Foaming in wastewater treatment plant (WWTP)

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Contents
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1 Introduction
2 Physical and chemical environment
o

2.1 Gas bubbles

2.2 Surfactants

2.3 pH and Temperature

2.4 Dissolved oxygen (DO)

2.5 Mean Cell Retention Time (MCRT)

3 Key Microorganisms

3.1 Microthix parvicella

3.2 Mycolata

3.3 Gordonia amarae

4 Microbial Processes

4.1 Substrate Storage

4.2 Cell Surface Hydrophobicity and Exoenzyme Activities

5 Foaming Control Strategy
6 Current Research

6.1 Identification of Filamentous Bacteria

6.2 Development of Effective Foaming-Control Chemicals

6.3 Mechanism of Foaming

7 References

Introduction
Foaming in activated sludge process is a common operational problem in many wastewater
treatment plants. The foam can occur in aeration tank, secondary clarifier, as well as in
anaerobic digester. Foam in WWTP is normally sticky, viscous and brown in color. It floats and
accumulates on top of the tanks, and can take up a large fraction of solids inventory and
reactor volume, thus decreasing the effluent quality and control of sludge retention time
(SRT). The foam can also overflow onto walkways and surrounding areas, posting severe
difficulties and risk to operation and environment.

Activated sludge foaming in aeration tank.http://web.deu.edu.tr/atiksu/toprak/ani4094.html

Many reasons are associated with foaming: presence of slowly biodegradable surfactants (eg.
household detergents) from industrial or municipal wastewater, excess production of
extracellular polymeric substance (EPS) by activated sludge microorganisms under nutrientlimited condition, proliferation of filamentous organisms and gas provided in aeration tank or
produced in anoxic zone of aeration tanks, secondary clarifiers and anaerobic digesters.
Stable foaming in WWTP is the product from interaction among gas bubble, surfactant and
hydrophobic particles. The hydrophobic particles congregate at the air-water interface and
strengthen the water film between air bubbles. Meanwhile, the particles also serve as
collector for surfactant which stabilizes the foam. Gas bubbles in WWTP are generated by
aeration, mechanic mixing and biological process like denitrification and anaerobic digestion;
Surfactants in WWTP come from the wastewater streams that contain slowly biodegradable
surfactants; hydrophobic particles are normally referred to the filamentous bacteria with a
long-chain structure and hydrophobic surface [1].

Physical and chemical environment

Gas bubbles
From the mechanism for foaming mentioned above, we know gas bubbles are essential in
foam generation. Gas bubbles are involved in many steps of activated sludge process. In
aeration tank, aeration and mechanic mixing is employed to ensure enough dissolved oxygen
for aerobic degradation of organic pollutants or nitrification. This thus creates abundance of
gas bubbles. Other than external introduction by aeration or mixing, gas bubbles can also be
produced from biological processes themselves. Both denitrification in secondary clarifier and
anaerobic digestion in digester produce gas like N2 or CH4, CO2. These gases favor the
generation of foam.

Surfactants
Most surfactants in WWTP originate from the detergents, oil and grease that used in
households or industry. The EPS produced by bacteria is also believed to contribute part of
the surfactants. Surfactant could stabilize the foaming and allow foam to accumulate. Ho and
Jenkins demonstrated the conducive effect of a slowly biodegradable nonionic surfactant in
foaming [1,2].

pH and Temperature
Filamentous bacteria are important in forming stable foam. The growth rate of filamentous
bacteria was found not affected significantly for range of pH from 6.7 to 8.0, only slightly
decreased at pH 8.4. Optimum temperature of Microthrix parvicella , a filamentous bacterium
associated with foam production, was reported around 25 oC, some growth was observed at
8 oC, and poor or no growth at above 35 oC [3].

Dissolved oxygen (DO)

Filamentous bacterium M. parvicella prefer low oxygen concentration, and were found
proliferate in WWTP with spatial or temporal low DO. In Ekamas study,M. parvicella was
eliminated by increasing DO to 2-3 mg/L[4]. As an effective control for sludge bulking and
foaming, aerobic selectors with high DO (>2mg/L) are often placed before aeration tank to
inhibit the growth of filamentous bacteria[3].

Mean Cell Retention Time (MCRT)

More filamentous bacteria were observed in WWTP and studies when increasing MCRT (1.5
to 20 d), while MCRT at around 1 d was effective in limit growth of filamentous bacteria.
Controlling MCRT sometimes could be difficult to achieve by increasing flow rate of water
since biomass could be retained in foam without moving with water[1,3].

