form saturated branched hydrocarbons i.e. isoalkanes. The PAO`s are therefore not biobased and It has the lowest bioavailability of the EAL based lubricants. The PAO`s are equivalent and even have some superior properties over the petroleum based fluids, such as excellent oxidation and hydrolytic stability. Compared to the petroleum based lubricants, the PAO`s are significantly more biodegradable (at least the lower viscosity PAO`s) even though alkanes are in general more biodegradable than branched alkanes. Based on the chemistry of the PAO, the initial biodegradation process involves enzymes related to hydrocarbon degradation such as monooxygenases and dioxygenases (van Beilen & Funhoff, 2005, van Beilen, et al., 2006). The microbial diversity capable of degrading the PAO`s may therefore be somewhat similar to the diversity able to degrade petroleum hydrocarbons, especially those organisms that are able to degrade alkanes, isoalkanes and cycloalkanes. 2.3.2. Triglycerides (plant oils) These EAL are based on plant oils, such as rapeseed or castor oils. The triglyceride based lubricants are the most biodegradable base stock. This is primarily due to a higher bioavailability of such oils, and a larger diversity of microorganisms able to degrade triglycerides compared to petroleum hydrocarbons. After all, fatty acids are found in every cell in the biosphere, and most species possess pathways for fatty acid degradation e.g. β- oxidation. Another challenge regarding vegetable oils is their low oxidation stability and low hydrolytic stability. Therefore, vegetable based lubricants is suitable only in low temperature applications and in open machinery such as chainsaws (Nagendramma & Kaul, 2012). Interestingly, from a totally environmental point of view, from crop production, using fertilizers, harvesters etc to use in machinery, with higher turnover of lubricants and spare parts, vegetable oils could have a higher environmental impact than petroleum based lubricants, except for CO2 emissions (McManus, et al., 2003).
  • 12. 12 FRAS Technology AS, Industrial Microbiology Reviews nr 1. 2013 The latter citation is nearly 10 years old, thus new additives could have been developed which could reduce some of the environmental impacts described in their work. 2.3.3. Synthetic esters Synthetic esters (SE) have been used as lubricants for more than fifty years and have many desirable properties, including good low temperature characteristics, high viscosity index, high thermo- oxidative stability, good hydrolytic stability and good anti-wear properties. As base stock for lubricants, two types of SE are available. These are the diesters and polyol esters and are composed of dicarboxylic acids and fatty acids attached to an alcohol, respectively (Nagendramma & Kaul, 2012). SE are readily degraded by microorganisms due to the ubiquitous nature of esters in the biosphere. The biodegradation involve esterase enzymes which splits the esters into an acid and an alcohol through hydrolysis (Kawai, 2012). 2.3.4. Poly glycols Poly glycols or polyalkylene glycols (PG), could be either water soluble or water insoluble based on molecular weight of the polymer i.e. what precursor it is derived from (EPA 800- R- 11- 002). The water soluble glycols are the most biodegradable. On the other hand, it is also the most toxic of the PG. If the recipient is small and lubricant loss is large, the concentration of glycols in water could be very high and toxic to organisms in its proximity. There is a wide variety of organisms able to degrade poly glycols, with the low molecular weight glycols being the most biodegradable (Kawai, 2012). PG is being successfully used in vessels using stern tubes for propulsion. This type of EAL can also tolerate significant higher percentage water contamination because it can be water soluble (Sada et al., 2008). However, for thruster application this type of EAL does not provide sufficient lubricity (Sada et al., 2009).
