Separation and Purication Technology 142 (2015) 813

Contents lists available at ScienceDirect

Separation and Purication Technology

journal homepage: www.elsevier.com/locate/seppur

Filtration characteristics of anaerobic fermented sewage sludge for fatty

acids production
Y.F. Zhu, H.B. Liu , H. Liu , S. Huang, H.J. Ma, Y. Tian
Laboratory of Environmental Biotechnology, School of Environmental and Civil Engineering, Jiangnan University, 1800 Lihu Avenue, Wuxi 214122, China

a r t i c l e

i n f o

Article history:
Received 6 March 2014
Accepted 29 November 2014
Available online 12 December 2014
Keywords:
Sewage sludge
Volatile fatty acid
Anaerobic fermentation
Filtration resistance

a b s t r a c t
The ltration characteristics of untreated sewage sludge, thermal alkaline pretreated and anaerobic fermented sewage sludge were investigated in the process of fatty acids production. The ltration resistance, viscosity, particle size, extracellular polymeric substances (EPS) and soluble microbial products
(SMP) were determined to identify the ltration characteristics of the sewage sludge. The results indicated that thermal alkaline pretreatment signicantly increased the ltration resistance and viscosity
of the sewage sludge, while anaerobic fermentation decreased the ltration resistance. The particle size
of the sewage sludge decreased after pretreatment and fermentation. Moreover, the EPS concentration in
the sludge sharply decreased after thermal alkaline pretreatment and then increased after the following
fermentation. The mechanism of the increase of sludge ltration resistance after pretreatment could be
attributed to the release of EPS from the microbial cells of the sludge. The degradation and conversion of
SMP into volatile fatty acids could explain the decrease in the ltration resistance after fermentation.
2015 Published by Elsevier B.V.

1. Introduction
The amount of sewage sludge produced in China has increased
signicantly because of urbanization and increased wastewater
treatment capacity. Because of land scarcity and increasingly stringent air pollution control regulations, sludge disposal by landll
and incineration may not be appropriate in the near future. Volatile
fatty acids (VFA) production from sewage sludge by anaerobic fermentation is a promising method for recycling of the sewage
sludge because it has a high organic matter (e.g. protein, carbohydrate) content [1,2]. The VFA can be used as an alternative carbon
source to improve biological nitrogen and phosphorous removal in
the wastewater treatment process [35]. Additionally, VFA have a
number of potential uses in various industries [6].
However, the separation of VFA from the anaerobic fermented
sewage is challenging [7]. Recently, several publications reported
membrane technology as a cost-effective method for VFA separation [8]. To date, a number of studies have investigated the ltration characteristics and mechanisms of activated sludge in
membrane bioreactors for the wastewater treatment [9]. However,
there is little information about sludge characteristics with high
solid concentrations during the process of anaerobic digestion,
Corresponding authors. Tel./fax: +86 85326670.
E-mail addresses: zhuyanfangjn@126.com (Y.F. Zhu), liuhongbo@jiangnan.edu.
cn (H.B. Liu), liuhe@jiangnan.edu.cn (H. Liu), huangshuai@126.com (S. Huang),
834876777@qq.com (H.J. Ma), tianyue1026@126.com (Y. Tian).
http://dx.doi.org/10.1016/j.seppur.2014.11.037
1383-5866/ 2015 Published by Elsevier B.V.

especially in anaerobic fermented sewage sludge for VFA production. Because the physical and chemical features of sludge inuence membrane ltration, it is necessary to investigate the
characteristics of fermented sewage sludge.
To achieve a high VFA yield, sewage sludge pretreatment before
fermentation, such as thermal or/and alkaline pretreatment, is usually used to improve the solubility of organic matter in solid
sludge. Ahn [3] reported that thermal pretreatment increased
sludge dewaterability through the destruction of the sludge oc
structure. Chen et al. [10] observed that acidic pretreatment
improved the dewaterability of sludge. However, to our best
knowledge, there is no information about how alkaline pre-treatment inuences the ltration characteristics of sludge. Earlier
results have indicated that extracellular polymeric substances
(EPS) and soluble microbial products (SMP) are the most important
substances inuencing membrane ltration resistance [2,11].
Organic matter, including EPS and SMP, will be solubilized during
the pretreatment process and consumed and converted into VFA
during acidogenic fermentation. Because the major components
of EPS and SMP are proteins and polysaccharides, the proles of
these substances should be studied during the processes of pretreatment and anaerobic fermentation.
We hypothesized that alkaline pretreatment would decrease
the dewaterability of sewage sludge, while anaerobic fermentation
would improve the dewaterability. The aims of this research were
to explore the ltration characteristics of acidogenic fermented