Key Microorganisms
Filamentous bacteria serve as hydrophilic particles which play important role in stabilizing
foaming in WWTP. There are two main groups of filamentous bacteria: the most commonly
observed group:Candidatus Microthix parvicella , and members of the Mycolata.

Microthix parvicella

Microthrix parvicella from foam in an activated sludge

plan.http://www.flickr.com/photos/45132912@N02/4185582040/

M. parvicella are gram-positive nonbranched filamentous bacteria. They are aerobic, nonfermentative and can reduce nitrate. Although M. parvicella can grow over a wide range of
oxygen concentration, they prefer microaerophilic conditions for good growth. The filaments
they produced at low DO (~ 0.4 mg/L) are long and regular without empty or deformed cells
that observed under high DO conditions[3,5].

Mycolata

Mycolata with right-angled branching pattern.http://www.environmentalleverage.com/what_is_in_a_name.htm

Mycolata with acute-angled branching pattern.http://www.environmentalleverage.com/what_is_in_a_name.htm

Mycolata, often referred to as "nocardia", are a group of filamentous bacteria that contain
mycolic acids in their cell walls. They are under the orderActinomycetales in
phylum Actinobacteria, isolates were identified as member in families Corynebacteriaceae,
Dieziaceae, Gordoniaceae, Mycobacteriaceae, Nocardiaceae,
Tsukamurellaceae and Williamsiaceae. They have two major morphotypes: one with rightangled branching pattern and the other acute-angled branching pattern. Mycolata were found
to uptake a wide range of organic compounds, and can use nitrate or nitrite as electron
acceptor. Many mycolata can store polyhydroxyalkanoate in cell and present high cell surface
hydrophobicity[6].

Gordonia amarae
Gordonia amarae belongs to right-angled branching mycolata , is one of the most common
filamentous bacterium found in foaming process.Gordonia amarae can utilize large number of
organic substrates, both hydrophilic and hydrophobic, and it was found capable in taking up
some substrates under aerobic, anaerobic and anoxic conditions[7]. Gordonia amarae cell
has very hydrophobic surface, and they can produce biosurfactants from a wide range of
substrates. Production of biosurfactants were believed beneficial for Gordonia amarae to
solubilize insoluble substrates which help Gordonia amarae survive in foam. It is generally
accepted that the high cell surface hydrophobicity and ability of biosurfactants production are
the two main reasons for Gordonia amarae to cause foaming[8].

Microbial Processes
Substrate Storage
M. parvicella and Mycolata were reported to be able to utilize various organic compounds as
carbon and energy source. The compounds contain organic acids, complex substrates and
fatty acids under aerobic, anoxic and anaerobic conditions. The substrates can then be
storaged intracellularlly in filamentous bacterium. Intracellular storage of poly hydroxyalkanoates(PHA) like inclusions were observed in aerobically-grown M.
parvicella under anoxic or anaerobic conditions[3]. Lipid storage granules were also observed
in some M. parvicella from activated sludge in nutrient removal WWTP [5]. Mycolata also can
form intracellular PHA inclusions for substrates storage[6]. The storage capability of
filamentous bacteria allows them survive in harsh conditions during operation(e.g. substrateslimiting in foam, alternating anaerobic-aerobic environment), and out-compete floc-forming
and other bacteria in activated sludge, most of which can not uptake and storage substrates
anaerobically.

Cell Surface Hydrophobicity and Exoenzyme Activities

Higher cell surface hydrophobicity were found in cells of M. parvicella and Mycolata than
other bacteria in activated sludge. The more hydrophobic cell surface enable filamentous
bacteria better attraction to hydrophobic substrates like lipids, long-chain fatty acid (LCFA).

Additionally, filametnous bacteria produce many exoenzymes such as lipases, which enhance
the degradation and utilization of substrates [3,6].

Foaming Control Strategy

There are no universal strategies effective for foaming control in WWTP. Specific measures
need to be taken according to the reason of foaming, organisms involved and operational
condition. Common strategies for foaming control include: Reduction of SRT (Sludge
Retention Time, similar to mean cell retention time, often used in wastewater treatment
operation) to wash out filamentous bacteria; removal of hydrophobic substances and
substrate that could enhance foaming or favor the growth of filamentous bacteria; introduction
of selectors before aeration tanks to suppress the growth of filamentous bacteria; addition of
oxidizing agents like chlorine to kill filamentous bacteria (chlorine also kills other bacteria)
[1,2,3].

