Alternative anti-fouling

strategies and the role of

monitoring

Hans-Curt Flemming

Biofilm Center

IWW Water Centre, Mülheim an der Ruhr

- Consulting - Research - Service -
1
 What is Fouling?
Definition adopted from heat exchanger technology:

Fouling is the undesired deposition of

material on surfaces (Epstein, 1981)

ƒ Inorganic fouling (precipitation of inorganic crystals), „Scaling“

ƒ Organic fouling (deposition of fat, oil, protein etc.)

ƒ Particle fouling (deposition of silt, clay, humic particles etc.)

ƒ Biofouling (deposition and growth of microorganisms on

surfaces) – particles which can multiply on the expense of
nutrients
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 Bright future for biofouling
An example: Washing machines
- Slime formation in washing powder fill-in box
- Slime formation in tubes
- Clothes are smelly if not dried fast enough
- Insufficient disinfection, infections spreaded

Cause: Biofouling – due to protection of environment and clothes

h Biologically readily degradable washing soaps
h Water saving & increasing washing soap concentrations
h Liquid washing soaps stick stronger to walls – higher local
concentrations at interfaces
h Low temperatures while washing
h No more peroxides or other bleaching agents in washing soap
h Water with washing soap remaining in tubes for extended periods
18.05.2006
of time because it is not completely drained after washing 3
An ideal biofouling environment: Paper mill, an
environment with free access of bacteria from air, water
and paper components (see Michael Meiers presentation)
Fouling chances increased by
- chlorine-free process
- neutral pH
- higher concentrations of nutrients
- temperatures between 30 and 45 °C
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- Use of contaminated additives 4
 How good intentions cause inexpected problems: Massive biofilm
development in a heat exchanger due to biodegradable corrosion
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inhibitor
 The Quest for the Silver Bullet
ƒ Get rid of biofouling once and forever!
ƒ The ultimate anti-fouling coating: Tributyltin paints
– a Silver Bullet too powerful

Normal oister shell

Oister shell,
18.05.2006 exposed to TBT 6
 Time scale for fouling protection
Kevin Marshall: „The organism always wins“ (Question: how fast!)

Anti-Fouling requirements in different time scales:

Hours to days
- Some Catheters
- Surfaces in contact with food

Days to weeks
- Environmental sensors
- Dialysis units

Weeks to months
- Heat exchanger systems
- Membranes and filters for water treatment

Months to years
- Ship hulls, oil rigs
- Drinking and process water pipes and reservoirs
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 What to do?
Steps in fouling cases
1. Detection
2. Sanitization
3. Prevention
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 Biofilm sampling:
On surfaces!

! Scratching, scratching, scratching (razor blades,

rubber scrapers, cotton swabs etc.)
! Removal of parts of support material with
biofilms ⇒ laboratory
! Expose test surfaces („coupons“), remove after
18.05.2006 given time ⇒ laboratory 9
 Verification of biofouling:
h Field tests:
- Visual observation: slime
- Smell
- Texture (slimy)
- Smolder test: heat sample, go for smell of burnt hair

hLaboratory:
- Microscopical observation, cells/cm²
- Cultural methods (cfu/cm²)
- Chemical analysis (water content, org./inorg.)
18.05.2006 - Spectroscopical analysis (e.g., FITR) 10
 The medicine-based strategy
ƒ Biofouling is considered a „technical disease“
ƒ Microorganisms cause this disease
ƒ Kill microorganisms and the problem is solved
ƒ Adoption of medical term: „Disinfection“ to cure the system

Drawback of this strategy:

- In many cases ineffective because biomass, not activity is the problem,
dead or alive
- Killing does not necessarily remove biomass: dead biomass can still
cause problems (heat exchanger, membranes, process waters etc.)
- „Disinfection“ means only „Inactivation of pathogenic organisms“
- Frequent failure of this strategy creates jobs and contributes to the
gross national product but not to sustainable solution of problems

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 Regrowth of P. aeruginosa on test surfaces after
application of H2O2 and Ag+
Disinfection 2

