DESALINATION

ELSEVIER Desalination 178 (2005) 115-124

www.elsevier.com/locate/desal

Filtration characterisation for assessing MBR performance:

three cases compared

H. Evenblij a*, S. Geilvoet a, J.H.J.M. van der GraaP, H.F. van der R o e s t b
°Section of Sanitary Engineering, Department of Water Management, Delft University of Technology
P O. Box 5048, 2600 GA Delft, The Netherlands
TeL +31 (15) 278-2258; Fax +31 (15) 278-4918; email: H.Evenblij@Witbo.nl
bDHV Water BV, PO. Box 484, 3818 EX Amersfoort, The Netherlands

Received 15 October 2005; accepted 24 February 2005

Abstract
Understanding of membrane fouling processes in membrane bioreactors (MBR) treating municipal wastewater
is essential for proper design and operation of full-scale MBR wastewater treatment plants. One of the tools in
investigating this subject must be a well-defined measuring protocol to quantify the filterability of activated sludge.
Such a method was developed and a filtration apparatus was built by Delft University of Technology [1]. This
article describes the results of applying the mentioned filtration characterisation method at three MBR pilot locations
in the Netherlands. The measurements were sustained by EPS analyses in the water phase of the respective activated
sludge broths. Each MBR installation produces an activated sludge with a specific filterability, which can be quantified
by the proposed measuring protocol quite well. Activated sludge from each installation could be filtered under sub-
critical conditions, i.e. without filtration resistance increase for at least thirty minutes of filtration. Photometric EPS
analyses of the water phase and obtained permeate seem to be too weak to explain the differences in filtration
behaviour.
Keywords: EPS; Fouling; Membrane bioreactor; Wastewater treatment

I. Introduction treatment with a membrane filtration step, offers

several advantages over the conventional activated
The membrane bioreactor process (MBR) for
sludge process, like limited footprint and superior
wastewater treatment, combining activated sludge
effluent quality [2]. However, it did not yet find
wide-spread application, mainly because of the
*Corresponding author.

Presented at the conference on Membranes in Drinking and Industrial Water Production, L 'Aquila, Italy, 15-17 November
2004. Organized by the European Desalination Society.

0011-9164/05/$- See front matter © 2005 Elsevier B.V. All rights reserved
doi: 10.1016/j.desal.2005.02.005
116 H. Evenblij et al. / Desalination 178 (2005) 115-124
poorly understood process of membrane fouling
and its causes. This is particularly a problem in
municipal wastewater treatment where usually
considerable daily and seasonal fluctuations occur,
both qualitatively and quantitatively. One of the
handicaps in attacking this problem is the diversity
of research approaches, which makes an honest
comparison of results by different research groups
difficult. The problem is complicated by the fact
that each installation treats a different influent.
All this hampers an efficient knowledge develop-
ment and most likely causes non-optimal design
and operation of MBR plants. Also the influence
of design and operating choices can only per
instance be evaluated and can hardly be gen~-
ralised.
Another problem is related to the type of
physical/chemical analyses that are carried out for
characterising the biomass, in order to identify
foulants. This is caused by the fact that there exist
contradictory reports on which substances are
exactly the foulants in MBR. Theoretical con-
sideration of cross flow filtration of suspensions
led to the hypothesis that only soluble and col-
loidal material are involved in fouling processes
of properly operated MBR systems. MLSS con-
centration as such turns out to be of minor im-
portance for filtration behaviour [3].
Recently, much attention has been given to the
analysis of extracellular polymeric substances
(EPS) since they were correlated to membrane
fouling in water treatment membrane systems [4-
6]. For MBR a distinction can be made with res-
pect to (EPS bound to) microbial flocs [7,8] and
(EPS in) the water phase [9-11 ]. Although there
is most likely a relation between the two, a direct
link with membrane fouling is exclusively ex-
pected from soluble matter.
Delft University of Technology started a re-
search programme into membrane fouling in MBR
for wastewater treatment, together with DHV
Water. First aim of this programme was the
development of a method to accurately quantify
filterability of activated sludge. This is done by
short term tubular membrane ultrafiltration
experiments under well defined circumstances [ 1].
The objective of the research described in this
article is to compare filterability of activated
sludge from different MBR installations. Acti-
vated sludge from three MBR-installations treat-
ing municipal wastewater with three different
types of membranes was evaluated with respect
to its filterability.
The objective of this work is two-fold:
• Test the developed measuring protocol and in-
stallation for filtration characterisation at three
pilot MBRs.
• Assessthe influence of soluble and suspended
EPS on membrane fouling. The hypothesis is
that each MBR installation provides boundary
conditions for the formation of a certain type
of activated sludge. All influencing factors lead
to a certain amount of polysaccharides in the
water phase, which can be correlated to its fil-
terability.
2. Material and methods
2.1. Filtration characterisation
A filtration apparatus and measuring protocol
for filterability measurements were developed.
The filtration apparatus consists of a single tubular
membrane module to be operated in a cross-flow
mode. The applied membrane, provided by X-
Flow, is an 8 mm tubular PVDF membrane of ap-
proximately 1 m length, with nominal pore size
of 30 nm (Fig. 1).
Activated sludge is circulated with a peristaltic
pump, and permeate extraction for constant flux
experiments is also performed with a peristaltic
pump. The apparatus is equipped with sensors and
pressure transmitters for monitoring TMP, cross-
flow velocity, temperature, dissolved oxygen
concentration and pH. Furthermore, permeate flux
is measured with a mass balance connected to a
PC. The filtration protocol consists of four steps,
alternated with clean water resistance measure-
ments:
 H. Evenblij et al. / Desalination 178 (2005) 115-124 117