Y.F. Zhu et al. / Separation and Purication Technology 142 (2015) 813

sewage sludge, and to identify how anaerobic fermentation

improves sludge ltration performance. The results can be used
to understand the mechanism of sewage sludge ltration and
develop membrane ltration technology for VFA separation from
sewage sludge after anaerobic acidogenesis.

collected in a measuring cylinder within 5 min [23]. The ltration

resistance of the sludge slurry was calculated using the following
formula:

2. Materials and methods

where F denotes the ltration resistance of the sludge slurry (m1);

DP denotes the trans-membrane pressure (Pa); l represents the
dynamic viscosity of the sludge slurry (Pa s); and J denotes the
instantaneous ltration ux, (m3/(m2 h)).
The particle size of the sludge was measured by a BT-2003 laser
particle size analyzer, which works on the principle of laser
diffraction. Viscosity was determined by a Brookeld viscometer
(DVEII+pro, Brookeld Engineering Laboratories, Middleboro, MA)
using the instructions provided by the manufacturer.

2.1. Sewage sludge and seeding sludge

Sewage sludge was obtained from the dewatered stage of the
Taihu Xincheng wastewater treatment plant (WWTP), Wuxi, China,
and had a water content of 85 3%. Sewage sludge was diluted to
total solids (TS) values of 75 g/L and 35 g/L. Thermal alkaline pretreatment of sludge was conducted as follows: the initial pH of
the sludge was adjusted to 12.0 using a 20 mol/L NaOH solution,
then the sludge was stirred for 2 h at 90 C. Seeding sludge inoculated for VFA fermentation was collected from the same site. Sewage sludge was diluted to a TS of value of 75 g/L and heated at
100 C for 2 h to kill non spore-forming methanogens [12,13].
Before use, the seeding sludge was re-activated to culture the acidogenic microorganisms as described elsewhere [14].
2.2. Batch acidogenic fermentation
The acidogenic fermentation was conducted in 1000 mL conical
asks. A sample (500 mL) of sewage sludge (75 g/L or 35 g/L), with
or without pretreatment, and 20 mL of seeding sludge (75 g/L)
were added to the ask for VFA production. The initial pH of the
sludge was adjusted to 10.0 using 20 mol/L NaOH solution or
20 mol/L HCl solution, and the pH was maintained at 10.0 throughout the fermentation process [15]. Sodium 2-bromoethanesulphonate (BES, 50 mmol/L) was used to inhibit methanogen growth in
the sludge [16,17]. Before anaerobic fermentation, oxygen in the
ask was removed from the head space by nitrogen gas sparging
for 3 min. The asks were capped with rubber stoppers and placed
in air-tight shakers with a shaking speed of 120 rpm and temperature of 37 1 C. Each batch underwent VFA fermentation for
15 days. Sludge samples were taken every other day for VFA determination [18].

DP

lJ

3. Results and discussion

3.1. VFA production
VFA production from sewage sludge with different total solid
(TS) concentrations is shown in Fig. 1. The VFA concentrations
increased as the fermentation process progressed and remained
stable after day 10. Fermentation of sewage sludge with a higher
TS content produced higher VFA concentrations, as did thermal
alkaline pretreatment of sewage sludge. In the thermal alkaline
pre-treated sewage sludge group, the highest VFA concentration