  • 13. 13 FRAS Technology AS, Industrial Microbiology Reviews nr 1. 2013 3. Cases where microorganisms are a probable cause of machinery malfunctioning in lubrication and hydraulic systems using mineral based lubricants. Case 1. Hydraulic control valves from an oil drilling platform (fig 1) were sent to FRAS technology AS for failure investigation. The unit was expected to have a duration period of approximately 10 years. After just 9 months the crane which controlled the drilling cone started to respond in an unpredictable manner. Upon arrival the unit was opened and the unit interior was found to be completely destroyed with the presence of elemental sulfur (S0 ) and aluminum oxide. Obviously, strong galvanic corrosion had taken place between the steel and aluminum parts of the valve, indicating the presence of a liquid with good electric conductivity in the valve (in lieu of the hydraulic oil), such as seawater. The presence of S0 must be considered as a hallmark of an underlying microbial process in such systems (Tang, et al., 2009). Most likely the sulfur originated from sulfate via sulfide, which is present in high amounts in seawater. Figure 1. A. Hydraulic control unit from an oil drilling platform, where microbial processes has deteriorated its interior. The unit was supposed to endure ten years of operation but this photograph was taken only after nine months of operation (photo taken by FRAS tech AS). B. Scanning electron micrograph, showing both coccoid and bacilli shaped microorganisms (with permission from Elin Ørmen at the Norwegian University of life sciences). A B
  • 14. 14 FRAS Technology AS, Industrial Microbiology Reviews nr 1. 2013 The mechanism behind the detrimental effects seen in this control unit is beyond the scope of this review, but it involves very complex chemical reactions initiated by bacteria. However, it is tempting to propose a short explanation, sulfate reducing prokaryotes (SRP) are strictly anaerobic organisms, and do not grow in the presence of oxygen. Interestingly, the microbial conversion of sulfide to S0 , requires oxygen. One of the probable scenarios therefore follows; SRP in anaerobic conditions causes the interior to deteriorate, sealing material is compromised and the organisms in the unit are exposed to oxygen due to air ingress. This leads to a shift in the microbial community where sulfide is oxidized to S0 by colorless sulfur bacteria, such as Thiobacillus sp (Tang et al., 2009). Also possible is the presence of a biofilm which contain both anaerobic and aerobic zones. This would sustain the simultaneous growth of both the anaerobic SRP and the aerobic colorless sulfur bacteria. Case 2. A ship’s thruster had a breakdown after a long period of downtime. According to the failure report (pers com CEO Sølve Fjerdingstad at FRAS technology AS and Dr. John Olav Nøkleby at DNV GL), the malfunctioning was caused by the lack of lubrication, resulting in massive stress on the mechanical parts. The damaged parts were sent in for mechanical inspection and a slimy substance was literally pouring out upon opening (fig 2). This slimy material is most likely not of mechanical origin, but most likely a result of microbial growth. Mechanically produced sludge is often highly viscous or semi solid and does not behave like a slimy substance, due to the presence of wear debris, oxidation products and sometimes water. A microbial survey was not initiated due to late recognition that this could be a microbial problem rather than a technical issue.
  • 15. 15 FRAS Technology AS, Industrial Microbiology Reviews nr 1. 2013 Figure 2. Slime of microbial origin and emulsified oil pouring out from the bearing part of a thruster (photo taken by FRAS tech AS) Case 3. A supply vessel had been in dry dock for maintenance due to unknown machinery breakdown. During this maintenance the lubrication oil was most likely not changed, nor was the system checked for the presence of any free water. Shortly after the vessel was put into operation after the maintenance procedure, the ship’s lubrication system stopped and the reason was found to be filter clogging. Since the vessel had been in dry dock for maintenance and no reasons for the previous breakdown were found, the lubrication oil was drained and the oil sump was thoroughly cleaned using a steamer. During steaming, flakes of paint loosened from the oil sump surface (fig 3).
  • 16. 16 FRAS Technology AS, Industrial Microbiology Reviews nr 1. 2013 Surface of the paint flake Yeasts Fungal spores Fungal hyphae EPS Bacteria Figure 3. Scanning electron micrograph showing various microorganisms and a thin biofilm, covering the surface of a paint flake sample (scale bar indicate 2µm). The engineers responsible for cleaning and maintenance observed material on the surface which they did not believe had any mechanical or chemical origin. The flakes of paint were sent to The Norwegian University of life sciences via Fras technology AS for microbial analysis. After cultivation, nine different bacterial genera were identified in addition to three fungal genera (Table 3). I will here give a short argumentation for the resulting filter clogging. During dry dock, the water in the lubrication system was stagnant. This leads to the formation of lose and patchy biofilms covered with oil emulsions, on surfaces and in the oil/ water interface (Fjeld, 2012 unpublished results). A possible reason for this unexpected occurrence of filter clogging is that these biofilms with low mechanical stability and bound emulsions will be disrupted when the oil again is in circulation. Consequently biofilm material and emulsified oil will follow the liquid flow, until it reached the filters, leading to filter clogging.