1,0E+8
Disinfection 1
1,0E+7
colony counts [cfu mL-1]

1,0E+6

1,0E+5

1,0E+4
-

1,0E+3

1,0E+2
without silver with silver
1,0E+1

1,0E+0
0 5 10 15 20 25 30
time [d]

Application of 600 mg L-1 H2O2 alone and with 300 µg L-1 AgNO3 added for 24 h at room
temperature
Determination of colony counts (PAP, 24 h , 36°C) at reactor outlet
(courtesy of S. Schulte)
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 Killing vs. removal
colony count biocide treatment total cell count biocide treatment
colony count control total cell count control
10
10 1. biocide- 2. biocide- 3. biocide- 4. biozide-
colony count and total cell count [cm ]
-2

10
9 treatment treatment treatment treatment

8
Cells/cm²
10
7
10
6
10
5
10

10
4 cfu
3
10
2
10
1
10
0
10
7 8 9 10 11
time [d]

Total cell count with DAPI

Colony counts on R2A agar, incubation 7 days at 20 °C

Repeated H2O2 treatment for 15 min

(Courtesy of Simone Schulte)
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 Five fundamental problems in
conventional anti-fouling strategies
1. Monitoring by process performance or product quality
Æ No early warning systems

2. Sampling of water phase and not on surfaces

Æ No information on biofilm site / extent

3. „Disinfection“
Æ No cleaning: dead biomass on surface, good
regrowth conditions

4. No nutrient limitation
Æ Nutrients = potential biomass; not reduced by
biocides

5. Efficacy control by process/product quality

18.05.2006 Æ No optimization of countermeasures 14
 Alternative: Strategies based on
better understanding of biofouling
Biofouling: „Undesired deposition and
growth of microorganisms at interfaces“
! Biofouling is a biofilm problem
! Dissolved substances are converted into
locally accumulated biomass
! Operational definition: Biofilm growth
exceeding „threshold of interference“
! Biofouling: A biofilm reactor in wrong
place & time
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 Elements of an integrated
Anti-Fouling Strategy:
Holistic approach, based on understanding
the principles of biofilm development
! Surface design and modification
! Cleaning friendly design
! Biofilm management
! Biofilm engineering
! Biofilm analysis
! Monitoring of deposit development
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 Anti fouling toolbox

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 The most obvious target: Surfaces
ƒ Surface modification to influence primary adhesion
(see sessions on Tuesday)
- Material selection (however, on a long term, all materials can be
colonized)
- Surface properties (hydrophobic, hydrophilic; hydrogels – selection for
specialized organisms)
- Surface topography (roughness, nanostructures, lotus effect - needs 3
phases, beware of surfactants!)
- Antiadhesive coatings (Can be fouled by abiotic matter)
- Surface bound biocides (Mechanism obscure; sometimes even effect
on water phase claimed; fate of dead cells; Fouling)
- Biocide releasing surfaces (Water contamination)
- Self polishing surfaces (Water contamination)
- Functional surfaces (H2O2+Catalyst; UV-light) (Fouling)
- Conductive coatings, application of electrical current (Fouling)
- Reversible change of surface properties (Fouling)

0 How long does the effect last?

0 Toxicity
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Most promising: Low adhesion surfaces, easy to clean18
 Biofilm Management

Threshold of
interference

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 Biofouling Potential:
1. Ubiquitous, always hungry biofilms
2. Nutrients from medium and support,
representing potential biomass
3. Surfaces inviting for colonization
4. Undisturbed growth conditions, e.g.,
infrequent cleaning
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 Strategy: Put the
biofilm into the
right place
Example:
membrane system
for treatment of
heat exchanger
water
Biofilter ahead of
the membrane

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 Biofilm

Biofilm

Membrane
Membrane Porous
Porous support
support

Before biofilter After biofilter

Thin cut of biofilm on membrane

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 Biofilm parameters before/after filter
Parameter unit before filter after filter