2.3. Analysis
Total solids (TS) and volatile solids (VS) measurements were
carried out according to standard methods [19]. For VFA measurement, the sludge samples were centrifuged at 7000 rpm for 10 min.
The supernatants were ltrated through 0.45 lm lter membranes. A gas chromatograph (GC-2010, Shimadzu Co., Kyoto,
Japan) equipped with an auto injector (AOC-20i, Shimadzu Co.)
was used to measure the concentrations of the VFA. The GC was
also equipped with a ame ionization detector and a fused-silica
capillary (PEG-20M, 30 m  0.32 mm, 0.5 lm, China). 4-Methylvaleric acid was added as an internal standard and the samples were
acidied with 3 mol/L phosphoric acid [20]. The GC column was
initially held at 80 C for 3 min, increased by 15 C/min to a nal
temperature of 210 C, and then held at this temperature for
2 min. The injection port and detector temperatures were both
250 C. The total VFAs concentrations were calculated as a sum
of the individual VFA concentrations.
The EPS and SMP were extracted and analyzed by the phenol/
sulfuric acid method [21] and Coomassie Brilliant Blue method
[22].
The ltration resistance of the sludge slurry was measured by
pouring 50 mL of sludge slurry rapidly into a glass funnel containing a folded lter paper, and measuring the volume of ltrate

Fig. 1. VFA proles during the anaerobic acidogenic fermentation of sewage sludge,
A: 35 g/L sludge with pretreatment; B: 75 g/L sludge with pretreatment; C: 35 g/L
sludge without pretreatment; D: 75 g/L sludge without pretreatment.

10

Y.F. Zhu et al. / Separation and Purication Technology 142 (2015) 813

with a TS of 75 g/L was 1.3 times higher (at 7.31 0.24 g/L) than
that with a TS of 35 g/L (at 5.55 0.35 g/L). When sewage sludge
was anaerobically fermented without thermal alkaline pretreatment, the highest VFA concentrations at TS values of 75 g/L and
35 g/L were 4.63 0.43 g/L and 3.15 0.54 g/L, respectively. Based
on the results in Fig. 1b, the VFA yields were 149.47
22.51 mg COD/g VS, 189.90 28.59 mg COD/g VS, 214.32 39.16
mg COD/g VS, and 289.00 52.80 mg COD/g VS for the 75 g/L
un-pretreated, 75 g/L pretreated, 35 g/L un-pretreated, and 35 g/L
pretreated sewage sludge, respectively. With acidogenic fermentation, the VFA yields for TS values of 75 g/L and 35 g/L with pretreatment were 27.05% and 34.86% higher, respectively, than those for
sludge without pretreatment. According to the results of Liu
et al. [18], total VFA yields of pretreated and un-pretreated sewage
sludge were 279.6 mg COD/g VS and 213.2 mg COD/g VS, respectively. The VFA yields of this study are very similar to our previous
research. Chen et al. [23] reported that pretreatment of sludge was
benecial for VFA production, and this agrees with the results in
the present study.

3.2. Variation in the ltration resistance and viscosity

Variations in the ltration resistance and viscosity of sewage
sludge during the anaerobic fermentation are presented in
Fig. 2a. Filtration resistance is one of the most important parameters for sewage sludge during the process of ltration, and it also
can be used to indicate the characteristics of the sewage sludge.