  • 17. 17 FRAS Technology AS, Industrial Microbiology Reviews nr 1. 2013 Table 3. Organisms identified from the cultivated oil phase, water phase and biofilm, size of query sequence, similarity with the closest match in the NCBI BLAST database and the proposed taxonomic affiliation of the organisms. Proposed taxonomic affiliation Query sequence (bases) Closest match in the BLAST database Max identity (%) Taxonomic rank Bacteria Acinetobacter 1500 Acinetobacter sp GXA5 (AY902243.1) 98% Gamma proteobacteria, pseudomonadales Shewanella ** 1493 Shewanella sp. W3-18-1 (CP000503.1) 99% Gamma proteobacteria, alteromonadales Delftia 1495 Delftia sp. LFJ11-1 (DQ140182.1) 99% Beta proteobacteria, burkholderiales Comamonadaceae Achromobacter ** 1492 Achromobacter sp. MT-E3 (EU727196.1) 99% Beta proteobacteria, burkholderiales Pseudomonas 1497 Pseudomonas sp. VKM B-2265 (DQ264636.1) 94% Gamma proteobacteria, pseudomonadales Variovorax ** 1488 Variovorax sp.RKS7-5 16S (EU934231.1) 96% Beta proteobacteria, burkholderiales Comamonadaceae Chryseobacterium * 1384 Chryseobacterium sp. B2 (EU109732.1) 96% Flavobacteria, Flavobacteriales, Flavobacteriaceae Serratia * 1492 Serratia sp. GU124497.1 95% Gamma proteobacteria, Enterobacteriales Bradyrhizobium * 1427 Uncultured Bradyrhizobium sp. clone PSC8 99% Alpha proteobacteria, Rhizobiales Fungi Phialophora ** 625 Phialophora sp. WRCF- AB8 (AY618680.1) 100% Ascomycota, Pezizomycotina, Sordariomycetes Fusarium ** 973 Fusarium solani strain ATCC 56480 FJ345352.1 98% Ascomycota, Pezizomycotina, Sordariomycetes Phoma ** 1024 Phoma sp. GS9N1a (AY465466.1) 100% Ascomycota Dothideomycetes Pleosporomycetidae * Organisms only found in the biofilm samples ** Organisms only found in the liquid (oil and water) samples.
  • 18. 18 FRAS Technology AS, Industrial Microbiology Reviews nr 1. 2013 Case 4. This case is not from the marine or offshore industry but still, it is important in the documentation of microorganisms that cause problems in lubrication oil systems. A project was initiated in 1996, called the baktiol (Bakterier I olje, or Bactiol; Bacteria in oil) project, ISBN 82- 576-9804-0. Senior researcher Jon Fredrik Hanssen and Dr. Aaslaug Lode at the Agricultural University of Norway (NLH), (now the Norwegian University of Life Sciences (NMBU)) initiated a thorough microbial investigation (Financed by the Norwegian research council and Statkraft engineering) based on traditional microbial techniques. During that time, new ash less lubricant additives (additives not containing metals) came into the market and industries were encouraged to use the more environmentally friendly lubricants. The hydroelectric industry in Norway noted a sudden increase in maintenance cost due to more frequent oil replacements and maintenance of machinery in some installations. The hydroelectric industry rarely replaces their turbine lubricants (pers com Dr. John Olav Nøkleby at DNV). Samples with high particle- and water content were received from Det Norske Veritas (DNV). The samples were heavily contaminated (opaque appearance, a lot of wear debris, high water content etc) oil samples from many different hydroelectric power plants in Norway. Of 67 oil samples, 24 contained microorganisms based on microscopic examination and culturing. The oil samples that contained microorganisms also contained free water. If polymerase chain reaction (PCR) based techniques had been used, the number of samples testing positive for microorganisms would probably be higher. The organisms isolated included hyphael fungi, yeasts and bacteria. Most of the isolates were able to grow between 20 and 40o C and some isolates grew well at 4o C. This study provided recommendations concerning reduction of especially water content but also the input of air in open oil systems. One drawback of their study was the lack of establishing the threshold of interference in such machinery. This threshold is very important if correct conclusions are to be drawn, thus microorganisms does not cause problems until they exceed a certain number of cells. Microorganisms will for sure be present in water that enters the systems and hydrocarbon degrading organisms are ubiquitous in nature. Therefore, the finding of microorganisms in water phase or in oil does not necessarily imply biofouling and MIP.