Pr~suregauge
A1 Ib_
v
Q Ftowmeter
2-wayvalve,operatedvia PLC

3-wayvalve,operatedvia PLC

[3~ 2-wayvalve,manuallyoperated

[~ 3-wayvalve,manuallyoperated

Membrane
X-Flow i Material: PVDF
Tube length mm 950
Tube diameter mm 8
Membrane area cm 2 240
Nominal pore size nm 30

®
Fig. 1. Schematic overview of filtration characterisation unit.

1. Clean water resistance measurement with clean
water (either permeate or demineralised water).
2. Filtration of activated sludge, with constant
flux o f 80 L/m2h and cross flow velocity 1 m/s,
until TMP = 0.70 bar.
3. Cleaning with permeate/DI water with forward
flush, 4 m/s
4. Clean water resistance measurement with clean
water.
5. Backflush with DI water, TMP = -0.75 bar
6. Clean water resistance measurement with clean
water.
7. Chemical cleaning, 500 ppm NaOCI for 30 min.
8. Clean water resistance measurement with clean
water.
In this work, the second part of the filtration
protocol is emphasised: activated sludge filtration.
A standard experiment is performed with flux of
80 L/m2h and cross-flow velocity o f I m/s. The
results are presented as total additional filtration
resistance against specific produced volume
(Fig. 2a). Each measured resistance value is re-
duced with the initial membrane resistance,
causing all filtration curves to start at 0 m ~(Fig. 2b).
To compare different curves, one value of each
curve was chosen, i.e. the value of additional re-
sistance after 25 I/m2 of filtration, AR25.
In some cases experiments under standard
conditions led to useless results, because fouling
rates were too high or too low, which necessitated
I 18 H. Evenblij et al. / Desalination 178 (2005) 115-124

4
E 4 too E

o
bo

8=
©Z ~ 8
~'2
1 • 2
I
o

20 40

-1 0 0 ,
Volume [l/m 2 0 20 4O 60 8O
Volume [ l / m 2]
(a) I ~,-R ~ p * J
(b) [ ~'- ~d,~ l

Fig. 2. (a) Typicalresults from filtration characterisation measurement. (b) Isolated filtration curve, with determination of
AR25.

another type of measurement, here indicated as 2.2. M B R pilots

critical flux determination. For this measurement
several filtration curves were made, with increas-
ing, or decreasing flux. All these curves were
summarised as AR25and plotted in one figure. The
highest flux value for which AR25 is 0 m -1 is held
to be the critical flux, J . The critical flux concept
was first reported by Field et al. [12] who supposed
the existence o f a flux below which no fouling is
observed. This was verified by measurements with
well defined solutions containing spherical par-
ticles. There was some scepticism regarding the
question whether such a critical flux would exist
in activated sludge filtration, bearing in mind the
wide range of particle sizes in MBR activated
sludge and their microbial origin [13].
In this comparative study, three MBR pilots
were used as a source o f activated sludge. All of
them were located at a municipal wastewater treat-
ment plant. They were all equipped with a pretreat-
ment step followed by biological treatment, de-
signed for COD, P and N-removal. The three sys-
tems were equipped with different types o f mem-
branes: Plate-and-frame membranes (Kubota),
hollow-fibre (Zenon) and rotating disk membranes
(Huber). For more information on the pilot plants
see Table 1.
During the measurement period, which lasted
from 6 to 8 days, the filtration characterisation
apparatus was located at the resepctive wwtp. For