As shown in Fig. 2a, the ltration resistances were

(1.04 0.22)  1011 m1 for pretreated sewage sludge and
(9.06 1.02)  108 m1 for fermented sewage sludge at 75 g/L TS.
The ltration resistance of sewage sludge with thermal alkaline
pretreatment was 522 times higher than that of the untreated sewage sludge. However, the ltration resistance after fermentation
decreased by almost 99% compared to the pretreated sewage
sludge. Similarly, for the 35 g/L TS sewage sludge, the ltration
resistances of untreated, pretreated and fermented sewage sludge
were
(3.52 0.68)  107 m1,
(9.90 0.49)  108 m1
and
(8.18 0.37)  108 m1, respectively. Similar trends to the 75 g/L
TS sewage sludge were observed for changes in the sewage sludge
ltration.
Fig. 2b shows the viscosity of the sewage sludge during the
anaerobic fermentation. The viscosity of the 75 g/L sewage sludge
after pretreatment was 5410 mPa s, which was 8.87 times higher
than that of the untreated sewage sludge (610 mPa s). The viscosities of the 35 g/L sewage sludge with and without pretreatment
were 104 mPa s and 49.2 mPa s, respectively. The above results
clearly indicate that thermal alkaline pretreatment increases the
viscosity of the sewage sludge. The reason for this increase may
be that the thermal alkaline pretreatment solubilizes the solid
organic matter and releases proteins and polysaccharides into
the liquid [2,24]. Some studies of the relationship between the
EPS and viscosity have demonstrated that composition and morphology will inuence the viscosity and ltration resistance of
the sewage sludge [25,26]. Changes in the EPS and SMP during pretreatment and fermentation will be discussed in detailed in Section
3.4.
In addition, the viscosity of the sewage sludge deceased significantly in the early stages of anaerobic fermentation. For the 75 g/L
sewage sludge, the viscosity decreased from 5410 mPa s to
189.2 mPa s after pretreatment. For the 35 g/L sewage sludge, the
viscosity decreased from 104 mPa s to 29.1 mPa s with pretreatment. Many factors will inuence the viscosity of the sewage
sludge, such as the concentrations of proteins and polysaccharides,
which usually cause high sludge viscosity [2]. In the present study,
because the protein and polysaccharides were consumed during
VFA production by the acidogenic microorganisms, the viscosity
decreased [27]. The ltration resistance and viscosity results for
the untreated, pretreated and fermented sludge veried our
hypothesis in the introduction.
3.3. The particle size distribution

Fig. 2. Changes in the ltration resistance and viscosity of sewage sludge with or
without pretreatment A: 35 g/L sludge with pretreatment; B: 75 g/L sludge with
pretreatment; C: 35 g/L sludge without pretreatment; D: 75 g/L sludge without
pretreatment.

The particle size distribution of sludge is an important parameter affecting the sludge dewaterability [28,29]. As shown in Fig. 3,
the particle size decreased after fermentation for both the 35 g/L
and 75 g/L sludges, regardless of whether the sludge was pretreated or not. For the 35 g/L sludge without pretreatment, the
cumulative percentage of fermented sludge with a particle size
below 100 lm shifted from 55.61% to 63.22%, while for the pretreated sludge it increased from 55.84% to 80.87%. The cumulative
percentage of thermal alkaline pretreated sludge with a particle
size below 100 lm increased by 44.82% and that for un-pretreated
sludge only increased by 13.68%. A similar trend was also observed
for the 75 g/L sludge. The cumulative percentages of sludge with
particle sizes below 100 lm increased by 56.32% and 46.01% for
thermal alkaline treated and un-pretreated sludge, respectively.
It should be noted that the peak of the particle size distribution
of the untreated sewage sludge was 149 lm with percentages of
3.25% and 3.28% for the 35 g/L and 75 g/L sludge, respectively.
However, the peak of the particle size distribution for the 35 g/L
sludge shifted to 30.6 lm with a percentage of 2.93% without pretreatment, and shifted to 27.9 lm with a percentage of 3.28% with
pretreatment. For the 75 g/L sludge, the peak moved to 27.9 lm

Y.F. Zhu et al. / Separation and Purication Technology 142 (2015) 813

11

Fig. 3. The particle sizes of the sewage sludge (a) 35 g/L sludge with pretreatment; (b) 75 g/L sludge with pretreatment; (c) 35 g/L sludge without pretreatment; (d) 75 g/L
sludge without pretreatment d original sludge; s fermented sludge.