Current Research
Identification of Filamentous Bacteria
Traditional identification of filamentous bacteria relies on their morphology under microscopy.
However, many filamentous bacteria may not have distinguishable morphology, therefore,
identification based on 16S or 23S rRNA genes are preferred. Nielsen group from Denmark
developed more effective permeabilization protocol for fluorescence in-situ hybridization
(FISH) that could enhance the hybridization and produce stronger signal. They performed a
various ecophysiology studies on different filamentous bacteria from foam and activated
sludge sample using MAR-FISH[6]. Other 16S-based techniques like PCR-DGGE were also
employed in detection of filamentous bacteria[9].

Development of Effective Foaming-Control Chemicals

Conventional oxidizing chemicals like chlorine that used to destroy filamentous bacteria also
affect the growth of other bacteria in activated sludge. More selective chemicals for controlling
filamentous bacteria are desired. Polyaluminium chloride (PAX-14) was found effective in
controlling the foaming caused by M. parvicella. Addition of PAX-14 did not affect the
nitrification and COD removal performance. However, the mechanism PAX-14 in
controlling M. parvicella is still not clear[10].

Mechanism of Foaming
Petrovski et. al closely examined the role of surfactant in foaming based on the data from 65
foaming Mycolata. They found the floatation theory can be applied in explaining the role of
surfactant in activated sludge foaming. Mycolata without surfactant could produce scum, while
presence of surfactant without hydrophobic particle created unstable foam. They also found
Bacillus subtilis, commonly culturable from foam could play important role in foaming with its
production of surface surfactant[11].

References
[1]Jenkins, D, Richard, MG, Daigger, GT. 2004. Manual on the Causes and Control of
Activated Sludge Bulking, Foaming, and Other Solids Separation Problems, 3rd edition..
Lewis Publishers, New York. http://www.iwapublishing.com/template.cfm?
name=isbn1843390469
[2]Hug T. 2006. Characterization and controlling of foam and scum in activated sludge
systems. ETH Ph.D. Dissertation. http://dx.doi.org/10.3929/ethz-a-005180592
[3] Rossetti S, Tomei MC, Nielsen PH, Tandoi T. 2005. Microthrix parvicella, a filamentous
bacterium causing bulking and foaming in activated sludge systems: a review of current
knowledge. FEMS Microbiology Reviews, 29(1): 49
64. http://www.sciencedirect.com/science/article/pii/S016864450400066X
[4]Ekama, GA Marais, GVR. 1984. Theory, Design and Operation of Nutrient Removal
Activated Sludge Processes, Water Research Commission. 5.15.18, Pretoria.
[5] Nielsen, PH, Roslev, P, Dueholm, T, Nielsen, JL. 2002. Microthrix parvicella, a specialized
lipid consumer in anaerobic aerobic activated sludge plants. Water Sci. Technol. 46: 73
80.http://www.ncbi.nlm.nih.gov/pubmed/12216691
[6] Kragelund C, Remesova Z, Nielsen JL, Thomsen TR, Eales K, Seviour R, Wanner J,
Nielsen PH. 2007. Ecophysiology of mycolic acid-containing Actinobacteria (Mycolata) in
activated sludge foams. FEMS Microbiol Ecol. 61(1):17484. http://www.ncbi.nlm.nih.gov/pubmed/17466023
[7]Carr EL, Eales KL, Seviour RJ. 2006. "Substrate uptake by Gordonia amarae in activated
sludge foams by FISH-MAR".Water Sci. Technol. 54(1): 39
45.http://www.ncbi.nlm.nih.gov/pubmed/16898135
[8]Pagilla KR, Sood A, Kim H.2002. "Gordonia (nocardia) amarae foaming due to
biosurfactant production". Water Sci Technol. 46(1-2):51924. http://www.ncbi.nlm.nih.gov/pubmed/12216680
[9] Shen FT, Huang HR, Arun AB, Lu HL, Lin TC, Rekha PD, Young CC. 2007. Detection of
filamentous genus Gordonia in foam samples using genus-specific primers combined with
PCR--denaturing gradient gel electrophoresis analysis. Can J Microbiol. 53(6):76874. http://www.ncbi.nlm.nih.gov/pubmed/17668037
[10]Roels, T, Dauwe, F, Van Damme, S, De Wilde, K,Roelandt, F. 2002. The influence of
PAX-14 on activated sludge systems and in particular on Microthrix parvicella. Water
Sci.Technol. 46(1-2), 487490. http://www.ncbi.nlm.nih.gov/pubmed/12216672

[11]Petrovski S, Dyson ZA, Quill ES, McIlroy SJ, Tillett D, Seviour RJ. 2011. An examination
of the mechanisms for stable foam formation in activatedsludge systems. Water Research.
45 (5): 21462154. http://www.sciencedirect.com/science/article/pii/S004313541000881X