Table 1
Characteristics of MBR pilots where activated sludge was sampled

Maasbommel w w t p Hilversum
wwtp Beverwijk wwtp
Membrane type Hollow-fibre(Zenon) Plate-and-frame (Kubota) Rotatingdisk (Huber)
Pore size, n m 40 400 40
Capacity, m3/h 10 1.5 4.2
pH (average) 7.58 7.73 7.70
Days of measurements 6 7 8
Tested fluxes, L/m2h 80, 100, 120 40, 80, 120, 160, 190 40, 80, 100, 120
 H. Evenblij et al. / Desalination 178 (2005) 115-124 119

a measurement, 25 L of activated sludge was

sampled from the recirculation from membrane .--- / /
tank to nitrification zone. The sludge was aerated -" / / fl,
and filtrated in the filtration characterisation unit.
During filtration characterisation the sludge was ti j
'o.s ................................... / ; 1 i 2 ; ..... i
recirculated and after the measurement the sludge
was discharged. This procedure was repeated three
or four times a day. ..Q

~ll
•

~ -
;; ;-;;
0.0
2. 3. Analyses ~

O
.................... ~

10
~ ~'°,

20
'

30
1

Additional to the filtration characterisation Volume ( L J m 2)

measurements several analyses were performed .... Maasbomrnel u i Beverwok ...................Hilversum

to gain insight in foulants and their character. The

activated sludge was fractionated in the water
phase and biomass by means of filtration with a
paper filter with a non-definite pore size of 7-
12 pm (Schleicher & Schuell, 5892). The water
phase, containing part of the colloidal and all
soluble material, was further analysed for proteins
and polysaccharides, these being the major con-
stituents of extracellular polymeric substances.
These analyses were carried out with photometric
methods, as described by Lowry et al. [14] and
Dubois et ai. [15]. Also permeate of the filtration
characterisation was analysed for EPS.
3. Results
3.1. Filterability of three installations
The three installations produce specific types
of activated sludge with totally different filter-
abilities.
Activated sludge from Hilversum wwtp showed
extremely good filtration behaviour. A standard
experiment with 80 I/m2h did not yield to a mea-
surable resistance increase even when filtrered
more than 1 h uninterruptedly. Activated sludge
from Maasbommel wwtp showed moderate filt-
ration behaviour, whereas activated sludge from
Beverwijk wwtp was the worst in these experi-
ments.
Fig. 3 shows the average filtration curves for
the three installations, including the bandwidth,
obtained with standard filtration experiments.
Fig. 3. Filtration characterisation curves from three MBR
pilot installations. From each pilot the obtained minimum,
average and maximum curves are plotted. Measurements
are carried out with d = 80 L/m2h for Maasbommel and
Beverwijk and J = 120 L/m2hfor Hilversum.
3.2. Daily variations
Results from earlier work indicated the occur-
rence of daily fluctuations in filtration behaviour.
These changes were ascribed to changes in
influent quality and quantity, as well as to the time
point of sampling in the filtration cycle of the
MBR pilot. All MBRs in this study are operated
in an intermittent filtration mode, where the
filtration can be alternated with backflushings or
a relaxation period. This may have its effect on
the filterability of the biomass in the membrane
tank. The measurements that were carried out in
this study do not very clearly indicate a daily
pattern in filterability. Of course the amount of
measurements is quite small, therefore conclu-
sions can only be drawn with great care.
Fig. 4 shows the variation of AR2s derived from
filtration curves at 80 L/m2h (Maasbommel and
Beverwijk) and 120 L/m2h (Hilversum). Activated
sludge from Maasbommel and Beverwijk was
sampled two or three times during a day and AR25
is presented per day, each bar representing one
measurement.
There is no clear development of AR25 during
a day. Sometimes filterability became worse
i 20 H. Evenblij et al. / Desalination 178 (2005) 115-124

Beverwijk J = 8 0 L/mZh during the Hilversum test period (Fig. 4c). The
2.0
E (a) first day of the Beverwijk measurements is
o exceptional in the sense that the first measurement
showed the highest AR25, followed by a sharp
decrease and a small increase, which was con-
.< 1.0 tinued on the following day.