with a percentage of 3.37% without pretreatment, and shifted to

30.6 lm with a percentage of 4.66% with pretreatment.
Liu et al. also reported that both thermo-alkaline and ultrasonic-alkaline pretreatment could reduce the particle size. Because
of the multi-level structure of sewage sludge oc, alkaline hydrolysis of the extracellular biopolymer, the highly porous ocs disintegrated into microocs or occuli and even primary particles. The
reduction in particle size was caused by cell wall damage and degradation. Alkaline pretreatment was more efcient at destroying
the cell walls than many other pretreatments such as with acid
or ultrasonic vibrations [30].
The particle size is very closely associated with the ltration
resistance of the sewage sludge, and the resistance is also dependent on the pore size of the lter membrane. When the particle
size of the sludge is bigger than or similar to the pore size of the
lter membrane, the ltration resistance will increase. Therefore,
our data indicate that the particle size of the sewage sludge should
be taken into consideration when selecting the pore size of the lter membrane.
3.4. Variations of EPS and SMP
To investigate the mechanism causing changes in the ltration
characteristics of the sewage sludge with pretreatment and anaerobic fermentation, the EPS and SMP concentrations were determined. The EPS declined sharply after pretreatment but increased
after fermentation for both the 35 g/L and 75 g/L sludges
(Fig. 4a). For the 35 g/L sludge, the EPS was 9.66 0.51 mg EPS/
g VSS for the untreated sludge, 1.49 0.67 mg EPS/g VSS for the
pretreated sludge and 6.32 0.55 mg EPS/g VSS for the fermented
sludge. For the 75 g/L sludge, the EPS concentrations of the
untreated,
pretreated
and
fermented
sludge
were
15.61 0.78 mg EPS/g VSS,
3.31 0.84 mg EPS/g VSS,
and

8.13 0.35 mg EPS/g VSS, respectively. EPS represents the organic

matter, usually proteins and polysaccharides, on the surface of
the suspended microbial cells in the sludge solid. Therefore, we
can conclude that the pretreatment results in solubilization and
release of organic matter from the sludge ocs into the liquid. It
should be noted that there was a 34.6% EPS decrease (from
untreated to fermented sludge) for the 35 g/L sludge and a 47.9%
decrease for the 75 g/L sludge with pretreatment. However, there
were only 20.7% and 37.8% EPS decreases for the 35 g/L and 75 g/L
sludges, respectively, without pretreatment. The results clearly
demonstrate that pretreatment greatly improves EPS solubilization
and release from the sludge into the fermentation liquid.
Fig. 4b shows changes in the SMP in the liquid from sewage
sludge fermentation. For the 35 g/L sludge, the concentrations of
SMP for the untreated, pretreated and fermented sludge were
140.47 15.26 mg/L,
2102.21 211.74 mg/L
and
1416.88
220.60 mg/L, respectively. While for the 75 g/L sludge, the values
were 209.21 11.29 mg/L, 4180.45 309.99 mg/L and 2745.24
362.83 mg/L for the untreated, pretreated and fermented sludge,
respectively. It can be seen that the SMP increased considerably
during pretreatment and fermentation, which is concomitant with
the EPS decrease. However, the SMP concentration decreased by
46.96 mg/L (from the untreated to fermented sludge) for the
35 g/L sludge and by 50.30 mg/L for the 75 g/L sludge. This was a
different trend to that observed with the pretreated sludge. The
reason for this is that pretreatment improves the release of the
EPS from the surface of the microbial cells into the liquid and
results in a large increase in SMP. Without pretreatment, the
SMP in the liquid was degraded and converted into VFA, resulting
in the decrease observed after fermentation.
The changes in EPS and SMP for the three kinds of sludge
explain the variations observed in the sludge ltration
characteristics, namely why ltration performance was worse after

12

Y.F. Zhu et al. / Separation and Purication Technology 142 (2015) 813

Acknowledgements
We gratefully acknowledge support from the National Natural
Science Foundation of China (Grant No. 51208231), Natural Science
Foundation of Jiangsu Province of China (Grant No. BK2012121),
and the Environmental Protection Department of Jiangsu Province
(Grant No. 2012035).