  • 19. 19 FRAS Technology AS, Industrial Microbiology Reviews nr 1. 2013 Additional factors that should be addressed if MIP is to be elucidated are the amount of water in systems, cellular numbers and the presence of events that cannot be explained by mechanical causes. Nevertheless, biocide treatment and short term follow up resulted in remission and maintenance cost was reduced, indicating that microbial growth was the cause of the problems experienced by the industry. 4. Future challenges with emphasis on environmentally acceptable lubricants EAL are now considered to be the new paradigm in lubrication technology. A large amount of lubricants are lost to the environment annually, which may have serious ecotoxicological effects (Michel, et al., 2004). The change into using such EAL has been pivotal for industries such as forestry and other terrestrial industries where lubricant loss is extensive. Because of environmental regulations, industries are encouraged into adapting EAL (forced in US waters). From a microbial and ecological point of view, they are, as already discussed, more biodegradable and could have less toxicological impact on their surroundings. This fact raises the question of whether these lubricants will deteriorate at a higher rate in systems during operation, if the systems have been contaminated with water. It is feasible to assume so, with reduced operational time and increased maintenance costs (downtime, replacement of spare parts, cleaning and disinfection) as a result. Biofouling and MIP in the marine industry are easily confused with mechanical causes and the incidence of MIP is therefore unknown, even with the use of petroleum based lubricants. Case nr 2 and 3 are good candidates for such confusion as they were initially reported as machinery malfunctioning of unknown causes. Therefore, there are reasons to believe that such problems could be underestimated even with the use of petroleum based lubricants. Bacterial infection of systems could coincide with other operational and mechanical occurrences. This could lead to a more rapid reduction in operational time of the systems and compromising of the integrity of the lubricating fluids, than if mechanical causes were the sole cause for detrimental effects.
  • 20. 20 FRAS Technology AS, Industrial Microbiology Reviews nr 1. 2013 If the EAL are going to successfully replace the petroleum based lubricants, the industries should implement a microbial surveillance protocol to monitor and detect any possible detrimental effects caused by microorganisms. If microbial problems are increasing dramatically in the future and future environmental regulations are not making an exception for the maritime industry, new systems design and construction materials should be considered. I would also recommend a higher degree of cross disciplinary cooperation where microbiology should be considered if reasons for machinery malfunctioning or other adverse events cannot be explained by a traditional approach. Indeed, the EAL have replaced petroleum based lubricants with great success in industries such as forestry with emphasis on direct environmental impact. Also, synthetic esters have been used extensively as jet engine lubricants with great success for several decades, due to many superior properties over petroleum based lubricant. In Forestry a rapid and complete degradation are desirable in combination with some superior properties of the EAL. It should be noted that care should be exerted regarding systems in which water is a common contaminant. 5. Discussion and Conclusions To understand MIP and biofouling in lubrication oil systems we need understanding of the underlying processes and to acknowledge that microorganisms, regardless of their small size, are able to cause heavy machinery to break down. The big unknowns in this respect are the threshold of interference and the extent of such problems in the maritime industry. The amount of free water is the driving force in exceeding the limit needed for interference. Microbial growth in systems must be avoided. The best practice to minimize biofouling and MIP is to have appropriate procedures for oil storage and minimize the amount of free water in systems. The four cases described were initially reported as unknown cause of machinery breakdown. This report illustrates that there is a pertinent need for cross disciplinary cooperation if correct mitigation strategies are to be adapted.
  • 21. 21 FRAS Technology AS, Industrial Microbiology Reviews nr 1. 2013 To my knowledge, microorganisms are not generally considered as a possible cause of adverse events in lubrication systems. However, Cases 3 and 4 are the few exceptions where microbial identification techniques were employed, after other causes had been ruled out. Some hallmarks of microbial activity in industrial applications include; the presence of slime, the odor of sulfide (H2S) produced by sulfate reducing prokaryotes (SRP), or other foul odor and precipitation of sulfur aided by for example Thiobacillus sp (Tang, et al., 2009). From the experiences with MIP and biofouling in systems using petroleum based lubricants, the problems usually occur after maintenance or other reasons for machinery downtime. I.e. biofilms with low mechanic stability is formed during low shear forces, and are easily disrupted when the system are put into operation with increased shear forces (Stoodley, et al., 2002). Biofilm material and emulsified oil will follow liquid flow and could lead to clogging of nozzles, valves and filters and friction increase when trapped between load carrying surfaces. Also, before adapting the EAL, I would recommend to determine the threshold of interference for any given system, i.e. how many organisms are required for problems to occur and how much free water does this require. Establishing early warning systems is also important to avoid MIP due to biofouling, especially in systems using EAL. Water control and water prevention actions will therefore be even more important with the use of the eco label lubricants.
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