3.3. Critical flux

The fact that the sludge from Hilversum wwtp
0.0
1 1 1 2 2 3 3
showed no fouling during any experiment at 40
Day [ # ] and 80 L/m2h hinted at a sub critical flux opera-
tion. The flux was increased to 120, 160 and even
Haasbommel J=80 L/m2h 190 L/m2h. From 120 L/m2h and upwards fouling
, ~ 1,0
E could be observed, but still relatively low, com-
(b) i pared to the other installations.
O
1,4
The sensitivity for changes in flux is plotted
~< 0.5
in Fig. 5, where from each filtration curve the addi-
tional resistance after producing 20 L/m 2 of per-
meate is presented. This leads to a critical flux
value for Maasbommel of somewhat lower than
0.0 80 L/m2h, for Beverwijk around 60 L/m2h and for
1 1 2 2 3 3 4 4 5 5 6 6 Hilversum between 80 and 120 L/m2h.
Day [#]
The filtration behaviour as presented in Fig. 3
is confirmed by the results presented in Fig. 5,
Hilversum J=120 L/m2h
*,'-" 0.10
see also Table 2. Furthermore, the order of filter-
E (c) ability does not change with applied flux. With
o increasing flux the differences in filterability be-
come more manifest.
~< 0.05
1""5 I t . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

S - " - - Hive mum Beverwijk - x - Maasbornmel

O

0.00
2
1 2 3 4 5 6 7 7 m
Day [ # ]

Fig. 4. Development of AR25 during the measurement

campaigns. Each bar represents one measurement. Note 1
the differences in the vertical axes!

(increasing AR25), like day 2 and 3 of Beverwijk 0 ~. X •

and days 2, 5 and 6 of the Maasbommel tests 0 SO 200 ].SO 200

Flux [L/m~h]
(Figs. 4a and 4b).
Sometimes AR25 decreased during a day, day 7 Fig. 5. Results of critical flux measurements.
 H. Evenblij et al. / Desalination 178 (2005) 115-124 121

Table 2
Comparison of results derived from Figs. 3 and 5

'Best' Medium ' Worst'

Filterability based on standard filtration curves, Fig. 3 Hilversum Maasbommel Beverwijk
Critical flux interpolatedfrom Fig. 5 Hilversum Maasbommel Beverwijk

3.4. Chemical analyses measurements are shown in this paper. Fig. 6

The water phase of the activated sludge and
permeate o f the filtration characterisation unit
were analysed for EPS, measured as polysaccha-
rides and proteins. The difference between per-
meate and the water phase concentration signifies
the amount o f EPS that is retained by the mem-
brane. In other words, this is the amount o f EPS
that is theoretically available for cake layer forma-
tion, since it is bigger than the membrane pore
size. Permeate also contains a rest of EPS, o f
course with a size range smaller than the pore size
of the membrane.
Other researchers found out that proteins are
less involved in fouling in MBR than polysaccha-
rides. Therefore, only results from polysaccharide
shows the results ofpolysaccharide measurements
in permeate of the filtration characterisation and
the water phase of the activated sludge under con-
sideration.
If polysaccharides were determining mem-
brane fouling in MBR, Hilversum activated sludge
would be expected to contain the lowest amount
in the water phase, followed by Maasbommel; the
highest amount o f polysaccharides should be
measured in the water phase o f Beverwijk acti-
vated sludge.
However, this is not the case. Polysaccharide
concentration in the water phase is highest in the
Beverwijk sludge, as was expected, but lowest in
the Maasbommel sludge. Hilversum is in between
those values (Fig. 6a).

~' lO
(a) (b)
.e.
Maasbommel
~D
.r.