References

Fig. 4. The concentration of EPS (a) and SMP (b) during the anaerobic fermentation.
A: 35 g/L sludge with pretreatment; B: 75 g/L sludge with pretreatment; C: 35 g/L
sludge without pretreatment; D: 75 g/L sludge without pretreatment.

pretreatment and better after fermentation. This demonstrates

that the EPS and SMP inuence the ltration characteristics of
the sludge [31].
The total amount of released EPS were 285.6 mg/L for the 35 g/L
sludge and 922.5 mg/L for 75 g/L sludge after pretreatment and fermentation. However, the determined SMP in the liquid were
1961.74 mg/L and 3971.24 mg/L for the 35 g/L and 75 g/L sludge,
respectively. The results indicate that the SMP is much higher than
the released EPS from the sludge ocs. This is because the SMP not
only comes from the EPS, but also from intracellular biopolymers
such as proteins and polysaccharides in the microbial cells.
4. Conclusions
Filtration characteristics were investigated for untreated, thermal alkaline pretreated and anaerobic acidogenic fermented sewage sludge. The hypothesis of this study was well veried by the
determination and analysis of the ltration resistance, viscosity,
particle size, and ow properties of the three kinds of sludge. The
most important conclusions were as follows: (1) fermentation of
sewage sludge with a high TS value gave a high VFA concentration,
and thermal alkaline pretreated sewage sludge also gave a high
VFA concentration; (2) thermal alkaline pretreatment led to a high
ltration resistance, but the ltration resistance after fermentation
decreased by almost 99% of the pretreated sewage sludge; (3) both
pretreatment and fermentation decreased the sludge particle size;
(4) the increase in the ltration resistance was attributed to the
release of EPS from the microbial cells of the sludge; and (5) degradation and conversion of the SMP into VFA explained the
decrease in the ltration resistance of the fermented sludge.