Cell number [#/cm2] 1,0 x 108 5,5 x 106

CFU [#/cm2] 1,0 x 107 1,2 x 106
Protein [µg/ cm2] 78 4
Carbohydrates [µg/ cm2] 26 3
Uronic acids [µg/ cm2] 11 2
Humic subst. [µg/ cm2] 41 12

Biofilm thickness [µm] 27 3

Flux decline [%] 35 <2

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 Biofilm engineering
Mitigating adverse effects of a biofilm
which cannot be removed
“Living with biofilms”

* Improved permeation properties

* Improved heat transfer
* Reduced friction resistance

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 McDonogh, R., G. Schaule and H.-C. Flemming (1994): The permeability of
18.05.2006 25
biofouling layers on membranes. J. Membr. Sci. 87, 199-217
 Confocal laser scanning microscopy (CLSM)
Vertical view of biofilm: change of structure

After 10 treatments
Control with H2O2
18.05.2006 108 cells/cm² Still 108 cells/cm² 26
 Natural Anti-Fouling Strategies I
Options:
- Tolerance
- Avoidance
- Defense
Organisms we can learn from (see Tuesday section):
" Microalgae
" Macroalgae
" Terrestrial plants
" Echinodermata (Stachelhäuter)
" Worms
" Water plants (e.g., seaweed)
" Fish
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 Natural Anti-Fouling Strategies II
" Mechanical defense
- Intense surface production of mucus
- Surface-bound cilia-driven migration of a mucus film
- Scraping of surface with specialized appendices
(„windshield wiper“)
- Surface renewal („scinning“)
- Cleaning by friction between body and sediment surface
- Specific surface structures

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 Natural Anti-Fouling Strategies III

" Chemical defense

- Surface-bound or excreted secondary metabolites
* Toxins (some are very toxic)
* Pheromones (AHL´s)
" Extrinsic defense
- Selected epibionts (inhibitory to others)
- Specialized epibiont predators (grazers)

" Combined strategies

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 Monitoring - the key to successful
anti-fouling strategies
! Early warning saves remedy
efforts
! Efficacy control of cleaning
procedures
! Optimization of cleaning
! Control of anti-fouling strategies
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 Biofilm monitoring:
Observation of biofilm development over time

What do we want to know?

! Site/location
! Quantity
! Thickness
! Distribution
! Nature of deposit
- organic/inorganic
- biological/abiological
- chemical composition
! Stability (how easy to clean?)
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! Kinetics of formation and removal 31
 The optimal monitoring method:
! in situ
! in real time
! on line
! non destructive
! fast and accurate
! Integrates over large surface areas
! easy to handle, stable
) Has predictive power
) Early warning capacity
18.05.2006 ) Saves value 32
 On-line Monitoring

System
System
System
Biofilm Cleaning procedures
Biofilm Biofilm
Sensors

Hard-/Software Fouling

Critical fouling boarder line

Defined alarm boarder line

Documentation/ Defined effectivity

Visualisation boarder line

time

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 The concept: use of monitors for
automatic fouling control

Modem

Dosage
Addi-
tives

Modem

Local data processing

-> control systems
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 3 levels of information of monitors:
Information Example for parameter
Level I Scattered light
„Detection of a deposit Differential turbidity
on a surface“ Optical fouling monitor
Pressure drop
Heat transfer resistance
Weight increase
Velocity of sound
Photoacoustic spectroscopy
Quartz crystal microbalance

Level II FTIR-spectroscopy
„Biological aspects of deposit“ Tryptophane fluorescence
Direct microscopy

Level III FTIR-spectroscopy

„Detailed information“ Raman spectroscopy
X-ray photoelectron spectroscopy
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 Conclusions: Five principles of
alternative anti-fouling strategies
1. Learn to live with biofilms and keep them under
control
2. Monitoring: crucial for timely detection of biofilms
and countermeasure optimization
3. Cleaning is more important than killing biofilm
organisms
4. Biofilm management: Nutrient limitation
(nutrients are potential biomass)
5. Biofilm engineering: Mitigating of biofilm effects
(hydraulic resistance)

It is possible to live with biofilms!

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