5
,,i,! Beverwijk

Hilversum

Permeate Retained Water phase )% 25% 50% 75% 101 ~%

13 Hilversum I::] Beverwijk ~ Maasbommel El Permeate polys ~ Retained polys

Fig. 6. Polysaccharide concentrations in the activated sludge from three pilot MBRs and permeate of filtration
characterisation. (a) EPS concentrations in permeate, the water phase and retained by the membrane. (b) Permeate and
retained EPS as fraction of the amount of EPS in the water phase.
! 22 H. Evenblij et al./Desalination 178 (2005) 115-124
Table 3
Comparison of results derived from Figs. 3, 5 and 6
Filterabilitybased on standardfiltrationcurves, Fig. 3
Critical flux interpolatedfrom Fig. 5
Polysaccharideconcentrations(waterphase and retention),Fig. 6a
Retainedfraction,relativeto concentrationin waterphase, Fig. 6b
If we consider normalised permeate and
retention values, i.e. as a fraction of the total avail-
able amount in the water phase, we see a somewhat
different picture. First of all, the differences are
quite small and the best filterable mixed liquor
(Hilversum) absolutely and relatively contains the
highest amount of polysaccharides smaller than
the membrane pore size(Figs. 6a and 6b). The
other two follow with decreasing relative fractions
in the permeate. This order is different from that
of Fig. 6a, see also Table 3.
4. Discussion
4.1. Filterability
The filtration characterisation unit provides a
useful tool for research into membrane fouling in
MBR. It was successfully applied to compare and
analyse filtration behaviour of activated sludge
from different installations. The results, expressed
as additional resistance against produced volume
of permeate, give a good impression of its filtra-
tion behaviour.
The observed differences in filterability be-
tween different MBR pilots are emphasised when
operating with higher fluxes.
For each installation subcritical filtration could
be performed, i.e. filtration for at least 30 min
without an increase in filtration resistance. The
observed filtration resistance in these cases equal-
led the clean water membrane resistance, as mea-
sured at the start of the experiment.
4.2. Comparing MBR pilots
Applied to MBR pilots treating municipal
'Good' Medium 'Bad'
Hilversum Maasbommel Beverwijk
Hilversum Maasbommel Beverwijk
Maasbommel Hilversum Beverwijk
Hilversum Beverwijk Maasbommel
wastewater, the method shows that there are
considerable differences with regards to filterabi-
lity of the biomass. Furthermore, the filterability
is quite stable. Measurements that were repeated
during 6-8 days showed the same trends within
quite a small range. Also changes in operating
conditions, like a temporary stop of influent flow
into the MBR pilot, were reflected as changes in
filterability (not presented here).
Differences in membrane type as applied in
the MBR pilot could not be linked to results with
the filtration characterisation. The MBR pilot plant
at Hilversum wwtp is equipped with a microfilt-
ration membrane, with a nominal pore size ten
times bigger than the other two (Table 1). It is
striking however that both the amount and the
relative fraction of polysaccharides smaller than
the pore size of the filtration characterisation
membrane (30 nm) were highest for the Hiiversum
sludge. In other words, the MBR pilot with the
biggest pore size (more 'escape possibilities' for
small substances) contained the highest amount
of'small' EPS!
4.3. EPS analyses
In order to explain observed phenomena the
following hypothesis was tested: high concen-
trations of EPS, measured as polysaccharides, are
responsible for membrane fouling in MBR. The
EPS analyses that were performed during the
measuring campaign do not indicate a direct
relationship between EPS in the water phase and
membrane fouling, contrary to Rosenberger et al.
[5]. An explanation for this is not easy to give,
and could originate from three directions:
 H. Evenblij et al. / Desalination 178 (2005) 115-124
• The EPS analyses are not appropriate for this
type o f measurement. Either, each installation
has its own type o f disturbing substance for
which the analysis has to be corrected; for
example the polysaccharide measurements
could interfere with nitrate-nitrogen [17]. Fur-
thermore, photometric methods tend to give
an underestimation of EPS.
• The EPS population in each MBR system con-