[1] F.Y. Garcia Becerra, E.J. Acosta, D.G. Allen, Alkaline extraction of wastewater
activated sludge biosolids, Bioresour. Technol. 101 (2010) 69836991.
[2] E. Neyens, J. Baeyens, A review of thermal sludge pre-treatment processes to
improve dewaterability, J. Hazard. Mater. 98 (2003) 5167.
[3] Y.H. Ahn, R.E. Speece, Elutriated acid fermentation of municipal primary
sludge, Water Res. 40 (2006) 22102220.
[4] A.S. Ucisik, M. Henze, Biological hydrolysis and acidication of sludge under
anaerobic conditions: the effect of sludge type and origin on the production
and composition of volatile fatty acids, Water Res. 42 (2008) 37293738.
[5] Y. Chen, A.A. Randall, T. McCue, The efciency of enhanced biological
phosphorus removal from real wastewater affected by different ratios of
acetic to propionic acid, Water Res. 38 (2004) 2736.
[6] M.F. Colmenarejo, E. Snchez, A. Bustos, G. Garca, R. Borja, A pilot-scale study
of total volatile fatty acids production by anaerobic fermentation of sewage in
xed-bed and suspended biomass reactors, Process Biochem. 39 (2004) 1257
1267.
[7] P. Gluszcz, T. Jamroz, B. Sencio, S. Ledakowicz, Equilibrium and dynamic
investigations of organic acids adsorption onto ion-exchange resins,
Bioprocess Biosyst. Eng. 26 (2004) 185190.
[8] B. Yordanov, L. Boyadzhiev, Pertraction of citric acid by means of emulsion
liquid membranes, J. Membr. Sci. 238 (2004) 191197.
[9] G. Skouteris, D. Hermosilla, P. Lpez, C. Negro, . Blanco, Anaerobic membrane
bioreactors for wastewater treatment: a review, Chem. Eng. J. 198199 (2012)
138148.
[10] Y. Chen, H. Yang, G. Gu, Effect of acid and surfactant treatment on activated
sludge dewatering and settling, Water Res. 35 (2001) 26152620.
[11] G.P. Sheng, H.Q. Yu, X.Y. Li, Extracellular polymeric substances (EPS) of
microbial aggregates in biological wastewater treatment systems: a review,
Biotechnol. Adv. 28 (2010) 882894.
[12] S.-E. Oh, S. Van Ginkel, B.E. Logan, The relative effectiveness of pH control and
heat treatment for enhancing biohydrogen gas production, Environ. Sci.
Technol. 37 (2003) 51865190.
[13] W. Park, S.H. Hyun, S.-E. Oh, B.E. Logan, I.S. Kim, Removal of headspace CO2
increases biological hydrogen production, Environ. Sci. Technol. 39 (2005)
44164420.
[14] Y. Nie, H. Liu, G. Du, J. Chen, Enhancement of acetate production by a novel
coupled syntrophic acetogenesis with homoacetogenesis process, Process
Biochem. 42 (2007) 599605.
[15] H. Yuan, Y. Chen, H. Zhang, S. Jiang, Q. Zhou, G. Gu, Improved bioproduction of
short-chain fatty acids (SCFAs) from excess sludge under alkaline conditions,
Environ. Sci. Technol. 40 (2006) 20252029.
[16] J. Wang, H. Liu, B. Fu, K. Xu, J. Chen, Trophic link between syntrophic acetogens
and homoacetogens during the anaerobic acidogenic fermentation of sewage
sludge, Biochem. Eng. J. 70 (2013) 18.
[17] K. Xu, H. Liu, J. Chen, Effect of classic methanogenic inhibitors on the quantity
and diversity of archaeal community and the reductive homoacetogenic
activity during the process of anaerobic sludge digestion, Bioresour. Technol.
101 (2010) 26002607.
[18] X.L. Liu, H. Liu, J. Chen, The Condition Optimization of Sewage Sludge for
Producing Volatile Fatty Acids and the Investigation of Acidogenic Mechanism,
Jiangnan University, Wuxi, 2008.
[19] A. Apha, WEF, Standard Methods for the Examination of Water and
Wastewater 20th Edition-4500-NO3-D Nitrate Electrode Method, American
Public Health Association, Washington, DC, 1998.
[20] W.N. Chan, Thermophilic Anaerobic Fermentation of Waste Biomass for
Producing Acetic Acid, Texas A & M University, 2002.
[21] B. Fr/olund, T. Griebe, P.H. Nielsen, Enzymatic activity in the activated-sludge
oc matrix, Appl. Microbiol. Biotechnol. 43 (1995) 755761.
[22] M.M. Bradford, A rapid and sensitive method for the quantitation of
microgram quantities of protein utilizing the principle of protein-dye
binding, Anal. Biochem. 72 (1976) 248254.
[23] Y. Chen, S. Jiang, H. Yuan, Q. Zhou, G. Gu, Hydrolysis and acidication of waste
activated sludge at different pHs, Water Res. 41 (2007) 683689.
[24] M.T. Agler, B.A. Wrenn, S.H. Zinder, L.T. Angenent, Waste to bioproduct
conversion with undened mixed cultures: the carboxylate platform, Trends
Biotechnol. 29 (2011) 7078.
[25] H.P. Yuan, X.B. Cheng, S.P. Chen, N.W. Zhu, Z.W. Zhou, New sludge
pretreatment method to improve dewaterability of waste activated sludge,
Bioresour. Technol. 102 (2011) 56595664.
[26] A. Sweity, W. Ying, M.S. Ali-Shtayeh, F. Yang, A. Bick, G. Oron, M. Herzberg,
Relation between EPS adherence, viscoelastic properties, and MBR operation:
biofouling study with QCM-D, Water Res. 45 (2011) 64306440.

Y.F. Zhu et al. / Separation and Purication Technology 142 (2015) 813
[27] A.S. Eissa, Newtonian viscosity behavior of dilute solutions of polymerized
whey proteins. Would viscosity measurements reveal more detailed molecular
properties?, Food Hydrocolloids 30 (2013) 200205
[28] P.R. Karr, T.M. Keinath, Inuence of particle size on sludge dewaterability, J.
Water Pollut. Control Fed. (1978) 19111930.
[29] J.H. Kang, D. Kim, T.J. Lee, Hydrogen production and microbial diversity in
sewage sludge fermentation preceded by heat and alkaline treatment,
Bioresour. Technol. 109 (2012) 239243.

13

[30] X. Liu, H. Liu, J. Chen, G. Du, J. Chen, Enhancement of solubilization and

acidication of waste activated sludge by pretreatment, Waste Manage. 28
(2008) 26142622.
[31] G.P. Sheng, H.Q. Yu, X.Y. Li, Extracellular polymeric substances (EPS) of
microbial aggregates in biological wastewater treatment systems: a review,
Biotechnol. Adv. 28 (2010) 882894.