tains different species with specific effects on
filtration behaviour. Since the photometric
determination gives a total amount o f a certain
type o f molecular bindings, it does not dis-
criminate between different species of poly-
saccharides.
• EPS as measured in this research are not the
only substances involved in membrane fouling.
5;. Conclusions
Filtration Characterisation is a useful tool for
the research o f fouling in MBR. Application to
three types o f MBR shows that three 'types' of
activated sludge are formed in the respective
installations, each with its specific filterability.
The filtration behaviour cannot be correlated
directly to amounts of EPS retained by the mem-
brane, i.e. the amount of EPS that is available for
membrane fouling.
Further research into the substances involved
in fouling will eventually lead to focusing on the
exact (group of) substances that cause membrane
fouling. Once this is accomplished, the most
important step is still to be taken: cultivation of
the biomass in order to minimise the release of
these substances to the water phase.
Acknowledgments
The authors would like to thank Dennis Piron
(Waterschap Rivierenland); Ferdinand Kiestra
(Royal Haskoning); Chris Ruiken and Ron
Corsten (Dienst Waterbeheer en Riolering) and
Darren Lawrence (DHV Water) for their support
123
and cooperation in providing locations for the
measuring campaigns described in this article.
DHV Water is acknowledged for their financial
support of and cooperation in this project.
References
[1] H. Evenblij and J.H.J.M. van derGraaf, Measuring
filtration characteristics of activated sludge from
MBRs. Afvalwaterwetenschap,2(4) (2003) 209-310
(in Dutch).
[2] T. Stephenson, S. Judd, B. Jefferson and K. Brindle,
Membrane Bioreactors for Wastewater Treatment.
IWA Publishing, London, 2000.
[3] S. Ltibbecke, A. Vogelpohl and W. Dewjanin,
Wastewater treatment in a bilogical high-perform-
ance system with high biomass concentration. Water
Research, 29 (1995) 793-802.
[4] I.-S. Chang, S.-O. Bag and C.-H. Lee, Effects of
membrane fouling on solute rejection during mem-
brane filtration of activated sludge. Process Bio-
chemistry, 36 (2001) 855-860.
[5] J-S. Kim, C.-H. Lee and I.-S. Chang, Effect of pump
shear on the performance ofa crossflow membrane
bioreactor. Water Research, 35(9) (2001) 2137-
2144.
[6] S. Rosenberger and M. Kraume, Filterability of
activated sludge in membrane bioreactors. Desali-
nation, 146 (2002) 373-379.
[7] H. Nagaoka, S. Ueda and A. Miya, Influence of
bacterial extracellular polymers on membrane separ-
ation activated sludge process. Water Sci. Technol.,
34(9) (1996) 165-172.
[8] 1.-S. Chang, P. Le Clech, B. Jefferson and S. Judd,
Membrane fouling in membrane bioreactors for
wastewater treatment. J. Environ. Eng., 128(11)
(2002) 1018-1029.
[9] S. Ognier, C. Wisniewski and A. Grasmick, Mem-
brane fouling during constant flux filtration in mem-
brane bioreactors. Membr. Technol., (7) (2002) 6-
10.
[10] S. Ognier, C. Wisniewski and A. Grasmick, Mem-
brane bioreactor fouling in sub-critical filtration con-
ditions: a local criticial flux concept. J. Membr. Sci.,
229 (2004) 171-177.
[11] W. Lee, S. Kang and H. Shin, Sludge characteristics
and their contribution to microfiltration in sub-
merged membrane bioreactors. J. Membr. Sci., 216
(2003) 217-227.
124 H. Evenbl~j et al. / Desalination 178 (2005) 115-124
[12] R.W. Field, D. Wu, J.A. Howell and B.B. Gupta,
Critical flux concept for microfiltration fouling. J.
Membr. Sci., 100 (1995) 259-272.
[13] P. Le-Clech, B. Jefferson, I.-S. Chang and S. Judd,
Critical flux determination by the flux-step method
in a submerged membrane bioreactor. J. Membr. Sci.,
227 (2003) 81-93.
[14] B. Jefferson, A. Brookes, P. Le Clech and S.J. Judd,
Methods for understanding organic fouling in
MBRs. Water Sci. Technol., 49(2) (2004) 237-244.
[15] M. Dubois, K.A. Gilles, J.K. Hamilton, P.A. Rebers
and F. Smith, Colourimetric method for determi-
nation of sugars and related substances. Analytical
Chemistry, 280 (1956) 350-356.
[16] O.H. Lowry, N.J. Rosebrough, A.L. Fan" and R.J.
Randall, Protein measurement with the folin phenol
reagent. J. Biological Chemistry, 1930 ( 1951 ) 265-
275.
[17] B. Frolund, T. Griebe and P.H. Nielsen, Enzymatich
acitivity in the activated-sludge floc matrix. Appl.
Microbiol. Biotechnol., 43 (1995) 755-761.